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2019 Vol. 34, No. 5
Published: 2019-10-28

Crop Genetics·Germplasm Resources·Biotechnology
Tillage and Cultivation·Physiology and Biochemistry
Resource and Environment·Plant Protection
0 Table of Contents for VOL.34 No.5
2019 Vol. 34 (5): 0-0 [Abstract] ( 18 ) 全文: HTML   PDF (257 KB)  ( 26 )
       Crop Genetics·Germplasm Resources·Biotechnology
1 Cloning and Expression Analysis of Aquaporin TaTIP1-4DL Gene in Wheat
ZHANG Shan, GUO Qiping, LIU Yuanyuan, DANG Renmei, WEN Shanshan
DOI: 10.7668/hbnxb.201751747
The TIPs family is a type of membrane intrinsic protein located on the tonoplast, which is responsible for regulating cell turgor and responds to external stress. To further explore the expression pattern and biological function of TIPs gene in wheat, the TaTIP1-4DL gene was amplified from flag leaves of wheat by homologous cloning. The coding region was 771 bp in length and encoded 257 amino acids. Bioinformatics analysis showed that the protein had a molecular weight of 26.3 ku and a theoretical isoelectric point of 6.82, belonging to a hydrophobic stability protein. The tertiary structure was a conservative hourglass structure containing six long alpha helices and two short alpha helices, corresponding to the classic model of aquaporin. The amino acid alignment and phylogenetic tree analysis showed that the TaTIP1-4DL protein of wheat was closely related to the homology of Aegilops Linn.and Brachypodium distachyou(L.) Beauv.The promoter cis-acting element contained the response element related to stress. Tissue-specific expression analysis showed that TaTIP1-4DL gene was expressed in different tissues and organs of wheat, and the highest expression level was in leaves and the least in stems. The stress expression analysis showed that the expression level of TaTIP1-4DL gene in leaves and seeds was induced in different degrees by drought, abscisic acid and high salt, and the expression level was generally up-regulated and then decreased. Among them, drought induced the highest level of the gene in leaves. After 6 hours of drought treatment, the expression of the gene in leaves reached 14 times of that before stress.
2019 Vol. 34 (5): 1-7 [Abstract] ( 56 ) 全文: HTML   PDF (1230 KB)  ( 65 )
8 Cloning and Expression Analysis of CmDWF1 Gene in Melon
ZHANG Jin, JIN Wuyun, CHEN Yingying, FAN Yunfei, HAO Jinfeng, HASI Agula
DOI: 10.7668/hbnxb.201751608
Brassinosteriod is a necessary for plant to regulate the plant physical activities, and DWF1 is one of most critical enzymes involved in brassinosteriod biosynthesis. In order to study the function of DWF1 in melon fruit development, the gene CmDWF1 was cloned from melon for analyzing the nucleotide and amino acid sequences. RT-PCR method was used to analyze the gene expression characteristics. The results showed that the open read frame length of CmDWF1 was 1 701 bp, encoding 566 amino acids. The predicted protein molecular weight of CmDWF1 was 66.115 11 ku, the theoretical isoelectric point was 6.96, the instability index was 48.18 and the GRAVY was -0.402. Signal sequence was not found and the location coefficient in the cytoplasm was 9, which suggested that the protein was localized in cytoplasm. The secondary structure of CmDWF1 protein contained 37.81% of alpha helix, which was relatively high. The CmDWF1 protein contained a FAD-binding-4 domain, which suggested that the protein acted as a FAD oxidoreductase. Phylogenetic analysis showed that Cucumis melo had closer relationship with Cucumis sativus. Real-time fluorescent quantitative analysis indicated that CmDWF1 could express in roots, stems and leaves, but the highest expression level was in roots. And the expression level increased significantly in the 25 d fruit after pollination, but decreased in 35-45 d fruit. These results demonstrated that CmDWF1 might be associated with the biosynthesis of brassinosteriods and then regulated the growth and development of melon.
2019 Vol. 34 (5): 8-14 [Abstract] ( 36 ) 全文: HTML   PDF (1419 KB)  ( 34 )
15 Cloning of LbCER1 Gene and Expression Analysis under Osmotic Stress in Lycium barbarum ssp. Bianguo
YUAN Huijun, MA Qianguo, GAO Ze, LI Xueyong, BAO Jingting, WANG Chunmei, LI Hujun
DOI: 10.7668/hbnxb.201751628
In order to identify the molecular characterization of fatty-aldehyde decarboxylase gene (ECERIFERUM1, CER1) involved in osmotic stress of xerophyte, CER1 gene was cloned from the cDNA of Lycium barbarum ssp. Bianguo by reverse transcription PCR (RT-PCR) and RACE method, which was a kind of salt-tolerant and drought-tolerant plant. Then the sequence was analyzed by bioinformatics software. Furthermore, the gene expression level of CER1 in L. barbarum ssp. Bianguo was detected in leaf, stem and root under osmotic stress treatment by quantitative Real-time PCR (qRT-PCR). The results showed that the gene sequence of CER1 in L.barbarum ssp. Bianguo contained 2 168 bp in full length and an open reading frame of 1 881 bp, which was named LbCER1, encoding 626 amino acids. Based on bioinformatics analysis, the protein contained four transmembrane regions, a fatty acid hydroxylase domain and a Wax2 domain of C-terminal. The calculated molecular weight, isoelectric point, aliphatic index, instability index and average hydropathicity (GRAV) of LbCER1 was 72.47 ku, 7.40, 91.28, 27.88 and -0.070, respectively. LbCER1 is a stable hydropathic protein possessing alkalinity and high-temperature tolerance, which is located in the membrane of plastic reticulum. Based on the secondary structure prediction of LbCER1, the majority of its amino acid compose alpha-helix (44.09%) and loop (45.21%), which was confirmed by tertiary structure prediction. In addition, LbCER1 was predicted to have 100% homologous with Solanaceae CER1 by phylogenetic analysis. Expression pattern of LbCER1 showed that it express in roots, stems, and leaves. The LbCER1 was also induced by 160 mmol/L sorbitol for 6 h in leaf, stem, and root to 1.8, 1.2, and 1.1 fold, respectively. On contrary, LbCER1 was only sensitive to 80 mmol/L sorbitol in leaves after 6 h-treatment. These results indicate that LbCER1 may play a role in osmotic stress response in L. barbarum ssp. Bianguo.
2019 Vol. 34 (5): 15-22 [Abstract] ( 52 ) 全文: HTML   PDF (1246 KB)  ( 16 )
23 Cloning and Expression Analysis of IiCYP79B2 from Isatis indigotica Fort.
HU Xiangyang, QU Xinyun, WU Gege, QIN Miaomiao, LIU Rui, GAO Tian, LI Tao
DOI: 10.7668/hbnxb.201751719
In order to explore the structure, characteristics and functions of IiCYP79B2 gene in Isatis indigotica, the IiCYP79B2 gene was cloned and the bioinformatics analysis and expression patterns under different conditions were carried out. The results showed that the full length of the gene was 1 827 bp with an intron, and its ORF was 1 629 bp, encoding 542 amino acids. IiCYP79B2 protein contained two transmembrane domains without signal peptide. The protein was localized in the chloroplast thylakoid membrane and mainly consisted of random coils and α-helix structures, having the closest relationship with Sinapis alba. The qRT-PCR analysis showed that the expression levels of IiCYP79B2 in different tissues were leaf > stem > fruit > flower > root, and the expression levels at different development stages were vegetative growth > flowering > seedling > germination. Moreover, IiCYP79B2 could be induced significantly by MJ and Glu, and repressed by SA and cold. The obtained results provided some experimental evidences for further studying the functions of IiCYP79B2 gene.
