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2018 Vol. 33, No. 5
Published: 2018-10-28

 
0 Table of Contents for VOL.33 No.5
2018 Vol. 33 (5): 0-0 [Abstract] ( 33 ) HTML (1 KB)  PDF (244 KB)  ( 35 )
1 Quantitative Trait Loci Analysis of Flag Leaf Size and Grain Relative Traits in Winter Wheat
LU Lahu, YANG Bin, ZHANG Ting, ZHANG Wei, YUAN Kai, SHI Xiaofang, PENG Huiru, NI Zhongfu, SUN Qixin
DOI: 10.7668/hbnxb.2018.05.001
To identify quantitative trait loci (QTL) for flag leaf length (FLL),width (FLW),area (FLA),thousand grain weight (TGW),grain number per spike (GNS),grain weight per spike(GWS) and understand the genetic relationship among those traits,we developed a doubled haploid(DH)population with 216 lines, derived from the cross between Nongda 3338 and Jingdong 6. The field experiments were carried out in Linfen and Yuncheng in 2015 and 2016, and QTL analysis was conducted by composite interval mapping method combining with genetic maps containing 469 markers. The results showed that a total of 105 additive QTLs were detected,forty of them were QTLs for flag leaf traits and sixty-five were for grain traits,with single QTL explaining 2.28%-39.91% of the phenotypic variances. Several QTLs were identified in the same region or adjacent regions on the 1B,2A,2B,2D,3A,4A,4B,4D,6A,6D and 7A chromosome, these QTLs were closely linked to each other or multi-effect, confirmed those phenotypic data were closely-related under QTL level. Seven QTLs (Qflw-4A.1, Qflw-4B.1, Qflw-4D.1, Qfla-4B, Qtgw-4A.1, Qtgw-4B.2, Qtgw-4B.3) had high LOD value were detected repeatedly under three or four environments. Those seven stable expression QTLs might have great significance for MAS breeding and gene cloning program.
2018 Vol. 33 (5): 1-8 [Abstract] ( 114 ) HTML (1 KB)  PDF (954 KB)  ( 71 )
9 QTL Analysis of Root Number and Root Length in Rice Seedling Stage under Salt and Alkali Stress
SUO Yining, ZHANG Chunke, YU Qiaoqiao, ZHANG Enyuan, XIE Dongwei, LENG Yue, WANG Liang, SUN Jian
DOI: 10.7668/hbnxb.2018.05.002
The number and the length of roots as well as its QTL would provide theoretical basis for genetic mechanisms and molecular marker assisted breeding of salt and alkaline tolerance. Recombinant inbred lines(RIL)of sensitive varieties Dongnong 425 tolerant varieties Changbai 10 cross were treated with 140 mmol/L NaCl and 0.15% Na2CO3 as salt and alkali stress,and normal condition as control. The number and the length of roots during seedling stage were measured,and using complete interval mapping(ICIM)of QTL IciMapping v3.3 software to analysis the QTL under salt,alkali and normal conditions. We detected eighteen additive QTL located on 1,2,3,4,5,7,8,9 and 10 chromosomes with LOD 2.01-3.35,and the contribution rate to phenotypic variation was 6.02%-20.06%.Under natural conditions,four QTL related to the root number were detected,among which,12.46% was the largest contribution rate of qNRN7-2,while no QTL were detected related to the root length. Under salt stress,five QTL were detected related to the number and length of roots,among which,20.06% was the largest contribution rate of qSRN3. Under alkali stress,three QTL related to the root number and root length were detected. The contribution rate of qARN2 was 12.99%,and the contributions of qARL3 and qARL5 were 7.04% and 8.88% respectively. We found four different number and length related QTL between normal and salt condition. Among them,the contribution rates of qN-SRN8-2 and qN-SRL1 were relatively large,14.01% and 14.12%,respectively.In normal and alkali conditions,two number and length related QTL were detected on 3 and 10 chromosomes. One QTL associated with root number, qN-ARN3,located on chromosome 3,with contribution rate 6.02%.One QTL were detected related to the number and length of roots,among which,7.45% was the largest contribution rate of qN-ARL10 on chromosome 10. Under alkali and salt stress conditions,the number and length of rice seedling roots were significantly affected,and compared with salt stress,rice was more sensitive to alkali stress.
2018 Vol. 33 (5): 9-15 [Abstract] ( 73 ) HTML (1 KB)  PDF (1156 KB)  ( 58 )
16 Cloning,Prokaryotic Expression and Ployclonal Antibody Preparation of Swine Viperin Gene
FANG Jianyu, ZHU Wenhao, LI Haili, GUO Xiaocan, BAI Xianxiao, WANG Keling
DOI: 10.7668/hbnxb.2018.05.003
To study the antiviralactivity of swine viperin in pigs,we extracted and cloned the swine Viperin genes from PK-15 cells. PK-15 cells were seeded onto a 24-well plate and treaded with IFN-α. After 24 h,the cells were harvested to extract the total RNA,and to amplify the Viperin gene by RT-PCR. Then the gene was cloned into the pEASY-blunt simple plasmid for sequencing. The antigenicity and hydrophobicity of swine Viperin were analyzed by DNA Star software,and the fragment with good immunogenicity were chosen and amplified,which was inserted into the prokaryotic expression vector pET-28a+. E.coli BL21 containing recombinant plasmid pET-sVIP was induced by IPTG. SDS-PAGE analysis demonstrated that the recombinant protein was expressed in the form of inclusion bodies in E.coli. Then inclusion body protein was purified. The BALB/c mice (9 day old) were immunized two times using the purified recombinants Viperin protein emulsified with equal amounts of Freund's complete adjuvant. Two weeks after final immunization,the serums were collected and the antibody specificity was identified by Western Blot and IFA assay. The result showed that the swine viperin polyclonal antibody could react with the sViperin protein expressed by eukaryotic expression vector pVAX-sVIP. Preparation of polyclonal antibody laid the foundation for further studying the antiviral activity of swine Viperin.
2018 Vol. 33 (5): 16-22 [Abstract] ( 64 ) HTML (1 KB)  PDF (1229 KB)  ( 43 )
23 Assembly and Analysis on Mitochondrial Genome Sequence in Inner Mongolia Cashmere Goat(Alashan Type)
SU Rui, YANG Zhong, LI Chun, LI Xiaokai, MA Yuhao, LI Jinquan
DOI: 10.7668/hbnxb.2018.05.004
In order to study the Inner Mongolia Cashmere goat mitochondrial genome sequence composition and the phylogenetic relationship of different varieties of goats,ensure the origin,evolution,differentiation of different groups and the formation mechanism of specific genetic characteristics of cashmere goats,we chose Inner Mongolia Cashmere goat (Alashan type) as object and sequenced the mitochondrial genome. The results showed that totally 2 551 175 100 bp base was got and Q20 was 95.40%. The construction quality of the library was good,the accuracy of the sequencing results was high,and the complete closed loop mitochondrial genome of 16 642 bp was obtained. The content of GC was 39.17% and K-mer was 53 bp. The mitochondrial genome sequence information of other varieties was downloaded from NCBI to carry out phylogenetic analysis. The results showed that the distance between 3 groups of Inner Mongolia Cashmere goats was close. The relationship between Inner Mongolia Cashmere goat (Alashan type) and Inner Mongolia Cashmere goat (Erlangshan type) was the closest,and the next one was Inner Mongolia Cashmere goat (Aerbasi type). Inner Mongolia Cashmere goat and Liaoning Cashmere goat could be divided into two groups. The outer range of San Clemente was the farthest.
