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2019 Vol. 34, No. 3
Published: 2019-06-28

Crop Genetics·Germplasm Resources·Biotechnology
Tillage and Cultivation·Physiology and Biochemistry
Resource and Environment·Plant Protection
0 Table of Contents for VOL.34 No.3
2019 Vol. 34 (3): 0-0 [Abstract] ( 29 ) 全文: HTML   PDF (253 KB)  ( 59 )
       Crop Genetics·Germplasm Resources·Biotechnology
1 Cloning and Functional Analysis of Cold Response Factor VcICE1 in Blueberry
SONG Yang, LIU Hongdi, WANG Haibo, ZHANG Hongjun, LIU Fengzhi
DOI: 10.7668/hbnxb.201751537
In order to excavate the role of blueberry cold response genes in cold stress and explore the regulation mechanism of cold stress response, a cold response factor gene VcICE1 (Inducer of CBF3 expression 1) from blueberry was isolated by PCR, and its expression pattern and response to cold stress were analyzed. The transgenic Arabidopsis thaliana were generated via Agrobacterium -mediated transformation, and the expression differences in response to cold stress were compared between the transgenic line and wild-type of Arabidopsis. The transient expression assays in tobacco leaves were carried out to test the transcriptional regulation of AtCBF3 by VcICE1. Sequence analysis showed that the open reading frame(ORF) of VcICE1 was 1 566 bp in length, encoding 522 amino acids. Protein structure analysis showed that VcICE1 contained a basic helix-loop-helix domain. The phylogenetic tree indicated that the blueberry VcICE1 exhibited the highest sequence similarity with the Arabidopsis thaliana AtICE1. Expression analysis showed that VcICE1 could express in roots, stems, young leaves, flowers and fruits, but the expression levels varied in different organs, the highest in young leaves and relatively low in fruits. The expression analysis result indicated that VcICE1 could be induced by cold stress treatment. The VcICE1 protein could activate the expression of AtCBF3 and favorably contributed to cold tolerance in transgenic Arabidopsis. VcICE1 could respond significantly to cold treatment. It was speculated that VcICE1 played a regulatory role in response to cold stress.
2019 Vol. 34 (3): 1-8 [Abstract] ( 85 ) 全文: HTML   PDF (1310 KB)  ( 99 )
9 Expression and Regulation Analysis of GhTFL1b Gene in Upland Cotton
ZHANG Xiaohong, WANG Hantao, WANG Congcong, ZHANG Fangfang, DENG Yan, WEI Hengling, HU Genhai
DOI: 10.7668/hbnxb.201751715
In order to study the response of cotton phosphatidylethanolamine-binding protein GhTFL1b gene to stress and physiological function, we cloned the GhTFL1b gene from upland cotton, and then analyzed its expression pattern and protein interaction. The results of promoter structure prediction using PlantCARE software revealed that GhTFL1b promoter contained MeJA responsiveness element, abscisic acid(ABA) responsiveness element, MYB binding sites involved in drought inducibility and light responsiveness elements. Meanwhile, Quantitative Real time PCR result indicated that the expression profile of GhTFL1b was induced by salicylic acid(SA) negatively, while it could be regulated by salt(NaCl) positively. Yeast two hybrid approach were under taken to elucidate the protein interaction of GhFD and GhTFL1 subgroup, and the experiments showed that GhFD could interact with GhTFL1b but not with GhTFL1a, GhTFL1b and GhTFL1c in cotton. Taken together, the result suggested that GhTFL1b might be involved in the regulation of response to abiotic stresses(SA and salt).
2019 Vol. 34 (3): 9-15 [Abstract] ( 46 ) 全文: HTML   PDF (1115 KB)  ( 54 )
16 Cloning,Bioinformatics and Expression Analysis of FvARF5 Gene in Strawberry
SU Liyan
DOI: 10.7668/hbnxb.201751324
Auxin response factor ARF is a new class of transcription factors. They play an important role during plant development as an activation or inhibition in auxin-related gene expression. In order to identify the function of FvARF5 gene during strawberry development, FvARF5 gene was cloned by RT-PCR from Fragaria vesca and analyzed by bioinformatics. Furthermore, Real-time quantitative PCR(qPCR) assay was performed to investigate the tissue-specific expression of FvARF5 gene, and its expression patterns under various phytohormone treatments. The results showed that the open reading frame of FvARF5 gene was 2 745 bp, encoding a protein of 914 amino acids. The molecular weight of FvARF5 protein was 100.65 ku, and the theoretical pI was 5.25. Multiple sequence alignment and phylogenetic tree analysis showed that FvARF5 was closely related to ARF5 from Rosa chinensis with 95% identity. FvARF5 was predicated to be the auxin response gene that was located in the nucleus. Promoter analysis revealed that the sequence of FvARF5 contained multiple hormone response elements. Real-time PCR analysis showed that FvARF5 had specific expression difference in different organs, and it had a higher expression in stem and green fruit. Also, FvARF5 had significant response to exogenous IAA and GA treatment. The results showed that the FvARF5 might play an important role in auxin and GA signal transduction during strawberry development. This research provided a theoretical basis for further study of the function and molecular regulation mechanism of FvARF5.
2019 Vol. 34 (3): 16-22 [Abstract] ( 48 ) 全文: HTML   PDF (1552 KB)  ( 46 )
23 Subcellular Localization and Expression Analysis of CiATAF1 Gene in Caragana intermedia
WANG Yanfei, HONG Geriqiqige, WANG Guangxia, WANG Ruigang, LI Guojing
DOI: 10.7668/hbnxb.201751243
To study the expression of NAC under abiotic stress, CiATAF1 gene from Caragana intermedia was cloned and analyzed. It had an ORF of 873 bp and encoded 291 amino acids, consisting of two introns and three exons, and presenting a typical NAC domain. The genome walking method was used to obtain the CiATAF1 promoter with a length of 826 bp, which contained various response elements to light, hormone and anaerobic induction. The subcellular localization was conducted by analysis of the transient expression of Arabidopsis leaf protoplasts, showing that the CiATAF1 gene was mainly located in the nuclei. Quantitative fluorescence RT-PCR analysis showed that the expression of CiATAF1 was tissue-specific, having the highest expression level in the leaves of Caragana intermedia and relatively low level in roots and stems. Under the treatments of drought, salt, cold and ABA, the expression level of CiATAF1 increased differentially. It was speculated that the CiATAF1 gene is related to the abiotic stress regulations. Multiple sequence analysis was carried out between CiATAF1 with homologues of Arabidopsis and other leguminous plants, and their consistency of the sequences was 81.14%, indicating that the ATAF1 was relatively conservative. Bioinformatics analysis showed that CiATAF1 protein was an unstable hydrophilic protein, and its secondary structure was composed of alpha helixes, beta sheets and random coils. This study had the significance for excavating genes resistant to abiotic stress in Caragana intermedia and for analyzing the mechanism of environmental stresses in depth.