2019 Vol. 34 (5): 23-29 [Abstract] ( 28 ) 全文: HTML   PDF (1501 KB)  ( 19 )
30 Cloning and Bioinformatic Analysis of RcPIP2 from Racomitium canescens
ZHANG Meijuan, XU Honghong, MA Tianyi, QIAN Pengzhi, SHA Wei
DOI: 10.7668/hbnxb.201751698
Aquaporins (AQPs) are small transmembrane proteins, and play an important role in tolerance of plants to abiotic stresses. To study the biological functions of aquaporin genes from Racomitrium canescens, a novel plasma membrane intrinsic protein gene of aquaporin family designated RcPIP2 was isolated from the drought transcriptome sequences. The bioinformation results showed that the cDNA full length of RcPIP2 gene was 1 267 bp,which contained an open reading frame of 807 bp and encoded a protein of 268 amino acid residues. It was predicted that the molecular weight of the protein was 28.3 ku, and its theoretical isoelectric point was 6.64, belonging to a stable and hydrophobic protein with no signal peptide. It was a non-secretory protein. RcPIP2 exhibited a typical structure with six transmembrane domains, and had the highly conserved sequence HINPAVTFG of membrane intrinsic protein (MIP) family and two highly conserved Asn-Pro-Ala (NPA) peptides. Its secondary structure contained 37.31% α-helixes, 20.9% β-sheets, and 38.43% random curls. The three-dimensional structure of the RcPIP2 protein was a typical tetramer consisting of four cylindrical subunits and the middle channel. Phylogenetic tree analysis indicated that the protein had the closest genetic relationship with PIP2 protein of Physcomitrella patens, being grouped to the same class. It was inferred that RcPIP2 was one of the aquapion family members.
2019 Vol. 34 (5): 30-36 [Abstract] ( 36 ) 全文: HTML   PDF (1303 KB)  ( 24 )
37 Genetic Mapping and QTL Analysis of Fruit Russeting in Malus domestica Borkh.
ZHANG Chaohong, CHEN Dongmei, YANG Fengqiu, ZHAO Tongsheng, LI Yang, ZHAO Guodong, ZHAO Yongbo
DOI: 10.7668/hbnxb.201751695
In order to effectively discover the apple fruit russet gene, based on SLAF-seq technology, the genetic map construction and QTL analysis for fruit russet gene were conducted using a population derived from a cross between Miyazaki Spur and Sakata Tsugaru. The results showed that 522 122 315 reads (110.32 Gb) of the sequencing data from seedlings was obtained, in which the average Q30 sequencing was 95.07%, and the average GC content and its parents was 40.11%. Then, 20 440 SLAF markers and 7 309 729 SNPs were developed. Based on these markers, seventeen linkage groups were constructed, of which 4 075 markers presented in the genetic map, and marker integrity was 99.92%. The total diagram spacing was 2 235.23 cM, and the average diagram spacing was 0.55 cM. A total of 9 QTLs related to fruit russet were detected, which were distributed on Chr3, Chr9, Chr11, and Chr15 chromosomes, with a marker distance of 0-4.9 cM and a contribution rate of 18.0%-85.5%. Among them, Qru-9, Qru-15 and Qru-3, located on Chromosome 9, 3, and 15, respectively, might be novel QTLs for fruit russeting of apple. A genetic map based on SLAF-seq method was constructed in apple progeny Miyazaki Spur crossing Sakata Tsugaru and nine QTLs related to apple russeting were obtained. These results provided the important information for further exploration of russeting QTL in apple.
2019 Vol. 34 (5): 37-44 [Abstract] ( 55 ) 全文: HTML   PDF (1315 KB)  ( 19 )
45 Identification of Differential Proteins in Roots of Tobacco with Overexpressing TaZAT8, a Zinc Finger Type Transcription Factor Gene from Wheat
GU Qian, DING Weiwei, JIANG Mingyue, SU Xiaoshuai, LI Xiaojuan, XIAO Kai
DOI: 10.7668/hbnxb.201751771
In order to further clarify the mechanism of over expression TaZAT8, a wheat C2H2 type zinc finger gene, to enhance the resistance of tobacco plants under P-starvation, the classical proteomics approach was applied to reveal the changes of protein expression between WT and transgenic tobacco plants. The results showed that there were 22 protein spots differentially expressed on 2-DE images, and among them, twenty protein spots were up-regulated and two were down-regulated respectively. Subsequently, these protein spots were identified by LC-MS/MS, which were classified into six functional groups, including primary metabolism, energy generation, stress response and defense, transcription and translation, protein turnover and unclear functions. Subcellular localization analysis indicated that these proteins were mainly located in five organelles, including cytoplasm, mitochondria, peroxisome, nucleus and chloroplast. Go analysis showed that the differentially expressed proteins participated in various metabolism processes or act as enzymes of different categories. Above results showed that over expression TaZAT8 laid the foundation for improving the plant tolerance to stress via regulating proteins involved in biological processes such as substance metabolism, energy production, stress response, etc.
2019 Vol. 34 (5): 45-51 [Abstract] ( 35 ) 全文: HTML   PDF (1148 KB)  ( 10 )
52 Screening and Identifying of Interaction Protein AtLBD15 in Arabidopsis thaliana
GUO Zhaolai, SUN Xudong, YANG Yongping, XU Huini
DOI: 10.7668/hbnxb.201751326
In order to further explore the role of LBD genes in Arabidopsis thaliana in response to plant abiotic stresses, we first inserted LBD15 gene into pGBKT7 vector and constructed the pGBKT7-LBD15 yeast expression vector. The pGBKT7-LBD15 was used to screen the interacted proteins of LBD15. The yeast two-hybrid technique was used to screen a full-length homogenized yeast library of Arabidopsis thaliana. There were 107 detected sequences in 146 samples of which 96 were identified as the Arabidopsis thaliana genes by NCBI Blast, accounting for 87.9% of the total. According to the gene accession number, 48 proteins were obtained, and 59% of them were duplicated. And then the protein sequences were searched via the website TAIR ( based on the gene accession number. The subcellular localization was analyzed on Through localization analysis, these proteins were mainly located in the chloroplast, cytoplasm and nucleus. Gene Ontology annotations using Blast2GO showed that the interacting proteins were involved in 14 biological processes, including cellular processes, metabolic processes, stimuli response, regulation of biological processes, single-tissue and multi-tissue processes, and developmental process regulation. Five of the possible interaction proteins were randomly selected and used to verify the interaction. The results showed that the colonies of AP19 protein and other proteins could grow normally on auxotrophic plates, further confirming that they were interacted proteins. The successful screening of yeast two-hybrid library laid the foundation for further study of the molecular mechanism of Arabidopsis LBD15.
2019 Vol. 34 (5): 52-58 [Abstract] ( 26 ) 全文: HTML   PDF (1155 KB)  ( 20 )
59 Construction of Plant Binary Expression Vector for OsPIP2; 7 with HA-tag and Its Transformation in Rice
WANG Xiaojing, SUN Linjing, SUN Yue, ZHANG Rongxue, LI Junling, YAN Shuangyong, SU Jingping
DOI: 10.7668/hbnxb.201750580
To analyze the function of OsPIP2; 7 under environmental stresses, specific primers were designed to clone OsPIP2; 7 from the complementary DNA of rice variety Zhonghua 11. The nucleotide sequence of 3×HA tag was connected to 3'-end of OsPIP2; 7 with removed terminator by nested PCR amplification, and then the DNA fragments were inserted into pHB vector to construct pHB-OsPIP2; 7-3×HA. The calli of of mature embryo from Zhonghua 11 were used to conduct the genetic transformation by Agrobacterium -mediated transformation method. After screening and transformation on a culture medium containing hygromycin, the positive transformants were obtained. Western-blot results revealed that OsPIP2;7-HA was over-expressed in transgenic plant at protein level when using HA as the first antibody. This study will help the subsequent research of OsPIP2; 7 function and mechanism.
