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  • JIA Weizhe, JIAO Bo, WANG Jiao, CHEN Wenye, YANG Fan, LIU Yongwei, DONG Fushuang, ZHAO Liqun, ZHOU Shuo
    Abstract (384) PDF (128) RichHTML (27)

    In order to explore stress resistance genes in wheat and study the molecular mechanism of calmodulin-like genes in plant stress resistance,a calmodulin-like gene TaCML8-A was cloned from wheat by electronic cloning combined with RT-PCR.Bioinformatics analysis showed that the open reading frame length of the gene was 519 bp,encoding 172 amino acid sequences,including PTZ00184 and FRQ1 superfamily,four EF-hand domains,including four calcium binding sites.The molecular weight of the encoded protein was 18.31 ku and the isoelectric point was 4.54.It belonged to acidic protein.Subcellular localization showed that TaCML8-A protein was distributed in the nucleus and cell membrane.Nucleotide sequence alignment showed that TaCML8-A had the closest genetic relationship with rice OsCML14,with a similarity of 79.17%.qRT-PCR results showed that the expression of TaCML8-A gene in shoot and root of wheat increased under salt,osmotic and cold stress,indicating that plants might respond to thoese stresses by increasing the expression of TaCML8-A.During heat shock,TaCML8-A gene was induced and inhibited in root and shoot,respectively,so as to play different functions.The calmodulin-like gene TaCML8-A was successfully cloned from wheat and analyzed for expression.It is preliminarily speculated that the gene might be involved in regulating abiotic stress of plants,so as to provide basic theoretical support for the follow-up study of its biological function.

  • TANG Long, ZHAO Yuwei
    Abstract (357) PDF (150) RichHTML (56)

    Application of some hormonal signaling compounds,as brassinolides and their derivatives,could significantly improve the salt stress resistance in plants.The purpose of present work was to test whether the over-expressing of Methylsterol monooxygenase gene(SMO),a key gene coding a bio-synthesizing enzyme of sterol in plants,could promote the salt-stress tolerance of target plants.PnSMO1.1,gene encoding of methylsterol monooxygenase in the plant species of Pharbitis nil was firstly cloned and then used as target gene for following genetic transformation process,while wild-type Pharbitis nil seedlings were used as the receptor plants for constructing of the transgenic lines which over-expressed PnSMO1.1 genes.In this work,the PnSMO1.1 gene transformed Pharbitis nil lines were constructed via an ovary injection transformation method.Plantlets from individual PCR identified transgenic plant lines were used as materials to detect vegetative growth figures and some pivotal physiological indicators,for instance,contents of malondialdehyde (MDA),relative conductivity,as well as castasterone and 6-deoxo-castasterone contents in cells under stresses with gradient NaCl conditions varied from 0—200 mmol/L.The results showed that the over-expression PnSMO1.1 significantly improved the relative growth of roots and hypocotyls in transformants than in wild-type (WT) or vacant plasmid transformed control (BL) plants under 100—250 mmol/L NaCl stresses.Compared to WT and BL seedlings,significantly higher accumulation of 6-deoxo-castasterone,but lower relative conductivity (rEC) values,castasterone accumulation or MDA contents were found in transgenic lines under various NaCl stresses.Stresses such as salinity,drought and freezing temperatures,had severely suppressed the vegetative growth of plants,as well as their yields.These results highlighted that over-expression of the PnSMO1.1 gene could significantly improve the salinity-stress resistance of transgenic plants by delicately adjusting the dynamic homeostasis of brassinolides in cells,and protecting the structural integrity of plasma membrane.

  • ZHAO Changjiang, DU Mengxiang, SONG Juqi, XU Shangyuan, HE Lin, XU Jingyu, YANG Kejun, LI Zuotong
    Abstract (229) PDF (51) RichHTML (167)

    NRL(NPH3/RPT2-Like)is a type of light-responsive protein unique to plants and plays a vital role in the phototropic signal pathway. To reveal the NRL gene maize genome's characteristics and expression,we analyzed them using bioinformatics methods combined with qRT-PCR technology. The property,structure,evolution of their encoded proteins,and growth period tissue expression and stress expression were analyzed. 31 ZmNRL genes identified were located in nine maize chromosomes,encoding protein amino acids 464-749 aa,which predicted to have chloroplast,nuclear and cytoplasmic locations. According to protein conservation,ZmNRL family was divided into four categories. Their gene structure also presented certain conservation,the most contained four exons. Analysis of the cis-elements of gene promoters revealed a large number of abscisic acids,jasmonic acid,light response,and anti-oxidation elements,among which G-box and Sp1 were two types of light-related elements. The expression of ZmNRL family genes in tissues during the growth period showed a temporal and spatial specificity,and the majority expression level was not high. Only ZmNRL2,ZmNRL4,ZmNRL24,and ZmNRL29 highly expressed. Furthermore,the characteristic modules were produced based on the data of the tissue co-expression genes. And the GO enrichment analysis of a particular leaf growth module containing six ZmNRL genes,mainly associated with the plastid organization biological processes and rRNA binding molecular functions. The expression of ZmNRL5,ZmNRL7,ZmNRL12,and ZmNRL19 genes were analyzed by qRT-PCR under salt,drought,high temperature,and Rhizoctonia solani inoculation treatments. The results showed that ZmNRL12 was significantly up-regulated in maize seedlings treated with high temperature,while ZmNRL5,ZmNRL7 and ZmNRL19 genes were down-regulated in drought,salt and pathogen treatments. In summary,31 ZmNRL genes were identified in the maize genome. They not only had apparent specific tissue expression but also participated in biotic and abiotic stress responses.

  • WANG Qingbiao, WANG Yanping, LINGHU Bo, QIAN Huihui, ZHAO Qiuju, ZHANG Li
    Abstract (226) PDF (186) RichHTML
    The effect of different concentrations salt stress on seedling growth and expression of related genes were explored in this study. Firstly, salt tolerant variety Yura Hama Daikon and susceptible variety Wujinhong were selected according to germination under salt stress conditions among 11 cultivars.And then the seedling height(SH) and the leaf scorch index(LSI) of Yura Hama Daikon and Wujinhong were investigated under salt stress treatment.The results showed that SH decreased and LSI increased under salt stress in both varieties.Compared with the salt-sensitive variety Wujinhong,the salt-tolerant variety Yura Hama Daikon had a smaller decrease in SH and lower LSI.Under 200 mmol/L salt stress,SH and LSI of salt-tolerant Yura Hama Daikon were 46.18% and 20.56,respectively,while those of salt-sensitive Wujinhong were 75.25% and 56.11.The transcription of RsCAT and RsSOD genes was studied in salt-tolerant and susceptible varieties under different salt concentrations by qPCR.The expression of RsCAT gene was first increased and then decreased under low salt concentration treatment,and reached the maximum at 7 day.When treated with high salt concentration,the transcription of RsCAT in Wujinhong was the highest at 48 h,while the expression level in salt-tolerant varieties increased gradually and maintained for a longer time,reaching the highest at 7 day.After high salt concentration treatment,the transcription of RsSOD gene reached the highest expression level at 24 hours,and then maintained a higher level in salt-tolerant variety.In salt-sensitive varieties,the maximum expression level of RsSOD appeared at 14 day.The above results will lay foundation for revealing the mechanism of salt stress in radish and provide technical support for radish salt-tolerant breeding.
  • TENG Haiyan, XU Dan
    Abstract (214) PDF (157) RichHTML
    The OsSP1 gene encodes a chloroplast localized protein in rice, and its expression clearly responds to drought stress.In order to elucidate the effect of OsSP1 gene on rice growth and drought stress tolerance, OsSP1 gene-edited and overexpressed rice plants were created by using CRISPR/Cas9 gene editing technology and gene overexpression technology, respectively. The mutation pattern of OsSP1 gene in the gene-edited plants was determined by sequencing analysis, and homozygous overexpression plants were selected by hpt sequence detection and hygromycin B screening assay. The results of phenotypic comparison, chlorophyll content and reactive oxygen species(ROS) determination and drought tolerance analysis showed that overexpression of OsSP1 gene in rice had almost no effect on rice growth and drought stress tolerance. However, after the OsSP1 gene was mutated through gene editing, the growth and fruiting of rice T0 and T1 gene-edited plants were severely affected with biallelic heterozygous or homozygous mutations, and all showed severe growth inhibition such as dwarf, etiolation, premature aging, and non-tillering, T0 plants produced very few seeds, while the T1 plants did not head at all, and the chlorophyll content in leaf of the gene-edited plants was significantly or extremely significantly lower than that of the wild-type and overexpressed plants. These results indicated the essential role of OsSP1 gene in rice growth and yield, and provided a theoretical and material basis for elucidating the underlying mechanism of OsSP1 gene in rice.
  • SHI Jiaqi, LIU Yuqing, WANG Yanling, YANG Zaiqiang
    Abstract (205) PDF (39) RichHTML (15)

    To reveal the effect process and mechanism of nitrogen application level on the chlorophyll fluorescence characteristics of cucumber in fruit stage under high-temperature stress,the Jinyou 101 cucumber was used as the tested material,the air temperatures were set at 35 ℃/25 ℃,38 ℃/28 ℃,41 ℃/31 ℃,in the same time the 28 ℃(daily maximum temperature)/18 ℃(daily minimum temperature)was as the control(CK);the nitrogen(N)application level was set at 0(N0),160(N1),240(N2),320 kg/ha(N3);total 16 treatments and three duplicates.The dynamic parameters of chlorophyll fluorescence of cucumber functional leaves were measured systematically after 9-days high-temperature stress in fruit stage,the differences between in various treatments were also discussed.The PSⅡ reaction centre of cucumber leaves were significantly damaged and the maximum fluorescence(Fm),maximum photochemical quantum yield(Fv/Fm),photosynthetic performance index(PIabs),area enclosed by Fm and fluorescence curve(Area)were significantly decreased after high-temperature stress in fruit stage.Under the 35 ℃ high-temperature stress,the values of ΔWO-J and ΔWO-K were negative in treatment N1—N3,and the oxygen evolution complex(OEC)was not inactivated.At the same time,the energy transfer between PSⅡ central thylakoids was unhindered.However,the ΔWO-J of leaves was positive in treatment N1 under 38 ℃ and treatment N2 under 41 ℃,which means the OEC was inactivated.Therefore,the energy transfer between PSⅡ central thylakoids was blocked due to the positive ΔWO-K in treatment N1—N3 under the 38,41 ℃.Under the high-temperature stress,the nitrogen application significantly increased the chlorophyll content,Fm,Fv/Fm,PIabs,PItotal,Area and Sm of cucumber leaves,and enhanced the OEC activity and smooth the energy transfer between the PSⅡ central thylakoids;the ABS/RC,TRo/RC and DIo/RC of cucumber leaves also decreased with the increase of nitrogen application level while the electron transfer energy(ETo/RC)increased.Nitrogen application rate and temperature had significant interactive effects on the fluorescence characteristics and yield of cucumber leaves.Under the 35,38,41 ℃high-temperature stress in fruit stage,when the nitrogen application rates were 236,283,177 kg/ha respectively,the photosynthesis of cucumber leaves was stronger and higher yield could be obtained.Therefore,the reasonable nitrogen application could improve the OEC activity of PSⅡ,promote energy transfer,and slow down the inhibition of electron receptor pool on the side of PSⅠ receptor of cucumber leaves under the high-temperature stress in fruit stage,and also improve the orderly progress of photosynthesis.

  • YI Qiang, YANG Zebing, TAN Jiaying, HOU Xianbin, LIU Yinghong, BAI Guangxiao, HUANG Yubi
    Mining the loci for stem diameter(SD) under different planting densities in maize could improve maize high-density tolerance and lodging resistance. SD was measured from 301 recombinant inbred lines(RIL) derived from the cross between the maize foundation parents Ye 478 and R08 under low(57 000 plants/ha) and high(114 000 plants/ha) planting densities in Jinghong, Yunnan Province, in 2014 and 2015. Quantitative trait loci(QTL) analyses were conducted by using inclusive composite interval mapping in QTL ICIMapping V4.1. QTL and environment interaction was analyzed by using the mixed-model-based composite interval mapping(MCIM) of QTLNetwork 2.0.The results showed that the difference of parental lines indicated Ye 478 did not differ for SD between two densities, but SD in R08 significantly decreased as the plant density increased. Considerable SD variation was observed in the RIL population. The range of phenotypic variation was decreased as the plant density increased. Heritability for SD in the RIL population under high and low planting densities were 48.01%, 65.03%, respectively. Correlation analyses revealed that SD was significantly correlated with plant height, ear height, and ear weight per plant(0.29> r ≥ 0.13, P <0.01), but had no significant correlation relationship with ear tip-barrenness and barrenness rate across both densities. QTL mapping by using joint analyses across both environments detected seven minor QTL for SD, with a range from 3.68% to 6.91%, and four QTL were observed under high density, namely qSD1-1, qSD3-1, qSD4, and qSD6. Only qSD6 was found across both planting densities. None of the four QTL(qSD1-1, qSD3-1, qSD3-2, and qSD4) was presented in the previous studies regarding QTL mapping for SD. Only one pair of loci for additive and additive interaction was found under high planting density. These results suggested different genetic regulations for SD across low and high densities. Among these QTL, qSD6 was simultaneously found across both densities and in the F2:3 population from the same cross reported in the previous study. Moreover, the QTL segments stated above are well descended and transmitted from Ye 478 and its descendants, which could be used for marker-assistance selection(MAS) in maize breeding.
  • YUE Yongqi, HUA Yonglin, JIA Yige, LI Jian, XIONG Yan, XIONG Xianrong
    Yak was used as the experimental object to clone the yak CCAAT/Enhancer binding protein alpha(CEBPα) gene, predicted its protein structure and function, isolated yak adipocyte, and detected the expression of CEBPα in different tissues of yak and yak fat expression during cell induction and differentiation. Collected the heart, liver, spleen, kidney, stomach, small intestine, subcutaneous fat, visceral fat and muscle tissue of an adult yak (4 years old), and used PCR technology to obtain the CDS sequence of the CEBPα gene;the method of bioinformatics predicted the sequence homology and protein structure of the gene;Real-time fluorescent quantitative PCR(qRT-PCR) was used to detect the expression of CEBPα in different tissues of yak;the yak preadipocytes were isolated by collagenase digestion, qRT-PCR was used to detect the expression pattern of CEBPα gene at 0, 3, 6, 9 d of induced differentiation;the results of the study showed that the CDS sequence of the yak CEBPα gene was 645 bp in length and encoded 214 amino acids. The yak CEBPα gene had relatively high homology with Bos taurus, with a nucleic acid homology of 99.73% and amino acid homology of 99.19%. The results of constructing a phylogenetic tree showed that the yak was more similar to ordinary cattle and sheep. CEBPα protein was an unstable hydrophilic protein with a transmembrane structure and phosphorylation/dephosphorylation. The secondary structure of the protein was dominated by α helix (27.10%) and random coil (67.29%). The results of qRT-PCR showed that the CEBPα gene was most highly expressed in the lower adipose tissue of the yak, and lowly expressed in the kidney, spleen and visceral adipose tissue. The expression of CEBPα gradually increased during the induction and differentiation of yak adipocytes. In this experiment, we cloned and obtained the CDS region sequence of CEBPα, determined its expression in various tissues of yak and clarified the expression pattern of this gene in the process of induced differentiation of yak adipocytes. Provide basic data to reveal the function of CEBPα gene in yak adipose tissue and adipocyte.
  • MEI Yu, LI Xiangrong, CAI Shike, GU Yan, WANG Jihua
    In order to in-depth understand the overall level of transcriptome and the flavonoid biosynthesis pathway genes of Pueraria thomsonii Benth. The mixed sample of root, stem and leaf of Pueraria thomsonii was used as materials for sequencing analysis using the single-molecule long read sequencing technology(SMRT) of Pacific Biosciences Sequel platform. A total of 10 994 967 high quality reads and 384 072 full-length non-chimeric(FLNC) were obtained. Sequencing data after quality control, 90 856 isoforms were obtained. There were 85 239 isoforms annotated by NR, SwissProt, KOG, KEGG and GO databases, and the number of NR annotation was 84 675, accounting for 93.2%;In KEGG database, only 22 330 genes were annotated in 132 pathways, and 9 368 genes were distributed more in metabolic pathway, accounting for 41.95%. A total of 3 507 transcription factors(TF) were predicted, and bHLH TF family was the most annotated genes. Besides, 14 127 genes were assigned to 17 R gene classes, mainly RLP. 33 660 SSR sequences were detected, most of them were AG/CT type. By analyzing the biosynthesis pathway of flavonoids, 110 genes were found, including 26 genes encoding HCT, 3 genes encoding CHS and 7 genes encoding CHI. Compared with Illumina HiSeqTM2500, PacBio platform obtained longer transcripts and higher annotation rates. This research intensively studied the Pueraria thomsonii at the full-length transcriptome level, which provided more reliable transcriptome data for molecular biology research and scientific basis for further development of molecular markers and mining excellent genes of Pueraria thomsonii.
  • GUO Yuhuan, YAMAMOTO Naoki, PENG Zhengsong, LIAO Mingli, WEI Shuhong, WU Yichao, YANG Zaijun
    In order to further locate the Pis1 gene, accelerate the molecular marker-assisted breeding of wheat, and obtain the wheat varieties with high quality, high yield and stable yield. Chuanmai 28(CM28) and its three pistils near-isogenic line CM28TP were used as research materials, the three stages of CM28 and CM28TP young spikes(spikelet length 0.2-0.5 cm, 0.5-0.7 cm, 0.7-1.0 cm) perform transcriptome sequencing, then, the GO database was used to classify and analyze the genes where the mutation site was located, and four SNP markers near the Pis1 gene were selected for verification. The results showed that there were 5 310 common SNP/InDel loci in CM28TP and CM28 young spikes at three stages, including 5 024 SNP loci and 286 InDel loci.In SNP sites, the conversion types(63.33%) were more than the transversion types(31.28%), and the ratio of the two types reached 2.02. There were more insert types(152) than deletion types(134) in InDel site. The SNP/InDel locus was most distributed in the A genome, followed by the B genome and the D genome was the least. The GO classification annotation of SNP/InDel genes showed that genes accounted for the highest proportion in biological processes, followed by cell components and molecular functions. The prediction of the impact of the SNP/InDel loci on protein function indicated that 36 mutant loci severely affected protein function, and 1 279 mutant loci moderately affected protein function. Four SNP sites were screened from the location interval of the Pis1 gene for PCR amplification and sequencing verification, and it was found that these four sites were consistent with the results of RNA-seq analysis, which indicated that the SNP/InDel sites excavated were accurate. This study enriched SNP/InDel markers in wheat and laid a foundation for high-density genetic map construction, gene mapping and marker-assisted selection breeding in wheat, at the same time, four SNP markers near the Pis1 gene were developed, which provided the possibility of map-based cloning of this gene.
  • JIA Liqiang, LIU Xun, DING Bo
    Abstract (145) PDF (69) RichHTML (15)

    In order to study the role of ZmbZIPs in the stress tolerance of maize,using maize inbreed line Zheng 58 for exprerimental material,expression analysis of 9 ZmbZIPs were performed under 200 mmol/L NaCl,20% PEG6000,4 ℃ low temperature and nitrate or ammonium deficiency stresses.Evolutionary tree analysis indicated that 9 ZmbZIPs were divided 3 subgroup.RT-qPCR analysis showed that 8 ZmbZIPs were detected in maize tissues,whereas ZmbZIP80 were not,indicated it could be pseudogene.Under simulated salt,drought,low temperature and nitrogen stress conditions,8 ZmbZIPs exhibited different expression pattern in response to various stresses,ZmbZIP37 and ZmbZIP53 were induced while ZmbZIP49 and ZmbZIP79 were downregulated obviously under NaCl stress,ZmbZIP37 and ZmbZIP53 of leaves significantly suppressed under nitrate deficit stress while ZmbZIP42 and ZmbZIP49 were upregulated in responsive to ammonium deficient stress.The result indicated 8 ZmbZIP played widely roles against stress conditions.Expression pattern of 9 ZmbZIPs gene differed in different tissues or under various adverse stresses.The study can offer scientific data for further reveal ZmbZIP biological function.