2019 Vol. 34 (3): 23-30 [Abstract] ( 34 ) 全文: HTML   PDF (1520 KB)  ( 23 )
31 Cloning and Bioinformatics Analysis of StNF-Y Nuclear Transcription Factor Gene in Potato
SHI Li, WU Yaru, WANG Xiaojuan, CHANG Yannan, PANG Pengxiang, GAO Gang
DOI: 10.7668/hbnxb.201751585
To study the role and function of potato StNF-Y gene under abiotic stress, a new NF-YC transcription factor(StNF-Y) was cloned from potato resistant to bacterial wilt genotype ED13. Bioinformatics analysis and biological function prediction showed that the full-length cDNA of StNF-Y was 1 132 bp, with an open reading frame of 693 bp, encoding 230 amino acids, with a formula of C1112H1735N317O339S10 and isoelectric point of 5.37. It contained 13 phosphorylation sites, and StNF-Y protein was distributed in the cytoplasm, without signal peptide.It was a hydrophilic protein. StNF-Y protein contained a highly conserved domain, existed in the BUR6 superfamily. Secondary structure showed that the protein consisted mainly of 45.65% α-helix, 5.65% extended chain, 45.22% random coil and 3.48% β-turn. Phylogenetic analysis showed that the StNF-Y sequence of potato was closely related to the NF-YC sequences of tomato, pepper and tobacco. In addition, StNF-Y played a certain role in a variety of plant stresses, so it was speculated that StNF-Y might be involved in the regulation of potato-related stress resistance. The results would provide a theoretical basis for further research on the function of potato StNF-Y and its resistance mechanism to stress.
2019 Vol. 34 (3): 31-37 [Abstract] ( 37 ) 全文: HTML   PDF (1194 KB)  ( 27 )
38 Construction and Induced Expression of Prokaryotic Expression Vector of T1084 Deletion and Substitution Mutations in NtTkr Tail of Nicotiana tabacum
TONG Wenyan, HU Mengke, XU Linna, QIAO Huicong, LI Fen
DOI: 10.7668/hbnxb.201751346
It is known that the threonine at position 1 084 of the third coiled-coil (CC3) of the new member of the tobacco-driven family NtTkr is crucial for the binding of the target protein. Multiple candidate proteins interacting with NtTkr were obtained by yeast two-hybrid screening of NtTkr tail. In order to determine the importance of T1084 deletion or replacement mutation between NtTkr and target protein vitro, first the pBI121-NtTkr plasmid was take as a template to obtain the tobacco NtTkr tail T1084 deletion and replacement tail by overlap extension PCR, and cloned T1084d,T1084A into pUC19, by the blue white spot screening, Sma Ⅰ-BamH Ⅰ double enzyme cuting the identification and gene sequencing, geting the right T1084 deletion and replacement NtTkr tail; Then restructured pUC19-T1084d, pUC19-T1084A and pMXB10 with Not Ⅰ-Nde Ⅰ double enzyme, the target fragment and the carrier fragment are recovered and connected, and the ligation product transforms DH5α. The recombinant was screened to Not Ⅰ-Nde Ⅰ double enzyme identification, managed to build the required pMXB10-NtTkr-T1084A and pMXB10-NtTkr-T1084d prokaryotic expression vector; Finally, pMXB10-NtTkr-T1084A and pMXB10 -NtTkr-T1084dd were transferred into BL21 (DE3), and the protein expression was detected by 12% SDS-PAGE after the induction of 0.05 and 0.06 mmol/L IPTG concentrations, respectively NtTkr-T1084A-1320 of about 77 ku and NtTkr -T1084d-1317 of 76.2 ku were highly expressed.
2019 Vol. 34 (3): 38-42 [Abstract] ( 32 ) 全文: HTML   PDF (1141 KB)  ( 16 )
43 Cloning,Expression Analysis and Vector Construction of CkCDPK Gene from Caragana korshinskii
JIA Huili, GUO Jicheng, WU Xinming, WANG Yunqi, LIU Jianning, FANG Zhihong, SHI Yonghong, DONG Kuanhu
DOI: 10.7668/hbnxb.201751471
Calcium dependent protein kinase is a key signal transductor, and plays an important role in plant resistance to envirenment stress by regulating and participating in metabolic pathways of plants. To explore the related function genes of caragana, we isolated an open reading frame of CDPK gene from caragana by homology cloning strategy, termed CkCDPK. Bioinformatic analysis showed that the gene had a 1 710 bp open reading frame(ORF), encoding 570 amino acids, with a molecular weight of 64.03 ku and PI of 9.12. Structural analysis showed that the CkCDPK gene contained the N-terminal variable region, a protein kinase domain, EF hand structure, calmodulin domains, etc. Quantitative real-time PCR technique was used to detect CkCDPK expression level under adverse situations. The results revealed that the CkCDPK presented a unimodal trend after treated with salinity(NaCl) and drought(PEG), of which the gene expression levels were higher at 6 h of salinity stress and 4 h of drought stress. Under ABA stress, the gene expression was up-regulated compared with the contrast. These results indicated that the CkCDPK gene might be involved in the regulation of environmental stress responses in Caragana korshinskii. The plant expression vector pCAMBIA3301-CkCDPK was also constructed. This study provided the theoretic and experimental basis for further study of the CkCDPK function in stress-tolerance regulation in Caragana korshinskii.
2019 Vol. 34 (3): 43-51 [Abstract] ( 35 ) 全文: HTML   PDF (1496 KB)  ( 18 )
52 Formation of Flesh Texture and Adhesion and Expression Analysis of Related Genes in Peach Fruit
HAN Qing, CAO Ke, ZHU Gengrui, FANG Weichao, CHEN Changwen, WANG Xinwei, LIU Kuozhan, YOU Shuanghong, WANG Lirong
DOI: 10.7668/hbnxb.201751738
The fruits of peach varieties Okubo, 08-9-106 and 08-9-107 were used as materials to analyze the fruit mesocarp firmness, flesh adhension, ethylene content, and related gene expression during the late fruit development period. The results showed that in Okubo, the flesh firmness decreased rapidly, and the accumulation of ethylene increased greatly, while the expression of PpPG, PpACS1, and PpYUC11 increased first and then decreased. In peach 08-9-106, the flesh firmness decreased rapidly, but there were no significant increase of ethylene and related gene(PpACS1 and PpYUC11) expression. Compared with Okubo and 08-9-106, 08-9-107 had no significant increase in ethylene and gene expression, which might result in the fruit firmness. Based on the flesh near endocarp, the Okubo was characterized as freestone, with higher ethylene content and increased gene expression obviously(mainly PpACS1). 08-9-106 was also classified as freestone peach, which had no obvious ehtylene release and gene expression activation. 08-9-107 was clingstone, and exhibited the similar characters with 08-9-106, except its higher gene expression of PpACO1 and PpYUC11. The above results suggested that Okubo and 08-9-107 were freestone-melting type and clingstone stony-hard type, respectively, which responded to ethylene. 08-9-106 was freestone melting flesh phenotype, with no response to ethylene. The changes of flesh firmness and the formation of adhesion trait in peach fruits were both related to gene expression of PpPG. PpACS1 (not the PpACO1) was the most essential gene in regulating the ethylene accumulation of the three peach germplasms.