2018 Vol. 33 (5): 23-28 [Abstract] ( 30 ) HTML (1 KB)  PDF (1102 KB)  ( 23 )
29 Analysis of OsRSR1 Sequence and Character Variation in Japonica Transgressive Variants and Varieties
WANG Haiwei, HAN Yunfei, ZHU Lin, QU Yue, HE Shenyu, ZHANG Zhongchen, JIN Zhengxun
DOI: 10.7668/hbnxb.2018.05.005
In order to compare and analyze the relationship between OsRSR1 expression,sequence and functional variation and trait genetic variation. Indica rice varieties with high amylose content(AC),glutinous rice varieties with very low AC,Japonica parents and hybrid progenies with similar genetic background and significantly different amylose content were used in this study. Cloning,expression assay and bioinformatics analysis of transcription factor gene OsRSR1 were carried out. The results showed that OsRSR1 expression level was high at the initial and late stage of grain filling,but low in the middle stage,like a V-shape. OsRSR1 expression level was high in varieties with low AC,vice versa in varieties with high AC. It indicated the negative relationship between OsRSR1 expression and grain AC,and transgressive variation in hybrid progenies. DNA sequence,transcript and protein sequence of OsRSR1 were not identical. DNA base changes were detected in exons and introns of sexual hybrid progenies,and single base substitution(SBS)occurred in transcript to produce code variation,this was an important approach to genetic variation and gene polymorphism. OsRSR1 is composed of eight exons and seven introns. The changes of SBS and amino acid did not alter OsRSR1 gene structure and relative motif as well as function,which implied AP2 domains was highly conserved. The results would be helpful for further study on the molecular regulation of gene transcriptional level and genetic variation mechanism.
2018 Vol. 33 (5): 29-39 [Abstract] ( 32 ) HTML (1 KB)  PDF (1990 KB)  ( 30 )
40 Transcriptomic Analysis of Rice Seedling Responsive to Low Temperature
ZHU Lin, YUAN Meng, GAO Hongxiu, QU Yue, HE Shenyu, WANG Jian, WANG Shan, WANG Siyu, YANG Ling, ZHANG Zhongchen, JIN Zhengxun
DOI: 10.7668/hbnxb.2018.05.006
Temperature is very important for growth and development of rice and yield production,and it is necessary to further define the molecular mechanism of rice response to low temperature provides a theoretical basis for crop breeding resistant to cold stress. In this study,japonica rice variety Zhonghua-11 was used as a test material under the condition of low temperature treatment at 17℃,a new generation of high-throughput sequencing methods-transcriptome sequencing (RNA-Seq) analysis. Through the transcriptome analysis,the clean high-throughput reads were 46 348 074 and 52 483 052,respectively,and 2 044 differentially expressed genes were identified and screened,of which 1 252 genes were up-regulated and 792 genes were down-regulated. GO annotation analysis showed that all the differential genes were divided into three major categories:biological processes,cellular components and molecular functions,each accounting for 63.29%,8.81%,and 27.90%. We used the KEGG database to analyze the number and location of differentially expressed genes in low-temperature-regulated two-pathway photosynthetic pathway and phenylalanine metabolic pathway,and predicted the function of novel genes and interaction proteins related to photosynthesis and phenylalanine metabolism. The results showed that the low temperature stress on rice plays an important role in biological processes,especially on photosynthesis,secondary metabolism biosynthesis,and phenylalanine metabolism. Also,differentially expressed WRKY transcription factors will provide direction and basis for further study of cold tolerance mechanism.
2018 Vol. 33 (5): 40-51 [Abstract] ( 38 ) HTML (1 KB)  PDF (1293 KB)  ( 46 )
52 Comparative Study on Differential Characters and Proteins Spot Identification of Resistance to Sugar Beet Rhizomania
ZHANG Hui, BAI Chen, WANG Huazhong, ZHANG Huizhong, LI Xiaodong, FU Zengjuan, ZHAO Shangmin, E Yuanyuan, ZHANG Ziqiang, WANG Liang, ZHANG Bizhou
DOI: 10.7668/hbnxb.2018.05.007
Sugar beet resistant and susceptible materials were used to compare the yield and quality in the field. Three different methods of protein extraction were compared,which included TCA/acetone precipitation method,soluble protein extraction method and phenol extraction method. After the treatment of viral stress by water culture,the different protein spots were analyzed by Image Master 2D Platinum Trial 5.0 software. The results showed that TCA/acetone precipitation method had better extraction effect. The comparison of field yield and quality showed that the growth of resistant materials were better than that of susceptible materials at seedling stage and late growth stage,and the yield and sugar content after harvest were higher than those of susceptible materials,thus the resistant materials performed well. The protein was analyzed by Image Master 2D Platinum Trial 5.0 software after two-dimensional electrophoresis,and 78 protein spots with obvious difference were deducted. A total of 34 active protein IDs were identified. Compared by NCBI and The Beta vulgaris Resource database Blast,the main functions of proteins expressed by disease 1 and disease 2 were photosynthesis,glycometabolism,respiratory antioxidation,signal transduction,lipid metabolism and some others. While disease 5 and disease 6 mainly expressed photosynthesis protein and respiratory antioxidant protein which were the proteins expressed by self stress reaction.
2018 Vol. 33 (5): 52-59 [Abstract] ( 25 ) HTML (1 KB)  PDF (1379 KB)  ( 17 )
60 Reference Genes Selection for Quantitative Real-time PCR in Chinese Cabbage-Cabbage Translocation Lines
CUI Feifei, MENG Chuan, WANG Yanhua, ZHAO Jianjun, CHEN Xueping, SHEN Shuxing, GU Aixia
DOI: 10.7668/hbnxb.2018.05.008
In order to analyze the gene expression more accurately in Chinese cabbage by quantitative Real-time PCR,Chinese cabbage translocation lines added No.4 chromosome fragments from cabbage under Chinese cabbage background were used as materials in this experiment. The leaves of Chinese cabbage-cabbage translocation lines in different growth stages,including seedling stage,rosette stage,folding stage and heading stage,floral buds of different sizes and leaves of folding stage treated with auxin(Indole-3-acetic acid,IAA)and auxin inhibitor(2,3,5-triiodobenzoic acid,TIBA)were selected to evaluate the expressions stability of genes. Expression stability of twenty one candidate reference genes were detected by qRT-PCR,including Apr, BcTIP41, U34559, EF1α, TUB4, CYP, DNAJ, HIS, TUA5, UKN1, SKIP16, CAC, ACTIN, ACTIN-1, ACTIN-2, GAPDH, UBC30, UBQ, PPR, PP2A, MDH. geNorm and NormFinder were used to analyze above these results,which showed that the qRT-PCR most suitable reference genes of Chinese cabbage-cabbage translocation lines had some differences in different development periods,different tissue of the material,and under condition of different hormone treatment. The UKN1 and TUB4 genes were most stable expression in vegetative growth stage(from seedling to heading)of Chinese cabbage-cabbage translocation lines. In folding stage of Chinese cabbage-cabbage translocation lines,the most stable expression reference genes were BcTIP41 and ACTIN after treatment with auxin IAA,and the most stable expression reference genes were UKN1 and BcTIP41 after treatment with auxin inhibitor TIBA. DNAJ, ACTIN and PP2A genes showed the most stable expression in the floral buds with six size levels of Chinese cabbage-cabbage translocation lines. These results will lay the foundation for the accurate analysis of the gene expression of Chinese cabbage-cabbage translocation lines,and further provided a reference for the selection of the reference gene in other plants of Brassica at different developmental stages and hormone treatments.
2018 Vol. 33 (5): 60-67 [Abstract] ( 22 ) HTML (1 KB)  PDF (1045 KB)  ( 24 )
68 Molecular Cloning, Temporal and Spatial Expression Patterns, and Subcellular Localization of Porcine TAK1 Gene
LI Rui, XU Haixia, WU Wei, WANG Suying, REN Mengke, ZHANG Pengpeng, XU Yongjie
DOI: 10.7668/hbnxb.2018.05.009
To investigate the biological role of transforming growth factor-β-activated kinase 1 (TAK1) in porcine skeletal muscle development, we cloned the porcine TAK1 cDNA sequences using rapid amplification of cDNA ends (RACE) method. We also detected the temporal and spatial expression patterns and subcellular localization of porcine TAK1 gene. The full-length cDNA sequence of TAK1 was 2 163 bp (GenBank No. KU504629), including a 1 740 bp open reading frame (ORF) that encoded a putative 579 amino acid, and the high sequence similarity of TAK1 between the pig and human, or monkey and or sheep were 98.8%. TAK1 contained a conserved serine/threonine protein kinase catalytic domain (STKc_TAK1), an activation loop (A-Loop), some ATP banding sites and TAB1(TGF-beta activated kinase 1)banding sites. The highly conserved domains indicated that TAK1 had a similar function. The tissue analysis showed that the TAK1 mRNA expression was the highest in pig spleen tissue and very low in pancreas and longissimus dorsi muscle tissue. The result of different developmental stages expression analysis indicated that TAK1 mRNA had the highest expression in embryonic and down-regulation during longissimus dorsi muscle development. Comparing the different breeds, the mRNA abundance of TAK1 in Large White was higher than in Meishan pigs at all development stages, whereas at 90 days, there was no significant difference (P >0.05). Moreover, the fluorescence co-localization showed that the expression of TAK1 was observed mainly in the cytoplasm. These results suggested that TAK1 was probably playing a key role in porcine skeletal muscle development, and made a foundation for the further study on the function of TAK1 in pigs.