2019 Vol. 34 (5): 59-64 [Abstract] ( 26 ) 全文: HTML   PDF (1185 KB)  ( 13 )
65 Genome-Wide Identification, Classification and Expression Analysis of Sugar Transporter Gene Family in Sorghum bicolor
XU Zhijun, LIU Yang, XU Lei, AN Dongsheng
DOI: 10.7668/hbnxb.201751467
The sugar transporter(STP)gene family plays critical roles in monosaccharide distribution and participates in diverse plant metabolic processes. To investigate the potential roles of STPs in sorghum growth and development, the genome-wide identification, classification and expression analyses of the STP gene family were performed in this study. A total of 19 SbSTP genes containing the Sugar_tr conserved motifs were identified from the sorghum genome, which distributed on 9 chromosomes unevenly, and 4 of them located in the genome segmental duplication region. According to their phylogenetic features, the 19 SbSTPs were further classified into 5 main groups, with high similar motif compositions within the same group, but they were different in exon-intron number. The synteny analysis and phylogenetic comparison of STP genes from sorghum, Arabidopsis and rice provided the valuable clues for revealing the evolutionary characteristics of SbSTP genes. RNA-Seq data analysis showed that 18 of the 19 SbSTP genes could expressed in various tissues with distinct expression patterns, among which SbSTP11 exhibited tissue-specific expression in flower, while SbSTP4 and SbSTP5 expressed in root or shoot by induction of ABA and PEG stress. The results provided a valuable resource for better understanding the biological roles and application potentiality of individual STP genes in sorghum.
2019 Vol. 34 (5): 65-73 [Abstract] ( 23 ) 全文: HTML   PDF (1205 KB)  ( 27 )
74 Functional Analysis of PmSOC1 -like Genes Related to Floral Organ Development in Prunus mume
LI Yushu, YANG Weiru, YANG Yujie, CHENG Tangren, WANG Jia, ZHANG Qixiang
DOI: 10.7668/hbnxb.20190196
The MADS-box gene SOC1/TM3 (SUPPRESSOR OF OVEREXPRESSION OF CONSTANS 1/Tomato MADS-box gene 3) integrates multiple flowering signals to regulate the transition from vegetative to reproductive development in plants. Three putative orthologs of SOC1 (PmSOC1-1, PmSOC1-2 and PmSOC1-3) were cloned from Prunus mume Changruilüe. The sense plant expression vectors driven by Cauliflower mosaic virus 35S promoter were constructed,and then were transformed separately into wild-type Arabidopsis plants, to study the function of PmSOC1-1, PmSOC1-2 and PmSOC1-3. Overexpression of PmSOC1-1, PmSOC1-2, and PmSOC1-3 in Arabidopsis caused early flowering. Early flowering also increased expression levels of four other flowering promoters, agamous-like 24 (AGL24), leafy (LFY), apetala 1 (AP1) and fruitfull (FUL). Moreover, the overexpression of PmSOC1-1 and PmSOC1-2 resulted in a range of floral phenotype changes such as sepals into leaf-like structures, petal color into green, petal into filament-like structures. These results suggested that the genes PmSOC1-1, PmSOC1-2 and PmSOC1-3 played an evolutionarily conserved role in promoting flowering in P. mume, and might have distinct roles during flowering development. These findings will help elucidate the molecular mechanisms involved in the transition from vegetative to reproductive development in P. mume.
2019 Vol. 34 (5): 74-82 [Abstract] ( 19 ) 全文: HTML   PDF (1216 KB)  ( 13 )
83 Backcross Breeding of Late Bolting and Detection of First Intron PCR Amplification of FLC2 Gene in Radish
LIU Chunxiang, SUI Ming, LIU Chunxia, XIANG Chunling, CAO Hongxiang, CHANG Tianli, YIN Chenlin
DOI: 10.7668/hbnxb.201751607
FLC2 gene is a key gene affecting the bolting and flowering and plays an important role in the process of transition from vegetative growth to flower development in cruciferous plants. In order to breed a late-bolting green radish cultivar suitable for early spring cultivation, based on the function of FLC2 gene effecting the bolting, the late bolting radish variety Baiyuchun was used as the donor to transfer the late bolting character to Weifang radish by backcross breeding, and the polymorphism of FLC2 was studied during the process of backcross breeding to explore the effects of FLC2 in late bolting radish breeding. The results showed that the first intron of FLC2 gene had an insertion mutation with a length of about 1 628 bp in the late bolting donor, which was important for late bolting, and the homozygous insertion mutation had greater effect on late bolting than the heterozygous type did. It was found that the length of vernalization period affected the PCR amplification results on the first intron of FLC2. The short-term vernalization mainly affected the amplification of the first intron of the wild type FLC2, and the long-term vernalization mainly affected the amplification of intron in the insertion mutant, while the amplification of the two genotypes in the bolting and flowering period was not affected greatly. The amplification efficiency could be improved by improving the primers, but it could not eliminate the effect of vernalization on the first intron amplification of FLC2. The PCR result of normal plants at seedling stage was not as good as that at bolting stage. In conclusion, late bolting trait could be transfer into the acceptor radish by backcross continuously. When the heterozygous genotype was created by backcrossing, it could transfer the gene to its offsprings, and then the homozygous genotype could be obtained by self-crossing. The homozygous mutants with FLC2 insertion sequence could be confirmed by PCR detection at flowering stage, which could ensure the genetic stability and avoid the separation of late bolting traits in the offsprings.
2019 Vol. 34 (5): 83-92 [Abstract] ( 15 ) 全文: HTML   PDF (1554 KB)  ( 6 )
93 Transformation and Application of One Molecular Marker from SCAR to KASP in Pepper CMS
ZHANG Qiang, ZHANG Tao, CHANG Xiaoke, HAN Yanan, CHENG Zhifang, LIU Wei, WANG Bin, YAO Qiuju
DOI: 10.7668/hbnxb.20190131
In order to narrow population range to be screened, reduce the workload and improve the selection efficiency of sterile and maintainer lines in the three-line cross breeding of pepper, the sequence polymorphism sites of cytoplasmic fertility marker SCAR130 was used to design KASP primer to transform SCAR130 into KASP130 molecular marker. The three lines material (male sterile line, maintainer line, restorer line) and F1 hybrids of pepper CMS were selected as the experimental subjects, and the KASP130 molecular marker were applied into detection of pepper cytoplasm type and its stability and reliability were tested through two detection platforms of real time PCR machine LC480 and the SNPline of LGC Company, respectively. The results showed that KASP130 marker, same as SCAR130, could accurately divide the tested pepper materials into fertile cytoplasm(N) and sterile cytoplasm(S). Furthermore, in the marker-assisted selection breeding, KASP130 was also successfully applied to the early identification of cytoplasmic fertility of pepper as well as the backcross breeding of pepper maintainer lines and male sterile lines. In conclusion, cytoplasmic fertility marker SCAR130 has been successfully transformed into KASP130 molecular marker. It can identify the cytoplasm type of pepper, which lays a foundation for application of KASP130 in three-line cross breeding programe of peppers.