  • YU Kun, LIU Ruili, LIU Xianxun, BAI Xuejin, DONG Yajuan
    The aims to explore the molecular mechanism of bta-miR-133a involved in the regulation of longissimus dorsi development in beef cattle. The longissimus dorsi muscle of Black cattle and Luxi cattle was collected for sRNA database construction, and the data was used to screen bta-miR-133a related to the development of beef cattle skeletal muscle. Use bioinformatics analysis software to identify its conservation and predict its target genes, perform GO enrichment analysis and KEGG pathway enrichment analysis for its target genes, and verify the predicted potential target genes through the dual luciferase report test. C2C12 cells were selected for functional verification, bta-miR-133a was overexpressed and knockouted. After 48 h, 2% horse serum was used to observe the myotube differentiation process, CCK-8 to detect cell proliferation rate, Real-time quantitative polymerase chain reaction to detect the expression of cell proliferation and differentiation marker genes PCNA, CCND1, Myh1, Myod1 and target genes. The results showed that the mature sequence of bta-miR-133a was highly conserved among various species. The target gene was predicted to be PAX7. The dual luciferase report test verifies that bta-miR-133a had a target relationship with PAX7.The result of GO enrichment and KEGG pathways analysis showed that the predicted target genes were significantly enriched in the regulation of actin cytoskeleton organization tissue, the negative regulation of the classical Wnt signaling pathway, actin filament binding and other GO components, and MAPK, Ras, FoxO and other signaling pathways related to muscle development in. Overexpression of bta-miR-133a promoted myotube differentiation(P <0.01), at 72 h, reduced cell proliferation rate(P <0.01), inhibited the expression of target gene PAX7 (P <0.01);knocked out bta-miR-133a inhibited myotube differentiation(P <0.01), increased cell proliferation rate at 72 h(P <0.05), and promoted the expression of target gene PAX7 (P <0.01). In terms of cell proliferation, overexpression of bta-miR-133a reduced the expression level of its marker genes PCNA and CCND1 (P <0.05), while the knockout group was the opposite. In terms of cell differentiation, overexpression of bta-miR-133a increased the expression level of the marker genes Myh1 and Myod1 (P <0.01), while the knockout group was the opposite. In summary, bta-miR-133a may participate in the development of longissimus dorsi by targeting and negatively regulating the expression of PAX7 to inhibit the proliferation of myoblasts and promote their differentiation.
  • DAI Liangxiang, DING Hong, SHI Xiaolong, XU Yang, ZHANG Guanchu, QIN Feifei, ZHANG Zhimeng
    To assess the peanut rhizosphere bacteria community structure in response to salt stress at different development stages, the peanut variety Huayu 25 was used as experimental material, and a pot experiment was set with three salt concentrations to study the effect of salt stress on peanut yield and analyze the changes of microbial community structure of peanut rhizosphere at flowering and harvest stage under salt stress by high-throughput sequencing technology. The results showed that the rhizosphere microbial composition of peanut were basically similar under different salt stress treatments, but the diversity and richness significantly varied between flowering and harvest stage. Under higher salt stress, the diversity and richness of rhizosphere bacterial community were increased at the flowering and needling stage but decreased at the harvest stage. The dominant bacteria phyla of all soil groups were Proteobacteria, Actinobacteria, Chloroflexi, Acidobacteria, Verrucomicrobia, Bacteroidetes, and Patescibacteria. The relative abundance of Cyanobacteria, Gammaproteobacteria, Verrucomicrobiae and Bacteroides significantly increased under salt stress, especially at the flowering and needling stage. Hierarchical clustering revealed that the microbial community diversity was markedly altered by the salt concentrations and growth stages, samples of the same growth period were clustered into one group under salt stress. KEGG functional prediction analysis indicated that sequences related to carbohydrate metabolism, amino acid metabolism, energy metabolism, and metabolism of cofactors and vitamins were enriched, whereas that of signal transduction mechanisms, lipid metabolism, replication and repair, xenobiotics biodegradation and metabolism, metabolism of other amino acids, and folding, sorting and degradation were decreased. Among them, salt stress increased the abundance of functional groups involved in substance and energy metabolism, membrane transport, translation, replication and repair, and signal transduction, but decreased the 100 fruit weight and 100 kernel weight of peanut resulted in the decrease of peanut yield. Therefore, salt stress had a great impact on the peanut rhizosphere bacterial community structure and peanut yield. The salt tolerance of peanut could be enhanced by improving soil microbial environment. The results provided a theoretical basis for the development of peanut production in saline-alkali areas.
  • ZHAO Man, QI Zhi
    To explore to study the effect of exogenous calcium on the tolerance of Leymus chinensis, the seeds which collected from Horinger County, Hohhot City, Inner Mongolia were used as experimental materials, and the method of aseptic culture was used to study the effect of CaCl2 on the growth and physiological characteristics of Leymus chinensis under four abiotic stresses of salt, alkali, osmotic and cold stress. The results showed that the growth of Leymus chinensis seedlings was significantly inhibited under 150 mmol/L NaCl stress. After adding 1 mmol/L CaCl2 to the culture medium, the germination rate of seeds was significantly increased, and the root length, leaf length and fresh weight of seedlings were also increased. Under pH 8.5 or 300 mmol/L Mannitol stress, the growth of Leymus chinensis seedlings was both significantly inhibited. The effects of alkali stress and osmotic stress on the growth of Leymus chinensis seedlings could not be effectively alleviated by adding 1-20 mmol/L CaCl2 to the culture medium respectively. Under 4℃ stress, the growth of Leymus chinensis seedlings was significantly inhibited. After adding 20 mmol/L CaCl2 to the culture medium, the leaf length and fresh weight of seedlings increased significantly, the content of malondialdehyde(MDA) in leaves decreased, the activity of superoxide dismutase(SOD), peroxidase(POD), catalase(CAT) and glutathione reductase(GR) increased, and the content of reduced ascorbic acid(AsA) increased significantly. In conclusion, the addition of exogenous CaCl2 could improve the salt and cold resistance of Leymus chinensis seedlings, but the ability to resist alkali and osmotic stress has not been improved.
  • LI Jingyi, WANG Ren, MENG Xiangjie, LIANG Yugang, GONG Xiangsheng, HUANG Huang, WEI Hailin
    In order to explore the effects of no tillage and semi-solid sowing mode on the physiological and photosynthetic characteristics of flag leaf of rice. Field comparative experiments were designed to study the physiological and photosynthetic characteristics of flag leaf of rice under different cultivation modes, including tillage semi-solid direct seeding(FZ), no tillage semi-solid direct seeding(MZ), rice transplanting(YZ) and traditional direct seeding(CK). In 2016, the flag leaf SPAD values of FZ and MZ treatments of early rice Zhongzao 39 and Xiangzaoxian 45 were significantly higher than those of CK treatment from milk stage to maturity stage. In 2017, the growth rates of FZ, MZ and YZ treatments of early rice Zhongzao 39 were 0.85%-15.41%, 1.60%-13.04% respectively compared with those of early rice and late rice of CK treatment, and the differences were significant at heading+28 d and heading + 35 d. The POD and SOD in flag leaves of FZ, MZ and YZ increased after heading, but decreased MDA. The average decrease of MDA content at each growth period of Zhongzao 39 was 13.35%, 9.35%, 12.49%, 9.43%, 9.53% and 7.12%, respectively, and that in Xiangzaoxian 45 was 10.22%, 7.75%, 10.01%, 6.72%, 8.44% and 6.01%, respectively. The soluble sugar content in flag leaves of FZ, MZ and YZ treatments was significantly higher than that of CK from 14 days to 35 days after heading. In the late rice season of 2017, the transpiration rate of FZ, MZ and YZ treatments was significantly higher than CK in the heading + 25 d, and the net photosynthetic rate was significantly higher than CK at heading stage and 10 days after heading by 10.56%, 11.13%, 10.29% and 12.40%, 14.46% and 15.65% respectively. Semi solid direct seeding and transplanting could improve SPAD value and chlorophyll content of flag leaves, which was beneficial to improve photosynthetic capacity of flag leaves, laid a foundation for accumulation of dry matter and organic matter after full panicle, and then improved rice yield.
  • WANG Zhenshan, YUAN Xilei, YAN Liuyan, YANG Pengjuan, SANG Luman, JIA Xiaoping, BAI Junyan
    In order to reveal the possible functional effects of SiCCT gene, and provide foundation for further research of the function of SiCCT gene in photo-thermal interaction-controlled flowering process of foxtail millet, the photoperiod sensitive millet variety Huangmaogu was used as experimental material to analyze the diurnal expression patterns of SiCCT gene under long-day and high temperature(LD, 27℃), long-day and low temperature(LD, 22℃), short-day and high temperature(SD, 27℃), short-day and low temperature(SD, 22℃), totally four photo-thermal combinational conditions by fluorescence quantitative PCR. Furthermore, based on the survey data of ten main agronomic traits for two consecutive years, candidate gene-based association analysis was performed. The results showed that the expression of SiCCT gene was not affected by temperature, and the expression level of SiCCT gene under long-day was wholly higher than that under short-day at light period. While at darkness period, the expression of SiCCT gene was affected by temperature, especially under short-day, the expression level of SiCCT gene at low temperature was obviously higher than that at high temperature. Based on the association analysis of candidate gene, it was found that 11 polymorphic loci detected in SiCCT gene were significantly associated with 8 major agronomic traits(P <0.05), among which, SNP-10 was associated with heading stage, leaf number and grain weight per panicle at Luoyang, Henan and Jilin, Jilin, SNP-100 and SNP-104 were associated with plant height, leaf number and panicle length at Luoyang. SNP-51 was detected at Ledong of Hainan, Luoyang and Jilin for two consecutive years, but the related traits were different, which were ear diameter, branch number per panicle and leaf number at Ledong, Luoyang and Jilin, respectively.The expression of SiCCT gene was controlled by photoperiod, and temperature also affected its expression at darkness period under short-day. SiCCT gene possessed pleiotropic effects, but its function was regulated by photoperiod condition.
  • LEI Qidong, SUN Xudong, XU Huini
    To investigate the function of AtTCP4 downstream target genes and in plant growth and development. Bioinformatics analyzed transcription factor TCP4 family and its protein sequence.The prokaryotic expression vector was constructed and the protein was induced and purified. EMSA was done to analyze AtTCP4 protein can bind to the cis-acting elements in the promoter region of LOX2 gene.The bioinformatics analysis showed that the transcription factor TCPs of Arabidopsis thaliana contain a conserved bHLH domain, and the phylogenetic tree showed that AtTCP4 belonged to the class Ⅱ TCP protein. The prediction of protein tertiary structure indicated that TCP4 could form homodimer or heterodimer. Specific primers were used to obtain the CDS sequence of AtTCP4 gene from Arabidopsis thaliana, whose open reading frame (ORF) was 1 263 bp. The prokaryotic expression vector pET-28a-AtTCP4 was constructed and transformed into Rosetta (DE3) competence. After IPTG induction, protein purification and Western Blot detection proved that the AtTCP4 protein was obtained. EMSA experiment verified that the purified AtTCP4 protein could bind to the cis-acting elements of gene LOX2. It affected biosynthesis of plant hormone jasmonic acid and regulated the process of plant growth and development.
  • LUO Xiaoyu, SHAN Chenyang, JIA Zhaozhao, ZHANG Xin, LIN Ling
    In order to further utilization of the endophytic antagonistic bacterium S258 isolated from cotton root for biological control, especially to use the volatile organic compounds(VOCs) produced by the strain to control plant disease, Verticillium dahliae and Fusarium oxysporum f. sp. niveum were used as the target pathogenic fungi, and Arabidopsis thaliana was used as the indicator plant, the antifungal activity of the VOCs from the strain against pathogenic fungi and the growth promoting activity against plants were determined by the dual culture on two-section of a Petri dish. The results showed that the VOCs from the endophytic bacterium S258 could inhibit the mycelial growth of pathogenic fungi and at the same time promoted the plant growth. Compared with the negative control E. coli DH5α, the growth inhibition rate of the VOCs against Verticillium dahliae and Fusarium oxysporum f. sp. niveum were 80.9% and 10.1%, respectively. Compared with the blank control without bacteria, the fresh weight per plant of Arabidopsis thaliana treated by the VOCs was 5.3 times that of the blank control. The VOCs from the endophytic bacterium S258 were collected by headspace solid-phase microextraction. Then the compositions of VOCs were analyzed by GC-MS. The VOCs were identified by NIST and WILIY mass spectrometry databases as 30 monomer compounds, belonging to alkanes, alkenes, alcohols, ketones, ethers, aldehydes, esters and heterocycles. According to the peak area of total ion current graph, dimethyl disulfide and indole were most abundant among these compounds. In conclusion, the endophytic bacteria S258 can produce volatile organic compounds with antifungal and growth-promoting activity, which is an important microbial resource for the development of biocontrol products, and its volatile organic compounds also have high research and application value.
  • YANG Ling, HAN Peipei, ZHANG Yuqin, DAI Jing, LI Yinshui, GU Zhiming, SHEN Xinjie, XIE Lihua, QIN Lu, LIAO Xing
    In order to explore nitrogen(N) or magnesium(Mg) deficiency on nutrient accumulation and N assimilation of rapeseed at the seedling stage, rapeseed cultivar ZS11 was applied in hydroponics and four treatments were set up, including control(CK), low N(-N), low Mg(-Mg), low N and low Mg simultaneously(-N-Mg). The effects of N or Mg deficiency on N and Mg uptake and accumulation, as well as key enzymes activity involved in N assimilation were analyzed. Results showed that, both N and Mg deficiencies significantly affected the growth of rapeseed shoots, compared with CK, shoots biomass were significantly decreased by 68.49%, 35.49%and 69.71% under -N, -Mg and -N-Mg treatments, respectively. While N starvation stimulated root growth and significantly increased root-shoot ratio, Mg deficiency interaction inhibited root growth and the root-shoot ratio of rapeseed seedling was significantly reduced. And N and Mg interaction also had significant effect on the absorption and accumulation of N and Mg of rapeseed, the Mg content and accumulation of rapeseed root were significantly increased under -N treatments, while the N content and accumulation of rapeseed shoots were dramaticlly reduced by -Mg treatment, whereas, Mg deficiency could lead to the significant enhancement of NH4+ content, and the effects on the N and Mg uptake and accumulation which occurred in -N-Mg treatment were mainly originated from the lack of N. Further comparison in the key enzymes activity refer to N assimilation pathway under different treatments found that, compared with CK, both the NR and GS activities in young and old leaf, together with NR, GS and GOGAT activities in root of rapeseed were all significantly repressed under -N treatments. The activity of GS were also significantly reduced under -Mg treatments, but the decrement was obviously less than that of -N treatment, however, the activities of NR and GOGAT were significantly increased under -Mg conditions. In conclusion, N and Mg deficiencies took significant interactive effects on N and Mg accumulation of rapeseed at the seedling stage, additionally, N deficiency could reduce N uptake and accumulation and then inhibit the N assimilation pathway, but rapeseed could promote the assimilation process between inorganic N and also the process involved in inorganic N to organic N through increasing ammonium N content and the activity of NR and GOGAT under Mg deficiency stress.
  • WU Xiaojun, LI Fengyuan, SHEN Yunxia, CHEN Xiangdong, HU Xigui, JIANG Hao, ZHANG Xuening, LI Xiaoli, HU Tiezhu, RU Zhengang
    In order to effectively utilize the introduced wheat germplasm resources and explore the breeding application value of some wheat grain weight genes, the 1000-kernel weight(TKW) of 48 introduced wheat germplasms were identified, and the allelic variation types and the distribution of high TKW alleles were detected by using the functional molecular markers of four grain weight genes(TaGW2-6ATaCwi-A1TaGS2-D1 and TaSus2-2B). The results showed that the average TKW of the introduced germplasm materials was 30.15 g, the variation range was 20.13-43.19 g, and the germplasm materials above the average accounted for 45.8%. There were 8 haplotypes in the 4 grain weight-related genes in the introduced wheat germplasms. The average TKW of high TKW haplotypes TaCwi-A1a of TaCwi-A1 gene and Hap-6A-A of TaGW2-6A gene were significantly higher than the corresponding low ones(TaCwi-A1b and Hap-6A-G). The average TKW of the low TKW haplotypes TaSus2-2Bb of TaSus2-2B gene and TaGS2-D1b of TaGS2-D1 gene were significantly higher than the corresponding high TKW haplotypes TaSus2-2Ba and TaGS2-D1a. Generally germplasm materials containing the same haplotype combination of each grain weight gene accounted for a relatively higher proportion of the tested materials, and their average TKW was relatively higher. Studies indicate that the grain weight-related traits of these wheat germplasms have good genetic improvement potential, and it is necessary to analyze the comprehensive genetic effects of related grain weight genes in wheat breeding.
  • ZHAI Lingxia, YU Song, HOU Yulong, QIN Meng, ZHU Xuetian, WANG Xiaoqin, YU Lihe
    In order to further reveal the role of kidney common bean NAC gene in regulating plant growth and development and in response to abiotic stress, this research was used the salt-tolerant common bean variety HYD as the test material and used the Illumina HiSeq technology, the transcriptome of common bean leaf tissue under the NaHCO3 and NaCl treatments was constructed. A total of 8 saline-alkaline related NAC transcription factors were screened from the transcriptomic data, and carried out analysis of physicochemical property, systematic evolution, phosphorylation site prediction, protein secondary structure prediction, promoter element analysis, gene structure analysis, chromosomal location analysis and expression analysis. The result showed:The molecular weight of 8 common bean saline-alkaline response NAC genes was 23 676.36-44 354.33 ku, and the protein codes were 203-394 amino acids. The isoelectric point was between 4.74 and 8.86, of which 3 genes encoded acidic proteins, and 8 genes were divided into 4 subfamilies (a-d), positioning in 8 chromosomes, including 3 membrane-bound proteins, the number of serine was the largest in 8 proteins, there were 10-22, and the random coil structure accounts for the largest proportion, 68.0% to 84.5%. All 8 proteins were located in the nucleus. Under NaHCO3 treatment, 4 genes were up-regulated expression (log2FC>2), 2 genes were down-regulated expression (log2FC<-2), Under NaCl treatment, 1 gene was up-regulated expression (log2FC>2), 4 genes were down-regulated expression (log2FC<-2), 3 genes respond to both NaHCO3 and NaCl. A total of 10 promoter elements were found in 8 gene promoters, and each gene contained 3-7 elements. It indicated that these genes might be involved in the stress response of common beans.
  • WU Honghong, DUAN Xuefen, GUO Yangdong, ZHANG Xichun
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    Tomato belongs to the cold sensitive model plant,which is vulnerable to chilling injury during its growth,thus affecting its yield.In order to provide a theoretical basis for further revealing the molecular mechanism of cold resistance of tomato plants and breeding tomato cold resistant varieties,tomato germplasm resource No.25 of vegetable genetics,Breeding and Biotechnology Laboratory was used as material,and tomato transcription factor SlMYB-related 2 was used as research object.Based on CRISPR/Cas9 gene knockout vector,tomato positive plants were obtained by Agrobacterium transformation.The VIGS transient silencing expression vector was constructed,and the wild-type and and virus transformed plants without inserted target gene fragment were used as control.The three groups of plants were treated with low temperature at 4 ℃(16 h day / 8 h night,60% humidity).The contents of physiological indexes related to cold resistance were measured and its cold tolerance was compared.The results showed that CRISPR/Cas9 gene knockout vector was constructed and two CRISPR silenced positive plants were obtained.Constructed the expression vector of VIGS silence,after low temperature treatment,it was found that with the extension of low temperature treatment time,the growth trend of proline and soluble sugar in SlMYB-related 2 gene transformation group was slower than that in control tomato group(wild type carrying TRV,CK)and wild group(WT),and the content was lower than that in CK group and wild group.However,the content of malondialdehyde was higher than that of CK group and wild group,which proved that the cold resistance of tomato plants with transient expression of VIGS was significantly lower than that of CK group and wild group.Through the analysis of cold resistance of tomato plants after gene transient silencing expression,it was found that SlMYB-related 2 gene played a positive regulatory role in low temperature stress.