2019 Vol. 34 (3): 52-58 [Abstract] ( 32 ) 全文: HTML   PDF (1074 KB)  ( 38 )
59 Cloning,Identification,Sequence Analysis and Spatio-temporal Expression Characteristics of Rs Gene in Kyoho Grape
CUI Tengfei, WANG Chen, TAN Hongyu, JIA Haifeng, BAI Yunhe, WANG Wenran, FANG Jinggui
DOI: 10.7668/hbnxb.201751391
To explore the potential role of Resveratrol synthase(Rs) in fruit development of Vitis vinifera cv. Kyoho, the structure and function of Rs sequence were analyzed, and the spatio-temporal expression specificity at different developmental stages of fruit was identified. The bioinformatics analysis and qRT-PCR method were used to analyze the sequence of Rs gene in Kyoho grape. The structure and subcellular localization of Rs gene were predicted, and its expression in different tissues of fruit at different developing stages was analyzed. The full-length cDNA of Rs gene was 1 539 bp, with an open reading frame(ORF) of 1 179 bp, encoding 392 amino acids. The predicted molecular weight was 42.88 ku, and the theoretical pI was 6.09. The gene contained chalcone, stilbene synthase activity sites and complete stilbene synthase family characteristic sites. The protein interaction predicted that Rs interacts with OMT2.1, which catalyzes resveratrol biosynthesis of Pterotilbene. The results of subcellular prediction showed that the Rs gene mainly expressed in the cytoplasm and chloroplast. The promoter analysis showed that the expression of Rs gene might be regulated by light, MYB, fungi and hormones, presenting a certain tissue specificity. qRT-PCR analysis showed that the Rs gene of grape expressed at different stages of the peel and flesh, but the expression level was highest in the 25th day after anthesis. Combined with the role of Rs gene accumulation in resveratrol, it could be inferred that there might be a high accumulation of resveratrol in the fruit peel during the early period of fruit development. The Rs gene of Kyoho grape had a high conservation during evolution, and its expression might be regulated by environment, fungi and hormone. It had the interaction with OMT2.1 and had a certain specificity in different tissues and development periods.
2019 Vol. 34 (3): 59-67 [Abstract] ( 42 ) 全文: HTML   PDF (1485 KB)  ( 16 )
68 Development of Novel SSR Markers on Wheat Chromosome 2B
LI Xiaojun, XU Xin, ZHANG Ziyang, LIU Mingjiu, RU Zhengang
DOI: 10.7668/hbnxb.201751761
In order to develop novel SSR markers for wheat breeding, simple sequence repeat(SSR) was identified from a 25 Mb region according to the Chinese Spring 2B chromosome reference sequence. In 2 852 SSR loci identified, 80.40% belonged to dinucleotides repeat.Six, thirty, seventeen, one kinds of SSR repeats were found in dinucleotide, trinucleotide, tetranucleotide, pentanucleotide repeats, respectively. For dinucleotides, GA/CT, AG/TC, AT/TA had the highest frequencies, occupying 20.44%, 19.00%, 17.15% of all SSR loci identified, respectively; for trinucleotides, CTT/GAA had the highest frequency, occupying 2.56% of all SSR loci identified; whereas the frequencies of tetranucleotide and pentanucleotide repeats were relatively low. Furthermore, the replication numbers were higher in dinucleotide than others, and 327 SSR loci which occupied 14.26% of all dinucleotides loci identified were with ≥ 10 replications. The numbers of SSR loci with ≥ 10 replications for AG/TC and AT/TA reached 98 and 97, respectively. The replication numbers in trinucleotide were relatively low, and 62 SSR loci which occupied 11.55% of all trinucleotides loci identified were with ≥ 8 replications. The number of TTC/AAG with ≥ 8 replications was the biggest, reached 12. For tetranucleotide, the numbers in ACAT/TGTA and TAGA/ATCT reached 16 and 14, respectively, and the replication numbers in other repeats of tetranucleotide varied between 5 and 7. Based on the found SSR loci, 135 SSR primer pairs were designed and synthesized. Of these, 101(74.81%) SSR primer pairs amplified clear products in wheat lines Cf5019-21 and Cf5240-41, and 17 SSR primer pairs showed polymorphism between them.
2019 Vol. 34 (3): 68-74 [Abstract] ( 30 ) 全文: HTML   PDF (938 KB)  ( 21 )
75 Genetic Diversity of HMW-GS and Its Relationship with Quality of Wheat Germplasms from Different Regions
ZHANG Ziyang, JIANG Xiaoling, WANG Zhiyu, ZHU Qidi, LIU Mingjiu, RU Zhengang
DOI: 10.7668/hbnxb.201751750
In order to clarify the relationship between high molecular weight glutenin subunit (HMW-GS) and wheat processing quality,148 wheat germplasms from different regions of the world were used to study the genetic diversity of high molecular weight glutenin subunits(HMW-GS) and its relationship with the quality of flour and steamed bread. Results showed that the tested materials had abundant polymorphisms at the three gene loci of Glu-A1, Glu-B1 and Glu-D1, of which, three, seven and seven types of subunits were detected, respectively. The highest frequency of allele was 1, 7+9 and 2+12 at each of the three loci, which accounted for 56.8%, 47.3% and 45.9%, respectively. A total of 35 HMW-GS combinations were identified in the tested materials, among which the combinations of(1/7+9/2+12), (1/7+8/5+10), (N/7+9/2+12) and(1/7+9/5+10) had higher frequencies. The compositions of HMW-GS were different in wheat varieties(lines) from different regions. Moreover, the most abundant type of subunit and the highest frequency of high-quality subunit were found in germplasms from aboard and Huanghuai winter wheat region. Furthermore, subunits 1, 2*, 7+8, 17+18 and 5+10 had positive effects on gluten strength, while subunits 2*, 17+18 and 2+10 had important influence on hardness and chewiness of steamed bread. The genotypes with combination forms of(1/14+15/2+12), (1/7+9/5+10) or(N/7+8/2+12) had good processing quality of steamed bread.
2019 Vol. 34 (3): 75-81 [Abstract] ( 36 ) 全文: HTML   PDF (950 KB)  ( 26 )
82 Structure Characteristics and Prokaryotic Expression Analysis of Soybean Glyma08g11030 Gene
WANG Ping, YU Yuehua, BAI Yucui, WAN Huina, NI Zhiyong
DOI: 10.7668/hbnxb.201751473
In order to further clarify the function of Glyma08g11030 gene, sequence analysis and protein domain analysis of Glyma08g11030 were performed by means of bioinformatics and PCR. Bioinformatics analysis showed that the full-length of Glyma08g11030 cDNA was 1 419 bp, with an open reading frame of 1 362 bp, encoding 453 amino acids. The molecular weight of the protein was about 51.288 ku, and the isoelectric point was 8.14. Sequence analysis indicated that the Glyma08g11030 genome contained two exons and one intron. Multiple sequence alignment revealed that the Glyma08g11030 protein contained one F-BOX conserved domain. Phylogenetic tree analysis showed that the protein had a homologous relationship with pigeonpea, kidney bean, peanut, cowpea and adzuki bean. In order to obtain the purified Glyma08g11030 protein, the amplified fragment was cloned into the prokaryotic expression vector pET28a, and the recombinant plasmid pET28a-Glyma08g11030 was constructed. The target protein was obtained by SDS-PAGE. The results showed that the protein mainly existed in the form of inclusion bodies. The results of this study laid the foundation for further purification and identification of Glyma08g11030 protein and investigation of its function.