2019 Vol. 34 (5): 93-98 [Abstract] ( 46 ) 全文: HTML   PDF (958 KB)  ( 18 )
99 Resistance of GhWRKY48 Negatively Regulated Cotton Against Verticillium dahlia
LIU Jianfen, LEI Yu, ZHANG Zhennan, HU Guang, TANG Ye, ZHANG Ning, SI Huaijun, WU Jiahe
DOI: 10.7668/hbnxb.201751632
Since Verticillium wilt causes great loss in cotton production every year, it is an economical and effective method to identify resistance genes for breeding new varieties to control Verticillium wilt. A WRKY transcription factor gene was cloned from upland cotton, named GhWRKY48. The full-length of GhWRKY48 cDNA was 882 bp,encoding 293 amino acid residues with a predicted molecular mass of 32.68 ku and an isoelectric point (pI) of 6.10. qPCR analyses showed that GhWRKY48 gene constitutively expressed in root, stem and leaf, and preferentially done in stem. In addition, GhWRKY48 expression could be induced by V. dahliae infection, jasminic acid and salicylic acid. To dissect the defense function, GhWRKY48 -silenced plants were developed by virus-induced gene silencing techniques, which were then inoculated with V. dahliae. The results showed that the resistance of these silenced plants significantly increased compared to the control. The rates of diseased plants and disease index in GhWRKY48 -silenced plants were lower than in the control. All together, GhWRKY48 was a negative transcription factor participating in plant resistance to V. dahliae. Therefore, the GhWRKY48 gene could be used as an ideal candidate gene for cotton defense breeding.
2019 Vol. 34 (5): 99-105 [Abstract] ( 18 ) 全文: HTML   PDF (1177 KB)  ( 4 )
       Tillage and Cultivation·Physiology and Biochemistry
106 Effects of Irrigation Methods on Physiological Characteristics, Dry Matter Accumulation and Yield of Rice in Double Cropping Rice Field
XU Yilan, LIU Tangxing, FU Aibin
DOI: 10.7668/hbnxb.201751562
Irrigation is an important management measure that affects the growth and development, grain yield of rice. In order to explore the effects of different irrigation methods on the physicochemical and biological characteristics, and grain yield of rice in double-cropping rice system, the changes physiological and biochemical characteristics, dry matter accumulation and distribution, grain yield of early rice and late rice were analyzed under 3 treatments of intermittent irrigation method(Ⅱ), wet irrigation method(WI)and water saving irrigation method(WSI). The results showed that, at main growth stages of early rice and late rice, the tillering number of rice in different irrigation methods was Ⅱ > WI > WSI and the leaf area index was WI > Ⅱ > WSI. The activities of SOD, POD and CAT of rice leaves increased in treatments Ⅱ and WI compared with that in treatment WSI. The Pn of rice leaves in different irrigation methods decreased following the order of WI > Ⅱ > WSI at main growth stages. The dry weights of root, stem and panicle were Ⅱ > WI > WSI, but the dry weight of rice leaves was WI > Ⅱ > WSI. There was no significant difference in panicle dry weight among different treatments. The results indicated that the number of effective tillers was higher in Ⅱ treatment than in WI and WSI, but there were no significant differences in spikelet number per panicle, seed setting rate and 1 000 grains weight of rice among different treatments. The grain yield of rice was higher in Ⅱ treatment than in WI and WSI. Compared with WI and WSI treatments, the grain yield of early rice in Ⅱ treatment increased by 107.9,461.7 kg/ha in 2016, and 98.8,422.2 kg/ha in 2017, respectively, while the grain yield of late rice in Ⅱ increased by 250.1,683.6 kg/ha in 2016, and 220.3,661.8 kg/ha in 2017, respectively. The intermittent irrigation method(Ⅱ)enhanced the protective enzyme activities and photosynthetic characteristics of rice leaves, increased the dry matter accumulation of rice plant and improved the yield components, thus increased the grain yield of rice.
2019 Vol. 34 (5): 106-115 [Abstract] ( 25 ) 全文: HTML   PDF (1371 KB)  ( 40 )
116 Effects of Illumination and Nitrogen Rate on N2O Emission in Phyllosphere and Rhizosphere of Rice
LIU Xiaolin, LIN Li, LIU Jiani, CHEN Zebin, YU Lei, LIU Guihua, XU Shengguang
DOI: 10.7668/hbnxb.201751246
This study aimed to clarify the effects of different N application rates and light intensities on N2O emission flux in phyllosphere and rhizosphere of rice and its mechanism. The experiment was carried out by hydroponic method in a small light-controlled incubator, using rice variety Peizataifeng as the material, and the above-ground and underground parts of rice were strictly separated in the inner and outer chambers of the test device. The amount of N2O emitted from the root and leaf interface of rice was determined by gas chromatography. First, we conducted the indoor low light (8:00-18:00, 4 000 lx) test using NH4NO3 as the nitrogen source, of which three N levels, low nitrogen (30 mg/L), medium nitrogen (60 mg/L) and high nitrogen (90 mg/L) at different developing stages were arranged. On this basis,we carried out the experiment under the conditions of low light (4 000 lx), strong light (8 000 lx) and natural light, including three N treatments:denitrification (0 mg/L), low nitrogen (30 mg/L) and high nitrogen (90 mg/L) at flowering and fruiting stages. The results showed that, under weak illumination (8:00-18:00, 4 000 lx), the mean rate of N2O emission was 34.9, 42.4 and 98.3 μg/(m2 · h) in rice phyllosphere, and was 29.6, 79.6, 246.1 μg/(m2 · h) in rice rhizosphere, respectively, for treatments of 1N (30 mg/L), 2N (60 mg/L) and 3N (90 mg/L). With the increase of N rate, the amount of N2O emission in rice phyllosphere and rhizosphere also increased (P<0.05), and the similar results were obtained at flowering/seeding stage and mature aging stage. The N2O emission rate in the phyllosphere of 0N (0 mg/L), 1N (30 mg/L) and 3N (90 mg/L) was 2.9, 29.1 and 116.3 μg/(m2 · h) for weak light treatment, was 23.6, 40.1 and 120.1 μg/(m2 · h) for strong light treatment (8:00-18:00, 8 000 lx), and was up to 10.9, 26.2 and 131.3 μg/(m2 · h) under natural light,condition, respectively. The N2O emission rate in the rhizosphere of 0N (0 mg/L),1N (30 mg/L) and 3N (90 mg/L) was 3.3, 77.1 and 308.4 μg/(m2 · h) for weak light treatment, was 14.1, 45.6, and 182.4 μg/(m2 · h) for strong light treatment, and was 19.3, 44.9 and 224.6 μg/(m2 · h) under natural light,condition, respectively. Under strong and natural light conditions, the N2O emission rate in the rhizosphere of rice was not significant between 0 N (0 mg/L) and 1N (30 mg/L) treatments (P>0.05). Under weak light condition, the N2O emission rate in the rhizosphere and phyllospher increased with the increase of N application amount. Under the same condition of N application (1N, 3N), the N2O emissions in the rhizosphere of rice could be inhibited by stronger light and sunlight compared with the results of weak light treatment. It was suggested that the combination of strong light and low N level (1N) could inhibit the N2O emissions in the rhizosphere and phyllosphere of rice.