  • XU Huan, DUAN Shengwen, FENG Xiangyuan, ZHENG Ke, YANG Qi, WANG Qiming, CHENG Lifeng, PENG Yuande
    In order to efficiently express the pectate lyase PelG403 from ramie biodegumming. This study replaced pel G403 from the recombinant plasmid pEASY-Blunt E1-G403 to the pET28a vector with a higher plasmid copy number, and then introduced Escherichia coli BL21(DE3) to induce expression. The enzyme activity was determined by DNS method, and the expression effect of PelG403 was tested by SDS-PAGE and Western Blot analysis, and the sequence was analyzed and identified. The results showed that the pectate lyase activity of pET28a-G403/BL21 was 335.8 U/mL, which was 3.05 times higher than that of pEASY-Blunt E1-G403/BL21;the molecular weight of the expressed product of pET28a-G403/BL21 was slightly smaller than that of the predicted fusion protein with His tag(43.6 ku);PelG403 contained 387 amino acids, and the first 35 amino acids at the N-terminal were signal peptides;the theoretical pI value was 7.64, 4 cysteines, and the instability coefficient was 27.42;its protein domain contained a typical right-handed β-helix structure and belonged to the Pec lyase C family. Therefore, replacing pelG403 from the recombinant plasmid pEASY-Blunt E1-G403 into the pET28a vector and introducing E.coli BL21(DE3) for inducible expression was a method and approach for high-efficiency expression of the ramie degummed pectate lyase gene pelG403, and its sequence was analyzed and identified. It laid a foundation for the purification of pectate lyase, key site analysis, molecular modification and the study of its degumming functional characteristics.
  • GAO Yanlong, ZHANG De, ZHANG Rui, ZHANG Zhongxing, ZHAO Ting, WANG Shuangcheng, WANG Yanxiu
    To explore the effects of spraying CaCl2 at different concentrations on physiological characteristics of apple seedlings under salt stress, two-year-old T337 seedlings were used as test materials, and the pot experiment was used to set 100 mmol/L NaCl(T1) and 100 mmol/L NaCl + four different concentrations of CaCl2 (5.0(T2), 7.5(T3), 10.0(T4), 12.5 mmol/L(T5)) treatment, watering with deionized water was used as the control(CK).Determine the chlorophyll content, photosynthetic and fluorescence parameters, antioxidant enzyme activity and osmotic adjustment substance content of each treatment group, and carry out correlation and principal component analysis. The results showed that with prolonged stress time, the catalase(CAT), peroxidase(POD), superoxide dismutase(SOD) of T337 leaves first increased and then decreased, malondialdehyde(MDA), intercellular CO2 concentration(Ci), proline(Pro) and relative conductivity(REC) showed a rising trend, initial fluorescence(F0), PSⅡ maximum light energy conversion rate(Fv/Fm) stomatal conductance(Gs) and the transpiration rate(Tr) showed a downward trend;compared with CK and T1, the variation amplitudes of T337 leaves after treatment with exogenous CaCl2 were all reduced, and there was a significant concentration effect. Among them, the relative chlorophyll content(SPAD), Pn, Tr, POD, F0 and SOD decreased the least under T4(10mmol/L) treatment, which was significantly higher than other treatment groups;MDA, REC and Ci increased the least, which was significantly higher than CK and lower than others processing group. Correlation analysis of 14 indicators showed that Pn was positively correlated with CAT, Tr, Gs, F0, Fv/Fm, ETR, SPAD, SOD and POD, and negatively correlated with Pro, MDA, Ci, and REC. Principal component analysis was performed on 14 related indicators after salt stress treatment, and 2 principal components with eigenvalues greater than 1 were extracted, and their eigenvalues were 11.190, 2.295, respectively;The first and second principal component variance contribution rates were 79.930%, 16.394%, respectively, the cumulative variance contribution rate reached 96.324%, which meets the analysis requirements. According to the ranking of principal component scores, the mitigation ability of exogenous CaCl2 to T337 salt stress was T4>T3>T2>T5 from high to low. Therefore, 10 mmol/L CaCl2 could improve the photosynthetic fluorescence characteristics of T337 rootstock leaves under salt stress, improve the antioxidant enzyme system and osmotic adjustment ability, and enhance the stability and permeability of biofilms to alleviate salt stress.
  • PENG Ligong, GONG Jing, LAN Yan, WANG Jin, SUI Xiaodong, DING Chunbang, LI Tian
    This study aimed to investigate the effect of different water stress stages and duration on rice starch synthesis and yield. With rice cultivars Yixiangyou 2115 as experimental materials, the content of soluble sugar and starch, the activities of enzymes related to starch synthesis, and yield of a rice grain under four water management modes traditional flooding(CK, 0 kPa, 1-3 cm water layer), light water stress(LD, (-20±5) kPa), moderate water stress(MD, (-40±5) kPa) and severe water stress(SD, (-60±5) kPa) at booting stage, flowering stage and filling stage respectively were analyzed in 2018-2019. The results showed that:the content of soluble sugar and sucrose were decreased and the content of amylose and total starch was increased under light water stress at the booting stage, flowering stage, and filling stage. The largest drop had reached 33.7%, 14.3% and the maximum raise had reached 2.4%, 4.4% respectively. On the contrary, moderate water stress and severe water stress had the opposite effect on them;The activities of enzymes related to starch synthesis such as AGPase, SSS, GBSS, SBE, and DBE all showed the variation tendency of falling after rising under four water management modes at different stages. Enzymes activities and their peak value were enhanced under light water stress at the middle and late filling stage, but moderate water stress and severe water stress had the reverse effect on them. Light water stress with a higher seed setting rate increased the yield by 3.1 percentage points only at the filling stage compared with traditional flooding. 1000-grain weight was increased at all stages under light water stress which the maximum raise had reached 3.7%. However, water stress at the booting stage was not favorable to the yield. Light water stress at the filling stage and flowering stage increased the yield by 0.8%, 9.3% respectively. In conclusion, light water stress at the flowering stage and filling stage could improve the activities of key enzymes for starch synthesis and the synthesis and accumulation of starch in grains of Yixiangyou 2115, thereby increasing the yield. Compared to light water stress at the flowering stage, light water stress at the filling stage had a higher increasing amplitude of yield.
  • YANG Zhongshuai, WU Jinzhi, HUANG Ming, LI Youjun, FU Guozhan, ZHAO Kainan, ZHANG Zhenwang, HOU Yuanquan
    In order to solve the main problems of dry land wheat production in China, such as low rainfall and uneven seasonal distribution, unreasonable fertilization method, the cultivation mode of dry land wheat was discussed.The experiment comprised four cultivation patterns, namely, conventional cropping with uniform fertilization(CK), furrow planting with uniform fertilization(FP), furrow planting with uniform fertilization but reduction of fertilizer application rate by 25%(FPRF), furrow planting with site-dressing fertilization but reduction of fertilizer application rate by 25%(FPOF). Tiller number, dry matter accumulation in the main growth period, soil water content of 0-200 cm at pre-sowing and maturity, yield, water use efficiency(WUE), partial fertilizer productivity of wheat were compared. Compared with CK, FP promoted soil water storage in the fallow season, thus increased soil water storage before sowing by 5.4%-5.5%, and increased the number of tillers and dry matter accumulation significantly in the main growth period, furthermore, the yield, WUE and partial fertilizer productivity were increased by 10.1%-11.2%, 7.2%-8.6% and 10.3%-11.4%, respectively. Compared with FP, FPRF significantly increased the fertilizer partial productivity by 22.9%-34.6%, had no significant effects on the yield and WUE in the first year, and significantly decreased them in the latter two years, but the above indicators were better than conventional plain cropping. Compared with the other three treatments, the yield, water and fertilizer use efficiency of FPOF were all the best, compared with FP, although the increase of yield and WUE was not significant, but the partial fertilizer productivity, were significantly increased by 35.2%-37.8%, respectively. Therefore, in the dryland of Western Henan, FP is beneficial to improve wheat yield and WUE, reducing fertilizer by 25% will reduce the yield to a certain extent, but it is beneficial to increase fertilizer partial productivity. FPOF is a suitable cultivation measure for winter wheat in arid areas due to synergistically improving yield, WUE and fertilizer partial productivity.
  • WEI Jiping, DAI Hang, LI Zhenyong, DING Zisu, TANG Ling, LUAN Xin, KE Shanwen, ZHANG Xiangqian
    Genetics analysis and fine mapping of osles3,a mutant with early senescent leaf in rice,were conducted to explore the molecular mechanism of rice leaf premature senescence.A leaf early senescence mutant osles3 was obtained from a natural mutant library of the Japonica variety Zhonghua 11.Phenotype and agronomic traits of osles3 and its wild-type were identified.The F2 population from mutant crossed to ZS97(Indica cultivar) was used to mapping the gene OsLES3 by the map cloning method,and the structure and function of OsLES3 were analyzed by bioinformatics method.Phenotypic observation showed that the mutant began to have water-soaked senescence spots in the late tillering period,and the whole plant senescence at maturity. osles3 mutant showed extremely significantly decreased in plant height,tiller number,seed setting rate,grain number per panicle and 1000-grain weight compared to the wild-type plant.Genetic analysis revealed that the mutant phenotype was controlled by a pair of nuclear recessive gene on the long arm of chromosome 3,between two markers RM15524 and RM15551 with 0.9,0.5 cM genetic distance,respectively.Morever,sequence polymorphorism and bioinformatic analysis revealed that 988 bp fragment insertion and 54 bp fragment deletion in 5' Čupstream of translation initiation ATG of OsLES3,which encoded a LSD1 zinc finger domain containing protein.This research completed the fine mapping of the rice premature leaf senescence mutant osles3,and revealed that the mutant phenotype might be caused by some DNA fragment deletion and insertion in OsLES3 gene.
  • JIANG Nan, ZHENG Qianqian, LI Qianwen, TU Jian, SONG Xiangjun, SHAO Ying, LIU Hongmei, QI Kezong
    Anhui Province Key Laboratory of Veterinary Pathobiology and Disease Control predicted a new ETT2 chaperone molecule YgeG in the early stage. To obtain a large number of active YgeG protein, identify its function, and screen out its related active ETT2 secretion effector protein, the prokaryotic expression conditions of the target protein were optimized.The ygeG gene was PCR amplified, with the APEC 81 strain(AE81) as the template. The amplified product was cloned into pGEX-6p-1 prokaryotic expression vector, and the recombinant plasmid pGEX-6p-1-ygeG was constructed and identified by sequencing. Then, the correctly identified recombinant plasmid was transformed into E. coli BL21(DE3), and was induced by IPTG. And optimizing the induction conditions of the recombinant protein to determine the optimal conditions for the expression of YgeG recombinant protein. The results of SDS-PACE and Western Blot showed that the optimal induction conditions were determined to be an induction temperature of 16℃, an induction time of 16 h, and IPTG final concentration of 0.25 mmol/L. The size of the recombinant protein was about 44 ku, which was highly expressed in BL21(DE3) and mainly in the form of soluble protein and has good immunoreactivity and antigenicity. The AE81-YgeG protein was successfully expressed, which laid the foundation for the further study of the structure and function of the protein and ETT2, and provided a theoretical basis for the prevention and treatment of avian colibacillosis.
  • YU Tianfei, XIE Pengyu, SUN Wanshu, LI Jing, YIN Haichang, LI Ming, YU Zhidan
    In order to express the light chain variable domain(VL) and heavy chain variable domain(VH) genes of the monoclonal antibody against Goose parvovirus(GPV) NS1 protein in E. coli, and determine their binding activity to the NS1 protein. The nucleotide sequences of the VL and VH genes of the monoclonal antibody against GPV NS1 protein were optimized according to the preferred codons of E. coli, and recombinant plasmids pUC57-VL and pUC57-VH containing variable region genes were artificially synthesized. Then pUC57-VL and pUC57-VH were double digested with Bam H Ⅰ/Xho Ⅰ, and 340 bp of VL gene and 370 bp of VH gene were recovered. The target genes were inserted into the prokaryotic expression vector pET-32a through the Bam H Ⅰ/Xho Ⅰ multiple cloning sites, and the recombinant plasmids pET-VL and pET-VH were obtained. The recombinant plasmids were identified by Bam H Ⅰ single restriction enzyme digestion and Bam H Ⅰ/Xho Ⅰ double restriction enzyme digestion and sequencing. The recombinant plasmids were transformed into E. coli Rosetta(DE3), and induced by IPTG. The recombinant proteins TRX-VL and TRX-VH were expressed with molecular weights of 30.3, 31.4 ku, respectively. The purified recombinant proteins could specifically bind to the His tag monoclonal antibody. And the identification results showed that 0.4 μg of TRX-VH could specifically bind to 25 ng of GST-NS1 protein, but 0.4 μg of TRX-VL could not specifically bind to each coating amount of GST-NS1 protein. The specific binding of TRX-VH and GST-NS1 protein could be completely blocked by 1:200 diluted GPV infected goose serum.
  • DONG Jiaxi, TIAN Xiuping, ZHAO Qiu, SHI Xinqian, YUAN Miaomiao
    In order to promote the further promotion of green manure planting in Tianjin, a field experiment was adopted, and 9 spring Brassica napus L. varieties were overturned with spring fallow fields as a control to study the effects of different spring Brassica napus L. on soil potassium content and maize nitrogen metabolism.The results showed that there were differences in the biomass and nutrient content of different spring Brassica napus L. varieties. Among them, Zhongyoufei 1901(7 716.50 kg/ha), 1804(6 577.02 kg/ha), and 1907(6 457.03 kg/ha) had higher biomass and nutrient content.The change trend of soil total potassium and available potassium in 2019 was the same as 2020. Among them, the content of total potassium and available potassium in the soil under the treatment of spring Brassica napus L. of Zhongyoufei 1901, 1804, and 1907 was higher than that of other tested varieties in different periods, and the total potassium and available potassium content of soil in 2020 were higher than those in 2019.The potassium uptake by the whole plant of maize and the total potassium uptake by the hectare of the three varieties of maize were higher than those of other tested varieties. Compared with 2019, the potassium uptake of the whole plant of maize treated with different spring Brassica napus L. in 2020 increased by 15.70%-24.34%;the NR and GS activities of the ear leaves in different spring Brassica napus L. treatments increased by 2.16-14.22 nmol/(min · g), 0.99-2.30 μmol/(h · g), respectively. The top 3 NR and GS activities of the maize ear leaves were the same as the potassium uptake of the whole maize plant. Compared with spring fallow, spring Brassica napus L. treatments' yield of maize increased by 10.02%-33.47%, which was 1.09-1.41 times of the spring fallow, and the highest yield in two years was 15 700.94 kg/ha (2020 Zhongyoufei 1901). It could be seen from the path analysis that the nitrate reductase activity of the leaf at the ear position had a direct effect on the protein content of corn grains.In the indirect effect, leaf protein content contributed the most to maize grain protein through leaf nitrate reductase activity.Correlation analysis showed that there were significant and extremely significant positive correlations among Brassica napus L. total potassium, soil potassium, maize potassium uptake, key enzymes in maize leaf nitrogen metabolism, maize leaf protein content, and maize grain protein content.
  • SUN Bin, TANG Lin, ZHANG Junfang, SUN Jianfu, CUI Yan, WANG Ying, WANG Enze, LI Qiang, LI Xiangzi
    The aim was to clone the CDS region sequence of Yanbian cattle UCP1 (Uncoupling protein-1), use RT-PCR technology and gene cloning to obtain Yanbian cattle UCP1 gene, conduct homology comparison with other species and construct a phylogenetic tree, using bioinformatics methods to predict the physical and chemical properties of UCP1 encoded protein, potential phosphorylation sites and other properties, and use Quantitative Real-time PCR (qPCR) method to detect the expression abundance of UCP1 gene in different tissues of Yanbian cattle. The results showed that the CDS region of Yanbian cattle UCP1 gene was 749 bp in length, encoding 249 amino acids. The homology comparison revealed that the UCP1 gene of Yanbian cattle had 99.47% homology with Indian cattle. The phylogenetic tree showed that Yanbian cattle and Indian cattle had the closest genetic relationship and the farthest related to South American alpaca. The UCP1 protein lacked stability, the fat solubility coefficient was 91.53, the half-life of reticulocytes in vitro was 30 h, and the total average hydrophilicity was equal to 0.212, which was predicted to have a certain degree of hydrophilicity. The secondary structure was a mixed protein composed of α-helix (54.03%), extended chain (12.90%), random coil (27.42%) and β-turn (5.65%). The analysis of phosphorylation sites and glycosylation sites showed that UCP1 had a total of 21 phosphate sites, 4 potential O-glycosylation sites, and 3 N-glycosylation potential sites. The encoded product of UCP1 gene had 0 transmembrane helix (TMHs) structure, the predicted value of the amino acid residues of the transmembrane helix was 19.008 57, and the predicted value of the transmembrane helix of the first 60 amino acids of the protein was 10.980 09, which was the total probability of being located on the cytoplasmic side of the membrane 22.995% and was a non-secreted protein. Real-time fluorescent quantitative PCR (RT-PCR) showed that the expression level of UCP1 gene in Yanbian beef fat and small intestine was relatively high, while the expression level in muscle and heart was low. The experimental results could provide a reference for further research on the function of this gene.
  • CHEN Dingshuang, WANG Ruilong, LIN Yaqiu, WANG Yong, ZHU Jiangjiang, LI Xin, ZHANG Hao, LI Yanyan
    Hoxa5 is a member of the HOX family and plays an important role in the process of growth,development and tumorigenesis.However,there is less research on it in ruminants.In order to obtain the goat Hoxa5 gene sequence,to clarify its expression characteristics and know its expression level in different tissues of goats.The Hoxa5 gene was cloned from the subcutaneous fat tissue of Jianzhou big ear sheep using RT-PCR technology,and its biological characteristics were analyzed by bioinformatics methods.Real-time fluorescent quantitative PCR technology was used to detect the expression of Hoxa5 gene in the heart,liver,spleen,lung,kidney,back,thigh,arm,etc.tissues of Jianzhou big ear sheep.The results showed that the open reading frame of the goat Hoxa5 gene was 813 bp,encoding a total of 270 amino acids.the relative molecular mass of Hoxa5 was 29.283 ku,the isoelectric point was 9.42,and there was no signal peptide and transmembrane domain.This protein was a non-secreted protein;the secondary structure prediction of Hoxa5 protein found that the content of random coils in Hoxa5 protein was the highest,followed by α helix,and β turn accounts for a relatively small number of amino acids;amino acid homology comparison results showed that the Hoxa5 amino acid sequence similarity between Jianzhou big ear sheep and Ovis aries,Bos taurus, Equus caballus, Sus scrofa, Rattus rattus, Cavia porcellus, Mus musculus and Homo sapiens were 100.00%,99.63%,99.26%,99.26%,98.89%,98.89%,98.89% and 98.15%.The results of Real-time fluorescent quantitative PCR showed that Hoxa5 gene was differentially expressed in selected tissues,with the highest expression level in the kidney;Fluorescence localization of pEGFP-Hoxa5 fusion vector transfected into goat subcutaneous adipocyte showed that the gene was mainly located in the nucleus.This study showed that the goat Hoxa5 gene was up-regulated during subcutaneous adipocyte differentiation and the protein was localized in the nucleus. The results suggested that this gene might act as a transcription factor to regulate goat subcutaneous adipocyte differentiation.
  • JIN Fengmei, XUE Jun, SUN Haibo, HAO Zhiyu
    In order to clarify the cause of tomato fruit browning in Tianjin, deep sequencing of small RNA and RT-PCR were used.The results showed that this symptom was caused by Tomato mosaic virus (ToMV). The sequences of CP(Coat Protein, CP), MP(Movement Protein, MP), and 126 proteins of the virus were further analyzed. Phylogenetic analysis showed that the three genes of the Tianjin isolate(ToMV-TJ) were the closest to the American isolate(USA_99-1).The similarity analysis showed that the average similarity of ToMV-TJ CP gene to other CP gene isolates was 95.53%.The average similarity of MP gene to other MP gene isolates was 95.21%.The average similarity of 126 protein gene to other 126 protein gene isolates was 92.72%.The proteins encoded by these three genes were analyzed. The results showed that the average similarity was above 99.0%. It is initially shown that ToMV-TJ isolate was imported through international trade.
  • GAO Guangliang, ZHANG Keshan, XU Guoyang, ZHAO Xianzhi, CHEN Zhuping, ZHOU Li, WANG Qigui
    To investigate the polymorphism of GnIH gene association with egg-laying or body measurement traits in Goose, we cloned the GnIH DNA sequence, and analysis the character of the genes with that of mammal and poultry. Moreover, we detected the GnIH DNA sequence in Sichuan White goose populations (208 individuals), and performed the association with the polymorphism of the GnIH gene with egg-laying and body composition traits. In this study, we cloned a 3 735 bp genomic sequence and a 522 bp mRNA sequence for GnRH, consisting with 3 exons and 2 introns, code 173 amino acid pre-protein. Genetic evolution analysis had shown that the similarity of GnIH gene sequence between Sichuan white goose and mallard duck was the highest (99%). Using a direct sequencing method, 26 single nucleotide polymorphisms (SNPs) of GnIH genomic sequence were obtained, and the association analysis has revealed that the 11 SNPs(g.811T>C, g.1892T>C, g.1320G>A, g.1809G>T, g.2164C>G, g.1708A>G, g.2164C>G, g.1809G>T, g.1892T>C, g.1929C>A和g.2164C>G) were significant or extremely significant associated with five traits, including first egg weight (FEW), body length (BL), neck length (NL), chest width (CW) and shank circumferences (SC) (P<0.05 or P<0.01). However, there was no significant correlation with egg number at 48 weeks and 64 weeks, birth weight, semi-submersible length, keel length and pelvic width (P>0.05). In conclusion, we cloned 3 735 bp goose GnIH DNA genomic sequence, and the 11 of the 26 SNPs of of the GnIH gene can be used as one of the potential molecular markers of the traits such as first-laying egg quality, body length, neck length, chest width and tibial girth.
  • SUN Lili, JIN Xiujuan, ZHAO Kai, Md Ashraful Islam, LU Juan, WANG Shuguang, SUN Daizhen
    Abstract (112) PDF (124) RichHTML (29)

    Cysteine protease,a kind of hydrolase in the process of protein degradation,participates in the senescence and maturation of plants and plays an important role in the growth and development of plants.In this study,a senesence-specific cysteine protease(SAG39)was screened from the previously obtained transcriptome data of wheat at senescence stage.In order to study the role of TaSAG39 gene in wheat at senescence,the gene structure and expression pattern of TaSAG39 were analyzed by bioinformatics analysis and Real-time fluorescence quantitative(qRT-PCR)technology.The results showed that the amino acid sequence encoded by the cysteine protease gene SAG39 had the closest relationship with the ancestral species of wheat,such as Aegilops tauschii,Triticum dicoccoides and Triticum durum,and it included the unique active site Cys-His-Asn and EFNIN structures of the papain sub-family.The full-length genomes of TaSAG39-5A,TaSAG39-5B and TaSAG39-5D were 1 455,1 435,1 439 bp,respectively,and the length of the coding sequence was 1 041,1 050,1 038 bp,respectively,with each containing two exons and one intron.The TaSAG39s proteins consisted of 346,349,345 amino acids.With relative molecular weight of about 37 ku and isoelectric point of 5.53—5.67,these proteins were stable negatively charged hydrophilic proteins.Its main components were random coils and α-helix,and there were signal peptides in the N segment of the protein.It had the conserved Inhibittor_I29 and Petidase_C1 domains,and contained 35—37 phosphorylation sites,mainly serine and threonine phosphorylation sites.TaSAG39 protein may interact with cysteine protease inhibitors and 14-3-3 protein.The promoter contained cis-acting elements relative to abscisic acid,methyl jasmonate,auxin,light,drought,low temperature and stress response.The results of qRT-PCR confirmed that gene TaSAG39 was induced to express by natural aging,darkness,drought,methyl jasmonate,high temperature and auxin,among which natural aging was the most significant.

  • TANG Yiping, LI Xiangfeng, WANG Hui, HU Wangqin, REN Chuting, HUANG Yaru, XU Peng, YOU Cuicui, KE Jian, HE Haibing, WU Liquan
    In order to improve grain yield and quality of large-panicle japonica rice, the two large-panicle japonica rice varieties, W1844 and CJ03 were grown in paddy field. The content of non-structural carbohydrate(NSC) in the stem and sheath before heading would be changed through the shading and removing tillering treatments, namely T0(control), S1(50% shading), S2(75% shading), D1(removing the whole row of rice plants in every other row), D2(removing the whole row of rice plants in every other row and the tillering of each hole, reserving only the main stem), to explore the effects of NSC on the grain-setting characteristics, grain filling characteristics and quality of superior and inferior spikelets in large-panicle japonica rice, and to clarify the correlation among carbohydrate-spikelet ratio(amount of non-structural carbohydrate in the stems and sheaths over the number of spikelets) with grains yield and quality formation. The results showed that D1 and D2 treatments significantly increased the carbohydrate-spikelet ratio at heading, filled-grain percentage and 1000-grain weight of inferior spikelets. Compared with the T0, the carbohydrate-spikelet ratio was increased by 47.84%-173.59%, filled-grain percentage of inferior spikelets was increased by 4.1-7.2 percentage points and 1000-grain weight of inferior spikelets was increased by 6.06%-14.29%. The carbohydrate-spikelet ratio at heading, filled-grain percentage and 1000-grain weight of inferior spikelets were significantly decreased under the S1 and S2 treatments. The carbohydrate-spikelet ratio was decreased by 33.28%-53.79%;filled-grain percentage of inferior spikelets was decreased by 6.8-32.8 percentage points and 1000-grain weight of inferior spikelets was decreased by 13.54%-45.02%. The D2 and S2 treatments significantly affected the early stage of filling for inferior spikelets. The filling time in the early stage for inferior spikelets was shortened by 6.80, 7.10 d under D2, respectively; and increased by 9.50, 8.26 d under S2, respectively. The mean grain-filling rate in the early stage for inferior spikelets was increased by 65.78%, 61.15% under D2, respectively; and decreased by 44.35%, 43.28% under S2, respectively. D2 treatments improved grain quality, while the S2 decreased grain quality. The correlation analysis indicated that carbohydrate-spikelet ratio had significantly or extremely significantly positive correlation with filled-grain percentage, 1000-grain weight, processing quality and amylose content of inferior spikelet, and had significantly or extremely significantly negative correlation with chalkiness, chalkiness degree and protein content of inferior spikelet. Together, the results showed that carbohydrate-spikelet ratio at heading stage could increase the grain filling in the early stage for inferior spikelets to enhance the filled-grain percentage and 1000-grain weight of inferior spikelet and improve rice quality.
  • HAN Zhenghong, DUAN Yuxuan, XU Shanbin, WANG Jingguo, LIU Hualong, YANG Luomiao, JIA Yan, XIN Wei, ZHENG Hongliang, ZOU Detang
    Abstract (111) PDF (88) RichHTML (38)

    In order to promote the breeding of long-grain japonica rice varieties,two knock-out vectors,pYLCRISPR/Cas9-GS3-RNA and pYLCRISPR/Cas9-GS3-GS9-RNA,were constructed with japonica rice varieties Dongfu 139,Longjing 31 and Dongnong 427,the genes of GS3 and GS9 were edited by Agrobacterium tumefaciens in vitro.In the end,GS3,GS9 and GS3,GS9 double-gene mutations without T-DNA elements were obtained in the T2.The results showed that the grain length and 1000-grain weight of gs3 mutant plants of three varieties were significantly increased compared with those of wild type,grain width,seed setting rate and grain number per panicle did not change significantly,grain length of gs9 mutant increased significantly,and grain width decreased significantly,while 1000-grain weight,seed setting rate and grain number per panicle did not change significantly,and grain length of gs3gs9 mutant increased more than gs3 and gs9,at the same time,grain width decreased significantly,1000-grain weight increased significantly,while seed setting rate and grain number per ear did not change significantly.To sum up,three japonica rice varieties,Dongfu 139,Longjing 31 and Dongnong 427 were improved by using CRISPR/Cas9 technique,which accelerated the breeding process of new long-grain japonica rice varieties.