2019 Vol. 34 (3): 82-88 [Abstract] ( 35 ) 全文: HTML   PDF (1460 KB)  ( 21 )
89 Development and Application of Gene Markers in the Exogenous Chromosome Segment of Chinese Cabbage-Cabbage Translation Lines
SU Jianhui, ZHANG Leqiang, WANG Ruying, WANG Yanhua, ZHANG Weiwei, MA Cong, XUAN Shuxin, SHEN Shuxing
DOI: 10.7668/hbnxb.201751153
In order to identify Chinese cabbage-cabbage translocation line by gene markers, the polymorphism primers were designed according to the nucleotide difference of collinear genes between Chinese cabbage and cabbage, and were screened with Chinese cabbage 85-1 and cabbage 11-1 as test materials. The applicability of the primers was detected using Chinese cabbage-cabbage translocation lines and their self-crossing progenies, Chinese cabbage varieties and cabbage varieties. The results showed that among 171 pairs of primers designed based on 263 collinear-gene sequences of Chinese cabbage and cabbage, 66 pairs of polymorphic primers were confirmed, the ratio being 38.60%. Among the 66 gene markers, two markers from Chinese cabbage gene sequences and 48 markers from cabbage gene sequences could be used as the specific gene markers to distinguish Chinese cabbage and cabbage, which were successfully applied to identify Chinese cabbage-cabbage translocation lines AT1-41 and AT1-47, and the self-crossing progenies of AT1-47. Furthermore, PCR amplification of the 66 gene markers was carried out in four Chinese cabbage varieties and four cabbage varieties, among which 52 markers could distinguished completely the Chinese cabbage varieties and the cabbage varieties, while other 11 markers presented the polymorphism in parts of the varieties. The results established the foundation for further studying the expression, regulation and interaction of exogenous genes in Chinese cabbage-cabbage translocation lines and for marker-assisted breeding.
2019 Vol. 34 (3): 89-95 [Abstract] ( 25 ) 全文: HTML   PDF (1628 KB)  ( 24 )
       Tillage and Cultivation·Physiology and Biochemistry
96 A Study on Grain Filling Characteristics and Yield in Different Wheat Varieties
JIANG Lina, ZHANG Yawen, ZHU Yalin, ZHAO Lingxiao
DOI: 10.7668/hbnxb.201751302
In order to provide the theoretical support for the cultivation and variety breeding of high-yield wheat in Henan Province, a field experiment with 6 wheat varieties was carried out to analyze the grain filling characteristics and yield of wheat. The cubic polynomial equation was used to fit the grain filling process, the correlation analysis was used to explore the relationship between grain weight and grain filling characteristics, and the difference of yield components among these varieties was discussed. The results showed that the 1000-grain weigh of Zhengmai 0943 and Xuke 316 was the highest(50.64 g) and lowest(45.02 g), respectively, among the 6 varieties. The grain filling rates were different among the varieties. The simulated curve of grain filling process presented a "S" shape, whereas the model parameters were different among these varieties. The 1000-grain weight had significant positive correlations with maximum grain filling rate(Vmax) and theoretical maximum grain weight(W), and also had higher positive correlations with the mean grain filling rate(V), effective grain filling period(Se), and mean grain filling rate during the effective grain filling period(Vs). It suggested that the grain weight could be greatly improved by optimizing the grain-filling characteristic parameters, including Vmax, W, V, Se and Vs. The grain yield of Yumai 49-198 was significantly higher than those of other five varieties, indicating that the improvement of grain yield should be in coordination with rational spike number and grain number per spike, in addition to raising the grain weight.
2019 Vol. 34 (3): 96-101 [Abstract] ( 52 ) 全文: HTML   PDF (1024 KB)  ( 59 )
102 Effects of Drought Stress on Photosynthesis and Chlorophyll Fluorescence Characteristics of Summer Maize
XIA Lu, ZHAO Rui, WANG Yizhen, JIN Haiyan, WU Xidong, GE Junzhu, ZANG Fengyan, LI Zifang, WANG Jinlong
DOI: 10.7668/hbnxb.201751385
In order to explore the photosynthetic physiological response mechanisms of three summer maize varieties (Xundan 20, Jinbei 288 and Dika 667) which were commonly cultivated in arid and semi-arid areas to drought stress, these three varieties were treated under different drought stresses (including non-drought treatment (CK), light drought (LD) and heavy drought (HD)). Responses of net photosynthetic rate to light intensity (Pn-PAR) and net photosynthetic rate to intercellular CO2 concentration (A-Ci) were estimated by the portable photosynthesis system Li-6400, and responses of chlorophyll fluorescence parameters to light intensity were measured by portable modulation chlorophyll fluorescence meter MINI-PAM, respectively. The results were as follows. As for all these three varieties, the response modes of Pn and stomatal conductance (Gs) to PAR were similar among these three maize varieties, and the responses of Pn to Gs shifted to left along the drought stress gradient. The initial chemical efficiency (CE) and photosynthetic capacity (Amax) were significantly lower under heavy drought treatment than under the other two treatments for each of the three varieties. Compared with the results under CK treatment, the non-photochemical quenching coefficient (qN) and the quantum yield of the regulatory energy dissipation (Y (NPQ)) were lower and the quantum yield of non-regulated energy dissipation (Y(NO)) was higher under LD or HD treatment for the two varieties of Jundan 20 and Dika 667; and the photochemical quenching coefficient (qP) was lower under HD treatment for the variety of Jinbei 288. Compared with the results under CK treatment, the maximum net photosynthetic rate (Pmax) and initial quantum efficiency (AQY) were a little higher under LD treatment for the variety of Jinbei 288. Compared with the other two varieties, the Pmax and AQY showed higher tolerance to drought stress by keeping relatively lower qQ and lower Y(NO) for the variety of Jinbei 288, and these results not only showed the greater tolerance of Jinbei288 to drought stress, but also provided reference for the screening of crop varieties for drought tolerance.
2019 Vol. 34 (3): 102-110 [Abstract] ( 65 ) 全文: HTML   PDF (1221 KB)  ( 100 )
111 Effects of Calcium Application on Peanut Yield and Physiological Characteristics under Different Planting Patterns
LIN Songming, ZHANG Zheng, NAN Zhenwu, MENG Weiwei, LI Lin, GUO Feng, WAN Shubo
DOI: 10.7668/hbnxb.201751319
In order to determine the response of peanut pod yield and physiological characteristics to calcium fertilizer under different planting patterns, a field experiment was designed with Huayu 25 as the test material, setting 2 planting patterns(3:4 maize-peanut intercropping and peanut monoculture) and 4 calcium treatments(Ca0:0 kg/ha, Ca1:150 kg/ha, Ca2:300 kg/ha and Ca3:450 kg/ha). The effects of calcium application on photosynthetic characteristics of peanut leaves, pod yield and its physiological characteristics under different planting patterns were systematically analyzed. The results showed that under the same calcium application condition, the intercropping decreased the chlorophyll relative content(SPAD), net photosynthetic rate, CAT, POD and SOD activities of peanut, but increased the MDA content of peanut leaves, and the yield of pods also decreased in a certain degree. Under the same planting pattern, calcium treatments increased the pod yield of peanut in different degrees compared with the Ca0, and the peanut yield in middle rows was higher than that of monoculture peanut. In addition, with the increase of calcium application, the pod yield of intercropping and monoculture peanut increased first and then decreased, and the highest came from Ca2 treatment. Moreover, the experiment results were basically consistent with the appropriate calcium amount applied in middle rows of monoculture and intercropping peanuts derived from the regression equation. The calcium application enhanced the photosynthetic capacity of peanut leaves under monoculture and intercropping patterns, and their net photosynthetic rate(Pn) increased by 3.1%-23.7% and 8.9%-21.8%, respectively. The Ca2 treatment could increase the chlorophyll relative content(SPAD), and the activities of CAT, POD and SOD in the leaves of monoculture and intercropping peanut, and reduced the MDA content of leaves, which was beneficial to improving the adaptability of peanuts to shade and increasing the production of intercropping peanuts. The Ca2 treatment was the suitable calcium amount in monoculture and intercropping shading patterns of the experiment.