2018 Vol. 33 (5): 68-75 [Abstract] ( 29 ) HTML (1 KB)  PDF (1292 KB)  ( 35 )
76 Transgenic of Brassica napus with Herbicide-resistant bar Gene and Insect-resistant cry1ab/ac Mediated by Agrobacterium tumefaciens
CHEN Zhangbin, ZHANG Zhenqian, CHEN Hao, LIU Zhongsong, XIONG Xinghua, WU Xianmeng, GUAN Mei, CHEN Sheyuan, GUAN Chunyun, XIAO Gang
DOI: 10.7668/hbnxb.2018.05.010
In order to obtain a new germplasm material of Brassica napus with both insect-resistance and herbicide-resistance properties,the hypocotyls of Xiangyou 15 were used as explants.The synthetic insect-resistant cry1ab/ac fusion gene was inserted into the expression vector pFGC5941 to form pFGC-Bt. pFGC-Bt was transformed into Brassica napus cultivar Xiangyou 15 by Agrobacterium tumefaciens infection of hypocotyls.In order to obtain novel materials of Brassica napus with insect-resistance and herbicide-resistance,the synthetic insect-resistant cry1ab/ac fusion gene was inserted into the expression vector pFGC5941 to form pFGC-Bt. pFGC-Bt was transformed into Brassica napus cultivar Xiangyou 15 by Agrobacterium tumefaciens infection of hypocotyls. After the callus differentiation and plant regeneration,seven T0 transgenic plants were obtained. The plants were harvested individually,and the seeds were then planted in pots,when seedlings were 4-leaf-stage,herbicide basta was applied. The surviving plants were then detected by PCR with bar gene primers. Eleven positive plants were selected for further test of Cry1Ab/Ac protein double antibody sandwich immunochromatographic assay. Cry1Ab/Ac protein was detected in all the eleven transgenic plants.The entomocidal activity was evaluated by insect feeding bioassays performed with larvae of Pieris rapae. The transgenic plants and non-transgenic control leaves were used in the insect bioassay. The Pieris rapae larvae that fed on transgenic plants were observed to stop growing and died after 7 days of feeding,whereas larvae fed on control plants grew normally.After 3 days, the leaves were basically eaten clean. The results of feeding identification indicated that the transgenic insect fusion gene cry1ab/ac was highly resistant to Pieris rapae. The results identified the transgenic plants had double resistance to herbicide basta and insect.
2018 Vol. 33 (5): 76-81 [Abstract] ( 18 ) HTML (1 KB)  PDF (1370 KB)  ( 24 )
82 Cloning and Expression Analysis of GhTGA1 and GhTGA9.2 in Cotton
ZHAO Zengqiang, LI Xiaoling, ZHANG Xi, ZHANG Wei, LIAN Wenming
DOI: 10.7668/hbnxb.2018.05.011
GhTGA1 and GhTGA9.2 of transcription factor genes were isolated by RT-PCR technology from Zhongmiansuo 12 cultivar of resistance to Fusarium oxysporum. Sequence analysis showed that the ORF of GhTGA1 and GhTGA9.2 were 1 281 bp and 1 461 bp,respectively.They encoded 405 and 486 amino acids,respectively. GhTGA1 and GhTGA9.2 all contained two conserved bZIP and DOG1 domains. They belonged to the TGA transcription factor gene of bZIP subfamily. The GhTGA1 and GhTGA9.2 expression feature were detected by Real-time quantitative PCR(qRT-PCR),the results showed that GhTGA1 gene was up-regulation expressed with the duration of the treatment of Fov,ethylene (ET) and jasmonic acid (JA). In each treatment time point,the expression of GhTGA1 gene was lower than that of Mock with the treatment of salicylic acid (SA). We speculated that GhTGA1 gene might be involved in ethylene (ET) and jasmonic acid (JA) signaling pathway and regulated resistance to Fusarium oxysporum in cotton; After the treatment of Fusarium oxysporum,the expression of GhTGA9.2 gene was lower than Mock at different treatment time points,and was down-regulated expression. In other hand,it was up-regulation expressed with the duration of the treatment of hormone (ET,JA,SA).It suggested that GhTGA9.2 might participate in stress response through hormone (ET,JA and SA) pathways. The analysis of gene expression characteristics of GhTGA1 and GhTGA9.2 provides a basis for further study.
2018 Vol. 33 (5): 82-88 [Abstract] ( 34 ) HTML (1 KB)  PDF (1237 KB)  ( 40 )
89 Analysis of the Relationship Between BcKMO Gene and MAPK Signaling Pathway in Botrytis cinerea
SU Youke, YUAN Xuemei, WANG Min, ZANG Jinping, CAO Hongzhe, ZHANG Kang, DONG Jingao, ZHANG Jing, XING Jihong
DOI: 10.7668/hbnxb.2018.05.012
To analyzed the relationship between BcKMO gene and MAPK signaling pathway in Botrytis cinerea,the specific inhibitor of MAPK signaling pathways U0126 was used to detect the sensitivity of the BcKMO gene mutants. It was found that the sensitivity of the BCG183 mutant to U0126 was remarkably weak compared to that of the BC22 and BCG183 /BcKMO. Expression patterns of BcKMO and MAPK signaling pathway key genes bmp1 and bmp3 were detected. The expression levels of BcKMO and bmp3 were stronger in 7 d mycelia and sclerotia of BC22. The expression levels of BcKMO, bmp1,and bmp3 were stronger in BC22 cultured on medium with sucrose or fructose. Real-time PCR technology was used to detect the expression levels of bmp1 and bmp3 in the BcKMO gene mutants,and the BcKMO expression levels in RNAi mutant of bmp1 and bmp3. The expression level of bmp1 was significantly upregulated in the BCG183 mutant,the expression level of bmp3 was significantly downregulated in the BCG183 mutant. The expression level of BcKMO in the RNAi mutants of bmp1 gene was obviously higher than that of BC22; the expression level BcKMO gene in the RNAi mutants of bmp3 was obviously lower than that of BC22. These results indicated that BcKMO negatively regulated the expression of bmp1 and positively regulated the bmp3 expression, bmp1 negatively regulated the BcKMO gene expression,and bmp3 positively regulated the BcKMO gene expression. Therefore,the results provide a foundation for clarifying the molecular mechanism of the BcKMO gene in growth,development and pathogenicity in Botrytis cinerea.
2018 Vol. 33 (5): 89-94 [Abstract] ( 28 ) HTML (1 KB)  PDF (1075 KB)  ( 23 )
95 Cloning and Analyzing the Function of the Promoter of an Effector Homologous of Chorismate Mutase in Sclerotinia sclerotiorum
CHANG Xue, SHENG Yinsheng, REN Aizhi, ZHAO Peibao
DOI: 10.7668/hbnxb.2018.05.013
In order to investigate the mechanism of the effector homologous of chorismate mutase in Sclerotinia sclerotiorum,the promoter was cloned and analyzed. The promoter element and the Cis element were analyzed through the online software,Promoter 2.0 and Promoter scan,then the DNA sequence of the promoter was cloned by the method of PCR and the GFP fusion vector were constructed,transformed in the protoplast of Sclerotinia sclerotiorum. In the end,the GFP fluorescence were detected through confocal microscope. As a result,TATA box and CAAT box were found in the upstream of the gene,and a 733 bp DNA sequence upstream of ATG was cloned by PCR using the primer XS1-1 and XS1-2,then the GFP fusion vector were constructed based on the plasmid of pBluntNAT-GFP,through transforming the protoplast of Sclerotinia sclerotiorum,some transformants were obtained in which the GFP fluorescence were detected through confocal microscope. All these indicated that the 733 bp DNA sequence upstream of ATG could act as the promoter of the gene.