  • ZHANG Jinyu, XU Xinjuan, CHAO Maoni, ZHANG Xiaohong, WU Xiangyuan, GAO Jitao, HUANG Zhongwen
    Abstract (111) PDF (17) RichHTML (117)

    Zinc finger proteins are transcription factors widely studied in eukaryotes,and play important roles in plant growth and development,and responses to stresses.In order to deeply understand the gene function of zinc finger protein in soybean,the full-length CDS sequence of GmZAT12 gene was cloned from Shangdou 1201,and the characteristics of coded protein by this gene was predicted by bioinformatics analysis.The subcellular localization of GmZAT12 protein was detected by the tobacco epidermal injection system, and the expression pattern of GmZAT12 gene in different tissues of soybean and abiotic stress was analyzed by Real-time PCR (qRT-PCR) technology. The results showed that GmZAT12 CDS contained 516 bp,encoding 171 amino acids and the molecular weight of the protein was 19.264 28 ku with a theoretical isoelectric point(pI)of 9.02.Its main components were random coils and α-helices and it contained 20 phosphorylation sites, mainly serine phosphorylation sites.Sequence analysis indicated that GmZAT12 possessed two conserved C2H2 zinc finger domains.The result of subcellular location indicated that GmZAT12 protein was localized in the nucleus.The results of qRT-PCR showed that CmZAT12 gene expressed mainly in roots,leaves and seeds of soybean,while low expression in flower and stem, and was induced by high temperature,low temperature,NaCl and ABA.The fact implied that this gene might be involved in abiotic stress signaling pathways.

  • CAI Zhaoming, CHENG Chunhong, FU Min, WANG Diandong
    To identify the candidate auxin response factors gene(ARF) responsive to salt and Plasmodiophora brassicae stresses in tuber mustard,and provide a basis for further studying their functions in the resistance of tuber mustard to stresses.Bioinformatic assay was used to analyze the cis-acting elements in the promoters of ARF family genes,and the fluorescent quantitative PCR method was used to analyze the expression patterns of ARF in different organs,as well as their expression levels under salt and Plasmodiophora brassicae stresses.The results showed that several cis-acting elements related to auxin,salt and pathogen stresses were found in the promoters of ARF genes. BjARF1A and BjARF4B were highly expressed in the leaves,and BjARF7C was highly expressed in the roots.After 3 h of salt stress treatment,the expression of BjARF1B and BjARF2B in root were severely induced;after 6 h of salt stress treatment,the expression of BjARF9A and BjARF13E were also significantly induced.Under the pathogen Plasmodiophora brassicae treatment,the expression of BjARF3A and BjARF3D were highly induced by the pathogen at 12,24 h,respectively;while the expression level of BjARF13B were continuously decreased,and the degree of decline gradually increased.In together,four salt stress responsive ARF genes and three Plasmodiophora brassicae responsive ARF genes were found,laying a foundation for the further study of ARF family gene functions.
  • WU Tingting, CHEN Hailong, HUANG Jun, LIANG Yi, ZHANG Ting, LI Bowen, LAI Yifan, ZOU Yuying, CHE Fanhao, WU Jun, LIU Jinling, LIU Xionglun
    To improve blast resistance of the CMS line Fengyuan A, a PCR-based co-dominant marker, ClonDF1R1, was developed according to sequence information of the broad-spectrum and durable blast resistance gene Pi9, and was employed in marker-assisted selection breeding practice.Resistance responses to 33 tested M.oryzae isolates showed difference between the Pi9 donor 75-1-127 and the receptors Fengyuan A and its maintainer Fengyuan B, presenting 87.9% and 33.3% resistance frequency respectively in greenhouse inoculation assay.75-1-127 showed high-level seedling blast resistance but highly susceptible for two receptors in field nursery.These results showed that the Pi9 gene was promising in rice blast resistance breeding, and blast resistance of two receptors needs to be improved urgently.The ClonDF1R1 was developed based on DNA sequence information of the Pi9 gene, this codominant marker primers amplified a 320 bp band from the donor genome while PCR product from two receptors was nearly 450 bp.The codominant molecular marker ClonDF1R1 was used in MAS backcross breeding practice for improving blast resistance of Fengyuan B and Fengyuan A respectively.Thirteen maintainer lines Fengyuan B-Pi9, and eleven CMS lines Fengyuan A-Pi9, carrying the Pi9 homozygous alleles and showing high-level seedling blast resistance, were developed.One improved CMS line, Fengyuan A-Pi9- 5, was bred through eleven-year consecutive MAS practice.Fengyuan A-Pi9- 5 showed high-level seedling blast resistance, stable male sterility performance, high exserted stigma percentage, excellent agronomic and yield traits.Thus, we achieved the goal to directionally improve blast resistance of rice CMS line Fengyuan A and its maintainer Fengyuan B, and created a set of new parents for three-line method hybrid rice breeding application.
  • MENG Fanlai, BAI Lei, GUO Huachun, ZHAO Dawei, YU Xinghua, WANG Yingmei, LI Yuxiang, TENG Juan
    In order to study the molecular mechanism of purple sweet potato leaf in response to enhanced UV-B radiation, the leaf of Dianziganshu 24(DZS24) was used as the test material and transcriptome sequencing analysis was performed by high-throughput sequencing technology under natural light and artificial supplementary the UV-B(T=7.2 kJ/(m2 · d)). The results showed that the expression of 477 genes(DEGs) were changed significantly, among which 382 and 95 were up-and down-regulated ones. GO analysis showed that these DEGs were mainly significantly enriched in oxidation-reduction process of biological process and oxidoreductase activity of molecular function. KEGG pathway analysis showed that a total of 9 pathways were significantly enriched, among which the biosynthesis pathway of secondary metabolites contained the largest number of DEGs. The DEGs of purple sweet potato leaf in response to enhanced the UV-B were mainly CHS (Tai6.1732, Tai6.53503, Tai6.45381) and DFR (Tai6.3019) in the flavonoid synthesis pathway as well as the CYP83B1 (Tai6.44115, Tai6.18064, Tai6.41206) in the glucosinolate biosynthesis and CYP85A1 (Tai6.2806, Tai6.19253, Tai6.15801) in brassinosteroid biosynthesis. The main transcription factors were C2H2, NAM, bHLH and RR-A-type. In summary, the key genes might be IbCHS, IbDFR, IbCYP83B1 and IbCYP85A1, as well as the main transcription factors might be C2H2, NAM, bHLH and RR-A-type in the purple sweet potato leaf response to the UV-B enhancement.
  • HAN Kaikai, YAN Ruofeng, LIU Qingtao, LIU Yuzhuo, LI Yin, ZHAO Dongmin, HUANG Xinmei, ZHANG Lijiao, YANG Jing, FU Chen
    The purpose of this study is to express the RdRp of DTMUV in eukaryotic cells and to analyze its structure and function by bioinformatics. Firstly,specific primers were designed and synthesized based on the sequence of the RdRp gene of DTMUV JS804 strain in GenBank,and then the gene was amplified by RT-PCR,subcloned into the pCMV-N-Flag eukaryotic expression vector,transfected into BHK-21 cells. Next,the expression of RdRp was determined by immunofluorescence observation and Western Blot. After that,bioinformatics software was used to analyze the sequence,structure and function of RdRp. RdRp gene was successfully amplified by PCR,and the constructed eukaryotic expression plasmid PCMV-Flag-RdRp was verified to be correct by double enzyme digestion and identification. Further,indirect immunofluorescence and Western Blot detection showed that the recombinant plasmid PCMV-Flag-RdRp was normally expressed in BHK-21 cells. Bioinformatics analysis found that,DTMUV RdRp consisted of 606 amino acids,with a formula of C3091H4810N870O894S41,an average hydropathicity of-0.536,and an instability index of 42.15. The RdRp was rich in glycosylation and phosphorylation sites. The ratios of α-helix,random coil and extending strand in the secondary structure were 46.37%,35.31%,12.54%,respectively. All of these results provided supporting data for the further research on biological functions of DTMUV.
  • DONG Li, SHI Haichun, ZHAO Changyun, YU Xuejie, KE Yongpei
    In order to explore new maize dwarf resources and study their genetic characteristics to provide a new material basis for maize dwarf breeding.The dwarf mutant K718d obtained from spontaneous mutation,and the wild-type K718 were used as materials to compare their phenotypic differences and the sensitivities to exogenous hormones;the genetic model of dwarf trait was analyzed based on the reciprocal F1,BC1,BC2 and F2 populations obtained from K718d mated with 5 high stalk inbred lines,the dwarf gene was mapped by BSA-SSR marker method,and the gene alleles were identified by allelic hybridization.The results showed that the plant height,ear height,number of internodes and internode length of K718d were reduced by 48.23%,75.57%,30.83% and 65.92% respectively compared with that of K718,and ear length was shortened by 28.57%,the yield was reduced by 36.44%,the differences reached extremely significant levels;the mutant K718d was not sensitive to GA3 and IAA.According to the experimental results of the two ecological sites,the reciprocal F1 populations were high stalks;the separation ratios of high plants and dwarf plants in the BC1 and F2 populations were accorded with 1:1 and 3:1 respectively,but the BC2 populations were high stalks;which indicated that the plant height of K718d was controlled by one pairs of recessive nuclear genes.The dwarf gene d718 was located between the SSR markers umc2569 and umc1278 on chromosome 1,and the genetic distances were 1.0,2.5 cM respectively,which was a br1 allele.The results lay the foundation for the further fine mapping and cloning of d718.
  • FAN Huifen, SUN Tianjie, SU Weihua, XIAO Fuming, ZHANG Jie, WANG Dongmei
    To elucidate the mechanism of the transcription factor GmWRKY50 in the interaction between soybean and Soybean mosaic virus (SMV),the full-length coding region of GmWRKY50 gene was cloned by PCR using RNA of soybean leaves as template,the similar amino acid sequences of GmWRKY50 in different species were compared by MEGA 7.0 and phylogenetic tree analysis was carried out.The recombinant plasmid pColdⅡ- GmWRKY50 was constructed through prokaryotic expression technology.After transformation of Escherichia coli BL21,the GmWRKY50 recombinant protein with His tag was induced by IPTG.The recombinant protein was purified by Ni-NTA column,and the purified recombinant protein was collected for polyclonal antibody preparation.Western Blotting was used to detect the expression level changes of GmWRKY50 before and after Soybean mosaic virus infection.The results showed that the CDS of GmWRKY50 gene was 495 bp in length,encoding 165 amino acids,and contained a WRKY domain at the N terminal,belonging to WRKY DNA-binding domain protein.Sequence alignment and phylogenetic tree analysis showed that soybean GmWRKY50 had the closest homologous relationship with Arabidopsis AtWRKY50. When IPTG concentration was 0.5 mmol/L and induction time was 4 h,the induced effect of recombinant protein was good,and the target protein fused with His protein tag was obtained by nickel ion affinity purification,which has high solubility.The polyclonal antibody prepared with fusion protein as antigen could specifically bind GmWRKY50,GmWRKY50 was up-regulated by SMV infection in incompatible combinations.This study lays the foundation for further investigation of the function of transcription factor GmWRKY50 in soybean resisting SMV infection.
  • WU Yue, CHEN Yang, WANG Xingzhe, SHAN Feibiao, ZHANG Yong, SUN Hongju
    Abstract (107) PDF (22) RichHTML (2)

    In order to obtain transcriptome information and functional genes expression characteristics of Astragalus complanatus,the leaves of A.complanatus were used as material,the high-throughput transcriptome sequencing and bioinformatics analysis were performed using Illumina HiSeq platform.A total of 19 280 Unigenes were assembled with total sequence length of 23 472 470 bp,of which the GC content was 42.74%,indicating a high quality of sequencing and assembly.Blast analysis showed 12 541,10 120,9 412,8 953,7 494,5 052 Unigenes got annotation in Nr,Swiss-Prot,GO,KEGG,KOG and COG databases,respectively.The annotated Unigenes of A.complanatus were mainly homologuous to leguminous plants,in particular,highly matched to the subfamily Pteriformis plant Cicer arietinum,Medicago truncatula and Abrus precatorius with a respective percentage of 31.49%,14.50% and 11.65%.Annotated Unigenes were divided into 3 GO classifications,354 KEGG metabolic pathways and 25 KOG function categories.The most abundant categories in each database were metabolic process,general function prediction only and purine metabolism,respectively.These results showed that the leaf cells of Astragalus membranaceus had active metabolism and abundant gene expression in the seedling stage.Additionally,a lot of Unigenes were annotated to the infectious disease category in KEGG database,suggesting a potential medicinal value of the plant part of Astragalus membranaceus.A total of 5 849 simple sequence repeats(SSRs)were identified by MISA software,with an occurrence frequency of 30.34%.The SSRs in the genome of Astragalus membranaceus were abundant and diversified,containing single base to six base repeats,among which the mono-nucleotide SSRs had the largest number with a frequency of 40.56%.The main types of SSR motifs involved A/T,AG/CT and AAG/CTT.

  • LEI Yang, QIAO Ning, BAI Yang, YANG Yuhua
    To explore the mechanism of epibrassinolide(EBR) on cadmium resistance in pepper, Capsicum annuum L. cv. Jinjiao 503 was used as experimental material, and various physiological indexes and stress-resistance related genes of pepper seedlings sprayed with different concentrations of EBR were compared under high cadmium(100 μmol/L) stress at seedling stage. The results showed that high concentration of cadmium ions could down-regulate the biomass, photosynthetic pigment content, antioxidant enzyme activity, the scavenging efficiency of ascorbic acid(AsA) -glutathione(GSH) cycle, and up-regulate the expression of DHAR, GR, MDHAR, CAT and WRKY25 genes in pepper seedlings. The application of 100 μmol/L epibrassinolide could activate the expression of DHAR, MDHAR and WRKY25 genes, increase the biomass of pepper seedlings, and increase the photosynthetic pigment content in leaves. The enzyme activities of peroxidase(POD), superoxide dismutase(SOD), catalase(CAT), ascorbate peroxidase(APX), dehydroascorbate reductase(MDHAR), dehydroascorbate reductase(DHAR) and glutathione reductase(GR) increased, and the ratio of reduced state to oxidized state in AsA-GSH circulatory system increased, thus reducing the O2 velocity and H2O2 content.The results showed that high concentration of cadmium ions could activate the expression of GR, CAT and DHAR genes, but inhibited the activities of corresponding enzymes, which hindered the normal response path of pepper to heavy metals, resulting in serious physiological stress. It indicated that EBR with appropriate concentration could effectively alleviate the toxicity of cadmium stress on pepper seedlings and improved the cadmium tolerance of pepper by regulating transcription factors and enzyme activities.
  • XIAO Jiachang, ZHENG Kaimin, MA Junying, ZHENG Yangxia
    To study the alleviating effect of exogenous Nitric oxide(NO) on the root system of watermelon under aluminum stress,watermelon variety Zaojia 84-24 was used as the experimental material,and the effects of different concentrations(50,100,200,500 μmol/L) of NO on the physiological and mineral content of watermelon root under aluminum stress were studied by applying nitroprune(NO donor) under high concentration of aluminum stress(1 200 μmol/L). The results showed that the root electrolyte leakage and the content of malondialdehyde increased under high Al ion stress,and the activities of superoxide dismutase(SOD),peroxidase(POD) and catalase(CAT) were significantly decreased in order to maintain the osmotic equilibrium by significantly increasing the content of proline and soluble sugar in the root. Aluminum stress inhibited the absorption of Ca,Mg,Fe and P,promoted the absorption of K,Zn and Al,but the content of Cu had not changed significantly. After the addition of exogenous NO,the root electrolyte leakage and the content of malondialdehyde decreased,the degree of plasma membrane peroxidation decreased,the content of soluble sugar in the root decreased significantly,and the content of proline increased significantly. The activities of SOD,POD and CAT increased first and then decreased with the increase of NO concentration. NO not only promoted the absorption and accumulation of nitrogen by increasing the activity of nitrate reductase,but also promoted the absorption of Ca,Mg,Fe,Cu and P by watermelon roots,and inhibited the accumulation of Zn,Al and K contents by roots. The addition of L-NAME,an endogenous inhibitor of NO,eliminated the alleviating effect of NO on aluminum stress in watermelon and even inhibited it. These results indicated that exogenous SNP could alleviate oxidative damage of watermelon roots under Al stress,and alleviate the effects of Al toxicity by affecting the absorption of nutrient elements by roots.
  • YE Fahui, CAO Dong, ZHAO Caixia, SHEN Jicheng, LIU Ruijuan, LIU Demei, CHEN Wenjie, ZHANG Huaigang
    In order to verify the function of Avenin-like b protein in wheat flour, thirteen Avenin-like b genes from different Aegilops tauschii plants were selected then they were carried out protein expression and purification in vitro. Through the micro-powder mixing experiment, the oxidized reducing agent(DTT+KIO3) and the purified Avenin-like b protein(TaALPb7A-M) expressed in vitro from 10 g Aegilops tauschii were added to the base flour.The formation time, stability time and breakdown time were measured and analyzed, respectively. The results showed that DTT+KIO3 in the base flour, had no significant effect on the formation time, stability time and fracture time of dough. Compared with control Gaoyuan448(GY448), the formation time decreased by 5.98%, and the others increased by 8.11%, 3.23%, respectively. Compared with the control GY448, the dough formation time, stability time and breaking time were increased by 12.82% -38.46%, 22.52% -44.14% and 16.67% -20.97% after the addition of Avenin-like b protein in vitro into basic flour. The formation time, stability time and breakdown time extremely increased significantly(P <0.01) the three parameters. However, different types of Aegilops tauschii Avenin-like b protein on the formation time and stability time of dough were different, the breaking time was relatively stable. In which, the dough formation time and stability time of TaALPb7D-F protein were increased by 38.46% and 44.14% respectively, which were extremely significantly higher than those of other types of purified Aegilops tauschii Avenin-like b protein(P <0.01). Thus, the Avenin-like b protein in Aegilops tauschii could significantly improve the flour processing quality of wheat, and the germplasm of Aegilops tauschii carrying excellent allelic variation of Avenin-like b gene could be considered to use for the selection of high-quality common wheat cultivar.
  • ZANG Hecang, CAO Tingjie, ZHANG Jie, ZHAO Qing, DI Jiaying, ZHANG Jiantao, ZHUANG Jiayu, CHEN Dandan, LIU Haijiao, ZHENG Guoqing, LI Guoqiang
    To evaluate the high yield,stable yield and adaptability for analyzing the genotype and environment interaction of new wheat cultivars under different ecological conditions,and to analyze the variation rule of yield of new wheat cultivars could provide scientific reference for the selection and breeding of wheat varieties with high quality.Regional trial was participated at Shangqiu, Luoyang and Xinxiang City in 2018-2020, with eight winter wheat cultivars as material, which were planted in Henan Province,mathematical statistics and GGE biplot were used to analyze the high yield,stability and adaptability of the new wheat varieties.Variance analysis showed that years,locations,cultivars and their interactions,except for years×cultivars,had extremely significant effects on wheat yield(P <0.01),and the years and locations had a greater contribution rate to wheat yield,38.63% and 32.86% respectively,while contribution rates of locations×cultivars,cultivars and years×cultivars to wheat yield was the smallest, only 1.31%. The yield of winter wheat in 2019-2020 was decreased by 9.71% compared with that in 2018-2019, the Shangqiu location had the lowest average yield for two years, which was significantly lower than the average yield of Luoyang and Xinxiang locations. The average yield of eight wheat cultivars from three regional test sites across two years were 8 049.04 kg/ha, yield of Taihe 896 had the highest, while yield of Bainong 207 had the lowest. In terms of high yield,Taihe 896,Nongkeda 888,Shengke 188,Hemai 53,Zhiyou 33,Yanbo 369 and Puda 1030 were varieties with better yield.In terms of stability,Bainong 207,Hemai 53,Nongkeda 888 and Zhiyou 33 were varieties with better yield stability.In terms of adaptability,Nongkeda 888,Taihe 896 and Shengke 188 were varieties with better adaptability.The research results provide a scientific basis for the rational utilization of new wheat varieties and have certain reference value.
  • TANG Wenfang, XU Shengsheng, DUAN Yanbi, LONG Wenhong, YANG Rongping, ZHANG Xuemei, MENG Jingui
    Abstract (101) PDF (28) RichHTML (8)

    The aim of this research was to clone IPT(Isopentenyl transferase,IPT ) genes in yam,predict the structure and property of DoIPT proteins and identify the expression of DoIPTs during bulbil sprouting.DoIPTs were cloned by homologous cloning techniques.The structure and property of DoIPT proteins were predicted by bioinformatics analysis.Real-time PCR technique was applied to identify the expression of these genes.Three nucleic acid sequences of 1 398,1 044,1 349 bp were obtained from yam.The three sequences were named DoIPT1,DoIPT5a and DoIPT5b,and the registration numbers of these genes were MW353160,MW353016 and MW353017,respectively.The reading frame lengths were 1 137,861,1 011 bp respectively,and the encoded amino acid lengths were 378,286,336 aa,respectively.Bioinformatics analysis showed that DoIPTs were unstable,hydrophilic and external family proteins.There were two signal peptides in DoIPT5b,and no signal peptide was found in the other two proteins.The secondary structures in the three proteins were consistent with the prediction models of the tertiary structures of three genes.The encoded regions of DoIPTs had high similarity with the isopentenyl transferase protein of other plants.Coding areas of DoIPTs showed that the N-terminal regions of DoIPTs contained the P-loop NTPase structural domain GXXGXGKS(T),which was able to catalyse cytokinin synthesis.The results of Real-time PCR technique showed that the expression of DoIPT5a in sprouting bulbils was highest under the condition of water shortage at 22 ℃;and the expression of DoIPT1 and DoIPT5b genes was the highest in a humid environment;the expression of DOIPTs genes raised with paclobutrazol treatment.These results laid a foundation for further study on the function of DoIPTs in yam.