2019 Vol. 34 (3): 111-118 [Abstract] ( 46 ) 全文: HTML   PDF (1100 KB)  ( 21 )
119 Effects of Brassinolide on Seed Germination and Seedling Physiology of Maize under Low Temperature Stress
SUN Yujun, WU Yue, MA Dezhi, LÜ Jinying, HE Yunhan, GONG Lei, LIU Zhe, GAO Lidan, LI Na, YAN Dong, ZHU Jinghuan, YANG Deguang
DOI: 10.7668/hbnxb.201751362
In order to explore the effects of brassinolide(BR) on seed germination and seedling physiological characteristics of maize under low temperature stress, the maize hybrids Tiannong 9 and Tianhe 1 were used as the materials to study the changes of agronomic traits and physiological properties after their seeds were soaked or sprayed with BR. The results showed that exogenous BR treatment could effectively alleviate the inhibition of maize growth under low temperature stress. The best treating concentration was 0.1 mg/L BR, which increased the germination rate, the biomass of overground and underground parts, the SPAD value, the activities of SOD, POD and CAT, and the contents of proline, soluble sugar and soluble protein, while decreased the MDA content and the relative conductivity in maize seedlings. Compared with the low temperature control, the maize hybrids Tiannong 9 and Tianhe 1 treated with exogenous BR of 0.1 mg/L initial germination rate increased by 18.89 and 11.10 percentage points respectively, the final germination rate increased by 6.67 and 18.89 percentage points, their aboveground dry weight increased by 55.92% and 49.15%, their underground dry weight raised by 38.79% and 14.26%, their relative electrical conductivity decreased by 7.33% and 6.80%, their CAT activity increased by 110.58% and 120.43%, and their proline content increased by 66.59% and 54.63%, respectively. It suggested that the relieving effects of 0.1 mg/L BR treatment on the two maize varieties under low temperature stress were different, Tiannong 9 being better than Tianhe 1. The maize varieties with different cold resistances had different adaptability to different BR concentrations.
2019 Vol. 34 (3): 119-128 [Abstract] ( 62 ) 全文: HTML   PDF (1115 KB)  ( 47 )
129 Effect of Melatonin on Rice Seedling Growth under Submergence Stress
CHEN Dong, LI Qiang, PENG Yan, WU Tianhao, ZHANG Xiuli, DONG Jiayu, MAO Bigang, ZHAO Bingran
DOI: 10.7668/hbnxb.201751428
The effects of melatonin-soaking seeds on physiological and biochemical characteristics of rice seedlings under submergence stress. Three concentration treatments of melatonin(10, 100, 1 000 μmol/L) were arranged. The agronomic traits such as Mesocotyl length, coleoptile length, shoot length, fresh weight and survival rate, and the physiological and biochemical indexes such as enzyme activity of peroxidase(POD), superoxide dismutase(SOD), catalase(CAT) and malondialdehyde(MDA) content were measured.The results showed that low concentration MT-soaking could alleviate the stress and high concentration MT-soaking aggravate the toxicity. Compared with short mesocotyl cultivars Huahang 31 and DH4040, long mesocotyl cultivars Ezhong 4 and DH4038 had higher survival rates under submergence stress. Among the four cultivars, Huahang 31 had the best effect of MT-soaking seeds in two treatments, i.e. 10,100 μmol/L MT. The treatment of 100 μmol/L MT-soaking seeds could greatly improve the seedling survival rate of all cultivars. When the MT-soaking concentration increased to 1 000 μmol/L, the survival rate of all cultivars was inhibited. MT-soaking seeds could reduce MDA content, increase POD, SOD and CAT activities, and promote agronomic traits such as mesocotyl length, coleoptile length, bud length and fresh weight, while 100 μmol/L MT-soaking seeds could also promote the related agronomic traits. While 1 000 μmol/L MT-soaking could also promote the related agronomic traits and enzyme activity,but the survival rate decreased significantly.It was concluded that soaking-seeds with appropriate concentration of melatonin could improve the vigor and emergence rate of rice seedlings under submergence stress, and significantly reduce the toxicity of submergence stress on rice seedlings.
2019 Vol. 34 (3): 129-136 [Abstract] ( 57 ) 全文: HTML   PDF (1257 KB)  ( 45 )
       Resource and Environment·Plant Protection
137 Effects of Different Soil Tillage Systems on Dry Matter Accumulation and Yield of Rice in Double Cropping Rice Field
TANG Haiming, LI Chao, XIAO Xiaoping, TANG Wenguang, GUO Lijun, CHENG Kaikai, PAN Xiaochen, WANG Ke, SUN Geng
DOI: 10.7668/hbnxb.201751185
In order to explore the characteristics of dry matter accumulation and grain yield of rice under different soil tillage systems in a Chinese milk vetch and double cropping rice system, the dry matter accumulation and grain yield of early rice and late rice were analyzed by using conventional experimental analysis methods from 2016 to 2017. The experiment included four different soil tillage treatments:conventional tillage plus residue incorporation (CT), rotary tillage plus residue incorporation (RT), no tillage plus residue retention (NT), and rotary tillage plus residue removement (RTO). The results showed that in different soil tillage treatments, the leaf area index was CT > RT > NT > RTO at the main growth stages of early rice and late rice. And the dry weights of roots, shoots, stems, leaves and panicles of population were also CT > RT > NT > RTO at the main growth stages of early rice and late rice. Meanwhile, the dry matter weight ratio of root to total plant at the heading stage and maturity stage of early rice and late rice had the trend of CT > RT > NT > RTO. The dry matter weight ratio of stem to total plant in RTO treatment was higher than that of other treatments at the heading stage and maturity stage of early rice and late rice. And the dry matter weight ratio of leaf to total plant in NT and RTO treatments was higher than that of other treatments at the seedling stage and tillering stage of early rice and late rice. In addition, the dry matter weight ratio of panicle to total plant in RT and NT treatments was higher than that of other treatments at maturity stage of early rice and late rice. The grain yield of early rice and late rice in different soil tillage treatments was CT > RT > NT > RTO. Compared with RTO treatment, the grain yields of early rice in CT, RT and NT treatments increased by 733.3, 534.1,300.5 kg/ha in 2016 and 731.1, 556.9, 276.2 kg/ha in 2017, respectively, while the grain yields of late rice in CT, RT and NT treatments increased by 582.5, 399.8,282.9 kg/ha in 2016 and 717.6, 558.9,345.1 kg/ha in 2017, respectively. The results indicated that the combined application of conventional tillage or rotational tillage with residue incorporation was an effective way for increasing the dry matter accumulation of rice plant and making the dry matter distribution more reasonable, so as to improve the grain yield and yield components of rice.