2018 Vol. 33 (5): 95-98 [Abstract] ( 12 ) HTML (1 KB)  PDF (1091 KB)  ( 19 )
99 Cloning and Expression Analysis of Potassium Channel NtSKOR Gene in Nicotiana tabacum
ZHUO Wei, CHEN Qian, YANG Shangyu, LI Jiahao, PENG Shuang, WANG Jing, LI Liqin
DOI: 10.7668/hbnxb.2018.05.014
Potassium channel is the main protein of plant uptake and translocation K+,and SKOR (Stelar K+ outwaed rectifier) belongs to Shaker channel family exogenous rectifier,which plays a key role in response to low potassium stress response in plants.In order to study the function and role of tobacco NtSKOR gene in abiotic stress response,one NtSKOR gene was cloned from Nicotiana tabacum K326 with homologous cloning strategy,gene expression patterns were analyzed by Real-time quantitative PCR,and their physicochemical properties,domains,phosphorylation sites and evolutionary relationships of proteins were predicted by bioinformatic analysis. Bioinformatics analysis showed that this gene contained 2 484 bp and encoded 827 amino acid. The predicted molecular weight was 94.75 ku and the isoelectric point (pI) was 6.52. The largest secondary structure element of this protein was α-helix,and the smallest was β-turn. NtSKOR contained six transmembrane domains(S1-S6),with three different kinase phosphorylation sites for serine,threonine and tyrosine. Phylogenetic analysis showed that the protein had 99% homology with Nicotiana sylvestris and 96% homology with Nicotiana tompentosiformis,thus it was named as NtSKOR. Expression patterns showed that the gene was expressed in roots,stems,leaves and flowers in mature stage,which had the highest expression in roots and the lowest expression level in flowers. Expression patterns under abiotic stress indicated the gene could respond rapidly to the adverse conditions of low potassium,high salt,drought,H2O2,ABA and 4℃. This suggests that NtSKOR gene plays an important regulatory role in tobacco abiotic stress,and the results provide a theoretical basis for further in-depth study of the NtSKOR function.
2018 Vol. 33 (5): 99-105 [Abstract] ( 24 ) HTML (1 KB)  PDF (1262 KB)  ( 25 )
106 Prokaryotic Expression and Purification of Hexon Protein of Fowl Adenovirus Serotype 4
LIU Na, LIU Qingtao, LI Yin, YANG Jing, LI Xiangrui
DOI: 10.7668/hbnxb.2018.05.015
To obtain the major structural protein hexon protein of serum type 4 avian adenovirus by prokaryotic expression; A pair of specific primers was designed according to the gene sequence of the hexon protein of Fowl adenovirus serotype 4 (FAdV-4) in GenBank. The full-length sequence of hexon gene was amplified by ordinary PCR. The serotype 4 avian adenovirus gene fragment obtained by PCR amplification was inserted into the vector pMD18T after inserting restriction enzyme sites at both ends of the gene. The PCR analysis and sequencing analysis showed that the gene was verified with the prokaryotic expression vector pET-32a. The constructed expression vector was transformed into BL21(DE3) and induced to express with 1 mmol/L IPTG. Analyzing the recombinant protein by SDS-PAGE and Western Blot; By the double-enzyme digestion and sequence analysis,the prokaryotic expression vector of pET-32a-hexon was successfully constructed. The size of the inserted hexon protein gene fragment was 2 814 bp. The recombinant protein expression was highest at 3 h after induction with 1 mmol/L IPTG,and the recombinant protein was mainly inclusion body forms exist in sediments. The SDS-PAGE analysis showed that the fusion protein had a molecular weight of 118 ku. Western Blot analysis showed that the fusion protein could be specifically recognized by anti-FAdV-4 serum and HIS tag antibody. This experiment successfully expressed the major structural protein hexon protein of serum type 4 avian adenovirus,and the fusion protein obtained had good reactogenicity,which could be used in the further research of serum type 4 avian adenovirus. It laid a good foundation for the prevention and treatment of serum type 4 fowl adenovirus disease.
2018 Vol. 33 (5): 106-110 [Abstract] ( 22 ) HTML (1 KB)  PDF (1029 KB)  ( 20 )
111 Prokaryotic Expression of PPRV Nigeria75/1 H Protein and Prediction of Antigen Epitope
LI Linjie, CHANG Qiuyan, MA Peng, WANG Yueying, MA Xiaoxia, BAI Jialin
DOI: 10.7668/hbnxb.2018.05.016
To clone the gene encoding hemagglutinin(H)protein of Nigeria 75/1 strain of attenuated Peste des petits ruminants virus(PPRV),express in prokaryotic cells and prepare its polyclonal antibody by bioinformatics. According to the encoding sequence of PPRV H gene(GenBank X74443),a pair of primers were designed,with which the overall length were amplified by RT-PCR and inserted into vector pET-32a. The constructed recombinant plasmid was transformed to E. coli Rosetta for expression under the induction of 1 mmol/L IPTG at 37℃. The expressed products were purified by affinity chromatography using the nickel ion protein purification volume with His tag,and identified by Western Blot. Preparing its polyclonal antibody by bioinformatics.Restriction analysis proved that recombinant plasmid pET-32a-H was constructed correctly. The expressed PPRV H protein,with a relative molecular mass of about 67 ku. The prepared polyclonal antibody recognized the whole viral antigen of PPRV and recombinant PPRV H protein expressed in baculovirus. Recombinant plasmid pET-32a-H was constructed successfully,and PPRV H protein was successfully expressed. Useing bioinformatics could find polyclonal antibody 5-8,14-16,73-75,83-90,125-131,142-147,170-177,236-245,281-285,312-317,360-363,370-379, 388-391,445-449,487-489,503-505,520-522,532-535,544-551,592-595. The positive plasmid pET-32a-H was successfully constructed,and the target protein was expressed. The potential epitope of H protein was successfully predicted. It is important for elimination of PPRV earlyer and preparation of novel subunit vaccines.
2018 Vol. 33 (5): 111-116 [Abstract] ( 18 ) HTML (1 KB)  PDF (1173 KB)  ( 21 )
117 Construction of Eukaryotic Expression Vector of DKK1,BMP4 Gene by Homologous Recombination and Its Expression in Fibroblasts
ZHANG Mengyao, YANG Feng, LIU Kaidong, LIU Jifeng, HE Jianning, LIU Nan
DOI: 10.7668/hbnxb.2018.05.017
The study aimed to use Homologous recombination build DKK1 and BMP4 genes expression vector study the interaction between DKK1 and BMP4 genes by change of expression. Firstly,a pair of primers that have homologous arm were designed by RNA extraction and referred DKK1 and BMP4 genes sequence information of Aohan Fine Wool Sheep in GenBank,the DKK1 and BMP4 genes fragment was amplified by PCR. We used SoSo kit connect the target fragment to pcDNA3.1 eukaryotic expression vector. Recombined plasimid pcDNA3.1-DKK1 and pcDNA3.1-BMP4 was constructed and transformed into E. coli DH5α competent cell. Then plasmid pcDNA3.1-DKK1, BMP4 was cotransfected into fibroblasts,and used qRT-PCR to detect the expression level of these genes. The results showed that,pcDNA3.1-DKK1, BMP4 plasmid cotransfected successfully and identified by enzyme and sequencing. The expression level of DKK1 gene in fibroblasts was lower than the control group and the expression level of BMP4 gene in fibroblasts was higher than the control group. The plasmid was constructed and cotransfected into fibroblasts successfully by Homologous recombination technique. The expression level of DKK1 gene decreased and the expression of BMP4 gene no variation. Therefore, BMP4 gene inhibited the expression of DKK1 gene,and the DKK1 gene had no effect on the expression of BMP4 gene. These results constructed the basis for further research.