  • LI Man, SHI Xiaolei, DI Rui, LIU Zhifang, MENG Qingmin, FU Cai, YANG Chunyan, WANG Dongmei, ZHANG Mengchen, ZHANG Jie, YAN Long
    Abstract (100) PDF (202) RichHTML (6)

    The objective of this work was to develop novel,convenient,and reliable molecular marker in soybean.Twelve soybean germplasm accessions from different geography region were used to develop large-size InDel markers based on 10 depth genomic re-sequencing.Developed InDel markers were used to construct DNA fingerprint of 96 varieties in Huanghuaihai Uniform Test in 2018.The results showed that a total of 66 561 InDel markers,with insertion/deletion fragment size larger than 20 bp,were discovered among the 12 accessions.The distribution pattern among the genome of these InDel markers were also illustrated.The ratio of the InDel in intron,upstream of the gene,downstream of the gene,intergenic region,intragenic region,5'-UTR and 3'-UTR were 12.35%,25.83%,20.44%,34.39%,0.19%,0.93%,1.51%,respectively.There were 42 453 InDels yield insertion/deletion fragment size between 20 to 40 bp,13 044 InDels between 41—60 bp,5 034 InDels between 61—80 bp,2 285 InDels between 81—100 bp,and 2 413 InDels larger than 100 bp.Then,160 InDel markers were randomly selected according to the insertion/deletion fragment size throughout the whole genome.As results,thirty-two InDel markers,that only yield 2 alleles in for each locus and could be distinguished easily on agarose gel,were developed and validated through primer design,PCR and agarose gel electrophoresis.Additionally,DNA fingerprint of 96 varieties in Huanghuaihai Uniform Test in 2018,were constructed using the 32 developed InDel markers.As results,the average purity of the tested soybean varieties was 96.84%,and no varieties shared the same name.The InDel markers developed in this study were stable,reliable and user friendly.They could be used to construct DNA fingerprint and test seed purity of soybean.

  • GAO Ruiyu, ZHANG Hua, XUAN Ning, LIU Xu, ZHANG Hao, ZHANG Mengqi, YAO Fangyin
    In order to study the reasons for the difference of rice heading date between SSSL (Single segment substitution line) and acceptor parent HJX74 (Huajingxian 74), used HJX74 and five SSSLs WY18 (W06-15-18-3-11), WY24 (W08-16-3-2), WY26 (W11-15-7-1-1), WY60 (W15-3-1-38) and WY61 (W17-10-5-5-35-9-2) with significant difference in heading date as materials, cloned the known heading date alleles on their substitution fragments, obtained their CDS sequence, and used a variety of bioinformatics methods to perform sequence alignment, including CDS sequences and amino acid sequences, compared the similarities and differences of amino acid physicochemical properties and expression differences. According to the phenotype of heading date in the field, WY18, WY60 and WY61 showed late heading date compared with HJX74, while WY24 and WY26 showed early heading date. Compared with HJX74, there were 4 SNP differences in the heading date allele DTH8 on the WY18 substitution fragment, 7 SNP differences in WY26 OsDof12, and 6 SNP differences in WY60 DTH8. However, none of the above SNP mutations changed the hydrophilicity and physicochemical properties of its amino acid sequence, indicating that the mutations of these heading date alleles might not be the cause of the heading date changes. Further expression analysis by qRT-PCR founded that the expression level of heading date alleles DTH8 of WY18 and OsDof12 of WY26 were higher than that of HJX74, indicating that their expression was regulated by the upstream regulatory region. The regulation of this expression might be the reason for the variation of heading date. The WY24 se14 and WY61 Hd1 had obvious sequence deletions, leading to loss function of se14 and Hd1 alleles which probably were the main reason for heading date variations of WY24 and WY61.
  • JING Fanli, ZHANG Peipei, MIAO Yongping, CHEN Tao, LIU Yuan, YANG Delong
    Abstract (98) PDF (128) RichHTML (24)

    In order to clarify the expression characteristics and biological functions of the sucrose phosphate phosphatase gene(SPP),and further understand the regulatory mechanism of SPP involved in sucrose biosynthesis.Herein,three TaSPP homologs,TaSPP-5A,TaSPP-5B and TaSPP-5D,located on the fifth homologous group,were cloned using the cDNA of wheat variety Jinmai 47 as the template.The physical and chemical properties,gene structure,cis-acting elements,phylogenetic tree and protein conserved domains of TaSPP were analyzed by the method of bioinformatics.The expression pattern of TaSPP was analyzed by qRT-PCR.The results showed that TaSPP-5A,TaSPP-5B and TaSPP-5D contained eight exons and seven introns.TaSPP-5A and TaSPP-5D encoded 422 amino acids,while TaSPP-5B encoded 413 amino acids.Phylogenetic analysis showed that TaSPP in wheat and its related species belonged to the same evolutionary branch with highly genetic similarity.The specific expression analysis showed that TaSPP genes were expressed in roots,stems,flag leaves,leaf sheaths,flower and seeds,whereas the higher expression levels were identified in flag leaves and stems.The expressions of TaSPP gene could be induced by ABA,PEG-6000,NaCl and IAA,indicating that TaSPP genes could play an essential role in abiotic stress tolerance in wheat.

  • JIN Xinxin, SONG Yahui, CHENG Zengshu, WANG Jin, LI Yurong, SU Qiao
    In order to make clear the specific time of peanut irrigation and make a reasonable peanut irrigation system,a experiment was conducted with different irrigation treatments to identify the specific irrigation time of peanut at anthesis under the condition of rain and drought shelter.The irrigation times were set at 20,25,30,35,40,45,50 days after sowing,respectively.The characteristics of main stem height,lateral branch length,leaf area,dry matter accumulation and yield of peanut were measured.The results showed that the earlier or later irrigation time at the flowering stage significantly affected the growth characteristics and dry matter accumulation of peanut.Under the influence of soil water stress,the main stem of peanut was shorter,the lateral branches were shorter and the dry matter accumulation was less,but grew rapidly after rewatering.There were no significant differences in the main stem height,lateral branch length and branch number among the treatments at harvest,the average were 59.06 cm,62.72 cm and 6.34,but the main stem height,lateral branch length,plant dry weight and pod dry weight were the highest in irrigation treatment of 30 days after sowing.The irrigation treatment of 30 days after sowing had the highest yield(6 415.25 kg/ha),100-pod weight(298.59 g) and 100-seed weight(112.41 g),other treatments were 4.59%-22.87%,10.55%-22.59%,4.92%-16.84% lower than those.The irrigation in the early flowering stage could ensure the higher soil moisture content in the thriving anthesis,leading to concentrated peanut bloom,more and full pods of single plant,higher hundred pod weight and hundred seeds weight,more yield.Irrigation at flowering stage should not be earlier than 7 days before anthesis,not later than 7 days after anthesis,that is,about 30 days after sowing.Irrigation in advance or in delay would result in soil water content deficit at flowering stage and affect the formation of final yield.
  • LU Siyu, YANG Zaiqiang, YANG Li, ZHANG Yuanda, ZHENG Han
    In order to explore the flowering effect of different photoperiods and its relationship with hormones and growth,Hongmian autumn chrysanthemum was used as test material,and five photoperiod treatments were set up.The natural photoperiod(long-day photoperiods) in summer in Nanjing was used as control.Photosynthetic pigments(chlorophyll a,chlorophyll b,carotenoids),growth indexes(plant height,stem diameter,leaf number),root activity,development duration of different periods and endogenous hormones(budding stage,bud breaking stage,petal formation stage,initial flowering stage,full blooming stage,early wilting stage),flower quality(flower diameter,flower number per plant,anthocyanin) were measured.The results showed that the chrysanthemum grew most vigorously under long-day photoperiods,the plant was tall and strong,and the photosynthetic pigment content was high.7 h/17 h,8 h/16 h plant photosynthetic pigment content was low,the worst growth,lodging,due to light stress,root activity was higher.The growth of plants under 10 h/14 h was dwarfing symmetrical and they had small plant height,thick stem and dense leaves.The flowering response of chrysanthemum was closely related to the endogenous hormone content in leaves.The flower development rate of chrysanthemum was inversely proportional to IAA and GA3,and was proportional to ZT and ABA.7 h/17 h,8 h/16 h and long-day photoperiods significantly delayed the flowering stage of chrysanthemum.In long-day photoperiods,due to sufficient growth,the cut flowers were large and more but wilted quickly.In 7 h/17 h and 8 h/16 h,due to insufficient vegetative growth,chrysanthemum was small and less,the anthocyanin content was low,and the flower quality was the worst.Flower bud differentiation started fastest at 10 h/14 h,and the development cycle of inflorescence was short,the flowering was early and lasted for a long time.The quality of flowers was also high due to good growth conditions.Photoperiod had the greatest influence on the initiation of flower bud differentiation and the formation of buds.Although the expansion of hyoid petals was also affected by photoperiod,it did not play a decisive role in the advance of flowering.Thus,under the control of 10 h/14 h chrysanthemum in growth morphology,inflorescence development cycle,cut flower quality performance was the best,most suitable for summer in Nanjing Area chrysanthemum flower cultivation,when chrysanthemum bud color after moving to long sunshine normal flowering.
  • ZHANG Hong, XU Yingli, WANG Chaonan, HUANG Zhiyin, FAN Weiqiang, LI Mei, ZHANG Bin
    In order to develop closely linked molecular markers for resistance to dry burning heart disease in Qingmaye Chinese cabbage and accelerate the cultivation of high-quality varieties resistant to dry burning heart disease.200 F2 and 100 BC1F1 populations were constructed simultaneously with the excellent Qingmaye high generation inbred line H227 and Baimaye high generation inbred line G83,the resistance and susceptibility phenotypes of the population were identified by in vitro leaf cutting method, and the genetic law of dry burning heart disease in Qingmaye Chinese cabbage was statistically analyzed. At the same time, combined with BSA method, the disease resistance gene of dry burning heart was preliminarily located and labeled by software JoinMap 4.0 and Mapchart.The results showed that the disease grading of F2 population and BC1F1 population constructed by parents with significant difference in resistance and susceptibility to dry burning heart disease showed obvious unimodal distribution, close to normal distribution, and the inheritance law of resistance had the genetic characteristics of quantitative traits.Based on the sequence information of Chinese cabbage genome database and the constructed molecular marker linkage map, the disease resistance gene in the experimental material was located on A07 chromosome, and a molecular marker BrIDCRT07 linked to the dry burning heart disease resistance gene was designed.The genetic distance between the molecular marker and BrID10343 and BrID10349 was 0.13, 0.78 cM respectively.It was verified that the coincidence rate of this marker in F2 population was 86.8%, and that in BC1F1 population was 94.9%. It could be used as an auxiliary screening marker for anti dry burning heart disease resources of Chinese cabbage in the future.
  • 2021, 36(5): 0-0.
  • 2021, 36(6): 0-0.
  • CHEN Ziqi, WANG Weiping, ZHAO Hongqiang, WANG Hao, HAN Xiao, YANG Luomiao, XIN Wei, LIU Hualong, ZHENG Hongliang, WANG Jingguo
    Abstract (95) PDF (78) RichHTML (6)

    To map QTLs that control cold tolerance at germination and bud stages,to explore the genetic mechanism,a recombinant inbred line(RIL)population was used as experimental material.The RIL population contained 189 lines derived from the cross between Caidao,a japonica rice variety that had strong cold tolerance and was adopted as the female parent,and WD,a indica rice variety that had weak cold tolerance and was used as the male parent.The QTLs that control cold tolerance at germination and bud stage were mapped between four phenotypic indexes and 978 Bin markers by IciMapping 4.2.The four phenotypic indexes included relative germination rate,average germination days,seedling rate and cold tolerance level.The results showed that two pairs of epistatic QTLs related to cold tolerance at bud stage were mapped.The LOD scores were 5.70 and 5.55 and the phenotypic contribution rates were 13.22% and 39.87%,respectively.In addition,five and five QTLs that were related to cold tolerance at germination and bud stage were mapped,respectively.These QTLs were located on chromosomes 1,3,5 and 8.The LOD score and phenotypic contribution rate of these QTLs ranged from 2.55 to 7.07 and from 6.41% to 17.17%,respectively.qLTG3 had the highest phenotypic contribution rate among the five germination stage QTLs and was also related to cold tolerance at germination stage.qCTP1-2 had the maximum of phenotypic contribution rate among the five bud stage QTLs.qCTB1,qCTP1-2 and qLTG8 had not been reported.The QTLs and epistatic QTLs detected will provide a reference for the genetic research and improvement of cold tolerance in rice.

  • WU Jianfei, LIU Yu, LI Heng, JING Tian, LU Jianyuan, ZI Xiangdong
    The objective of this study was to investigate the sequence and tissue expression characteristics of DNA damage inducible transcript 3(DDIT3) in female yaks. The yak DDIT3 gene was cloned by RT-PCR. The structure and function of DDIT3 protein were analyzed by bioinformatics software. The expression levels of DDIT3 gene in different tissues, and different reproductive organs and oocytes at different physiological stage of female yaks were detected by Real-time quantitative PCR(RT-qPCR). The results showed that the full-length CDS region of yak DDIT3 gene was 507 bp, which encoded 168 amino acids. Nucleotide sequence alignment showed that yak had the highest homology with bison(99.71%) and more than 88% homology with other species, indicating that DDIT3 gene was highly conservative in the process of evolution. The expression of DDIT3 gene in ovary was extremely significantly higher than that in heart, liver, kidney, spleen, lung, uterus and fallopian tube(P <0.01). In the ovary, the expression of DDIT3 gene was significantly higher during pregnancy than that during follicular phase, luteal phase and fetus stage(P <0.05), and it was significantly higher during luteal phase than that in fetus stage(P <0.05). In the uterus, the expression of DDIT3 was significantly higher than that in follicular stage and fetus stage(P <0.05). In oviduct, its expression was not significantly different at different physiological stages. The expression of DDIT3 gene in M Ⅱ stage oocytes was significantly higher than that in GV stage and M Ⅰ stage(P <0.05). Its expression in M Ⅱ granulosa cells was also significantly higher than that in GV stage and M Ⅰ stage(P <0.05). There was no significant difference in DDIT3 expression between oocytes and granulosa cells in GV stage and M Ⅰ stage. In conclusion, yak DDIT3 gene possible plays an important role in regulating ovary function and pregnancy maintenance, follicle development and maturation.
  • WANG Xiaoya, BU Ruifang, HU Haiyan, LONG Qiang, SUN Lianxuan, QI Lu, LIU Zheng, LI Xiaoyao, LI Chengwei
    To explore the regulation mechanism of Jasmonic acid(JA) on growth,maturation and yield in wheat,the gene TaJIP2 was cloned from wheat Bainong 207 by homologous cloning method.Analysis of tissue differential expression showed that TaJIP2 gene was expressed in all tissues and the expression level was the highest in wheat hull.The TaJIP2 gene RNA interference(RNAi) vector was constructed to conduct genetic transformation and functional analysis in wheat.The results showed that silencing TaJIP2 gene increased the key enzymes activities of jasmonic acid biosynthesis,allene oxide cyclase(AOC) and 12-oxo-phytodienoic acid reductase(OPR),and JA content,and the difference in JA content was mainly reflected in the roots.At the same time,it significantly promoted the growth of roots and above ground parts in wheat,and the difference was also mainly in root growth.In addition,observation of the ultrastructure of root tip cells showed that the root tip cells of TaJIP2 gene silenced plants were complete in structure,rich in variety and number of organelles,and strong in cell activity,thus promoting root growth.Silencing TaJIP2 significantly improved the photosynthetic efficiency of wheat leaves,made the heading time earlier,and significantly increased the grain length and width,spike number per plant,grain number per spike,1 000-grain weight and yield per plant.The results showed that RNAi-induced silencing of TaJIP2 significantly promoted the growth,early maturity and high yield of wheat,which indicated that TaJIP2 was a key negative regulator of wheat growth,maturity and yield.These findings provided genetic resources and research basis for promoting the photosynthetic efficiency,growth,early ripeness and high yield in wheat.
  • ZHANG Jinjin, WANG Yunping, WANG Xiao, ZHANG Shuxing, WANG Xuemin
    To explore the biological function of alfalfa MsWRKY33 in stress tolerance at the protein level,by using Medicago sativa cv.Zhongmu No.1 as the material,MsWRKY33 antigen sequence was cloned and its properties and functions were analyzed.The structural characteristics analysis and immunogenicity prediction of MsWRKY33 protein were performed by using bioinformatics.The structure and characteristics of MsWRKY33 protein analysis results showed that the full length of MsWRKY33 protein was 511 aa,which was mainly composed of random coil and extended chain structure.MsWRKY33 protein had no signal peptide and was a hydrophilic protein.The immunogenicity prediction results indicated that the protein was highly immunogenic.According to the specificity of the binding sequence,the appropriate segment 247-457 aa was selected,and the MsWRKY33 polyclonal antibody was prepared by using this segment through protein purification,immunization and other steps.Subsequently,Western Blotting method was used to analyze the expression of MsWRKY33 protein under four abiotic stress treatments:salt,cold,PEG and ABA,all could enhance the MsWRKY33 protein expression.Among them,NaCl brought the highest expression level,indicated that MsWRKY33 might play an important role in salt stress regulation.The test results provided a reference for further exploration of the function of MsWRKY33 and the mechanism regulating the stress tolerance of alfalfa.
  • ZHANG Zhenwang, WU Jinzhi, HUANG Ming, LI Youjun, ZHAO Kainan, HOU Yuanquan, ZHAO Zhiming, YANG Zhongshuai
    Abstract (94) PDF (94) RichHTML (31)

    In order to clarify the effects of drought stress on the photosynthetic characteristics in flag leaf in the afternoon during grain filling stage and grain yield of winter wheat with different drought resistance,an experiment with two winter wheat cultivars and four water levels was conducted under the condition of rainproof pond cultivation in 2018—2019 and 2019—2020.The two winter wheat cultivars were Jinmai 47(JM47,strong drought resistance)and Yanzhan 4110(YZ4110,weak drought resistance).The four water treatments included severe drought(W1:65% MFC(maximum field water capacity)before sowing + 45%—55% MFC after jointing),moderate drought(W2:75% MFC before sowing + 55%—65% MFC after jointing),mild drought(W3:75% MFC before sowing+65%—75% MFC after jointing),suitable water supply(W4:75%MFC before sowing+75%—85% MFC after jointing).The net photosynthetic rate(Pn),stomatal conductance(Gs),intercellular CO2 concentration(Ci),transpiration rate(Tr),instantaneous water use efficiency(IWUE),maximum photochemical efficiency of PS Ⅱ (Fv/Fm)and actual photochemical efficiency of PS Ⅱ(ΦPS Ⅱ)in flag leaf from 14:00 to 16:00 during the early,medium and medium-late grain filling stage and the grain yield and its components at maturity were investigated.The results showed that both water and cultivars had significant effects on the photosynthetic and fluorescence characteristics in flag leaf in the afternoon during the grain filling stage and the grain yield at maturity of winter wheat.From the two-year average,compared with W4,the Pn,Gs and ΦPSⅡ in flag leaf in the afternoon during the grain filling stage under drought stress(W1,W2 and W3)respectively decreased by 2.07%—68.92%,-3.23%—50.00% and -1.89%—30.19% in JM47,and decreased by 7.71%—80.19%,11.11%—59.26% and 0—73.47% in YZ4110;the flag leaf Tr values in the afternoon during the medium grain filling stage in JM47 and YZ4110 respectively decreased by 6.30%—32.87% and 6.49%—41.74%,and the flag leaf Fv/Fm values in the afternoon during the medium-late grain filling stage decreased by 1.20%—18.52% and 2.50%—30.00%.In general,for all the above indexes,the decreasing amplitude for the same index was JM47<YZ4110.Compared with YZ4110,the Pn,Gs,ΦPSⅡ and Fv/Fm in flag leaf in the afternoon during the grain filling stage under drought stress(W1,W2 and W3)of JM47 respectively increased by 0.86%—64.89%,8.33%—36.36%,1.96%—184.62% and 1.25%—17.86%,and the grain yields of JM47 were respectively increased by 28.91%,8.06% and 5.40%.Except for IWUE,the flag leaf photosynthetic parameters in the afternoon during the grain filling stage were significantly and extremely significant correlated with grain yield,but the correlation indexes varied with variety and grain filling stage.For JM47,the correlation indexes between the grain yield and flag leaf photosynthetic parameters were highest for Pn,Gs and Fv/Fm during the medium-late grain filling stage,for ΦPSⅡ during the medium grain filling stage,and for Tr during the early grain filling stage.For YZ4110,the correlation indexes were highest for Pn,Gs and Tr during the early grain filling stage,for ΦPSⅡ during the medium grain filling stage and for Fv/Fm during the medium-late grain filling stage.In summary,drought stress decreased the photosynthetic function of flag leaf in the afternoon during grain filling stage and thus decreased the grain yield of winter wheat,the strong drought resistance variety could maintain better flag leaf photosynthetic characteristics in the afternoon during the grain the filling stage under drought stress condition,and significantly improved the ΦPSⅡ of flag leaf in the afternoon during the medium grain filling stage and the Pn,Gs and Fv/Fm of flag leaf in the afternoon during the medium-late grain filling stage,thereby increasing the grain yield.