2019 Vol. 34 (5): 116-125 [Abstract] ( 20 ) 全文: HTML   PDF (1136 KB)  ( 35 )
126 Effects of Zinc Fertilizer on Root Growth and Yield of Winter Wheat under Drought Stress
ZHANG Jun, LIANG Zhenkai, WANG Xueping, HU Lihong, LI Youjun
DOI: 10.7668/hbnxb.20190387
Luohan 6 was used as the experimental material in a pot experiment to investigate the effects of zinc fertilizer on root growth and grain yield of winter wheat under drought stress. Four treatments were set up:T1 (normal irrgation+0 mg/kg Zn), T2 (normal irrgation+1 mg/kg Zn), T3 (drought stress+0 mg/kg Zn), and T4 (drought stress+1 mg/kg Zn). The results showed that, compared with T1, the total root length, root mean diameter, total root surface area and total root volume of wheat were noticeably reduced in T3 treatment. The density and diameter of root hair were dramatically decreased, but the difference of root hair length was not significant. The root hair structure was damaged by drought stress, of which the cell wall of root hair became thinner, no nucleus was seen, most of the organelles disintegrated, the inclusion disappeared, and the root hairs decayed. The contents of zinc and indoleacetic acid (IAA) in the root, as well as the root activity, decreased, which resulted in the marked decrease of grain yield per plant. Under T4 treatment, the total root length, root mean diameter, total root surface area and root volume increased observably compared to T3 treatment, and the density and diameter of root hair also increased greatly. The root hair structure was intact, the nucleus was clearly visible, the inclusion was sufficient, and the root hair developed well. In addition, the contents of zinc and IAA in the root, as well as the root activity, likewise ascended remarkably. Compared with T3, the spikes per plant, grain per spike, thousand kernel weight and grain yield per plant of T4 increased by 6.25%, 8.01%, 6.87% and 50.00%, respectively. Comprehensive analysis showed that under the ecological conditions of dryland in Western Henan, the application of zinc fertilizer can enhance the stability of wheat root growth and improve the development of wheat root. As a result, the application of zinc fertilizer has an important role in alleviating wheat yield reduction under drought stress.
2019 Vol. 34 (5): 126-136 [Abstract] ( 24 ) 全文: HTML   PDF (1577 KB)  ( 15 )
137 Effects of Different Drought Stresses on Growth and Physiological Properties of Soybean
JIA Sichun, WANG Na, HAO Xingyu, ZONG Yuzheng, ZHANG Dongsheng, LI Ping
DOI: 10.7668/hbnxb.201751770
To study the physiological effects of different degrees of drought on soybean cultivars in different development periods. Two soybean cultivars with different drought resistances were taken as the experiment materials to study the effects of drought stress on the resistance index and physiological characteristics of soybean in different growth periods, and to explore the physiological mechanism of soybean response to different drought intensities at different developing stages in pot culture. Four treatments were set up:continuous mild drought stress (T1), continuous moderate drought stress(T2), drought stress in full seed stage (T3) and without drought stress(CK). The results showed that under drought stress, the chlorophyll contents of both soybean cultivars increased, and Jindazaochun 2 was more obvious at early stage, but it was less than that of Jinda 74 at grain-filling stage. Under continuous mild drought stress and continuous moderate drought stress, the MDA content of Jindazaochun 2 at different growth stages significantly increased, while the POD and MDA contents of Jinda 74 significantly increased under continuous moderate drought stress. At the branching stage, the content of reducing sugar in Jinda 74 decreased significantly during moderate drought. In the Granulation stage, the reducing sugar content of soybean leaves in Jindazaochun 2 was significantly increased, Jinda 74 was only reduced under drought treatment in the Granulation stage. At flowering stage, reducing sugar content reduced in Jindazaochun 2, but increased significantly in Jinda 74. It was found that drought had a great influence on reducing the plant height, node number, stem diameter of both cultivars. The decrease in plant height and stem diameter of Jinda 74 was more obvious. The resistance of Jinda 74 to drought was higher than that of Jindazaochun 2, which was related to the reduction of plant height and water consumption under continuous drought conditions. The results will provide a basis for comprehensive understanding of the soybean resistance to drought stress.
2019 Vol. 34 (5): 137-144 [Abstract] ( 26 ) 全文: HTML   PDF (1030 KB)  ( 22 )
145 Effects of Regulated Drip Irrigation at Different Growth Stages on Yield, Quality and Water Use Efficiency of Potato in Oasis Region
ZHANG Wanheng, ZHANG Hengjia, LI Fuqiang, WANG Zeyi, GAO Jia, BA Yuchun
DOI: 10.7668/hbnxb.201751588
In order to explore the effects of irrigation treatments at different growth stages on the growth dynamics, water consumption characteristics, quality, yield and water use efficiency of potato in Hexi Oasis, a field experiment was carried out, taking Qingshu 168 as the material and setting up two irrigation treatments:mild and moderate water deficits. The plant height, leaf area, yield and tuber organic matter content of potato were determined. The results showed that the yield of potato treated with mild water deficit in tuber forming period did not decrease significantly compared with CK, while the water use efficiency, commercial potato rate and bunched potato proportion increased by 22.07%, 10.09 percentage point and 8.20 percentage point respectively, and the dry matter, starch, protein and reducing sugar of potato tubers did not significantly reduced.The mild (WD5) and moderate (WD6) water deficits at tuber bulking stage reduced the potato yield by 15.77% and 18.93% respectively, reaching to a 0.05 significant level, and they were not beneficial to organic matter accumulation of potato tubers and decreased the contents of dry matter, protein and reducing sugar in potato tubers by 1.62 percentage point and 3.38 percentage point, 28.50% and 24.35%, and 34.38% and 46.88%, respectively. The mild (WD7) and moderate (WD8) water deficits at starch accumulation stage had less effect on yield, however, WD7 treatment increased the commercial tuber rate, protein content and reducing sugar content by 8.61 percentage point, 3.11% and 15.63% respectively, while the water use efficiency was not improved significantly. Therefore, in pursuit of higher yield and water use efficiency of potato, mild water deficit treatment at tuber formation stage is the best irrigation strategy in this experiment, but for seeking higher quality of potato, mild water treatment in starch accumulation period is the best.
2019 Vol. 34 (5): 145-152 [Abstract] ( 17 ) 全文: HTML   PDF (1109 KB)  ( 13 )
153 Effects of Nutrient Solution Concentration on Quality, Yield and Photosynthetic Characteristics of Cucumber Grown in Enclosed Circulation Trough
MENG Xianmin, JI Yanhai, WU Zhanhui, CHU Zhaosheng, LIU Mingchi
DOI: 10.7668/hbnxb.201751708
In order to screen the suitable concentration of nutrient solution for enclosed circulation trough of Cucumis sativus L., taking advantages of the characteristics of nutrient solution recycling in enclosed circulation trough, the effect of different concentrations of nutrient solution on the growth, quality, yield, photosynthetic capacity and microstructure of leaves were studied. Using Zhongnong 26 as experimental material, the basic EC value of nutrient solution was 2.7 mS/cm, the five treatments were T1, T2 (CK), T3, T4 and T5 (EC value 2.3, 2.7, 3.1, 3.5, 3.9 mS/cm respectively) and the growth status, photosynthetic capacity of leaves, fruit quality and yield were screened. The results showed that with the increase of EC value, the content of soluble sugar increased and VC, free amino acid and soluble protein of cucumber first increased and then decreased. Compared with T2, T3 had a better blade structure with closely arranged palisade tissue and relatively loose sponge tissue, and the qP,ΦPSⅡ and ETR values were higher than those of other treatments, which the open degree of PSⅡ reaction center, the efficiency of the original photochemical reaction and the electron transfer of the PSⅡ was significantly improved.And the light energy was mainly distributed to the photochemical reaction P and the antenna heat dissipation D, and reduced the part of non-photochemical dissipation Ex. The value of β/α-1 for T3 was the lowest and qP was the highest, the PSⅠ and PSⅡ were relatively balanced, the light energy utilization efficiency was higher, and the photosynthesis of leaves was enhanced, which was also the material basis for improving fruit quality and yield. Therefore, T3 treatment was more suitable for cucumber growth and development. The content of soluble sugar and soluble solids of T4 was higher than T2, there was no significant difference with T3 yet. The net photosynthetic rate and the photosynthetic pigment content in T4 leaves were the highest, Pn was increased by 58.85% compared with T2.In addition, the T3 and T4 treatments were lower in Fo and the Fv/Fm was higher than the T2 treatment, the PSⅡ reaction center activity of both were higher. Under the condition of closed trough culture, the photosynthetic electron transfer efficiency of cucumber leaves with EC value of 3.1 and 3.5 mS/cm was higher, and the photosynthetic capacity was enhanced, which improved the yield and quality of cucumber.