2018 Vol. 33 (5): 117-124 [Abstract] ( 16 ) HTML (1 KB)  PDF (1573 KB)  ( 15 )
125 Screening of Pear Viral Disease Detection Methods and Detection of Pear Virus Disease in Tianjin Region
YANG Rui, HAN Xuechun, LI Qing, CHEN Zhaorong
DOI: 10.7668/hbnxb.2018.05.018
In order to investigate the incidence of known viral diseases of pear in Tianjin Region,the total RNA of snow pear leaves was extracted by improved CTAB method. And the latent virus species of the 70 pear trees from three regions of Tianjin were detected through RT-PCR identification,including Apple cholortotic leaf spot virus (ACLSV), Apple scar skin viroid (ASSVd), Apple stem pitting virus (ASPV), Apple stem grooving virus (ASGV) and Apple dimple fruit viroid (ADFVd). The results showed that the improved CTAB-mercaptoethanol method was a kind of comparatively ideal method for total RNA extraction from snow pear,the RNA had less impurity, the rate of total RNA extrate was higher, and the mean concentration was 73.880 μg/g. The obtained RNA meted the requirements for the next analysis. By RT-PCR technique the infection rates of the 70 pear trees with the above five viruses were 47.1%,11.4%,10.0%,8.6%,and 1.4%,respectively,and the mixed infection rate was 28.6% on average. The higher the age of pear trees, the higher the virus carrying rate. And the sensitivity of different varieties to viruses was different. The results showed that the pear virus disease in Tianjin was common, in which apple chlorotic leaf spot virus was the main type of virus, and had a certain proportion of multiple virus complex infection. The low level of pear planting management and the lack of standardized detection and monitoring of pear introduction among regions could lead to the occurrence and spread of pear virus disease. It was very unfavorable for the healthy development of Tianjin pear industry.
2018 Vol. 33 (5): 125-129 [Abstract] ( 30 ) HTML (1 KB)  PDF (953 KB)  ( 20 )
130 Cell Transport of DOFH10 in Arachis hypogaea L. Is Possibly via Vesicle and Specifically Recognized in Clathrin Related Pathway
YAN Haiyan, HUANG Zhenling, SUO Hui, HUANG Jiaquan, LIAO Boshou
DOI: 10.7668/hbnxb.2018.05.019
Study unknown DOFH10 (DNA binding with one finger) genes in peanuts,cloning of DOFH10 genes with PCR methods according to cDNA of DOF transcription factor GW391728 with primers s162-a1469 and primers s30-a1469 from 16 different peanut cultivars of Arachis hypogaea L.,sequences comparison,analysis on enzyme site and 3D structure and Western Blot on DOFH10 proteins were conducted. Two types of DOFH10 genes were separated from genome DNA of 5 cultivars with primers s162-a1469,type A DOFH10 was only existed in four cultivars with multiple seeds per fruit and one cultivar with two seeds per fruit. They were identical to XM_016100960.2 from Arachis duranensis chromosome A03 and to XM_016334585.2 from Arachis ipaensis chromosome B03 respectively. With primers s30-a1469,only type B DOFH10 was cloned from all the 16 cultivars used including all the six cultivars with multiple seeds per fruit. Type B DOFH10 had identical RNA and protein sequences at DOF zinc finger region to that of putitive clathrin assembly protein XP_016199412.2 encoded by gene on chromosome B01 of Arachis ipaensis. Multiple possible myristoylation sites scattered on DOFH10 protein,both results suggested that transportation of DOFH10 was probable via vesicle membrane traffic related to clathrin. DOFH10 protein was specifically expressed in developing cotyledons displayed by Western Blot. A and B two types of DOFH10 genes existed in cultivated peanut. The type B was present in all 16 peanut cultivars,and was an essential gene,was possibly transported into nucleus through clathrin dependant vesicle pathway. The type A DOFH10 tended more appeared in cultivars with multiple seeds per fruit,probably related to cell division in early fruit development.
2018 Vol. 33 (5): 130-143 [Abstract] ( 17 ) HTML (1 KB)  PDF (3939 KB)  ( 15 )
144 Comparison of Physiological Characteristics and Differential Expression of FtNHX1 Gene among Twelve New Fagopyrum esculentum Varieties under Salt Stress
ZHANG Jiasong, LU Qihuan, LI Faliang, YANG Hongying, XU Chan, SONG Jinnan, WANG Yaqi, YANG Hongbing
DOI: 10.7668/hbnxb.2018.05.020
In order to study the effects of salt stress on physiological characteristics and relative expression of FtNHX1 in Fagopyrum esculentum,twelve new Fagopyrum esculentum varieties (Pingqiao 2,Dingtianqiao 3,Chitianqiao 1,TQ11-3,T407-8,Tongqiao 2,Yuntianqiao 1,Meng 0825,Tianqiao 1307-179,Pingxuan 03-122,TQ11-6 and Chitianqiao 2) were used as experimental materials. The indexes of the seeds germination rate,seedlings fresh weight,roots vigor,leaf plasmalemma permeability,MDA content,SOD activity and relative expression of FtNHX1 in roots were determined under salt stress to compare the physiological characteristics and relative expression of FtNHX1 among twelve new Fagopyrum esculentum varieties. The results showed that the seeds germination rate and seedlings fresh weight of Dingtianqiao 3 and Chitianqiao 1 were decreased more than that of Pingqiao 2 and T407-8,while the seeds germination rate of Pingqiao 2 and T407-8 had no significant difference with the control,and had obviously promoting effects of salt stress on seedlings growth of the two Fagopyrum esculentum varieties. The leaf plasmalemma permeability and MDA content of Dingtianqiao 3 and Chitianqiao 1 were increased greatly,while the leaf plasmalemma permeability of Pingqiao 2 and T407-8 had no significant difference with the control,and the leaf MDA content of Pingqiao 2 and T407-8 increased to a lesser extent. The roots vigor and leaf SOD activity of Dingtianqiao 3 and Chitianqiao 1 were decreased greatly,while the roots vigors of Pingqiao 2 and T407-8 were decreased little,and the leaf SOD activities of Pingqiao 2 and T407-8 were increased significantly,indicated that had strong stress resistance. The relative expression of FtNHX1 gene of Dingtianqiao 3 was decreased significantly,while that of Pingqiao 2 and T407-8 were increased significantly,indicated that more salt tolerance. Fagopyrum esculentum varieties of Pingqiao 2 and T407-8 were determined salt tolerant varieties,while that of Dingtianqiao 3 and Chitianqiao 1 were determined salt sensitive varieties.
2018 Vol. 33 (5): 144-150 [Abstract] ( 27 ) HTML (1 KB)  PDF (1124 KB)  ( 20 )
151 Response of Nutrient Accumulation in Quinoa to Source-sink Adjustment
REN Yongfeng, HUANG Qin, WANG Zhimin, ZHAO Peiyi, GAO Yu, LIAN Haifei
DOI: 10.7668/hbnxb.2018.05.021
During the quinoa cultivation,the yield and quality of quinoa are reduced due to the high rate of branch breakage and ineffective panicle.In this experiment,the randomized block design was adopted to study the effects of 6 treatments,which are cutting leaves (CLE),cutting side ears (CSE),cutting leaves and cutting side ears (CLS),topping(TOP),package the top ear (PTE) and control (CK),and to find the responses of plant nutrient accumulation to different regulation measures. The results showed that:the yield of quinoa was significantly different with the control,the unbroken spike yield of CSE treatment was highest,the yield,the stem diameter and the 1 000-grain weight of CSE were 5.2%,14.0% and 6.0% higher than that in CK,the number of branches of CSE were 10% lower than that in CK,the branches of topping was 8.0% higher than that in CK;The effect of top panicle more than that of lateral branch spike on leaf area index increasing,CSE was significantly higher than CK in photosynthetically active radiation,and there was a significant positive correlation between yield and grain weight/leaf dry weight, grain weight/leaf area and kernel per ear/leaf area. The effects of source-sink organs on the grain filling rate and dry matter accumulation was leaf > top ear > lateral ear. From the filling stage to the ripening stage of quinoa,the contribution of top and lateral branch spikes were rising trend. The contribution rate of CSE,TOP and CLE were 100.5%,87.6% and 49.0% of the control. Therefore,in the late growth period of quinoa,spike is one of the main source organs,and the top panicles have obvious advantages. It is on the basis of selection of fine varieties,appropriate management of water and fertilizer and rational close planting,and support with topping or side branches spike measures to remove the top advantage or invalid library organs,so as to achieve the purpose of regulating the flow of nutrients and improve the efficiency of nutrient accumulation,and promote the harmonious development of the source-sink,to achieve quinoa quality and yield improvement.