2019 Vol. 34 (3): 137-146 [Abstract] ( 43 ) 全文: HTML   PDF (1048 KB)  ( 37 )
147 Effects of Different Planting Modes on Nitrogen Uptake and Accumulation of Late Rice under Water-saving Conditions
ZHOU Wentao, LONG Wenfei, DAI Wei, FU Zhiqiang
DOI: 10.7668/hbnxb.201751069
In order to investigate the effects of different rice planting modes on the nitrogen utilization of double-cropping late rice under the water-saving condition of "early water and late use", three planting modes of artificial transplanting, throwing seedlings and transplanting seedlings by machine were compared. The results showed that nitrogen accumulation per plant increased gradually with time during the main growing stages of rice. Nitrogen accumulation per plant in different stages of rice accounted for the proportion of total nitrogen accumulation in the whole growth stages showed the following order (from high to low):tillering to full heading stage (25.18%-68.01%), sowing to tillering stage (23.47%-58.47%), milky to maturity stage (5.39%-27.25%),full heading to milky stage (0.52%-12.51%). The accumulation amounts of nitrogen per plant followed the order of tillering to full heading stage (148.28* *),milky to maturity stage (105.05* *), sowing to tillering stage (71.65* *), and full heading to milky stage (57.26* *). The nitrogen accumulation as a proportion of total plant biomass in stem sheaths and leaves of rice decreased with the growth of rice as a whole, while the nitrogen accumulation increased first and then decreased, reaching to maximum at heading stage. Nitrogen accumulation as a proportion of total plant biomass in panicle increased first and then decreased with the growth of rice, and nitrogen accumulation increased gradually. There was a significant positive correlation between yield and nitrogen accumulation per plant during the milky stage(0.87*), but no significant correlation was founded during tillering to full heading stage and full heading to milky stage. There was no significant negative correlation between yield and nitrogen accumulation as a proportion of total plant biomass in maturity stage. Among different treatments, the nitrogen accumulation of rice and the nitrogen accumulation as a proportion of total plant biomass of leaves,stem sheaths and panicle was the highest in the treatment of "The early rice ploughs were planted and reaped+Late rice harvest by no-tillage and throwing machine+seedling-increase and nitrogen-reduction" in the maturity stage.
2019 Vol. 34 (3): 147-154 [Abstract] ( 45 ) 全文: HTML   PDF (940 KB)  ( 26 )
155 Effects of Different Nitrogen Application Patterns on Leaf Physiological Characteristics and Grain Yield of Double Cropping Rice
XU Yilan, FU Aibin, LIU Tangxing
DOI: 10.7668/hbnxb.201751226
In order to investigate the physiological and biochemical characteristics of rice leaves and grain yield with different N application patterns in double-cropping rice system, the protective enzyme activities and photosynthetic characteristics of rice leaves, grain yield of early rice and late rice were analyzed under 5 models of N application:120.0 kg/ha of early rice plus 210.0 kg/ha of late rice(N120:210), 150.0 kg/ha of early rice plus 180.0 kg/ha of late rice(N150:180), 180.0 kg/ha of early rice plus 150.0 kg/ha of late rice(N180:150), 210.0 kg/ha of early rice plus 120.0 kg/ha of late rice(N210:120), and taking zero N of early and late rice as the control(CK). The results showed that the activities of SOD, POD and CAT in rice leaves increased with different N fertilizer treatments compared to CK at main growth stages of early and late rice. Meanwhile, compared with CK, the MDA and Pro contents in rice leaves decreased with different N fertilizer treatments at main growth stages of early and late rice. The Pn and Tr of rice leaves with different N fertilizer treatments decreased in the following order:N150:180 > N180:150 > N210:120 > N120:210 > CK and N150:180 > N180:150 > N120:210 > N210:120 > CK at main growth stages of early and late rice, respectively. Meanwhile, the leaf Gs of early and late rice with different N fertilizer treatments decreased in the following order N210:120 > N180:150 > N150:180 > N120:210 > CK and N120:210 > N150:180 > N180:150 > N210:120 > CK at main growth stages, respectively. The results also indicated that there were no significantly difference in spikelet number per panicle and 1 000-grain weight of rice among different N fertilizer treatments. The grain yields of early rice and late rice in N150:180 and N180:150 treatments were higher than in other treatments. Compared with CK, the grain yields of early rice and late rice in N150:180 and N180:150 treatments increased by 1 849.5, 1 879.5 kg/ha, respectively. The results indicated that the N150:180 treatment could improve the protective enzyme activities and photosynthetic characteristics of rice leaves, so as to increase the grain yield of rice.
2019 Vol. 34 (3): 155-163 [Abstract] ( 48 ) 全文: HTML   PDF (1511 KB)  ( 34 )
164 Effects of Nitrogen Fertilizer Management Pattern and Planting Density on Yield and Diseases of Summer Corn in Southwest China
LEI En, GENG Yongke, CHEN Lufa, YANG Yongbing, WANG Yuedong, TANG Qiyuan
DOI: 10.7668/hbnxb.201751125
The objective of the study was to investigate the effects of nitrogen fertilizer management pattern and planting density on the yield formation, stalk rot and ear and kernel rot of summer corn in field condition of Southwest China. Two levels of nitrogen fertilizer management pattern were arranged, i.e. conventional nitrogen fertilization(CNP) and nitrogen fertilizer reduction plus deep application in holes between plants and rows(RDNP). And three levels of planting density were designed:commonly used rare-planting(D1, 52 500 plants/ha), densification Ⅰ(D2, 67 500 plants/ha) and densification Ⅱ(D3, 82 500 plants/ha). The yield and its composition, dry matter accumulation, photosynthetic performance and other factors that affected the mechanical harvest quality such as the incidence and severity of maize stalk rot, ear and kernel rot were analyzed. The results revealed that nitrogen fertilizer management pattern had no significant effect on the yield, but the nitrogen partial productivity was 33.7% higher in RDNP than in CNP. RDNP restrained the declining extent of net photosynthetic rate(Pn), leaf SPAD value and leaf area index during the grain-filling stage, which made up for the insufficience of grain numbers per ear formed before flowering, and stabilized the dry-matter accumulation and yield. The RDNP significantly reduced the incidence and severity of maize stalk rot by 4.8 percentage points and 26.8%, respectively, but had little effect on the incidence of ear and kernel rots compared to CNP. The grain yields of D2 and D3 were greater than that of D1, but the sustaining yield increase rate of D3 was significantly lower than that of D2. The incidence and severity of stalk rot and ear and kernel rot were relatively high in D3 than in D2 and D1, while there was no significant difference between D2 and D1. The combination of RDNP and D2 was able to increase the maize yield, improve nitrogen partial productivity, and control the occurrence of maize stalk rot and ear and kernel rot effectively, so as to reduce the risk of lodging and stem breaking. The results could provide the theoretical basis for increasing the production and facilitating the mechanized harvest of summer corn in Southwest China.
2019 Vol. 34 (3): 164-172 [Abstract] ( 40 ) 全文: HTML   PDF (1136 KB)  ( 37 )
173 The Response of Potassium Forms to Long-term Located Fertilization in Black Soil
DU Yan, WANG Tianye, WU Yangsheng, GAO Qiang, GAO Yunhang, LIU Shuxia
DOI: 10.7668/hbnxb.201750976
In order to understand clearly the effect of long-term fertilization on the potassium forms in black soil, the field trials were carried out through different long-term positioning experiments. The trial treatments included three categories:single application of chemical fertilizer, low-volume corn stalk with chemical fertilizer, and high-volume corn stalk with chemical fertilizer. The results showed that under long-term fertilization conditions, the treatments of low-volume corn stalks combined with chemical fertilizer (except NP+S2.5) and high-volume corn stalks combined with chemical fertilizer increased the total potassium content compared to CK (without fertilization), among which the effects of S5.0, N+S5.0 and NPK+S5.0 treatments were significant. The treatment of single fertilizer was inconsistent to CK treatment in the change of total potassium content.The total potassium content in N and NP treatments was lower than that of CK, but it increased significantly in K and NPK treatments. The content of slow-acting potassium increased significantly under the influence of high-content corn stalks combined with chemical fertilizers, and the highest came from the K+S5.0 treatment.The mineral potassium was relatively stable in soil, so it was less influenced by long-term fertilization in black soil. Under long-term fertilization conditions, the treatments of low-volume corn stalks combined with chemical fertilizer and high-volume corn stalks combined with chemical fertilizer significantly increased the content of available potassium compared to CK, and NPK+ S2.5, K+S5.0 and NPK+S5.0 treatments had the highest content of available potassium. K+S5.0 treatment had the most effect on the contents of water-soluble potassium and exchangeable potassium for long-term fertilization. The mineral potassium in black soil accounted for 90%-98% of total potassium(the highest), and the water-soluble potassium was the lowest. It was concluded that the application of chemical fertilizers with corn stalks in different fertilization treatments had the greatest influence on various forms of potassium in soil, followed by NPK fertilizer application, and the potassium forms in black soil were different due to different fertilization treatments.