2019 Vol. 34 (5): 153-162 [Abstract] ( 17 ) 全文: HTML   PDF (1063 KB)  ( 20 )
163 Effects of NaCl Stress on Growth and Physiological Characteristics of Sugar Beet at Different Seedling Stages
HUANG Chunyan, SU Wenbin, FAN Fuyi, GUO Xiaoxia, LI Zhi, JIAN Caiyuan, TIAN Lu, REN Xiaoyun, GONG Qianheng
DOI: 10.7668/hbnxb.20190029
In order to investigate the adaptability and physiological response of sugar beet to NaCl stress, the effects of NaCl stress on growth and physiological characteristics of sugar beet seedlings were studied by pot culture. The results showed that with the increase of NaCl concentration at the same seedling stage, seedling reservation rate began to decrease at 0.6% NaCl concentration, and plant height decreased gradually. The dry weight of sugar beet showed a single peak curve change. Low salt stress was beneficial to the increase of sugar beet biomass, and the chlorophyll content increased. The plasmalemma permeability and MDA content of leaves increased, and the cell membrane system was damaged seriously. SOD and POD activities increased, and antioxidant enzyme activities enhanced. With the advance of seedling age, the differences of seedling reservation rate, plant height, dry weight, chlorophyll content, plasmalemma permeability of leaves and MDA content gradually decreased among NaCl stress treatments, while the differences of SOD activity and POD activity gradually increased, and the salt tolerance of sugar beet increased. In conclusion, with the increase of NaCl concentration, SOD and POD activities, chlorophyll content and plasmalemma permeability and MDA content of leaves increased, but seedling reservation rate, plant height and dry weight decreased. With the advance of seedling age, the inhibition of salt on the growth of sugar beet weakened.
2019 Vol. 34 (5): 163-169 [Abstract] ( 23 ) 全文: HTML   PDF (1100 KB)  ( 18 )
170 Effects of Different Rootstocks on the Growth and Fruit Quality of Marselan Grapes
GAO Zhan, PENG Yuanyuan, DONG Kaixiang, ZHOU Long
DOI: 10.7668/hbnxb.201751612
In order to provide a theoretical basis for the selection of suitable scion-stock combinations of wine grape in Xinjiang Manas River Basin, we studied the growth, photosynthetic property and fruit quality of different combinations. 2-year-old rootstock scion combinations of Marselan/SO4 and Marselan/5BB were selected as the treatments, and Marselan seedling was used as control. The growth, photosynthesis, chlorophyll fluorescence characteristics and fruit quality of Marselan were measured. The inter-segment thickness, inter-segment length, SPAD value and the fruit setting of Marselan/5BB were significantly higher than those of Marselan. The net photosynthetic rate(Pn), stomatal conductance(Gs)and PSⅡ actual photochemical efficiency(ΦPSⅡ)of Marselan/5BB were the highest, while the intercellular CO2 concentration(Ci)and the non-photochemical quenching coefficient(NPQ)was the lowest. Matheran/SO4 had the largest panicle quality, peel thickness and seed quantity, while grape skins had the smallest total tannin content. The total sugar content, total flavonoid content and total phenol content of grape skin in Marselan/5BB were the largest, but the seed weight was the smallest. According to the principal component analysis, the comprehensive scores were from high to low:Marselan/5BB > Marselan/SO4 > Marselan self-root seedlings. It suggested that the resistance of 5BB was better than that of SO4. Marselan/5BB was a suitable combination in Xinjiang Manas River Basin, possessing the great growth potential, high photosynthetic capacity and good fruit quality.
2019 Vol. 34 (5): 170-176 [Abstract] ( 22 ) 全文: HTML   PDF (944 KB)  ( 20 )
       Resource and Environment·Plant Protection
177 Effects of Organic Fertilizers on Root Growth and Soil Nutrition of Grape
WANG Xiaolong, LIU Fengzhi, SHI Xiangbin, WANG Xiaodi, JI Xiaohao, WANG Zhiqiang, WANG Baoliang, ZHENG Xiaocui, WANG Haibo
DOI: 10.7668/hbnxb.201751420
In order to explore the application effects of different organic fertilizers in vineyards and to provide a reliable basis for scientific fertilization of grape, the effects of three organic fertilizers(bio-organic fertilizer, sheep manure and pig manure)on root growth and soil nutrient status of grape were studied in a three-year-old organic vineyard of Xingcheng, Liaoning. The results showed that the application of bio-organic fertilizer increased the total root length and root surface area. The bio-organic fertilizer and sheep manure treatments could increase the total nitrogen content of roots(P<0.05), and the former increased more. Bio-organic fertilizer treatment significantly increased the contents of most mineral elements, especially in the harvest period except B element, and the peaks of other tested elements all appeared in the bio-organic fertilizer treatment group. Fertilization treatment significantly improved the soil organic matter content in the color change period and harvest period of grape(P<0.05), with the order of bio-organic fertilizer > sheep manure > pig manure > control, which increased by 70.72%, 53.67% and 52.31% respectively in color changing period, and by 51.61%, 36.14% and 34.84%(P<0.05)respectively in harvest period, compared to the control. The application of organic fertilizer could increase the content of alkali nitrogen and total nitrogen of soil at color-changing and harvesting stages, with the order of bio-organic fertilizer > sheep manure > pig manure > control, but only the organic fertilizer treatment had significant difference with the control in different periods(P<0.05). The soil available phosphorus contents in bio-organic fertilizer and sheep manure treatments was significantly higher than that of the control in the color change period and the harvest period, but there was no significant difference between the pig manure and the control(P<0.05). Fertilization treatment could lead to the increase of the content of most mineral elements in soil to a certain degree, while the B and Mo contents of soil in the color change period was lower than that of the control. The reason might be that the fertilization treatment enhanced the absorption of trace elements of grape and the trace elements in organic fertilizers released slowly. Compared with the control, the three organic fertilizers could improve the growth of grape roots, increase the content of mineral elements in roots, and increase the content of organic matter, total nitrogen, available phosphorus and mineral elements in soil. The effect of bio-organic fertilizer application was the best.