2018 Vol. 33 (5): 151-159 [Abstract] ( 24 ) HTML (1 KB)  PDF (1138 KB)  ( 27 )
160 Qualitative and Quantitative Study on Lodging Resistance of Wheat Varieties in Henan Province
HU Weiguo, CAO Tingjie, WANG Xicheng, ZHANG Yu'e, CHANG Ping, ZHAO Hong
DOI: 10.7668/hbnxb.2018.05.022
In order to measure the lodging resistance of wheat effectively,84 new wheat varieties from the Regional Trial of Henan Province in 2015-2016 were analyzed using lodging index,cluster analysis and principal component analysis. The result showed that the average lodging index was 7.06, 40 varieties were below 3.50,accounting for 47.6% of all tested varieties,and 8 varieties had the lodging index value above 20,accounting for 9.5%,which indicated that the lodging resistance of most new varieties tested in Henan Regional Trial was good. The correlation coefficient of lodging index with lodging area,lodging level and ratio of severe lodging pilot was 0.977* *,0.912* * and 0.906* *,respectively,reaching to 0.01 significant level,which suggested that the lodging index could quantify the lodging resistance well. Principal component analysis showed that lodging index,lodging area,lodging level and ratio of severe lodging pilot (based on 2017 criterion) were the best indicators to measure the lodging resistance of wheat. The combination of phenotypic clustering with lodging index and lodging parameters (including ratio of severe lodging pilot,ratio of lodging pilot,lodging area and lodging level) was better than lodging index or lodging parameter alone to evaluate the lodging resistance. It was concluded that this method could measure the lodging resistance qualitatively and quantitatively in single spot or multi-spot trials,and it could be extended to other cereal crops to identify lodging resistance. The proposed method provided a feasible way for estimating and classifying the lodging resistance of wheat varieties effectively.
2018 Vol. 33 (5): 160-167 [Abstract] ( 27 ) HTML (1 KB)  PDF (999 KB)  ( 31 )
168 Alleviating Effects of Exogenous CO on Germination and Growth of Cucumber Seeds under Drought Stress
MU Xuejiao, ZHANG Qiang, WU Yan, WANG Xuejuan, ZHANG Yuanbing
DOI: 10.7668/hbnxb.2018.05.023
In order to explore the intrinsic mechanism of exogenous carbon monoxide (CO) improving the germination and growth of cucumber seedlings under drought stress. Cucumber Xinjinyan 4 seeds were pretreated with the CO donor,hematin (0.01 mol/L),and their germination,nutrient absorption ability,plasma membrane stability,osmotic adjustment substance content,hydrolase activities and isoenzyme expression under drought stress simulated by 10% polyethylene glycol 6000 were investigated. The results showed that,under PEG stress,exogenous CO improved the germination potential,germination rate,root length,hypocotyl length and fresh weight of cucumber seeds,promoted imbibition of cucumber seeds,enhanced root activity,significantly decreased plasma membrane permeability and malondialdehyde (MDA) content,and increased the contents of soluble protein and soluble sugar. Meanwhile,the results also demonstrated that exogenous CO apparently up-regulated the activities of amylase and esterase and their isozyme expression. Compared with the PEG treatment,the root activity,soluble protein and soluble sugar content,amylase and esterase activity of PEG+CO treatment increased by 12.57%,5.84%,32.56%,9.87% and 48.87%,respectively,MDA content decreased by 25.81%. These results suggested that exogenous CO could improve the adaptability of cucumber seeds to drought stress by enhancing the nutrient absorption ability of cucumber seeds,protecting the stability of cell membrane system,improving osmotic adjustment ability,enhancing the hydrolase activities and their isozyme expression,and consequently induce the germination and growth of cucumber seeds under drought stress.
2018 Vol. 33 (5): 168-173 [Abstract] ( 28 ) HTML (1 KB)  PDF (1017 KB)  ( 22 )
174 The Effect of Enhanced UV-B Radiation on Soybean Root's Activate Oxygen Metabolism and Antioxidative System
YANG Lu, ZHAO Tianhong
DOI: 10.7668/hbnxb.2018.05.024
In order to explore the regulation of active oxygen metabolism and the response of antioxidant system in soybean root under enhanced UV-B radiation. Contrast with enhanced UV-B radiation through natural state,those was tested that the soybean roots' lipid peroxidation (malondialdehyde content and relative electrolytic conductivity),reactive oxygen species metabolism (super oxygen anion production rate and Hydrogen peroxide content),protective enzyme activity (superoxide dismutase activity,peroxidase activity and catalase activity) and protective substance content (ascorbic acid content,carotenoid content and proline content) of different growth stages of the soybean,and a series of different indexes were measured. The results showed that:compared with the control,enhanced UV-B radiation could cause O2· and Hydrogen peroxide accumulate. For specific performance,compared with CK,O2· producing rate and Hydrogen peroxide accumulating went down 11.5%,and went up 9.9% in florescence. The UV treatment was higher than CK in other reproductive period. Compared with CK,UV treatment was significantly different in all growth stages. And could lead MDA content and relative electrolytic conductivity to increasing. For specific performance,compared with CK,UV treatment was higher and significantly different in all growth stages for MDA content and relative electrolytic conductivity. And enhanced UV-B radiation conditions also could lead to protective enzyme activity reducing,such as SOD,POD and CAT. For specific performance,compared with CK,SOD,POD and CAT activity went up 15.5%,went down 15.3% and 12.8% for UV treatment in branch stage, other stages were lower. Compared with CK,UV treatment was significantly different except CAT activity in branch stage. Cause protective substances' content to decline,such as AsA,Car and Proline. For specific performance,compared with CK,AsA,Car and Proline content went down 11.4% and 15.4%,went up 10.5% for UV treatment in branch stage. And went up 31.2%,went down 17.9%,went up 20.7% in florescence. Other stages were lower. Compared with CK,UV treatment was significantly different except Car content in podding stage or proline content in branch stage. Therefore,enhanced UV-B radiation could cause the active oxygen metabolism and lipid peroxidation to speed up,membrane permeability to increase,and cause the antioxidant system to damage. And with the soybean growth period goes on,the effect moves obviously.
2018 Vol. 33 (5): 174-180 [Abstract] ( 18 ) HTML (1 KB)  PDF (1059 KB)  ( 20 )
181 Effects of Long-term Fertilization on Potassium Q/I in Cropping Maize
WU Junnan, LIU Yuxin, WANG Tianye, GAO Qiang, WU Haiyan, LIU Shuxia
DOI: 10.7668/hbnxb.2018.05.025
In order to understand the effect of long-term fertilization on the dynamic balance of potassium nutrients in black soil corn fields. We based on the long-term fertilization experiment (1984-2015) in black soil corn field,the effects of different fertilization treatments on the characteristics of Q/I curve and the parameters (potassium supply capacity,potassium supply potential and potassium buffer capacity) of black soil in maize field were studied. The results showed that long-term different fertilizer treatments of black soil potassium Q/I curve and typical Q/I curve was similar. Q/I curve among different fertilizer treatments presented a similar shape. It consisted of two parts:curve and straight line. Fertilization could increase in the black soil-ΔK0 value and AReK value,reduced the black earth PBCK value,treatment of NPK with high straw content (NPK+S0.5) black soil-ΔK0 value of up to 0.167 9 cmol/kg,AReK value was also the highest under NPK+S0.5 treatment 1.718 5×10-3(cmol/kg)-1/2.However,the PBCK value was the smallest,which was 97.7 (mg/kg)/(cmol/kg)1/2. Applying potassium fertilizer and straw was more effective in increasing the-ΔK0 and AReK values and decreasing the PBCK value in the corn field soil. The correlation analysis showed that there was a significantly positive correlation between black soil-ΔK0 and AReK values and soil organic matter,total nitrogen,total phosphorus,total potassium,and available potassium content (P <0.01),and PBCK values were exactly the opposite. In general,chemical fertilizers combined with straw had a great influence on the physicochemical properties of the soil and the-ΔK0 value,AReK value and PBCK value. It could not only increase soil nutrients,but also increase soil potassium supply capacity. Therefore,reasonable fertilization will help increase the supply capacity of soil potassium nutrition and increase crop yield and quality.
