ACTA AGRICULTURAE BOREALI-SINICA
 
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2018 Vol. 33, No. 6
Published: 2018-12-28

 
1 Cloning and Expression Analysis of Transcription Factor Tamyb59 in Wheat
ZHANG Pengyu, LIU Yuxia, CAO Liru, YUAN Zhen, WANG Guorui, WANG Tongchao, YIN Jun, WEI Li
DOI: 10.7668/hbnxb.2018.06.001
In order to further excavation of the role of wheat stress response genes in plant abiotic stress response,explored the regulation mechanism of abiotic stress response, a MYB gene was screened from a transcriptome result and named Tamyb59, temporarily.The full length sequence of Tamyb59 was cloned by PCR. Sequence comparison and conserved domain were analyzed by NCBI and DNAMAN. Amino acid composition and hydrophilic coefficient were analyzed by online software Expasy and TMHMM. Phylogenic tree was constructed by MEGA 6.0 according to the NJ method. And a fusion expression vector pMDC83-GFP was constructed to identify the subcellular localization. The Real-time PCR (qRT-PCR) was used to analyze the expression characteristics of different tissues in different abiotic stress treatments. The significance was analyzed by SAS. The results showed that:Tamyb59 gene contained a typical conserved SANT domain. The full-length CDS of Tamyb59 was 522 bp,which encoded 173 amino acids with a molecular weight of 19.7 ku and the theoretical isoelectric point was 7.61;Phylogenetic analysis showed that Tamyb59 had 52.0%-85.6% homology with MYB transcription factors of 9 other plants (Aegilops tauschii, Oryza sativa Japonica and Zea may, etc.). Tamyb59 had the highest homology with the MYB sequence of Setaria italica. The protein of Tamyb59 encoded specifically located in the nucleus.qRT-PCR was used to analyze the expression characteristics of different tissues in different abiotic stress treatments. The results revealed that the expression level of Tamyb59 was the highest in the root, and lower in stem, leaf and young spike. Under the PEG and NaCl treatment stress, the expression of Tamyb59 showed a trend of rising first and then decreasing, indicating that Tamyb59 had different responses under different stresses.
2018 Vol. 33 (6): 1-7 [Abstract] ( 73 )   全文: HTML   PDF (1108 KB)  ( 114 )
8 Bioinformatics Identification of SBP Genes and Expression Profiles in Wheat
LÜ Liangjie, CHEN Xiyong, HU Mengyun, LIU Yuping, SUN Lijing, MA Le, LI Hui
DOI: 10.7668/hbnxb.2018.06.002
The squamosa promoter binding protein (SBP) gene family is specific transcription factor in plant, containing 79 amino acid residue conserved domains, and is mainly involved in plant growth, development and physiological and biochemical processes.However,genome-wide analysis of the SBP gene family has not been reported in wheat.To further explore the function of the wheat SBP gene, 50 SBP genes, named TaSBPs, were obtained by analyzing the latest genomic data of wheat, and were ranked according to the chromosome number as TaSBP1-TaSBP50. Combined with the published genome data of cultivar Chinese Spring, genes structure, chromosome distribution, conserved domain of proteins, phylogenetic trees and gene expression profiles of the wheat cultivars were analyzed using bioinformatics methods.The results showed that 50 TaSBP genes were distributed on the remaining 19 chromosomes except 4B and 4D chromosomes. The genes encoded 192-1 104 amino acids in length and the number of gene exons was from 2 to 11.The results of tandem repeat analysis showed that fragment replication and tandem repeats were the main patterns of gene expansion in the TaSBP family. The phylogenetic tree of seven crops SBP proteins were divided into four categories, and the structure of the same class was similar; The 50 TaSBP gene families in wheat contained 10 motifs, and it was presumed that the TaSBP gene family should contain motif1, motif2 and motif4.50 TaSBP genes were all detected in 13 tissues and organs, and the expression of TaSBP genes in different tissues were significantly different.