2019 Vol. 34 (5): 177-184 [Abstract] ( 18 ) 全文: HTML   PDF (1048 KB)  ( 17 )
185 Effect of Long-term Fertilization on Phosphorus (P) Balance and Critical Value of Soil Olsen-P in Fluvo-aquic Soil of Shandong Province
SUN Cuiping, ZHANG Yingpeng, LUO Jiafa, ZHONG Ziwen, SUN Ming, LI Yan, LIU Zhaohui, JING Yongping, BO Luji
DOI: 10.7668/hbnxb.201751350
To reveal the effects of long-term fertilization on P balance and critical Olsen-P value of fluvo-aquic soils in Shandong Province, a split-plot experiment was designed, taking the winter wheat-summer maize rotation as the research object for 33 years. The main treatments included single inorganic fertilizer and organic/inorganic fertilizer combination. The results showed that the P deficit amount of soil was 23.27-29.20 kg/(ha · a) and 12.10-22.27 kg/(ha · a), respectively, for no P application and only manure application. The P surplus amount was 68.34-85.99 kg/(ha · a) and 88.84-107.64 kg/(ha · a), respectively, for single phosphate fertilizer treatment and organic/inorganic phosphate fertilizer combination. The positive response of Olsen-P to P balance was higher in organic/inorganic phosphate fertilizer combination than in single inorganic fertilization. With an average surplus of 100 kg/ha, the Olsen-P increased by 0.76,1.46 mg/kg, respectively, in the two main treatments. The critical Olsen-P value for wheat and maize yield was 12.60,8.90 mg/kg, respectively, for single inorganic fertilization, but there was no obvious correlation between crop yield and Olsen-P in organic/inorganic fertilizer treatment. Under the long-term tillage, the application of phosphate fertilizer and organic fertilizer was needed for no P fertilization and only manure treatment in order to increase the soil phosphorus, but for P fertilization treatment, the phosphate fertilizer amount should be reduced appropriately to regulate the P balance. The amount of phosphorus application could be determined reasonably based on the critical Olsen-P value for crop yield and effective Olsen-P conversion rate.
2019 Vol. 34 (5): 185-191 [Abstract] ( 23 ) 全文: HTML   PDF (1121 KB)  ( 13 )
192 Effects of Biochar and Humic Acid on Chemical Properties and Chromium Contents of Chromium-contaminated Soil in Potted Rape
GUO Ru, HONG Jianping
DOI: 10.7668/hbnxb.201751758
The surface soil from the sewage-irrigation area in Liujian village of Taiyuan city was taken as the research object to study the effects of different biochar and humic acid treatments on Cr-contaminated soil in potted rape. The experiment was arranged with three biochar treatments:corn straw biochar (MBC), rice husk biochar (RBC) and wheat straw biochar (WBC), each of which had three doses (0.3%, 0.6% and 0.9%), taking no biochar as the control (CK). The basic chemical properties, enzyme activity and total Cr content of soil were measured. The results showed that different biochar treatments significantly increased the contents of nutrients, and the activities of sucrase, urease and phosphatase in the soil contaminated with heavy metal chromium. With the increase of WSB/RSB/MSB amounts, the contents of nutrients, sucrase and urease increased significantly. Among them, organic matter, available potassium and sucrase changed significantly with the increase of application amount, while total nitrogen, alkali-hydrolyzed nitrogen, total phosphorus and total potassium had no significant difference between the treatments due to the small application gradient. Under different biochar treatments, the available phosphorus content and the activities of phosphatase and catalase in soil all showed the trends of RSB > MSB > WSB. The content of available phosphorus and the activity of phosphatase in soil increased first and then decreased with the increase of dosage. The soil catalase activity increased significantly only in the treatment of 0.3% RSB, but decreased significantly when the dosage increased to 0.9%. Under different biochar treatments, the fixation effect of chromium in soil was obvious, and the content of chromium in rape reduced significantly, but it had no significant difference between different kinds of biochar and between different treatments of doses. The applied amount of biochars was relatively low, and their gradient difference was small, so they could not reach to the minimum limit of adsorption fixation. The corn straw biochar had the best effect, so the further study should pay more attention to the application amount of biochar.
2019 Vol. 34 (5): 192-200 [Abstract] ( 23 ) 全文: HTML   PDF (1098 KB)  ( 14 )
201 The Effects of Intercropping Green Manure on Functional Diversity of Microbial Community in Tobacco Planting Soil
ZHAO Dongxue, WANG Panpan, CHANG Chunli, LI Jinqiu, WANG Jingying, AO Hong
DOI: 10.7668/hbnxb.201751398
The effects of green manure intercropping on the functional diversity of soil microbial communities in tobacco growing soils were studied by Biolog technique to provide the theoretical basis for application of green manure intercropping with tobacco.The average well color development (AWCD) of soil microbial carbon metabolic activity at the stages of rosette and maturity were intercropped with Melilotus officinali > Lolium > control, indicating that intercropping with green manure had a significant effect on soil microbial carbon utilization. The Shannon index,Simpson index and Mclntosh index of intercropping green manure were higher than those of control, while the soil microbial Shannon index, Simpson index and Mclntosh index of intercropping Melilotus officinalis had the highest value. The results showed that intercropping green manure affected the microbial community structure of tobacco growing soil and improved the microbial community diversity of tobacco growing soil. Metabolic intensity analysis of different carbon sources showed that the utilization of amino acid, carboxylic acid and carbohydrate by soil microorganisms in different green manure and different growth stages was significantly higher than that of ammonia/amine, miscellaneou and polymer carbon sources. Principal component analysis showed that the functional diversity of soil microbial community was significantly different, and carboxylic acids, amino acids and sugars were the three main carbon sources, which led to the difference in soil microbial metabolism. Intercropping green manure enhances the ability of soil microorganism to utilize carbon source, and improves the diversity of soil microbial community structure and community function in tobacco growing soils. The effect of intercropping Melilotus officinalis is more obvious.
2019 Vol. 34 (5): 201-207 [Abstract] ( 23 ) 全文: HTML   PDF (1012 KB)  ( 17 )
208 Spatial-temporal Dynamics of Physiological Races and Virulence Structure of Phytophthora sojae Population in Eastern Heilongjiang Province
JIA Mengzhen, ZHANG Bin, TIAN Miao, JIE Yan, ZHAO Yuqi, GAO Xinying, GU Xin, DING Junjie, CHEN Yufei, WEN Jingzhi
DOI: 10.7668/hbnxb.201751427
In order to study the spatial-temporal dynamics of physiological races and virulence structure of Phytophthora sojae, the soil samples were collected from four destructive fields in Eastern Heilongjiang Province for four years, and the P.sojae isolates were isolated by modified leaf disc baiting method. The isolates were acquired after morphological and DNA molecular identification, and their physiological races and virulence structure were identified using the universal hypocotyl inoculation method. The results showed that the 497 isolates included 135 pathotyps with 29 races, 1 IRT and 105 new pathotyps, of which races 16, 18, 22, 28 and 35 were first reported in China, while races 4, 33, 38, 39, 43, 48 and 49 were not detected. Race 13 had the highest isolation frequency of 3.2%, and race 1 was only 2.4%. It suggested that the virulence of P.sojae population in Eastern Heilongjiang was weak. The virulence frequencies to eight Rps genes varied from 27.8% to 53.1%, of which the virulence frequencies to Rps1k, Rps3a, Rps1c and Rps6 genes were less than 35%, but that to Rps7 gene reached to 53.1%, indicating that the soybean cultivars containing Rps1k, Rps3a, Rps1c or Rps6 were safer in Eastern Heilongjiang and that containing Rps7 was more dangerous. The races and pathotypes of P.sojae in Eastern Heilongjiang differentiated rapidly with time, their virulence and population pathogenicity had enhanced, and the virulence frequencies to majority of Rps genes increased. The number of races and new pathotypes had no significant difference between the production field and the experimental field, but their types were quite different and the virulence of single isolate and population in experimental fields was stronger.