  • TIAN Zhigang, ZHANG Shuwei, CHEN Fang, CHANG Lifang, JIA Juqing, ZHANG Xiaojun, LI Xin
    Nucleotide binding site(NBS) genes are a type of disease resistance genes widely present in plants.In order to screen the powdery mildew-resistant genes for wheat breeding,the changes in the expression level of wheat NBS genes after infection with Blumeria graminis f.sp.tritici(Bgt) were analyzed.Bioinformatics methods were used to assemble the original transcriptome data of a disease-resistant wheat variety infected by Bgt at 0,24,48,72 h,and 1 283 TaNBS with expression data were screened out,which were distributed on 21 chromosomes.Results of expression difference analysis showed that the expression levels of 395 TaNBS were significantly changed after being infected by Bgt,and they were divided into five types,including down-up-up,up-up-down,up-down-up,down-down-up as well as up-up-up,according to the change trend in three time periods(0-24 h,24-48 h,48-72 h).Using qRT-PCR technology,ten TaNBS with the most significant differences were selected from the up-up-up type for verification.The results showed that three genes Ta7dlLoc004854, Ta7dlLoc000139 and Ta3asLoc007663 remained significantly up-regulated in a self-breeding disease-resistant wheat germplasm CH7124 after being infected by Bgt. Using virus-induced gene silencing technology,the expression levels of the above three genes in CH7124 were down-regulated to obtain the silent plants.The results of the inoculation test showed that only the Bgt-infection type of Ta7dlLoc000139-silent plants changed from immune to high resistance or medium resistance,indicating that the decrease in the expression level of Ta7dlLoc000139 caused the plant resistance to weaken.Sequence analysis results showed that Ta7dlLoc000139 had a total length of 4 042 bp,contained three exons and two introns,and contained a large number of regulatory elements such as CGCG-Box,GATA-Box and CAAT-Box in the 2 000 bp region before its start codon.The research results could provide a theoretical basis for wheat disease resistance molecular breeding.
  • WANG Dafeng, LU Shuchang, WANG Wei
    In order to improve nitrogen utilization in facility farmland and reduce soil nitrogen accumulation,waxy corn and feeding sweet sorghum was selected as the catch crops,and six different levels of biochar were designed,namely C1(0),C2(0.5%),C3(1%),C4(2%),C5(4%),C6(8%),to study the effects of biochar application on nitrogen uptake by summer crop and soil carbon and nitrogen changes.The results showed that with the increase of biochar application rate,the nitrogen uptake of the two crops first increased and then decreased.Among them,the nitrogen uptake of the aboveground part of the waxy corn C3 treatment was the largest(142.69 kg/ha),feeding sweet sorghum was the best C4 treatment(132.43 kg/ha);the soil total nitrogen decreased by 2.5%-11.0% among different treatments,and the C3 treatment decreased by the most(15.4%),except for C6 treatment, the decrease range of other treatments for feeding sweet sorghum was 1.4% to 15.5%, and the decrease was the most in C3 treatment. After planting feeding sweet sorghum,different treatments and different soil layers had reduced nitrate nitrogen content compared with before planting;The C/N value of the facility soil could be increased by combining different amounts of biochar with catch crops;the application of biochar was beneficial to increase the organic carbon content,the planting of catch crops could reduce the soil urease activity in the early growth period of the subsequent crops,and reduce the nitrogen supply capacity of the subsequent crops during the growth period.With the increase of the application amount of biochar,the soil urease activity increased firstly and then decreased in each treatment.The highest urease activity was 3.33 mg/(g·d) in NC3(preceding waxy corn+C3) treatment after planting for 15 days.After planting for 45 days,the highest urease activity was in TC3(preceding sweet sorghum+C3) treatment,followed by TC2(preceding sweet sorghum+C2) treatment.The combination of biochar application and crop cultivation could increase the soil microbial biomass carbon content at the beginning of the subsequent crop,which was beneficial to the improvement of soil fertility.Under the conditions of this experiment,it was concluded that planting waxy corn with 1%-2% biochar application rate had a better effect on nitrogen absorption,and planting filling feeding sweet sorghum with 0.5%-2% biochar application rate had a better effect on nitrogen absorption,which was beneficial to reduce soil nitrogen accumulation.Planting feeding sweet sorghum has a better effect on reducing soil nitrogen movement than planting waxy corn.
  • LONG Yanxi, LUO Yanru, ZHU Zhenghang, LIAO Zhiyi, WANG Lan
    In order to explore the relationship between different genetic composition and quality of the high-yield tetraploid rice,and the reasons for the changes in protein content,the national high-quality varieties Meixiangzhan 2(MXZ2) and Xiangyaxiangzhan(XYXZ),Aijiaonante diploid(AJNT-2x),high-yield autotetraploid Aijiaonante(AJNT-4x),and neo-tetraploid Huaduo 1(H1) were used as the test materials. The endosperm protein content was determined by BCA method,the quality analysis was performed by the near-infrared grain analyzer,the amino acid content was determined by the automatic amino acid analyzer,and the expression difference of the glutelin genes was analyzed by qRT-PCR. The results were as follows:The protein contents of brown rice and polished rice of AJNT-4x were respectively for 182.70 and 115.70 mg/g,which was much higher than that of MXZ2,XYXZ,and AJNT-2x,and the nutritional quality of AJNT-4x was greatly enhanced;the gel consistency of AJNT-4x was only for 44.70 mm,which was much lower than that of MXZ2,XYXZ,and AJNT-2x,and the difference was significant,and eating quality of AJNT-4x was significantly reduced. The protein content of polished rice of H1 was the lowest of the samples,which was only for 71.30 mg/g,but its gel consistency was for 112.70 mm,which was between the national high-quality varieties MXZ2 and XYXZ.At 30 days after flowering, the total contents of 17 amino acids and essential amino acids of AJNT-4x were higher than those of AJNT-2x and H1,and which of H1 was the lowest;the total content of 17 amino acids for H1 reached a peak at 15 days after flowering,while the total content of 17 amino acids for AJNT-4x and AJNT-2x reached a peak at 20 days after flowering,the accumulation of amino acid content stopped prematurely in H1, which resulted the lowest total content of 17 amino acids in H1 at mature period.During the whole endosperm development process,the accumulation trend of the four storage proteins was basically the same in AJNT-4x,H1,and AJNT-2x,but there were differences in the rapid accumulation time;rapid accumulation of glutelin in AJNT-4x was earlier than that in AJNT-2x and H1,and the rapid accumulation time in AJNT-4x was longer than those in AJNT-2x and H1,which resulted in highest protein content in AJNT-4x. The nine glutelin subfamily genes were expressed at 5 days after flowering. With the development of the endosperm,the expression of each glutelin gene gradually increased,then gradually decreased and tended to stabilization. Different glutelin genes had different expression abundances at the same time;except the expression peaks of GluB-1 and GluB-4 were higher in H1 than those in AJNT-4x and AJNT-2x,the expression peaks of the other seven glutelin genes were much lower in H1 than those in AJNT-4x and AJNT-2x,and the difference was significant;the expression quantities of nine glutelin genes in AJNT-4x were all the highest in the three materials at 30 days after flowering. The above results provided a theoretical basis for high-quality breeding of rice polyploidy.
  • SU Xiaoyu, GAO Tongmei, LI Feng, WEI Libin, TIAN Yuan, WANG Dongyong, ZHU Songtao, WEI Shuangling
    In order to explore the physiological response to high temperature stress in sesame,the seedlings of two different sesame genotypes,Zhengtaizhi No.3(heat tolerance) and SP19(heat sensitivity),were treated at 45℃ as high temperature stress and 30℃ as control for 10 days.The growth phenotype,antioxidant capacity,photosynthetic parameters and microstructure of stomata and chloroplast of the seedlings were analyzed.The results showed that with the extension of stress time,the plant height,leaf length,leaf width,relative water content,chlorophyll content,Fv/Fm,φPSⅡ,ETR(Ⅱ),net photosynthetic rate(Pn),transpiration rate(Tr) and stomatal conductance(Gs) decreased,while the relative electrical conductivity,reactive oxygen species and malondialdehyde content increased significantly,and the change range of SP19 was larger than that of Zhengtaizhi No.3.The antioxidant enzymes(superoxide dismutase,peroxidase and catalase) activities in the plants treated with high temperature were higher than that of control.Compared to SP19,the antioxidant enzyme activity was higher in Zhengtaizhi No.3.Microstructure observation showed that under high temperature stress for 10 days,the stomatal aperture was decreased for both varieties,and the stomata of SP19 were completely closed,while Zhengtaizhi No.3 stomata were not completely closed.Meanwhile,chloroplasts became round and moved to the middle of cells,grana accumulation thickened,lamella fractured and osmiophilic granules increased in Zhengtaizhi No.3,and the structural stability of chloroplasts of Zhengtaizhi No.3 was higher than that of SP19.In conclusion,the antioxidant enzyme activity,membrane lipid peroxidation,chloroplast structural stability and the photosystem Ⅱ(PSⅡ) activity are the key factors to affect heat tolerance of sesame seedlings.
  • WANG Bohan, GUO Doudou, HUANG Shaomin, XU Qihao, ZHANG Keke
    This study compared the effects of superphosphate and monoammonium phosphate as phosphate fertilizer sources on the availability of phosphorus in fluvo-aquic soil,and analyzed the reasons why phosphate fertilizer varieties changed the availability of phosphorus in soil.Using a micro-area experiment,two phosphate fertilizer varieties,superphosphate and monoammonium phosphate,were set on the fluvo-aquic soil,and the effects of three different phosphate fertilizer rates on wheat and corn yield,phosphate partial productivity,soil available phosphorus,and soil inorganic phosphorus were analyzed after three consecutive years.Results showed that under the three levels of phosphorus application(P1,P2,P3),compared with the application of superphosphate,the average yield of wheat in monoammonium phosphate treatment increased by 24.9%,19.7%,22.0%,and the average partial productivity increased by 31.0,16.2,13.6 kg/kg,respectively;the average yield of corn increased by 29.6%,28.7%,32.2%,and the average partial productivity increased by 42.1,24.2,25.4 kg/kg,and the average content of soil available phosphorus increased by 9.6%,-3.4%,5.4%.The inorganic phosphorus of the soil treated with monoammonium phosphate increased mainly by Ca2-P and Ca8-P,Ca2-P increased by 13.4%,18.5%,26.1%,and Ca8-P increased by 17.2%,21.3%,and 15.8%.Compared with superphosphate treatment,the relative content of Ca2-P in soil treated with monoammonium phosphate increased by 0.25, 0.30, 0.33 percentage points,and the relative content of Ca8-P increased by 1.73, 0.80, 3.60 percentage points under the three phosphorus application levels.Superphosphate treatment was mainly based on the increase of Al-P and Fe-P,Al-P increased by 19.1%,23.7%,23.1%,and Fe-P increased by 24.9%,23.3%,32.6%.The relative content of Al-P in soil treated with superphosphate was increased by 1.27, 1.19, 1.26 percentage points,and the relative content of Fe-P was increased by 2.12, 2.87, 2.49 percentage points,compared with monoammonium phosphate treatment.Monoammonium phosphate and superphosphate had no significant effect on soil Ca10-P and O-P content,but monoammonium phosphate could reduce the relative content of Ca10-P and O-P by 2.47, 2.07 percentage points respectively.Compared with superphosphate treatment,monoammonium phosphate can increase the yield of wheat and corn,the partial productivity of phosphate fertilizer,and the soil available phosphorus content in fluvo-aquic soil.Monoammonium phosphate improves the availability of soil phosphorus by increasing the content and relative content of Ca2-P and Ca8-P,which are highly available in fluvo-aquic soil.
  • HE Jie, SUN Shaoguang, GE Changbin, SONG Danyang, QIAO Jiliang, LI Suoping, SU Yarui, LIAO Pingan
    Abstract (90) PDF (28) RichHTML (5)

    In order to explore the key factors affecting stem strength and screen the valuable parent resources for wheat "strong stem breeding" ,72 wheat varieties(lines)were used as materials.Firstly,the strength of stems in flowering stage,filling stage and milk stage of wheat was comprehensively analyzed and all the materials were grouped into clusters.On this basis,the genetic variation of lignin content,cellulose content and microstructure of wheat stem and their relation with stem strength were further analyzed,and the key characters related to stem strength were defined.In addition,principal component analysis(PCA)was used to extract the main factors of stem cross section microstructure,and calculate the score of comprehensive factors,and further cluster analysis was carried out combined with lignin and cellulose content,so as to comprehensively evaluate the performance of stem strength related traits of different wheat varieties(lines).The results showed that the mechanical tissue thickness decreased significantly or extremely significant at the same time as the strength of wheat stem weakened from flowering stage to filling stage.Moreover,parenchyma thickness,mechanical tissue thickness,large vascular bundle area,lignin and cellulose content in high stem strength wheat were much higher than those in low stem strength wheat.Correlation analysis also showed that mechanical tissue thickness,large vascular bundle area,lignin and cellulose content were significantly of extremely significantly correlated with stem strength.Finally,the lignin content,cellulose content and the main component comprehensive factor scores of the stem microstructure were used as three variables for cluster analysis,and a total of 18 wheat materials with outstanding performance in the stem microstructure and biochemical indicators were screened out.These wheat materials can be used as important genetic resources for wheat "strong stem breeding".

  • HAN Xiaoyong, JIANG Lu, YIN Jianmei, JIN Lin, ZHANG Peitong
    Abstract (89) PDF (25) RichHTML (20)

    In this study,the transcription level of resistance genes to the infection of Colletotrichum gloeosporioides were explored at the molecular level and provided candidate genes for further study of disease resistance mechanism.Suyu 8 is a new yam variety highly resistant to anthracnose,which was bred by tissue culture mutagenesis of the highly susceptible strain 024.In order to find out the anthracnose resistance genes,Suyu 8 and strain 024 leaves were inoculated for different time by Colletotrichum gloeosporioides strain 4-2,the differentially expressed genes were analyzed by transcriptome sequencing.Concurrently,the uninoculation leaves of Suyu 8 were used as control.The number of differentially expressed genes,inoculation for 24,48,72,96 h,were 197,132,187 and 313,respectively.After removing the common differentially expressed genes at each time point,we obtained 711 differentially expressed genes.Go enrichment showed that the differential genes were mainly related to response to biological stimulation,defense response,cell wall metabolism and oxidation-reduction process.KEGG enrichment analysis revealed that many metabolic pathways related to plant disease resistance were changed.The expression of multiple genes varied which defense-related plant hormone signal transduction pathways,such as auxin,jasmonic acid,and ethylene.Among them,five early auxin-responsive genes,the key genes of jasmonic acid and abscisic acid biosynthesis were up-regulated,while ERF036 was down-regulated,which might negatively regulate the infection of Colletotrichum gloeosporioides.Several cytochrome P450 genes,ubiquitin ligases involved secondary metabolite modification and phytosterol synthetases,defensins and lectins were up-regulated.WRKY,MYB and TIFY transcription factors positively or negatively regulate the expression of disease resistance genes.Under the regulation of transcription factors,PR protein,NBS-LRR disease resistance genes,receptor kinases were highly expressed,the expressions of CAT and SOD genes of antioxidant protective enzyme systems were activated by reactive oxygen species.In addition,the enzymes related to starch and sucrose synthesis were up-regulated,and enzymes related to starch degradation were down regulated.The expression trends of LAX4,IAA4,IAA21 were consistent with that of transcriptome sequencing.The relative expression of LAX4,IAA4 and IAA21 in resistant varieties were significantly higher than susceptible varieties,suggesting that auxin signaling pathway is beneficial to the immune response to anthracnose for yam.

  • PAN Gen, TAO Jie, NIE Rong, ZHOU Bing, HUANG Siqi, CHEN Anguo, LI Jianjun, TANG Huijuan, LI Defang, ZHAO Lining
    In order to reveal the functions of cannabidiolic acid synthase genes(CBDAS), the CBDAS genes in cannabis were identified using bioinformatics methods, and their physical and chemical characteristics, evolutionary relationship, gene structure, the cis-acting elements and transcript patterns were analyzed.Totally, 5 CBDAS genes were identified, and they distributed only on 2 chromosome and 7 chromosome.Their amino acid sequence size, molecular weight and isoelectric point were 541-545 aa, 61.49-62.41 ku, and 6.90-9.00, respectively.Evolution analysis showed that 33 CBDAS gene family members from plants, animals and microorganism were divided into three subclasses, and most of CsCBDAS belonged to the same subclass.The promoters of all CsCBDAS genes were rich in cis -acting elements related to light response.In addition, tissue-specific expression analysis showed that CsCBDAS1 and CsCBDAS2 preferred expressed in female flower and CsCBDAS4 and CsCBDAS5 preferred expressed in root.Moreover, the CsCBDAS members showed different expression patterns between the high-CBD cultivar and low-CBD cultivar, and the expression level of CsCBDAS1 in a high-CBD cultivar was higher than that in a low-CBD cultivar.Exogenous cadmium treatment could increase the exprssion levels of CsCBDAS4 and CsCBDAS5 with cadmium treatment were significantly higher than those without cadmium treatment.The transcript levels of CsCBDAS1, CsCBDAS2 and CsCBDAS5 exhibited significant difference between in light treatment and in darkness treatment, the transcript levels of CsCBDAS1 in light treatment was significantly lower than that in darkness treatment, but the expression levels of CsCBDAS2 and CsCBDAS5 were higher than those in darkness treatment.The CBDAS may involved in the growth and development of cannabis and cannabinoid synthesis, and this study laid the foundation for the further elucidation of the functions of CBDAS genes in cannabis.
  • LIU Junfeng, LI Yimeng, LIANG Chao, ZHOU Chanchan, WANG Shu, JIA Baoyan, HUANG Yuancai, WANG Yan, WANG Yun
    Abstract (88) PDF (38) RichHTML (8)

    Using rice variety Shendao 9 as an entry,a field experiment was conducted in 2019—2020 to study the effect of nitrogen application pattern and row spacing on canopy structure and yield of rice under split-plot design.The main plot treatments were zero nitrogen(A0),farmer's pattern(A1),low basal nitrogen(A2),and basal nitrogen backward(A3).The subplot treatments were conventional method(row spacing of 30 cm,B1),reducing row spacing(row spacing of 25 cm,B2),and narrow-wide row(row spacing of 40 cm+20 cm,B3).The layer upon layer cut method was used to investigate the leaf area index(LAI),photosynthetic active radiation(PAR),photosynthetic characteristics of flag leaves,and grain yield of rice.The results showed that nitrogen application pattern and row spacing had extremely significant interaction effects on grain yield,the maximum value(9.85 t/ha)appeared in A3B2.Compared with A1 and A2,the effective spikelets per unit area,kernel-setting rate,net photosynthetic rate(Pn),stomatal conductance(Gs)and transpiration rate(Tr)of A3 increased by 4.31%—10.55%,2.87—4.09 percentage points,4.84%—9.12%,14.08%—15.71% and 11.33%—15.83%,respectively.Meanwhile,rice obtained higher interception rate of PAR,population leaf area index under A3.Compared with B1 and B3,the spikelets per unit area,and population leaf area index(LAI)of B2 increased by 7.57%—9.97% and 4.29%—20.43%,respectively.And interception rate of PAR of B2 was 88.99%.Given the results of grain yield,PAR,and photosynthetic characteristics in rice of two years,it could be concluded that the basal nitrogen backward pattern combined with 25 cm row spacing could enhance spikelets per unit area,kernel-setting rate,1000-grain weight,improve the population structure and obtain higher yield.

  • FAN Yafang, GAO Julin, SUN Jiying, LIU Jian, SU Zhijun, WANG Zhigang, YU Xiaofang, HU Shuping
    Twenty-one maize inbred lines were used to study the effects of potassium fertilizer on stem phenotype,stem lodging resistance,ear traits,yield-related traits and grain quality of maize inbred lines with different yield types.The results showed that compared with 0K,the stem diameter of high-yield,middle-yield and low-yield maize inbred lines under 45K treatment increased by 14.21%,11.60% and 8.41%;the bending strength of stem increased by 30.58%,27.52% and 24.59%;the ear diameter increased by 3.61%,2.57% and 1.72%;the ear tip barrenness decreased by 10.19%,7.41% and 4.81%;the 1000-grain mass increased by 4.97%,3.55% and 2.23%;the yield increased by 7.50%,5.57% and 4.45%;the grain protein content increased by 0.62,0.40,0.20 percentage points respectively.Application of potassium fertilizer could improve maize stem morphology and mechanical index,improve maize lodging resistance,promote maize plant growth and development,and then affect maize yield and quality.The results showed that high-yield type>middle-yield type>low-yield type of maize inbred lines with different yield types.The increase of plant height,ear height,stem diameter,stem puncture strength,stem compressive strength,stem bending strength,grain number per ear and 1 000-grain mass,as well as the decrease of ear tip barrenness of maize inbred lines could significantly improve maize yield. Therefore,the above indexes can be used as screening indexes of yield of maize inbred line under potassium fertilizer application.This experiment provided a theoretical basis for breeding high-yield and high-quality maize materials,selecting suitable basic materials for studying the physiological characteristics of maize inbred lines with different yield types,and provided a reference basis for high-yield and high-efficiency maize breeding and rational utilization of potassium fertilizer in China in the future.
  • ZHANG Rui, WANG Tengfei, ZHANG Meihua, ZHANG Yongsheng, YANG Changyu, CHEN Zhipi
    In order to reveal the response relationship between potato yield and rhizosphere soil microbiological characteristics under plastic film mulching and supplementary irrigation technology in semi-arid area,field experiments of potato mulching and supplementary irrigation were carried out in Dingxi irrigation experimental station of Gansu Province,which included six treatments,drip irrigation under plastic film(PFID),drip irrigation under semi mulching(PHID),furrow irrigation with full film mulching(PFIF),furrow irrigation without film mulching(PNIF),border irrigation with plastic film mulching(PFIB),and no irrigation in border field covered with plastic film(PFIBN).Flat planting without film and no irrigation(PNIN) was used as control.The effects of different treatments on potato yield,rhizosphere soil organic matter(TOM),organic carbon(TOC),microbial biomass carbon(MBC),microbial biomass nitrogen(MBN),urease and catalase were analyzed.The results showed that mulching methods had significant effects on rhizosphere soil microbiological properties under drip irrigation and furrow irrigation.MBC and MBN of full film mulching treatment were significantly higher than that of half mulching treatment,while urease and catalase activities of semi mulching or no mulching treatment were significantly higher than that of full mulching treatment(P <0.05).On the other hand,the activity of urease and catalase of all supplementary irrigation treatments(PFID,PHID,PFIF,PNIF,PFIB) were significantly higher than control treatment(PNIN).The potato yield,number of tubers per plant and tuber weight per plant of supplemental irrigation treatment were also significantly higher than control treatment.In particular,the yield of semi mulched drip irrigation treatment(PHID) was 44 603.70 kg/ha,which was 51.00% higher than control.Correlation analysis showed that potato yield was highly significant correlated with MBC,urease activity and catalase(P <0.01),and positively correlated with TOM and TOC(P <0.05).
  • LI Ju, GAO Chengfei, MA Ning, WANG Shuya, LUO Shilei, Lü Jian, FENG Zhi, HU Linli, XIAO Xuemei, YU Jihua
    To determine the best method of fertilizer reduction,high yield and stable yield in the production area of cauliflower in Northwest Plateau.The Lihe Green stem 100 days cauliflower was used as material,six treatments were set as follows:the non-fertilization(CK1) and conventional fertilization(CK2) as control;chemical fertilizer reduction by 30% added balanced fertilizer(T1);chemical fertilizer reduction by 30% added balanced fertilizer and biological organic fertilizer 3 000(T2), 6 000(T3),12 000 kg/ha (T4).Compared with CK2 treatment,the economic yield of cauliflower decreased by 3.8%,and the utilization rate of phosphorus fertilizer increased from 3.7% to 11.4% in treatment T1;the economic yield of T2,T3 and T4 increased by 9.5%,11.3%,18.8%,respectively. Meanwhile,the accumulation of dry matter and nutrients,and the each element distribution proportion in flower ball were increased. Compared with CK2,T2,T3 and T4 treatments increased dry matter accumulation by 0.5%,8.3% and 7.2%,nitrogen accumulation by 9.1%,19.7% and 19.1%,phosphorus accumulation by 3.1%, 12.1% and 11.9%,and potassium accumulation by 2.4%,11.2% and 10.9%,respectively. Among them,T3 and T4 treatments were significantly different(P < 0.05). In addition,compared with CK2,chemical fertilizer reduction combined with biological organic fertilizer significantly increased the utilization rate of phosphorus and potassium fertilizer. The utilization rate of phosphate fertilizer increased from 3.7% to 26% and 25.9%,and that of potassium fertilizer increased from 43.5% to 73.4% and 72.9%,respectively.Chemical fertilizer reduction by 30% added appropriate amount of biological organic fertilizer could promote the absorption,accumulation and rational distribution of nutrients on cauliflower,improve the fertilizer utilization rate,and then increase the yield.
  • LIAN Kaiqi, JI Shuang, ZHOU Lingling, SHI Zhiqi, WANG Fei, ZHANG Yuanchen
    Abstract (85) PDF (6) RichHTML (8)

    To clone the full gene sequence of Mythimna separate serine protease,and express it in vitro,Mythimna separata was used as the research object,the partial gene of serine protease was amplified by RT-PCR,and further the whole gene was obtained by RACE technology.The obtained gene sequence and deduced protein sequence of serine protease were analyzed using Blast software and other softwares.The gene of serine protease was expressed using the Escherichia coli expression system.The results showed that the full serine protease gene from Mythimna separate was successfully cloned and named as MsPG,with the sequence length of 2 010 bp,and the open reading frame of 1 593 bp,encoding 530 amino acids.Its protein had the molecular weight of 67.6 ku,and the isoelectrical point of 7.63,and owned a clip-type domain containing 6 cysteine,which was speculated as a clip-type serine protease.The amino acid sequence of MsPG had identities of 70.00%—91.92% to those of other Lepidoptera insects,and showed the highest identity(91.92%)to that of Heliothis virescens(GenBank No.PCG78401.1).The recombinant plasmid pET30a(+)-MsPG was successfully constructed and expressed by 0.1 mmol/L IPTG induction in E.coli prokaryotic expression system,with the highest expression at 25 ℃.In conclusion,MsPG has conserved functional sites in the serine protease family and can be prokaryotically expressed in vitro.