2018 Vol. 33 (6): 8-16 [Abstract] ( 70 )   全文: HTML   PDF (1405 KB)  ( 56 )
17 Preparation of Polyclonal Antibody of Maize AGPL2 Protein and Analysis of Its Spatiotemporal Expression Analysis
LÜ Yanan, YU Guowu, HUANG Yubi
DOI: 10.7668/hbnxb.2018.06.003
In order to investigate the functional mechanism of maize AGPL2 in starch accumulation during grain development,this study cloned AGPL2 gene and constructed the GST-tagged prokaryotic expression vector PGEX-6T-1-AGPL2. Using to the inducing expression system of Escherichia coli,GST-AGPL2 fusion protein was able to be obtained expression at high levels by inducing with 0.5 mmol/L was opropyl-β-D-thiogalactoside (IPTG) at 28℃ for 6 h. Then GST (Glutathione S-transferase) tag protein purification medium was used to obtain higher quality GST-AGPL2 fusion protein by affinity chromatography. The AGPL2 polyclonal antibody was prepared by immunizing New Zealand white rabbit with purified GST-AGPL2 recombinant protein,and the antiserum was isolated and purified. Then the GST tag of the antigen GST-AGPL2 fusion protein was removed by rTEV Protease,and Western Blot assayed with the purified AGPL2 polyclonal antibody showed that AGPL2 polyclonal antibody had high specificity and sensitivity and could be used to detect nanogram antigen protein. The distribution and expression patterns of AGPL2 protein in different tissues and pollination stages of maize was studied by Western Blot. The results showed that the expression of AGL2 protein in different tissues of maize was tissue-specific and the highest in endosperm. The expression level of AGPL2 protein increased first and then decreased during the different pollination period of maize endosperm,and reached the maximum in the middle pollination. The results were consistent with the accumulation of starch during the development of maize kernels,and indicated that AGPL2 was mainly present in maize endosperm and might be involved in the synthesis of starch. It also indicated that AGPL2 polyclonal antibody was highly specific and could recognize AGPL2 antigen in corn.
2018 Vol. 33 (6): 17-23 [Abstract] ( 47 )   全文: HTML   PDF (1200 KB)  ( 29 )
24 Cloning and Bioinformatics Analysis of BjSnRK2C Gene from Potherb Mustard
ZHANG Chunlan, MAN Lili, XIANG Dianjun, LI Xuxin, BAO Wurina, LIU Peng
DOI: 10.7668/hbnxb.2018.06.004
The SnRK2 family genes, which are key regulators of plant resistance to abiotic stresses in the ABA pathway, encode a class of serine/threonine protein kinases in plants. Based on the conservative cDNA sequences of SnRK2 members, a full-length cDNA sequence of SnRK2, named as BjSnRK2C, was cloned from Brassica juncea by RT-PCR and SON-PCR methods. The sequencing results showed that the length of full-length cDNA (GenBank accession No. MF983711)was 1 380 bp containing a 3'-UTR of 137 bp, a 5'-UTR of 214 bp, and a complete open reading frame of 1 029 bp with 342 amino acids. The putative molecular weight and theory isoelectric point of the BjSnRK2C protein were 38.2 ku and 5.61, respectively. The BjSnRK2C was a hydrophilic protein with 30 phosphorylation sites, contained two significant transmembrane domains, contained no signal peptide, and might played a physiological role in the cytoplasm. The secondary structure of the BjSnRK2C protein contained 130 α-helixes, 126 random coils, 59 extended strands, and 27 β-turns. The BjSnRK2C protein contained a serine/threonine enzyme activity domain and an ATP binding site, and was at the same evolutionary branch with AtSnRK2.8 from Arabidopsis. The three-dimensional structure of the BjSnRK2C protein kinase was very similar to that of AtSnRK2.8 protein kinase from Arabidopsis, suggesting that they had similar functions. The C terminal of the BjSnRK2C protein kinase contained only the domain Ⅰ, and lacked the domain Ⅱ, suggesting that the BjSnRK2C gene was probably independent of ABA regulatory pathway in abiotic stress process, which will lay a theoretical foundation for further studying the BjSnRK2C gene function.