2019 Vol. 34 (5): 208-215 [Abstract] ( 43 ) 全文: HTML   PDF (961 KB)  ( 12 )
216 Isolation and Identification of Maize ear rot Pathogens and Phylogenetic Analysis of Dominant Species in Shandong Province
DING Mengjun, YANG Yang, SUN Hua, MA Hongxia, LIU Shusen, SHI Jie
DOI: 10.7668/hbnxb.201751508
The objective of this study was to determine the composition of pathogen causing Maize ear rot in Shandong Province and to analyze the epidemic trend of dominant pathogen. A total of 138 Maize ear rot samples were collected from Shandong Province during 2015-2017 and were used for pathogen isolation.The isolates were identified by morphological and molecular biology methods, and the occurrence regularity of dominant pathogen was clarified by phylogenetic analysis. The results of isolation and identification showed that a total of 18 fungi were obtained. The isolation frequency of Fusarium verticillioides was 67.39%, which was the dominant pathogen in Shandong Province, followed by F. proliferatum, F.graminearum, F.equiseti and F.oxysporium with separation frequencies of 10.87%, 7.97%, 5.80% and 0.72%, respectively; And the separation frequencies of Trichoderma harzianum, Penicillium oxalicum, Aspergillus flavus and A. niger were 7.91%, 6.52%, 2.17% and 5.07%, respectively. Based on the EF-1α gene sequence, the phylogenetic tree of F. verticillioides isolates in Shandong Province from 2015 to 2017 was constructed, and the genetic evolution relationship of isolates between Shandong Province and Hebei Province was analyzed. Phylogenetic analysis showed that 89 isolates of F. verticillioides could be divided into two large groups with a small genetic distance of 0.002-0.016, indicating that the genetic relationship between these isolates was relatively close. The genetic distance of F. verticillioides isolates in Shandong Province and Hebei Province was small and showed a close genetic relationship, indicating that there were frequent gene exchanges. The results of this study would provide a scientific basis for resistance breeding and integrated control of F. verticillioides.
2019 Vol. 34 (5): 216-223 [Abstract] ( 24 ) 全文: HTML   PDF (1077 KB)  ( 19 )
224 Prokaryotic Expression of Affinity Peptides to Membrane Protein of Transmissible gastroenteritis virus and Analysis of Viral Affinity Activity
YU Tianfei, DONG Huiying, XIE Pengyu, SUN Wanshu, YIN Haichang, LI Ming, YU Zhidan
DOI: 10.7668/hbnxb.201751554
In order to investigate the viral affinity of Transmissible gastroenteritis virus of swine (TGEV) membrane protein (M protein) affinity peptide and establish a rapid and effective TGEV detection method, a tandem gene containing the TGEV M protein affinity peptide was designed. The nucleotide sequence was optimized according to the E. coli preference codon. A recombinant plasmid pUC57-MQHT containing M protein affinity peptide gene was obtained by artificial gene synthesis. According to the sequence of the synthetic gene, a specific primer was designed and synthesized. And then, using pUC57-MQHT as a template, TGEV M protein affinity peptide gene (about 150 bp) was obtained by PCR. The affinity peptide gene was respectively inserted into the Bam H Ⅰ/Xho Ⅰ multiple cloning sites of the prokaryotic expression vectors pET-32a and pGEX-6p-1 to obtain the recombinant plasmids pET-32a-MQHT and pGEX-6p-MQHT. The recombinant plasmids were identified by single enzyme digestion of Bam HⅠ, double enzyme digestion of Bam HⅠ/Xho Ⅰ and sequencing. The recombinant plasmids were transformed into E. coli Rosetta (DE3) and induced by IPTG to obtain the expression of recombinant proteins TRX-MQHT and GST-MQHT, respectively. The molecular weights of the two recombinant proteins were 25, 31 ku, respectively. After purification, the recombinant proteins TRX-MQHT and GST-MQHT were identified by His-tag monoclonal antibody and GST-tag monoclonal antibody, respectively. The results indicated that the purified proteins obtained were the target proteins. Western Blot analysis showed that the two recombinant proteins could specifically bind to TGEV. Dot-ELISA analysis indicated that the two recombinant proteins showed positive reaction to TGEV at 1×103, 5×102 and 2.5×102 TCID50/mL, but no visible spot was found at 1.25×102 TCID50/mL. The results showed that the two recombinant proteins had good affinity to TGEV virions. Dot-ELISA analysis showed that the minimum binding titer of the two recombinant proteins to TGEV virions was 2.5×102 TCID50/mL. Blocking experiments showed that TGEV-positive serum diluted 1:100 or 1:200 could completely blocked the binding of the two recombinant proteins to TGEV H strain (1×105 TCID50/mL). The results showed that the specificity of the two recombinant proteins was good. This study laid a certain theoretical and material basis for the establishment of TGEV diagnostic methods.
2019 Vol. 34 (5): 224-230 [Abstract] ( 20 ) 全文: HTML   PDF (1147 KB)  ( 26 )
231 Cloning and Expression of Goat CPT1A Gene and Its Correlation with Intramuscular Fat Content
LIANG Jijun, LIN Yaqiu, YU Yuyang, WANG Yong, ZHU Jiangjiang
DOI: 10.7668/hbnxb.201751529
The aim of this study was to obtain the gene sequence of goat carnitine palmitoyl transferase I liver subtype CPT1A (Carnitine palmitoyltransferase 1A, CPT1A), analyze its biological characteristics, and elucidate its tissue expression and cell differentiation temporal expression. Besides, the relationships between the expression of CPT1A gene and intramuscular fat content (IMF) in the longissimus dorsi, biceps femoris and triceps were revealed. Seven one-year-old healthy Jianzhou Big-Eared goats were used as the test animal. After slaughter on an empty stomach, the heart, liver, spleen, lung, kidney, longissimus dorsi, biceps femoris, triceps, subcutaneous fat and abdominal fat tissue were collected rapidly. Total RNA was extracted using the TRIzol method. The goat CPT1A gene was cloned by RT-PCR for biological analysis. Real-time PCR was used to detect the expression level of CPT1A gene in different tissues and the temporal expression of CPT1A in the differentiation of goat preadipocytes. The correlation between CPT1A gene expression and IMF content was analyzed by Pearson correlation coefficient. The cloned C PT1A sequence (MH345735) was 2 380 bp, containing the full-length CDS 2 319 bp, 5'UTR 38 bp and 3'UTR 23 bp, encoding 773 amino acids. The results of tissue expression showed that the expression level of CPT1A in the liver and kidney of Jianzhou Big-Eared goat was significantly higher than in other tissues (P<0.01). Quantitative results of cell temporal expression showed that the expression level of CPT1A gradually increased in 0-5 d, reaching to the highest at 5 d (P<0.01), and then decreased gradually in 5-9 d. Correlation analysis between CPT1A gene expression and IMF content showed that CPT1A gene expression was significantly positively correlated with intramuscular fat content in each muscle tissue. These results indicated that CPT1A might play an important regulatory role in intramuscular fat deposition, and facilitate further study on the mechanism of CPT1A regulation in goat lipid metabolism.
2019 Vol. 34 (5): 231-238 [Abstract] ( 24 ) 全文: HTML   PDF (1379 KB)  ( 15 )
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