2019 Vol. 34 (3): 173-179 [Abstract] ( 22 ) 全文: HTML   PDF (963 KB)  ( 12 )
180 Effects of Saline-alkali Land Improver and Fertilizer on Soil pH and Maize Growth and Development
DU Kangrui, DUAN Ximing, ZHAO Jinzhong, HU Xiaofang, JIA Junxian, ZHANG Jiangang, ZHANG Yongpo, YUE Aiqin, DU Weijun
DOI: 10.7668/hbnxb.201751586
The salinization of land in China is becoming more and more serious, and the sustainable development of agriculture is seriously hindered.Taking corn Xianyu 335 as the research object in this experiment, the effects of different combinations of acetylated glucose, urea, phosphate fertilizer and organic fertilizer on soil character improvement effect and effects of maize growth and development were investigated under the saline and alkaline land conditions in Xugou Town, Qingxu County, Shanxi Province. The results showed that the combination of acetylated glucose, urea, phosphate fertilizer and organic fertilizer had the most significant effect on the improvement of saline-alkali land. The pH of the soil was 0.32 lower than that of the control (without any treatment). The SPAD value of maize leaves increased by 50.834 and the net photosynthetic rate increased by 21.8 μmol/(m2 · s). The plant height and stem diameter of maize at seedling uprooting stage and harvest stage increased by 45.99, 0.36 cm and 77.13,0.50 cm, respectively. The ear length, ear weight, 100-grain weight and grain number per ear of maize increased by 5.06 cm, 118.216 g, 11.123 g and 271.8 grains, respectively. The combination of acetylated glucose and fertilizer can promote crop growth and improve soil properties, which can be used as one of the important measures to improve saline-alkali land in the future.
2019 Vol. 34 (3): 180-185 [Abstract] ( 38 ) 全文: HTML   PDF (941 KB)  ( 25 )
186 Cloning and Bioinformatics Analysis of Elastinolytic Metalloproteinase Gene from Puccinia helianthi Schw
LI Xinchun, SONG Yang, LU Yan, JING Lan
DOI: 10.7668/hbnxb.201751261
In order to study the pathogenic mechanism of Puccinia helianthi Schw, bioinformatics method was used analyze the mature part of KU994904 gene that had a high expression level in the transcriptome of Puccinia helianthi Schw. The protein ORF region was analyzed by Interpro which can determine its mature region. Using the bioinformatical databases and softwares, its function was analyzed and predicted. Total RNA was extracted from Puccinia helianthi Schw and used to amplify the mature gene of the metalloproteinase by RT-PCR. Finally, the gene sequence of mature protein was cloned. The results showed that the sequence full-length of the gene was 1 335 bp, coding 445 amino acids. And the protein molecular weight was 49.5 ku, it was found that the gene had high similarity with five sequences of M36 family and the gene had 53.77% similarity with the extracellular metalloproteinase (mep). So it was deduced that the gene coded an elastinolytic metalloproteinase.
2019 Vol. 34 (3): 186-193 [Abstract] ( 13 ) 全文: HTML   PDF (1598 KB)  ( 9 )
194 Comparative Transcriptome Analysis of Phytophthora blight Resistance of Pepper at Seedling Stage
LEI Yang, CHENG Yan, QIAO Ning, JIAO Yansheng, MIAO Ruyi, YANG Yuhua
DOI: 10.7668/hbnxb.201751450
The objective of this study was to study the molecular mechanism of pepper phytophthora blight and to explore the functional genes related to phytophthora blight resistance. Based on the identification of phytophthora blight resistance in 114 natural populations of pepper, one resistant material and three susceptible materials were selected as materials to perform high-throughput transcriptome sequencing by means of Illumina RNA-seq sequencing platform. The clean reads were selected to map the reference sequences of Pepper_Zunla_1_Ref_v1.0. The results showed that 78.95%-85.11% clean reads could be matched to unique genome locus. The gene expression level was calculated using RPKM method. The false discovery rate ≤ 0.001 and the absolute value of|log2 Ratio| ≥ 1 were used as the threshold to judge the significance of gene expression differences. Finally, the functions and pathways of differentially expressed genes(DEGs) were annotated by comparing them with the Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG). According to the top 10 GO classification, the main functions included the oxidoreductase activity, carboxylolyase activity, peroxidase activity, metabolic process and adverse reaction etc. Among them, 117 DEGs could be classified into the KEGG pathway, including 25 up-regulated DEGs and 92 down-regulated DEG. These pathways were associated with the ubiquitin-mediated proteolysis, oxidative phosphorylation, plant hormone signaling, phospholipase D signaling, and phosphatidylinoinositol signaling systems. The disease resistance of pepper was mainly related to enzymatic activity, stress response, protein metabolism pathway, plant hormones and other regulations. It was found that the disease resistance of pepper was a highly complex process, which was composed of multiple cross channel regulation, including metabolic processes, defense responses and hormone regulation, etc. The results laid the foundation for further studying the molecular mechanism of pepper disease resistance.
2019 Vol. 34 (3): 194-202 [Abstract] ( 33 ) 全文: HTML   PDF (1068 KB)  ( 19 )
203 Mapping of Clubroot Resistance Gene and Developing Linkage Markers in Brassica rapa L. ssp. pekinensis
WANG Chaonan, ZHANG Bin, ZHANG Hong, WEN Juanjuan, WANG Tao
DOI: 10.7668/hbnxb.201751417
In order to identify the clubroot resistance gene of Chinese cabbage in G6 and develop linkage markers, high-generation inbred line G4 of Chinese cabbage with high-susceptible to clubroot disease, high-generation inbred line G6 of Chinese cabbage with high-resistant to clubroot disease, F1 obtained by crossing G4 and G6, and the F2 generation separation populations constructed by F1 selfing were used as materials. By artificial inoculation, phenotypic identification and genetic analysis, it was found that clubroot disease resistance in the disease resistance materials were controlled by a dominant single gene. Further expanding the number of F2 generation populations to map the genes of clubroot disease and screening molecular markers linked to disease resistance genes, the linkage analysis of polymorphic markers was performed using JoinMap 4.0 software, and five InDel markers linked to the resistance gene of Chinese cabbage were obtained. The most recent markers on both sides were BrID10727 and BrID10867, the genetic distance from the disease resistance genes were 4.6,2.5 cM, respectively, and the disease resistance gene was located on the chromosome A08 of Chinese cabbage. In addition, it was found that the newly developed polymorphic marker BrID10381 based on the Crr1 gene sequence was located outside the initial localization range of the newly located clubroot resistance gene, so it could be inferred that the newly located clubroot resistance gene locus was not the same locus as Crr1, and the marker BrID10381 could be used for molecular marker assisted selection of Crr1 gene.