  • ZHAO Yajie, ZHAO Xuanwei, TIAN Zhendong, HU Shuping, BAO Haizhu
    In order to understand the differential gene mining and molecular regulation mechanism of abscisic acid metabolism of oil sunflower in response to drought stress, high-throughput transcriptome sequencing and physiological characteristics were measured on oil sunflower under normal water supply (CK) and drought stress (T) treatments. The results showed that the content of abscisic acid (ABA) was the highest at 8 d of drought stress, at 23.55 ng/mL, and higher than 0 d at 4,8 d of drought stress. The differentially expressed genes were screened and compared with CK on 8 and 16 days of drought stress, the number of differentially expressed genes in the treatment group was 940, 1 743, and the number of genes that were significantly down-regulated was 1 752, 3 037, respectively. According to the functional annotations of KEGG and GO, there were more gene types under drought stress at 16 d than at 8 d. Among them, the most involved biological processes, the second was involved in cell components, and the least involved in molecular functions. The main metabolic pathways annotated by KEGG were plant MAPK signal transduction pathway, plant hormone signal transduction, glycerophospholipid metabolism and so on. Analyzed the hormone-related genes under drought stress for 8 days. A total of 39 differentially expressed genes were found, and 7 genes were related to abscisic acid metabolism. The transcription factors involved in abscisic acid metabolism were ABF2, SAPK2, PP2C, AHG1, PYL2 and SAPK3 were up-regulated, and the expression level of drought stress was higher than that of CK. The conclusion of this study was that 7 different genes related to the up-regulation of ABA metabolism in oil sunflower under drought stress were discovered. The transcription factors involved in the metabolism of abscisic acid (PYR/PYL→PP2C→SnRK2→ABF) were all up-regulated, indicating that the plant when suffering from drought stress, it would resist drought stress by accumulating hormones such as abscisic acid.
  • CHEN Longzheng, LIU Jing, LIU Zhiyang, XIA Pengfei, YUAN Xihan, NING Yu
    Abstract (82) PDF (11) RichHTML (4)

    MLO gene,acting as susceptibility factor,play an important role in regulating responses of host plants against powdery mildew.To determine the gene function of bitter gourd McMLO1 gene,cloning,bioinformatics and expression analysis were carried out in the present study.The results suggested that the full-length of McMLO1 was 4 019 bp,among which the CDS sequence was 1 707 bp,encoding 568 amino acids.ProtParam prediction indicated McMLO1,whose molecular weight and theoretical isoelectric point was 65.40 ku and 9.36,respectively,was an unstable hydrophilic protein which located in cell membrane.McMLO1 protein,harboring a conserved MLO domain of 477 amino acids,was consisted of random coil and alpha helix on secondary structure.Multiple sequence alignment and phylogenetic analysis revealed high homology between McMLO1 and cucumber MLO protein.qRT-PCR analysis suggested that the expression level of McMLO1 in leaf was much higher than that in other tissues.Moreover,the expression level of McMLO1 was significantly up-regulated and reached peak at 6 hours after inoculation with powdery mildew pathogen,indicating McMLO1 participate in the process in response to powdery mildew in bitter gourd.

  • ZHANG Aoshen, YONG Xiaoyu, HAN Qiaoxia, YAO Yongwei, SHI Huanting, LI Gezi, KANG Guozhang
    Abstract (82) PDF (89) RichHTML (23)

    In order to investigate the effects of exogenous brassinolide on physiological characteristics,transcriptional level and growth of wheat at jointing stage under freezing stress,the semi-winter wheat variety Bainong 207 was used as material,and irrigated with 0.1 mg/L 2,4-Epibrassinolide(EBR)to study its chlorophyll content(SPAD value),malondialdehyde(MDA)content,relative conductivity,free proline(Pro)content,stress resistance related gene expression,young panicle freezing rate,grain yield,1 000 grains weight and dry matter quality.The results showed that before freezing stress,there was no significant difference in SPAD value,MDA content and relative conductivity between the control group and the treatment group,but the Pro content in the treatment group was significantly lower than that in the control group;after freezing stress,compared with the control,external application of EBR significantly reduced the MDA content and relative conductivity of wheat leaves,and increased the SPAD value and Pro content.At 48 hours of freezing stress,the relative expression of stress resistance related genes SOD,POD,CAT,P5CS,and WCS120 in the treatment group were significantly higher than those in the control.After freezing stress,compared with the control,the freezing rate of young panicles in the treatment group decreased significantly by 20.93 percentage point,while the grains yield,1 000 grain weight and dry matter mass increased significantly by 19.38,2.37,54.40 g,respectively.In conclusion,external application of EBR at jointing stage of wheat can improve the frost resistance of wheat and reduce the yield loss of winter wheat caused by low temperature and freezing injury by reducing the content of MDA,increasing the content of Pro,chlorophyll and the expression of stress resistance related genes SOD,POD,CAT,P5CS,and WCS120.

  • MA Jie, SUN Xiangyang, SUO Linna, WANG Lei, SUN Na, XU Ning, LI Jia
    Abstract (82) PDF (21) RichHTML (16)

    To investigate the effects of zeolite and Ca-Mg-based bentonite on the passivation of cadmium in calcareous soil in Northern China and the growth of pakchoi,a pot experiment was conducted to study the effects of two modifiers(mass fraction of zeolite was 0.5%,1.0%,2.0% and mass fraction of calcium-magnesium bentonite was 0.2%,0.3%,0.4%,0.5%,0.6%,0.8%)on pH value,available cadmium content of calcareous cadmium-contaminated soil in Northern China and cadmium content,dry matter accumulation and chlorophyll content of aboveground part of pakchoi.The results showed that compared with the control,adding different doses of zeolite could increase the soil pH value.With the increasing of zeolite application dose,the available cadmium content of soil,the aboveground cadmium content,the dry matter accumulation and chlorophyll content of pakchoi decreased gradually.The germination rate of pakchoi increased gradually,but the effects of zeolite treatments on the indexes determined in the experiment were not significant.The application of Ca-Mg-based bentonite significantly increased the soil pH value(0.70—1.07)and decreased the Cd content in the aboveground part of pakchoi(63.83%—93.62%).The aboveground dry matter accumulation(5.56%—29.22%)and chlorophyll content(5.42%—11.72%)of pakchoi increased,and the soil available cadmium content significantly decreased by a higher dose(≥0.4%).However,the germination rate of pakchoi decreased significantly and inhibited germination.The research showed that Ca-Mg-based bentonite was more suitable for the passivation of cadmium in calcareous soil in Northern China than zeolites.The content of available cadmium in calcareous soil and the content of cadmium in the aboveground part of pakchoi could reduce by applying Ca-Mg-based bentonite.However,the amount of application should be strictly controlled to avoid affecting the germination of pakchoi and reducing yield.The comprehensive analysis of quality and yield factors showed that the addition of 0.3% Ca-Mg-based bentonite could effectively reduce the aboveground cadmium content of pakchoi and increase the aboveground dry matter accumulation and chlorophyll content of pakchoi.

  • LI Fujian, XU Dongyi, LIU Kaili, ZHU Min, LI Chunyan, ZHU Xinkai, DING Jinfeng, GUO Wenshan
    Abstract (81) PDF (60) RichHTML (9)

    To investigate the cultivation approach achieving robust seedlings and high yields of wheat in rice stubble,the effects of three tillage methods,including plow tillage followed by rotary tillage(PR),rotary tillage twice(RR),and no-tillage(NT),on the growth and physiology of main stem and tillers of wheat seedlings and their single spike yield at maturity were studied under the conditions of rice straw retaining with full amount.The results showed that PR increased the stems number per plant at the beginning of over-wintering by 1.5%,12.8%,respectively,compared with RR and NT,with significant differences(P<0.05).Leaf area and dry matter weight per plant under PR were 94 cm2 and 787 mg,respectively,which were significantly higher than those under NT(P<0.05).Compared with NT,PR significantly improved activities of nitrate reductase(NR),glutamine synthase(GS),and glutamate synthase(GOGAT)of the top full expanding leaves in the main stem and the first,second,and third tillers(P<0.05).PR also significantly promoted the activities of diphosphate carboxylase(Rubisco),pyruvate phosphate dikinase(PPDK),triosephosphate isomerase(TPI),sucrose synthase(SS-Ⅱ),and sucrose phosphate synthase(SPS),and enhanced SPAD and soluble sugar content(P<0.05).And compared with NT,PR significantly increased the nitrogen content in seedling leaves of main stem,second and third tillers(P<0.05).Moreover,PR and RR increased leaf area and dry matter weight of the main stem and the first and second tillers.Therefore,compared with RR and NT,PR was more beneficial to the synergistic improvement of nitrogen metabolism,glucose metabolism,and light energy utilization at the seedling stage,increasing photosynthetic productivity and forming robust seedlings.The improvement of carbon and nitrogen metabolism and photosynthetic capacity by RR was higher than that by NT but was weaker than that by PR.There were no significant differences between PR and RR in the grain number per spike,single grain weight of the main stem(P>0.05),but they were significantly higher than those under NT(P<0.05).The grain number per spike in the first and second tillers under PR was significantly higher than that under RR and NT(P<0.05),but single grain weight and grain yield per spike of the first,second,and third tillers were not significantly different between PR and RR(P>0.05).In conclusion,PR could improve the activity of glucose and nitrogen metabolism and photosynthetic physiology of the main stem and the first and second tillers of wheat in rice stubble,resulting in the increases of the grain number per spike and the productivity per spike through forming strong seedlings.

  • CHEN Wenxun, ZHOU Ting, YAN Qiongxian, TIAN Lina, CAO Manhu, TAN Zhiliang, ZOU Aihua
    Abstract (81) PDF (26) RichHTML (5)

    The aim of this study was to investigate the effects of low protein diet on meat quality,amino acid and fatty acid composition of skeletal muscle and blood,and mRNA expression of myogenic regulatary factor genes of black goat.A total of 24 female Xiangdong black goats were randomly divided into control group(CON,10.77% CP,n=12)and low protein group(LP,5.52% CP,n=12).The pre and post-trial lasted for 25 d and 45 d,respectively.The results were as follows:Low protein diet increased L* 24 h of meat color,but had no difference on pH value and water loss rate.The percentage of type Ⅰ muscle fiber and type Ⅱ muscle fiber had no difference between two groups,only the average fiber cross sectional areas of semitendinosus had a tendency to reduce.Valine,leucine,lysine,phenylalanine,glutamate and total essential amino acids of vastus lateralis significantly reduced,isoleucine,methionine,aspartate,arginine and total amino acid tended to decrease.Glutamate of semitendinosus in the LP group significantly reduced,threonine and serine tended to decrease.Compared with the control group,C20:2 and C22:1n9 of semitendinosus in the LP group significantly decreased.Total PUFA,C18:1n9c,and C20:4n6 of vastus lateralis in the LP group significantly decreased;C18:1n9c of longissimus dorsi in the LP group significantly decreased.Compared with the control group,the mRNA expression of MYOD in semitendinosus were significantly down-regulated in the CON group.In conclusion,low protein diet didn't affect type conversion and cross sectional area of muscle fiber.Dietary protein deficiency reduced the deposition of amino acid in muscle,while reduced the deposition of polyunsaturated fatty acids in muscle,which affected on muscle flavor of goat.

  • YANG Jinrui, WANG Wenqian, WANG Yue, YAO Yao, JI Ze, LI Junyi, CHEN Bin, XIAO Guanli
    In order to clarify the diversity and differences of endophytic bacteria in leaves of different potato varieties and provide basis for studying the mechanism of potato stress tolerance and the development of endophytic bacteria.The endophytic bacteria from the leaves of four main potato varieties in Yunnan Province were isolated and cultured on LB medium,all those strains were identified based on the morphological characteristics and the 16S rDNA sequence.There were 10 species of endophytic bacteria belonging to 9 genera,8 families and 3 phyla were isolated from Qingshu No.9 leaves. Staphylococcus and Exiguobacterium were the dominant genera,and the relative abundance of both genera were 18.18%.There were 12 species of endophytic bacteria belonging to 11 genera,11 families and 4 phyla were isolated from Hui-2 leaves. Acinetobacter and Pseudomonas were the dominant genera,and the relative abundance of both genera were 15.38%.There were 13 species of endophytic bacteria belonging to 10 genera,9 families and 4 phyla were isolated from Hezuo No.88 leaves. Acinetobacter were the dominant genera,with a relative abundance of 23.53%.There were 17 species of endophytic bacteria belonging to 8 genera,7 families and 4 phyla were isolated from Lishu No.6 leaves. Staphylococcus were the dominant genera,with a relative abundance of 22.22%.There were 37 species of culturable bacteria were isolated from the leaves of four potato varieties.There were 4,8,4,10 unique endophytic bacteria from the leaves of Qingshu No.9,Hui-2,Hezuo No.88 and Lishu No.6,respectively.The species composition and dominant genera of endophytic bacteria in leaves of different potato varieties were different,the results would provide bacteria resources for studying the function of endophytic bacteria in potato leaves.
  • LI Qi, YANG Changheng, WANG Yong, LIN Yaqiu, XIANG Hua, ZHU Jiangjiang
    Abstract (79) PDF (18) RichHTML (2)

    The aim of this study was to obtain the CDS of CIDEB and CIDEC gene,detect the expression of CIDEB and CIDEC in different tissues of goats,predict their interaction proteins.It would provide basial data for revealing the regulatory role of CIDEs in lipid metabolism.The CDS of CIDEB and CIDEC gene were cloned by RT-PCR,and the biological characteristics and interaction proteins of CIDE proteins were analyzed by online tools.Real-time quantitative PCR(RT-qPCR)was used to detect the expression of CIDEs and the genes,encoding the interaction proteins,in different tissues of goats,following the correlation analyses by a SPSS software.The CDS of CIDEB was 660 bp,encoding 219 amino acids,while the CDS of CIDEC was 714 bp,encoding 237 amino acids.Phylogenetic tree analysis showed that goat CIDEA,CIDEB and CIDEC were closely related to corresponding genes from sheep.RT-qPCR detection showed that CIDEA,CIDEB and CIDEC were highly expressed in the rumen,liver and small intestine of goat,respectively.The expression of CIDEA was significantly correlated with the expression of DFFB (P<0.01),ELOVL3 (P<0.01),DIO2 (P<0.05),TMEM26 (P<0.05),PRDM16 (P<0.05)and PLIN1 (P<0.05)in rumen.The expression of CIDEB was significantly correlated with that of DFFA and SDR39U1 in liver(P<0.05).The expression of CIDEC was obviously correlated with that of PLIN2,GPLD1 and ADIPOQ in small intestine(P<0.05).Through the analysis,we found genes related to the expression level of CIDEs,which could provide a theoretical basis for further revealing the role of CIDE gene in lipid metabolism and its regulatory network.Moreover,the proteins of CIDEs were all lipid-droplet related proteins,and could also provide a reference for further studying the precise mechanism of lipid droplet formation.

  • SHI Jianlei, XIONG Zili, SU Shiwen, WANG Kelei, ZAI Wenshan
    Abstract (79) PDF (43) RichHTML (19)

    To explore bacterial wilt resistance genes,RNA sequencing was used to characterize the transcriptomes of resistant and susceptible tomato inbred lines with Ralstonia solanacearum inoculation(RsI).The results showed that a total of 75.02 Gb high-quality data were generated in 12 libraries.With the fold change(FC)≥2 and false discovery rate(FDR)<0.01 as the standard,970 and 695 differentially expressed genes(DEGs)were identified in the two tomato lines,respectively.The 1 312 DEGs accounted for 3.71% of the total.Among them,the numbers of up-regulated genes were 457 and 450,respectively,totaling 693;the numbers of down-regulated genes were 513 and 245,respectively,totaling 621.Among these DEGs,836 genotype-specific DEGs were highlighted.These DEGs were mainly divided into 47 functional groups such as metabolism,regulation,response,binding,and catalysis,and 88 metabolic pathways such as DNA replication,secondary metabolite synthesis,plant-pathogen interaction,and signal transduction by GO and KEGG annotation.Specifically,4 NBS resistance genes,6 plant-pathogen interaction genes,11 plant hormone signal transduction genes,22 defense response genes,32 protein kinases,65 transcription factors,and several other important functional genes were involved,indicating that they played important roles in response to Rs.Promoter analysis revealed that these genes possessed multiple defense and stress response elements.The output was confirmed using RT-qPCR for 50 representative genes.It was found that more than half of the genes were consistent with RNA-seq in expression.Solyc02g086980.3 and Solyc04g011670.3 might be involved in the resistance response,whereas Solyc01g073985.1,Solyc09g092580.4,Solyc09g098100.4,and Solyc10g081300.1 might be the opposite.Together,these gene expression profiles serve as fundamental information to understand the potential molecular basis in the response to Rs in tomato,and facilitate the application of related resistance genes in breeding.

  • SONG Shanshan, ZHANG Xinting, WANG Qi, LU Yuanjing, HOU Chunyan, WANG Dongmei
    Abstract (77) PDF (5) RichHTML (3)

    The aim to study the role of TaRanGAP2 in HR induction of wheat resistance to Puccinia triticina,so as to lay a foundation for elucidating the mechanism of Puccinia triticina resistance and breeding for disease resistance in wheat at the molecular level.The wheat near-isogenic line TcLr26 and its recurrent parent Tatcher(Tc)were used to form incompatabile(TcLr26×260)and compatabile combination(Tc×260)with Puccinia triticina race 260,respectively.Bioinformatics analysis showed that the total length of CDS of TaRanGAP2 was 1 665 bp,encoding 554 amino acids.Analysis of its conserved domains revealed that the protein encoded by TaRanGAP2 gene belonged to the Ran GTPase family;RT-qPCR was used to detect the expression of TaRanGAP2 in the two combinations.It was found that the expression of TaRanGAP2 was significantly up-regulated in the incompatible combination;the subcellular localization detection of TaRanGAP2 by tobacco transient transformation system showed that TaRanGAP2 was localized in the cytoplasm;Virus induced gene silencing(VIGS)technology were used to silence TaRanGAP2 gene in TcLr26.After inoculation with Puccinia triticina race 260,it was found that HR area of TaRanGAP2 gene-silencing plants was significantly increased,and the number of HMCs were also significantly increased compared with the unsilenced plants control plants.These results proved that TaRanGAP2 played a positive regulatory role in the induction of HR in wheat resistance to Puccinia triticina.

  • ZHAO Huiying, YANG Guangkai, GAO Yan, ZHANG Xiaojun, HAO Yanyan
    Abstract (77) PDF (58) RichHTML (15)

    Gene STAYGREEN(SGR)plays an important role in the process of plant chlorophyll degradation and metabolism.To explore the biological function of gene SGR2,MdSGR2 gene sequence was cloned from the peel of Granny Smith by RT-PCR.The homology,physicochemical properties,protein structure and cis-acting elements of the promoter were predicted and analyzed by bioinformatics software.The plant overexpression vector was constructed by double enzyme digestion.The results showed that the complete open reading frame(ORF)cDNA of gene MdSGR2 was 840 bp.Encoding 279 amino acids in total,it belonged to the Staygreen superfamily.Physicochemical analysis showed that the total molecular weight of the protein was 31.27 ku,and the isoelectric point pI was 8.52,indicating that the protein was an unstable hydrophilic protein.Homology analysis showed that MdSGR2 encoded amino acid sequence was of the highest homology with Pyrus ussuriensis,reaching 84.12%.The protein structure analysis showed that the secondary structure and tertiary structure of MdSGR2 was determined structurally to be 40.55% α-helix and 44.44% random coil.Promoter analysis showed that the promoter region of the gene contained cryogenic cis-acting elements,photo-responsive elements and induced cis-acting elements such as abscisic acid,salicylic acid and methyl jasmonate response elements.These results suggest that gene MdSGR2 may be regulated by many factors such as environment and hormones.The plant overexpression vector pCAMBIA2301-MdSGR2 was successfully constructed.