2018 Vol. 33 (5): 181-187 [Abstract] ( 17 ) HTML (1 KB)  PDF (1086 KB)  ( 20 )
188 Effects of Long-term Fertilization on Reclaimed Soil Aggregate Stability and Distribution of Carbon in the Southeastern Shanxi Mining Area
LI Jianhua, LI Hua, GAO Chunhua, ZHANG Qiang, JIN Dongsheng, LU Jinjing
DOI: 10.7668/hbnxb.2018.05.026
To explore the aggregate stability and organic carbon distribution characteristics of reclamation soil in the Southeastern Shanxi Mining Area under different fertilization,the long-term located trials with eight years were conducted to study the amount, stability and organic carbon distribution of soil aggregate. The test consisted of four fertilizer treatments,no fertilization (CK),chemical fertilizer (NPK),combination of low organic fertilizer and chemical fertilizers (LOF) and combination of high organic fertilizer and chemical fertilizers (HOF). The results showed that the application of fertilization significantly increased the number of large aggregates (> 0.25 mm),HOF treatment significantly increased the proportion of aggregate at each particle size, the effects on the particle size of 2-1 mm and 0.50-0.25 mm aggregates were highest with the HOF treatment. The average weight diameter (MWD) and geometric mean diameter (GMD) of the aggregates were significantly increased and the fractal dimension (D) of soil aggregates was significantly reduced by the HOF and LOF treatments,which improved the stability of soil aggregates. There was not significant difference between HOF and LOF,and NPK had no significant influence on the fractal dimension. The organic carbon content of each particle size aggregate of>0.25 mm was higher than that of<0.25 mm,and the large aggregate was rich in organic carbon. The application of HOF significantly increased the contribution rate to soil nutrient of 5-2 mm and 2-1 mm particle size aggregate. There was a significant positive correlation between soil organic carbon (SOC),large aggregate number (R0.25),MWD,GMD and aggregates >0.25 mm particle size,respectively. SOC and R0.25,MWD,GMD were a very significant positive correlation. Therefore,in the Southeastern Shanxi Mining Area reclaimed,the application of high organic fertilizer and chemical fertilizers (HOF) is the effective method to improve soil organic carbon content and enhance the stability of soil aggregates.
2018 Vol. 33 (5): 188-194 [Abstract] ( 17 ) HTML (1 KB)  PDF (970 KB)  ( 17 )
195 Effect of Different Fertilization Treatments on Soil Microbial Function Diversity in Rhizosphere of Sorghum
CUI Peipei, WU Ailian, WANG Jinsong, DONG Erwei, NAN Jiangkuan, BAI Wenbin, JIAO Xiaoyan
DOI: 10.7668/hbnxb.2018.05.027
Rhizosphere soil microorganisms play an important role in the regulation of soil rhizosphere environment and nutrient transformation. Sorghum has been widely used as forage and strong barren tolerance.Study of rhizosphere microorganisms on the ecological response of different nutrients,which can provide the theoretical basis for nutrient management of different soil nutrients under conditions of sorghum. This paper was based on the long-term field test of sorghum/corn rotation,combined with the pot experiment of artificial greenhouse. BIOLOG micro plate was applied to test the effect of NPK,PK,NK,NP fertilizers and CK (without fertilization) on sorghum rhizosphere soil microbial functional diversity. The results showed that the field test was in agreement with the pot experiment. PK treatment increased the metabolic activity of rhizosphere microorganisms,and improved the utilization of amino acids,carboxylic acids,amines,sugars and polymers,and reduced the utilization of amphiphilic compounds. The Shannon-Wiener index(H),Richness index(S) and Simpson index(Ds) were higher than other treatments. The effects of NK and NP on the functional diversity of rhizosphere microorganisms were not significant,but NK treatment reduced the microbial metabolic activity in the rhizosphere of the late culture. CK treatment significantly decreased the rhizosphere microbial utilization of carbon sources,the Shannon-Wiener index(H),Richness index(S) and Simpson index(Ds) were lower than other treatments,Substrate evenness(E) higher than other treatments. Principal component analysis showed that there was no significant difference in rhizosphere soil microbial carbon source utilization capacity and type between NPK and NP treatments,and the rest treatments showed significant difference in the field test. In pot experiment,the difference of carbon source ability and type of NPK,NK and NP treatment was not significant,and the difference between the other treatments was significant. In summary,fertilization was conducive to improving soil microbial characteristics. The responses of sorghum rhizosphere microorganisms to nitrogen,phosphorus and potassium stress were different. When nitrogen stress was applied,the increase of soil microbial activities in sorghum rhizosphere was probably the cause of nitrogen tolerance in sorghum.
2018 Vol. 33 (5): 195-202 [Abstract] ( 20 ) HTML (1 KB)  PDF (1000 KB)  ( 37 )
203 Difference of Nitrogen Response in Different Genotypes of Potato
JIAO Jiaojiao, WEI Qiaorong, PEI Jianing, GUO Yuxin, TANG Liyang, SHI Ying
DOI: 10.7668/hbnxb.2018.05.028
The differences in nitrogen(N) uptake and utilization of different potato varieties at seedling stage in Northeast China were evaluated,and potato genotypes with high responsiveness to N were screened to provide a theoretical basis for the selection and mechanism study of potato N efficient genotypes. Pot experiment was used to compare the differences in the ability of different genotypes of potato to obtain N at the seedling stage under the three levels of N supply and their causes. The results showed that different N supply levels had significant differences in the ability of different genotypes to auumulate N. With the increased of N supply levels,the N accumulation of four potato varieties such as Dongnong 310,Yanshu 4,Kexin 13 and Kexin 22 increased significantly. The response rate was high,while the N accumulation of seven potato varieties such as Dongnong 308,Dongnong 311,Dongnong 312,Dongnong 316,Yanshu 9,Yanshu 7 and Kexin 19 had no significant increase,and the response to N was low. For the genotypes with high nitrogen responsiveness,the Nitrate reductase active(NRA),Glutamine synthetase active(GSA),and root activity a little bit increased significantly with increasing N supply levels,while the chlorophyll(SPAD)values increased less. The increase in N accumulation,NRA,GSA,root activity,and SPAD values were significantly greater than those with low N response. The correlations between N accumulation and NRA,GSA,SPAD,and root activity of potato under low N and high N conditions were analyzed. NRA and GSA activity could better reflect the N accumulation ability of potato during seedling stage,the ratio of SPAD and root activity. The reliability is better,and the strength of NRA and GSA activity can be used as an important indicator for evaluating the N uptake accumulation of potato seedlings.