2019 Vol. 34 (3): 203-208 [Abstract] ( 38 ) 全文: HTML   PDF (1136 KB)  ( 13 )
209 Extraction Method of Total Protein from Trichothecium roseum Suitable for Proteomic Analysis
WANG Peng, WANG Junjie, LI Zhenbiao
DOI: 10.7668/hbnxb.201751366
In order to optimize the protein extraction method suitable for two-dimensional electrophoresis of Trichothecium roseum and further study the pathogenic mechanism of T. roseum on the proteomic level, the isolated and purified T. roseum were chosen to extract the total protein by 5 methods:ultrasonic-TCA-acetone, ultrasonic-phosphoric acid-TCA acetone, ultrasonic-SDS, ultrasonic-TCA/acetone-phenol/SDS combined extraction and TCA/acetone-phenol/SDS combined extraction,respectively. Meanwhile, two better extraction methods were screened by comparative analysis of protein content and purity. Under the appropriate gel concentration of polyacrylamide electrophoresis, the best protein extraction method was obtained by two-dimensional electrophoresis. The results showed that 12% polyacrylamide gel concentration had clear background and no serious tailing. Moreover, the concentration and content of protein samples obtained by the method of ultrasonic-TCA/acetone-phenol/SDS combined extraction were 6.650 mg/mL and 2.993 mg/g respectively. Furthermore, clear bands were obtained in the extracted protein by SDS-PAGE analysis and 1 238 independent clear protein spots were obtained by two-dimensional electrophoresis analysis. It is suggested that the T. roseum protein obtained by the combined extraction method of ultrasonic-TCA/acetone-phenol/SDS was suitable for two-dimensional electrophoresis analysis and further proteomic research.
2019 Vol. 34 (3): 209-216 [Abstract] ( 23 ) 全文: HTML   PDF (1200 KB)  ( 23 )
217 Development of a IPMA Method Applicable for Screening mAb Targeting PRRSV
YANG Yanyan, QIAO Songlin, LI Rui, GUO Junqing, LI Qingmei, TENG Man, WANG Li, ZHAO Dong, LI Xuewu, DENG Ruiguang, ZHANG Gaiping
DOI: 10.7668/hbnxb.20190199
To develop a high-throughput cell culture detection method for screening specific and sensitive monoclonal antibodies against Porcine reproductive and respiratory syndrome virus(PRRSV). A single layer of Marc-145 cells covered with 96-well plates was inoculated with a virus amount of about 100 cells infected per well. After 12 h, cells were fixed with 3% H2O2 in methanol to prepare an immunoperoxidase monolayer cell assay(Immunoperoxidase monolayer assay, IPMA) reaction plate. The culture supernatant of the hybridoma cells cultured for 10 days after the fusion was added to the IPMA reaction plate in an amount of 100 μL/well, and horseradish peroxidase-labeled goat anti-mouse IgG-HRP was used as the secondary antibody and 3-amino-9-ethylcarbazole(AEC) was used as a chromogenic substrate, then observed under an inverted microscope. The results showed that 39 PRRSV monoclonal antibodies, such as 1D1 and 7G8, were screened, and these 39 monoclonal antibodies were able to specifically stain Marc-145 cells infected with high-viral strain HN07-1 and classical strain BJ-4 PRRSV. There was no cross-staining of Marc-145 cells infected with Classical swine fever virus, Pseudorabies virus, and Porcine epidemic diarrhea virus. Therefore, this IPMA method constructed by the study can capture PRRSV monoclonal antibodies sensitively and accurately.
2019 Vol. 34 (3): 217-223 [Abstract] ( 34 ) 全文: HTML   PDF (1010 KB)  ( 14 )
224 Molecular Characterization and Tissue Expression Analysis of HCRT and HCRTR2 Genes in Goose
OUYANG Qingyuan, LIANG Jinmin, WANG Yushi, GAN Xiang, HU Shenqiang, WANG Jiwen
DOI: 10.7668/hbnxb.201751344
The aim of this study was to clone the complete coding sequences of HCRT and HCRTR2 genes in goose by designing the specific primers according to their respective reference sequences of humans, chicken and ducks on NCBI website, and then they were analyzed by corresponding bioinformatic method. The expression levels of HCRT and HCRTR2 mRNA in nineteen tissues of Sichuan White Goose were detected. The complete coding regions of HCRT and HCRTR2 genes in goose were 456,1 506 bp, respectively, encoding 151 and 501 amino acids. Among them, the precursor orexin protein encoded by HCRT gene was hydrolyzed to produce OXA and OXB polypeptides. Sequence analysis showed that goose OXA and OXB contained 34 and 28 amino acid residues, respectively. The sequence alignment showed that the amino acid sequences of OXA, OXB and HCRTR2 were highly conserved among the nearest species, suggesting that OXA, OXB and HCRTR2 might play an important role in the evolution of species. However, the signal peptide conservatism of orexin protein among species was low, suggesting that the signal peptide sequence of orexin protein might be important to the functional characteristics of species. The evolutionary tree analysis showed that the evolution of HCRT and HCRTR2 were similar. Goose had the closest relationship with chicken and the farthest with zebra fish. RT-qPCR results showed that the expression levels of HCRT and HCRTR2 genes in the hypothalamus were the highest, though they were also expressed in other tissues to a certain degree. It is speculated that orexin system might participate in the regulation of animal reproductive function through hypothalamus-pituitary-gonad axis (HPG). The results of this study laid the foundation for further studying the role of HCRT and HCRTR2 genes in goose reproductive activities.
2019 Vol. 34 (3): 224-231 [Abstract] ( 37 ) 全文: HTML   PDF (1450 KB)  ( 15 )
232 Cloning of Goat FGF9 Gene and Its Expression Pattern in Myoblast Differentiation
HUANG Kai, LIN Yaqiu, ZHU Jiangjiang, MA Jieqiong, WANG Yong
DOI: 10.7668/hbnxb.201751384
The aim of this study was to clone the goat FGF9 gene sequence and perform bioinformatics analysis, so as to elucidate the tissue expression characteristics of FGF9 and its differential expression during myoblast differentiation. The experimental animal of this study was the Jianzhou Big-eared sheep. The FGF9 gene sequence was cloned by RT-PCR. Real-time quantitative PCR (qRT-PCR) was performed to detect the expression of FGF9 in different tissues of goat and at different stages of myoblast differentiation. The results showed that the sequence of goat FGF9 gene was 818 bp, with an ORF region of 627 bp, encoding 208 amino acids. The nucleotide sequence of the CDS region was 99% homologous to Bostaurus and Ovisaries. The FGF9 protein had a transmembrane domain and a FGF family homology domain, which was an unstable hydrophilic protein. The FGF9 gene could express in all tissues of goats, and the expression level in the kidney was the highest, which was significantly higher than in other tissues (P<0.01). The expression level of FGF9 gene in the 2nd day of induced differentiation was significantly higher than that before differentiation (P<0.05), and reached a very significant level in the 4th day (P<0.01). This study provided the important data for further exploring the role of FGF9 gene in goat muscle growth.
2019 Vol. 34 (3): 232-238 [Abstract] ( 56 ) 全文: HTML   PDF (1146 KB)  ( 29 )
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