  • YAN Liuyan, LI Jianfeng, ZHANG Shiwen, ZHANG Bo, WANG Yongfang, ZHANG Xiaomei, ZU Chaofan, WANG Zhenshan, SANG Luman, HE Zhanxiang, JIA Xiaoping, DONG Zhiping
    Abstract (77) PDF (26) RichHTML (69)

    The SiPRR73 gene was cloned from Yangu 11 using RT-PCR technology,and through analyzing tissue-specific expression,responsive features of SiPRR73 to different photoperiods,photo-thermal combinational treatments and five abiotic stress treatments,the regulation mode of photoperiod and temperature on SiPRR73,and the responsive pattern of SiPRR73 to abiotic stresses in foxtail millet were explored. The results showed that totally 2 928 bp cDNA sequence of SiPRR73 was obtained from Yangu 11,which included 2 283 bp CDS region,encoding 760 amino acids. The SiPRR73 proteins of C4 crops including Panicum miliaceum,Panicum hallii,Sorghum bicolor and Zea mays showed relatively close relationship with SiPRR73. The second parietal leaf was the highest expression tissue of SiPRR73,but the expression level at root,stem and panicle tissues was relatively lower. The expression level of SiPRR73 was higher at light period than that at dark period under both short-day and long-day conditions,and during the whole vegetative growth phase,SiPRR73 showed higher expression level under long-day compared to short-day,which indicated that the expression of SiPRR73 was induced by light and controlled by photoperiod. The temperature determined expression peak number of SiPRR73 and the photoperiod determined occurrence time of expression peaks,so temperature and photoperiod participated in regulating of SiPRR73 expression mutually. PEG and low temperature stresses induced SiPRR73 expression totally,NaCl induced SiPRR73 expression at early stress stage,but inhibited it at later stress stage. Fe stress inhibited SiPRR73 expression at early stage,but induced it at later stage. ABA stress caused the close responsive feature of SiPRR73 to NaCl. This study indicated that SiPRR73 showed light-dependent expression feature,and photoperiod and temperature regulated SiPRR73 by interaction pattern,suggesting that SiPRR73 participated in adaptability regulation process to different photo-thermal conditions and might play a certain role in coping with drought,low temperature,ABA,NaCl and Fe stresses in foxtail millet.

  • ZHOU Yuanhui, MA Shangyu, WANG Yanyan, HUANG Zhenglai, ZHANG Wenjing, FAN Yonghui, HOU Junyou, GAI Panpan, SHAN Yajing
    In 2015-2016 wheat growing season, Wankenmai 076 and Wanmai 52 were used at the Wanzhong Experiment Station of Anhui Agricultural University to study the effect of waterlogging after anthesis on the grain filling characteristics and yield of winter wheat. Four treatments, 0(W0), 3(W3), 6(W6), and 9(W9)days of waterlogging after anthesis, were established. The results showed that 14 days after flowering, no significant differences were observed in net photosynthetic rate, SPAD value, malondialdehyde(MDA)content and peroxidase(POD)activity in flag leaf between W0 and W3 treatments. Moreover, W6 and W9 treatments showed significant lower values than that of W0. However, superoxide dismutase(SOD)and catalase(CAT)activity of flag leaf were different in different varieties, SOD and CAT activities of flag leaf in Wankenmai 076 of W9 treatment were significantly lower than those of other treatments, SOD and CAT activity of flag leaf in Wanmai 52 of W3 and W6 treatment were significantly lower than those of W0 treatment, but higher than those of W9 treatment. In addition, W6 and W9 showed significantly lower average grain filling rate than W0 and W3 treatments, and different wheat varieties had different effects on the grain filling process under waterlogging conditions. There was no significant difference in spike number of wheat under waterlogging after anthesis. Moreover, there was no significant difference in grain number between W0 and W3 treatments, while W6 and W9 treatments were significantly lower than that of W0 in Wankenmai 076. However, the grain number and weight of W3, W6 and W9 in Wanmai 52 showed significantly lower values than those of W0 treatment, and W9 treatment was the lowest. The 1000-grain weight and yield of both varieties ranked as W0>W3>W6>W9. After 9 days of waterlogging, the yield of both varieties decreased by 34.99% and 40.44%. Under the conditions of this experiment, waterlogging during flowering period reduced the photosynthetic capacity of flag leaves of wheat, affected the grain filling process, reduced the average grain filling rate, and led to the decrease of 1000-grain weight, thus reducing the final yield.
  • PING Huaixiang, CUI Jianyu, CHEN Shuo, WEI Lulu, CHEN Qing, ZHANG Delong
    Abstract (76) PDF (22) RichHTML (3)

    Stoichiometry characteristics of soil CNP and MBC,MBN,MBP,and the changes of enzyme activity with different fertilization can help to understand the relationship between fertilization and soil nutrient balance,and thus formulate more reasonable fertilization measures.Published literature data on the changes of CNP status and enzyme activity in fertilized soil were collected,and the changes of CNP stoichiometry and enzyme activity in cultivated soil nutrient/biomass with different fertilization were analyzed by Meta-analysis.The results showed that the C/N under single fertilizer,compound fertilizer and the application of chemical fertilizer and manure was 14.1,14.8 and 23.2,the value of C/P was 20.4,15.3 and 14.8,the value of N/P was 5.03,4.43 and 3.10,respectively.There was no significant difference in MBC/MBN and MBN/MBP with different fertilization.The ratio of MBC/MBP was:application of fertilizer and manure>single fertilizer>compound fertilizer.Different fertilizations could improve the activities of soil sucrase,phosphatase,urease and catalase,especially the application of chemical fertilizer and manure(which increased urease activity by 100%).It showed that the application of chemical fertilizer and manure could significantly affected the stoichiometric ratio of CNP in soil,enhanced microbial activities and promoted microorganisms to release more enzymes conducive to soil nutrient mineralization.It was an effective way to improve crop yield and ensure soil health in agricultural production.Soil enzyme activity was restricted by soil fertility and microbial process.A certain correlation between CNP stoichiometric ratio and soil enzyme activity was observed,which was also affected by land use type,soil pH and rotation system.The study of soil nutrient-microorganism-enzyme activity system can provide a theoretical basis for optimizing fertilization management in farmland system.

  • XU Lingqi, GUO Xiaohong, ZHANG Jianing, ZHAO Yang, LI Xiaolei, LIU Shaofeng, CUI Zhiyuan, AN Yiliang, LÜ Yandong
    Abstract (76) PDF (23) RichHTML (3)

    In order to explore the changing law of dry direct-seeding rice quality under different organic fertilizer treatments,this experiment was conducted from 2018 to 2020.Longjing 31 was used as the test material.Treatments includes:zero fertilizer(N0),conventional fertilization(NPK),Biochar+conventional fertilization(OF1),seaweed bio-organic fertilizer+conventional fertilization(OF2),Jishiwang bio-organic fertilizer+conventional fertilization(OF3),attapulgite organic fertilizer+conventional fertilization(OF4).Changes in the processing quality,appearance quality,nutritional quality,and cooking and eating quality of dry direct-seeding rice were investigated.Compared with NPK,the head rice percentage of OF1,OF2,OF3,OF4 increased by 2.64 ,1.78,1.06,2.53 percentage points,respectively.The chalkiness degree of OF2 was the highest(1.82%),which was 0.11 percentage points higher than that of NPK.However,the chalkiness degree of OF1,OF3 and OF4 were lower than that of NPK,with an average decrease of 0.26,0.41,0.51 percentage points,respectively.There was no significant difference in grain length,grain width and length width ratio among the treatments.From 2019 to 2020,compared with NPK,organic fertilizer treatment increased protein content,but decreased amylose content;the protein contents of OF1,OF2,OF3 and OF4 were 8.99%,9.13%,9.08% and 9.16% respectively,which were 0.54,0.68,0.63,0.71 percentage points higher than those of NPK;the amylose content of OF1,OF2,OF3 and OF4 decreased by 0.20,0.64,0.69,0.38 percentage points,respectively;compared with NPK,the application of organic fertilizer at the initial stage could improve the luster,taste,flavor and taste value of dry-direct seeding rice,but long-term application of organic fertilizer would lead to poor cooking and eating quality;from 2019 to 2020,the average taste value of NPK treatment was 72.70,compared with NPK,OF1,OF2,OF3 and OF4 reduced the aroma,luster,taste,flavor and taste value of dry direct-seeding rice by 5.53%,6.40%,3.71% and 3.23%,respectively,but organic fertilizer increased the integrity of rice.In conclusion,long-term application of organic fertilizer can improve the processing quality,appearance quality and nutritional quality of dry direct-seeding rice,but it is not conducive to the formation of cooking and eating quality and reduce amylose content.

  • ZHENG Mengjing, LI Yan, JIA Xiuling
    Diversified rotation system is an important agricultural technical measure for the sustainable development of modern agriculture, which is of great significance to improve the efficiency of regional resource utilization, meet the diversified demand of the market and promote the reform of agricultural supply side. The implementation of diversified crop rotation system plays a key role in reducing the input of chemical products, protecting the ecological environment, increasing the biodiversity of farmland and reducing the damage caused by diseases, insects and weeds. In view of the current development status of monoculture system and a series of ecological environment problems, this paper put forward the necessity of implementing diversified crop rotation mode. On the basis of introducing the development process of crop rotation system, the yield effects, ecological environment benefits, increasing biodiversity, crop cover effects and research methods of diversified crop rotation system were analyzed in detail. At the same time, it also put forward the future development direction of diversified rotation system, such as integrating management technology of agricultural machinery and agronomy technologies to developing intensive rotation system. And establishing comprehensive evaluation system of diversified crop rotation mode was contributed to give full play to the comprehensive effect of crop rotation. Also, research methods of crop rotation system should be innovated to explore the internal mechanism of crop rotation effect. The development of diversified crop rotation system had become an important way to realize the healthy, green and sustainable development of agriculture worldwide.
  • HE Qiulan, ZHANG Hang, WANG Zhengwei, WAN Liqiang, HAI Meirong
    Abstract (75) PDF (32) RichHTML (5)

    In order to explore the effects of different concentrations of exogenous melatonin on the growth of potato seedlings under low temperature stress after cold acclimation,using Yunnan main potato variety Hezuo 88 as material,the effects of different concentrations of exogenous melatonin(50,100,150 μmol/L)on the growth and active oxygen metabolism system of potato seedlings under low temperature stress(4 ℃ 14 d ,-2 ℃ 12 h)were studied.The purpose of this study was to explore the feasibility of exogenous MT in alleviating the injury of potato seedlings caused by low temperature stress.The results showed that compared with potato seedlings under low temperature stress,the survival rate of seedlings treated with 50 μmol/L MT increased by 38.89 percentage point,and the plant height and stem diameter increased by 2.92% and 1.86%,the contents of SS,SP and Pro increased by 12.66%,13.80% and 1.96%,the activities of POD and CAT increased by 29.24% and 351.62%.The plant height and stem diameter of seedlings treated with 100 μmol/L MT increased by 16.97% and 2.82%,the contents of MDA and H2O2 decreased by 19.35% and 0.48%,the activities of POD and CAT increased by 55.44% and 213.56%.The survival rate of seedlings treated with 150 μmol/L MT increased by 27.78 percentage point,the plant height and stem diameter increased by 5.62% and 13.20%,the content of MDA decreased by 9.45%,the content of Pro increased by 38.70%,SOD,POD and CAT activity increased by 52.61%,25.25% and 300.94%,and the content of AsA and GSH increased by 3.48% and 3.97%.The results showed that exogenous MT could promote the growth of potato seedlings,reduce the content of MDA and H2O2 in leaves,promote the accumulation of SS,SP and Pro,increase the activities of SOD,POD and CAT,and increase the content of AsA and GSH.Thus,it can alleviate the damage of low temperature stress to potato seedlings,and then improve the ability of potato seedlings to resist low temperature stress.In summary,50 μmol/L MT treatment could better improve the stress resistance of potato seedlings under low temperature stress.

  • WANG Yong, MENG Qingfeng
    Abstract (75) PDF (20) RichHTML (28)

    In order to explore the effects of cattle manure on soil salinity and sodicity on the sodic soil in long-term experiments,the experiments were performed in a randomized complete block design with four treatments;soils that received manure applications for 8,12,18 years were used as the experimental treatments,and soil that did not receive cattle manure application was used as the control treatment(CK).The results showed that the application of cattle manure to saline-sodic soil resulted in a reduction in the bicarbonate ion(HCO3-)contents,the elimination of carbonate ions(CO32-),the decrease in soil bulk density(ρb),the increases in soil porosity(ft)and soil organic matter(SOM),the decreases in the exchangeable and soluble sodium ion (Na+)contents associated with increases in the exchangeable calcium ion(Ca2+),soluble potassium ion(K+),and magnesium ion(Mg2+)contents compared to those in untreated soil.The soil exchangeable sodium percentage(ESP)and pH were both significantly and positively correlated with the exchangeable Na+ and HCO3-,and CO32- contents,and soil pH was significantly and negatively correlated with SOM.Regression analysis showed that the dominant factors affecting the sodium absorption ratio(SAR)were the soluble Mg2+ and Na+ contents in the soil.Pearson correlation analysis showed that there were significantly negative correlation between the accumulated amount of cattle manure among the indicators of soil salinization degree,such as pH,EC,ESP and SAR.It was concluded that long-term manure application significantly decreased the soil pH,ESP,electrical conductivity(EC)and SAR due to the replacement of soil colloidal Na+ with Ca2+,the leaching of soil soluble salts from the topsoil and changes in the soil soluble salt ion composition.These outcomes were likely due to the decrease of ρb associated with increase of ft and Ca2+ and Mg2+ contents caused by annual manure application.

  • WAN Xuejie, ZHANG Yan, SHI Changhai, ZHOU Xuancai, SUN Xinling, LI Songjian, ZHANG Shoufu, LIU Yiguo
    In order to explore the effects of drought priming on antioxidant properties of wheat seedlings, control treatment and drought training treatment were set up in this experiment. PEG6000 nutrient solution was used for drought training when one heart and one leaf, and then Hoagland's total nutrient solution was restored. When three leaves are one heart, the drought stress is treated separately, and the dynamic changes of the substances related to the antioxidant properties in the young leaves were observed. The results showed that with the increase of drought degree and stress time, SOD, POD activity and soluble protein decreased, but MDA and soluble sugar content increased significantly. Compared with the control, the leaf SOD, POD activity and soluble protein decreased less after drought exercise, the soluble sugar content increased more, and the MDA increased less. Under the different drought stress, the antioxidative capacity of drought-stressed wheat under 25% PEG6000 stress was higher than that of 35% PEG6000. This showed that a certain concentration of drought training in wheat can significantly enhance its ability to adapt to drought conditions, which was beneficial to the growth of the seedling stage and provides theoretical support for the research on wheat resistance.
  • WEI Jinjian, QIN Xiaomin, NONG Yuqin, LUO Yanfei, LU Jinmei, CHEN Yuanquan, WEI Chizhang
    To explore the metabolic functional diversity of root soil microbial community in tea and soybean intercropping contributes to further understand the mechanism of high yield and good quality of tea garden. A field trial with three planting patterns(tea monocropping, soybean monocropping, tea and soybean intercropping)was conducted to analyze the changes of metabolic function diversity of soil microbial community by Biolog technology. Results showed that intercropping increased the average well color development(AWCD)of soil microbial community of tea and soybean by 11.52% and 12.99% at 120 h, respectively, compared with monocropping pattern. Intercropping enhanced the utilization of carbohydrate, amino acid, carboxylic acid and amine by soil microbial community of tea and soybean and improved the soil microbial diversity indices. Principal component analysis demonstrated that tea and soybean intercropping significantly improved the metabolic diversity of soil microbial community by changing the utilization patterns of carbohydrates and carboxylic acids by soil microorganism. Under this experiment, intercropping of tea and soybean changed the composition of rhizosphere microbial community and improved the functional diversity of microbial community.
  • FAN Fangfang, JIAO Xiaoyan, LIU Jiaqi, GUO Jun, WANG Jinsong, WU Ailian, BAI Wenbin, PING Junai
    Abstract (74) PDF (19) RichHTML (3)

    To study the decomposition characteristics of sorghum and maize residues and the functional diversity of microbial community in straw decomposition,under different residual types(sorghum stalk-leaves,maize stalk-leaves,sorghum root,maize root),soil type(cinnamon soil,yellow loam soil),nitrogen treatment(adjust C/N ratio,not adjusted),the culture experiment method was used to explore the decomposition characteristics of residues under different decomposition conditions,and to analyse the metabolic functional diversity of microbial community used the BIOLOG-ECO plates method in straw decomposition process.The results showed that the decomposition rate of sorghum and maize residues was faster in the early stage than the late stage.The residual dry matter degradation rate and CO2 release rate had the following pattern of decomposition rate was shown,sorghum stalk-leaves>maize stalk-leaves>sorghum root>maize root,under the same soil and nitrogen treatment.According to the second experiment, the dry matter degradation rates of sorghum stalk-leaves and maize stalk-leaves were 55.50% and 48.00%,respectively,however,the dry matter degradation rate of sorghum root and maize root were 31.25% and 16.75%,respectively at 60 d of incubation,under the condition of cinnamon soil+N treatment.The degradation rate of hemicellulose and cellulose in root were lower than that in stalk-leaves,under the same soil and N treatment conditions,the decomposition of hemicelluloses and cellulose was decreased by 23.70,18.80 percentage point,in sorghum root treatment than sorghum stalk-leaves treatment at 90 d of incubation,under the condition of cinnamon soil+N treatment.The microbial metabolic activity of straw was the highest at 30 d and lowest at 90 d during decomposition.The metabolizable ability of microbial community of straw decomposition to amines and phenolic acids was lower on the 1st day of decomposition,and the metabolizable ability to carbohydrates,amino acids and polymers was significantly decreased on the 90th day,compared with 30 d.Collectively,the decomposition of sorghum residues was easier than maize,and adjusting C/N ratio could accelerate the decomposition of residue to a certain extent.The microbial metabolic diversity was the highest at 30 d under the experimental conditions.

  • LIU Zhiping, MA Yu, LI Jianbo, GUO Chengyu, LÜ Ersuo, WANG Haize, WANG Wendi, XU Shoujun
    Abstract (74) PDF (54) RichHTML (13)

    In order to explore the relationship between soluble sugar content and grain starch content of spring barley,Mengpi 3 and Mengpi 5 were used as test materials,and four density treatments of 3.75,4.50,5.25,6.00 million plants/ha were set.The dynamic changes of soluble sugar and starch content in leaves,stems and grains during grain filling of barley under different planting densities were studied from 2018 to 2019.Its effect on the formation of grain starch components was analyzed.The results showed that the soluble sugar content in leaves,stems and grains of 2 years and 2 varieties showed a unimodal curve with the grain filling process,and the peak value appeared on 21 days after anthesis.With the increase of planting density,the soluble sugar content in leaves,stems and grains was increased first and then decreased,with the increase of planting density,and the peak value was 5.25 million plants/ha treatment.The contents of total starch,amylose and amylopectin in grains of 2 varieties in 2 years increased gradually with the filling process,increased first and then decreased with the increase of planting density,and the peak value was 5.25 million plants/ha treatment.Correlation analysis showed that there was a very significant positive correlation between the soluble sugar content of leaves,stems and grains at each filling stage and the starch component content of grains at maturity;the path analysis showed that the soluble sugar content of leaves,stems and grains at 21 days.After anthesis had the greatest effect on the total starch content of grains at maturity,and the path coefficients reached 1.002 3,0.580 4 and 0.745 5,respectively.The soluble sugar contents of leaves,stems and grains at 35 days after anthesis and 7 days after anthesis were the most affected on amylose content,and the path coefficients were 0.776 6,0.469 7 and 0.715 6,respectively.The soluble sugar content of leaves at 21 days after anthesis,stems at 7 days after anthesis and grains at 14 days after anthesis had the greatest effect on the amylopectin content of grains at maturity.The path coefficients were 1.046 9,0.638 2 and 0.775 6,respectively.5.25 million plants/ha was the most suitable planting density to increase the content of starch components in barley grains.The soluble sugar content of stem and grain at the beginning of grain filling 7 d after anthesis is of great significance to improve the yield and quality of barley.

  • LI Hui, WEN Chunxiu, LIU Lingdi, WEN Saiqun, TANG Yinghong, JIANG Tao
    Abstract (73) PDF (24) RichHTML (23)

    In order to investigate the function of 5-phosphomevalonate kinase PMK gene in biosynthesis pathway of terpenoids in Perilla frutescens L.,we analyzed the transcriptome data of Perilla frutescens and mined the reference sequence of PMK.The PMK gene was cloned from Perilla frutescens by gene cloning technique,and was analysed using bioinformatics and Real-time PCR(qRT-PCR).The results showed that the ORF of PfPMK gene was 1 524 bp,encoding 507 amino acids.Bioinformatics analysis showed that the molecular weight of PfPMK was 54.73 ku and the isoelectric point was 5.20,which was a hydrophilic protein.The amino acids of PfPMK were higher homologous with SmPMK,SsPMK,SbPMK and PvPMK,which indicated PfPMK protein was highly conserved in the evolutionary process.The phylogenetic analysis showed that PfPMK was closely related to PMK of Salvia miltiorrhiza and Salvia splendens.WoLF-PSORT predicted that PfPMK protein might be located in the plasma membrane or endoplasmic reticulum.Real-time fluorescence quantitative PCR results showed that PfPMK gene was expressed in roots,stems and leaves of Perilla frutescens,and the expression level in roots was higher than that in leaves and stems.The fluorescence quantitative PCR analysis showed that the expression level of PfPMK gene was higher in the middle and late September.The full length of PfPMK gene was first cloned from Perilla frutescens.Bioinformatics analysis showed that PfPMK gene belonged to 5-phosphomevalonate kinase gene and participates in the biosynthesis of terpenoids in Perilla frutescens.

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