2018 Vol. 33 (5): 203-208 [Abstract] ( 38 ) HTML (1 KB)  PDF (933 KB)  ( 27 )
209 Effects of Exogenous γ-aminobutyric Acid on Grain Nitrogen Formation and Yield in Cold-region Japonica Rice under Drought Stress at Booting Stage
GU Haitao, JIA Yan, ZHANG Bo, SUN Bin, WANG Zhuoqian, ZHAO Hongwei
DOI: 10.7668/hbnxb.2018.05.029
To investigate the regulation effect of exogenous γ-GABA on the formation of nitrogen and yield of Japonica rice in cold region under drought stress at booting stage,Dongnong 425 and Songjing 6 were used as experimental materials,and 6 GABA concentrations(0,0.25,0.50,1.00,2.00 and 4.00 mmol/L)were set up. The effects of exogenous GABA on dry matter amount,total nitrogen content and activity of nitrogen metabolism enzyme of Japonica rice were analyzed under drought stress at booting stage with normal irrigation as control(CK). The results showed that compared with the control,the yield and the yield components,dry matter accumulation of grains(in addition to 7 days after full bloom)decreased significantly,the activity of straw transaminase(GOT)and glutamine pyruvic transaminase(GPT)decreased,the activity of glutamine synthetase(GS)increased(7 and 14 days after full bloom),the content of total nitrogen(in addition to 21-35 days after panicle of Songjing 6)and endogenous GABA increased significantly. Compared with the drought treatment,using different concentrations of GABA could significantly improve grain yield,yield components and dry matter accumulation of grains,nitrogen metabolism key enzyme activity,total nitrogen and endogenous GABA content,and with the increased of GABA concentration. The yield and yield components of Dongnong 425 and Songjing 6,and the accumulation of grain dry matter quality,the activities of nitrogen metabolism key enzymes,and the content of total nitrogen and endogenous GABA increased first and then decreased. And the optimum concentrations of exogenous GABA in Dongnong 425 and Songjing 6 sprayed by drought stress at booting stage were 0.54,1.22 mmol/L.
2018 Vol. 33 (5): 209-217 [Abstract] ( 27 ) HTML (1 KB)  PDF (1172 KB)  ( 22 )
218 Impact of Substitution of Green Manure under Different Mixed Cropping Modes to Chemical N Fertilizer on Yield Formation Characters of Double Cropping Rice
LIU Sichao, TANG Lizhong, LI Chao, YANG Jing, SHI Quan, CHEN Pingping, TU Naimei, YI Zhenxie, ZHOU Wenxin
DOI: 10.7668/hbnxb.2018.05.030
To probe the optimal green manure mixed cropping modes under condition of chemical fertilizer reduction,the effects of different green manure mixed cropping modes(Astragalus smicus monoculture,AS; Astragalus smicus+Ro-phanus sativus L. mixed cultivation,AR; Astragalus smicus+Lolium perenne L.,AL; Astragalus smicus+Ro-phanus sativus L.+ Lolium perenne L. mixed cultivation,ARL; Winter fallow,CK)and nitrogen reduction application on yield formation characters of early and late rice were studied with Zhongzao 39 and Taiyou 390 as early and late rice materials,respectively. The results showed that:the substitution proportion of green manure to chemical nitrogen in treatments of green manure return to field was 28.98%-39.68% in 2016 and 30.55%-39.47% in 2017 respectively,which in AS treatment was bigger and which in AL treatment was the lowest. The substitution of green manure to basal chemical nitrogen had not clear affect on tillering dynamics,while all treatments of green manure return to field had the better yield-increasing effect,and the difference among different treatments was observed. In 2016,the yield increasing effect of AS treatment on early and late rice was the best (increased by 7.3% and 8.5%,respectively),and the next was AR treatment (increased by 6.0% and 7.1%,respectively),while in 2017 the yield increasing effect of AR treatment on early and late rice was the best(increased by 7.2% and 5.5%,respectively),and the next was AS treatment(increased by 5.5% and 4.7%,respectively).The substitution of green manure to basal chemical nitrogen was advantageous to heighten the leaf area index and leaf SPAD value at filling stage and dry matter accumulation,and then improve the effective panicles and seed setting rate,while the effect was different among different treatments,and AS and AR treatment performed well in common.So,the substitution of green manure to basal chemical nitrogen fertilizer was advantageous to heighten yield of double cropping rice,and astragalus smicus monoculture and astragalus smicus+Ro-phanus sativus L. mixed cultivation performed well.
2018 Vol. 33 (5): 218-225 [Abstract] ( 25 ) HTML (1 KB)  PDF (1002 KB)  ( 22 )
226 Effects of Potassium Levels on Storage Root Yield and Nutrient Utilization of Purple Sweet Potato
ZHAO Fengqin, LIU Qing, DU Zhiyong, SHI Yanxi
DOI: 10.7668/hbnxb.2018.05.031
In order to find out the effect of different potassium doses on the yield and nutrient utilization of purple sweet potato,it provides theoretical and practical basis for the rational application of potash fertilizer. Using design of randomized block experiment,field experiment and chemical analysis method,the different amount of potassium application on low fertility soil accumulation of dry matter,yield and its components purple sweet potato factor,NPK factor accumulation,the effects of potassium use efficiency were studied. The results showed that dry matter accumulation in roots of purple sweet potato was higher than that of K0 treatment during the growth of sweet potato,when the amount of potassium was 180 kg/ha,the accumulation of root dry matter reached the maximum. Potassium had effect on the accumulation and distribution of dry matter,increased the number of potatoes and single potato weight,and could increase the yield of sweet potato and obtain the largest yield increase when the rate of potassium application was 180 kg/ha. Compared with the K0,the yield increased by 20.78% and 26.19%,respectively,in 2016 and 2017,and the difference was significant(P <0.05).Compared with K0 treatment,potassium could promote the absorption of nitrogen and phosphorus in purple sweet potato,significantly increased the accumulation of whole plant nitrogen and phosphorus,and increased the maximum growth rate of K3. The accumulation amount of the root and the whole plant nitrogen increased by 75.38% and 65.75% respectively,and the cumulative phosphorus accumulation increased by 45.24% and 60.28%,respectively,and difference was significant(P <0.05). Besides,potassium fertilizer treatments could also increase the utilization ratio of potash fertilizer and agronomic efficiency. Compared with K0,apparent utilization and agriculture utilization increased by 47.82 percentage points and 158.11%,respectively,and the difference was significant(P <0.05). Therefore,the optimum application dosage of potassium fertilizer was confirmed to be 180 kg/ha.
2018 Vol. 33 (5): 226-231 [Abstract] ( 31 ) HTML (1 KB)  PDF (980 KB)  ( 26 )
232 Studies on the Efficient Utilization and Coupling Effect of Nitrogen-water in Wheat
LI Wuchao, LI lei, WANG Wei, LI Jingjing, YIN Jun
DOI: 10.7668/hbnxb.2018.05.032
Semiwinter wheat variety Zhoumai 22 was used as material to study the effects of different water-nitrogen combinations on photosynthetic characteristics,water use efficiency,water consumption and yield during the grain filling period of wheat,in order to determine the optimal combination of high yield and high efficiency utilization of water-nitrogen for wheat.The experiment was carried out using pot cultivation under rain shelter,setting up five water level:the relative soil water content of 55% (H1),65% (H2),70% (H3),75% (H4) and 85% (H5) from jointing to mature period on the basis of the relative soil water content of 55% from returning green stage to jointing stage,and 3 nitrogen level:no nitrogen (0 kg/ha,N0),medium nitrogen (195 kg/ha,N1) and high nitrogen (270 kg/ha,N2). The results showed that nitrogen fertilizer had a extremely significant effect on yield,water consumption coefficient and fertilizer use efficiency,and the interaction between water and nitrogen was extremely significant. Under the conditions of 65%-70% soil relative water content and medium nitrogen,the net photosynthetic rate and transpiration rate of wheat were significantly higher than that of other treatments during the whole grain filling period,of which the transpiration rate of 10 d after flowering was over 11 mmol/(m2·s),the grain filling rate increased 15%~64%,the water use efficiency was over 1.7 g/kg,while the partial productivity and the agricultural utilization of nitrogen fertilizer were over 15.1,7.5 kg/kg,respectively. It was the best combination for wheat to obtain high efficiency utilization of water-nitrogen,maximum grain filling rate and grain yield. Thus,it is concluded that the combination of 195 kg/ha nitrogen fertilizer and 65%-70% soil relative water content from jointing stage to maturity stage not only give the highest yields,but also have the highest WUE,nitrogen partial productivity(NPEP) and nitrogen agronomic use efficiency(NAUE).
2018 Vol. 33 (5): 232-238 [Abstract] ( 18 ) HTML (1 KB)  PDF (1262 KB)  ( 35 )
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