2018 Vol. 33 (6): 24-32 [Abstract] ( 30 )   全文: HTML   PDF (1546 KB)  ( 38 )
33 Mapping of Quantitative Trait Locus and Developing of Closely Linked Markers for Tillering in Foxtail Millet
DU Xiaofen, WANG Jun, LI Yunfei, WANG Zhilan, YUAN Guobao, DU Guohua, HAN Fang, PENG Jianxiang, ZHANG Wenna, CAI Wei, YUAN Feng, CUI Juduo, GUO Erhu, ZOU Hongfeng, ZHANG Linyi, PENG Shuzhong
DOI: 10.7668/hbnxb.2018.06.005
Tillering is an important trait related to yield in many crops,and is the same for yield improvement of foxtail millet. However, the genetic mechanism of tillering in foxtail millet (Setaria italica) remains largely unknown. To accelerate mapping of relevant QTLs or genes, a restriction site-associated DNA sequencing (RAD-seq) approach, MSTmap and WinQTLCart 2.5 were employed to identify QTLs responsible for tillering in two F2 populations called Cross AJ (543 F2 individuals) and Cross HC (131 F2 individuals). A total of six QTLs including qAJTN1, qAJTN5, qAJTN7-1,qAJTN7-2, qAJTN7-3 and qAJTN9, were identified in Cross AJ, explaining 0.7%-9.8% of the phenotypic variance, and two QTLs including qHCTN5 and qHCTN7 were identified in Cross HC, explaining 1.4%-8.3% of the phenotypic variance. Of which qAJTN1, qAJTN5, qAJTN7-2, qAJTN9 and qHCTN5 were newly identified QTLs, and qAJTN7-1, qAJTN7-3 and qHCTN7 were consistent with previous reports. Meanwhile, we screened the insertions and deletions in the QTL regions compared with the reference genome, insertion-deletion (InDel) markers linked with tillering were developed. These results of this study will facilitate the revelation of the genetic mechanism of tillering and molecular marker-assisted breeding of foxtail millet.
2018 Vol. 33 (6): 33-41 [Abstract] ( 65 )   全文: HTML   PDF (1011 KB)  ( 29 )
42 Cloning and Disease Resistance Verification of Chitinase Gene in Peanut
GUO Yue, JIANG Pingping, PAN Leilei, ZHOU Wenjie, XU Lei, ZHANG Ruqin, SUI Jiongming, GUO Baotai, WANG Jingshan, QIAO Lixian
DOI: 10.7668/hbnxb.2018.06.006
The chitinase gene was cloned and its function was further analyzed by genetic transformation, aiming at exploring the role of chitinase gene in resisting fungal diseases in peanut.The DNA (1 779 bp) and cDNA (795 bp) of peanut chitinase gene were obtained successfully by PCR and RT-PCR amplification, with the genomic DNA and cDNA of peanut variety Huayu 23 as templates respectively.The sequence alignment result between DNA and cDNA showed that peanut chitinase gene contained three exons and two introns, conforming to "GT……AG" rule in intron splicing. The cDNA coding sequence contained 795 bp, coded for a 265-amino acid protein, and was named by Ah-Chi, which was registered in NCBI (GenBank accession No. HQ439775). As determined by Blast analysis, the Ah-Chi protein had homology with proteins from Oryza sativa (83%), Zea mays (83%), Medicago sativa (72%), Glycine max (58%) and Arabidopsis thaliana (49%). The over-expression vector pCAMBIA1301-Ah-Chi was constructed successfully by substituting Gus of pCAMBIA1301 for Ah-Chi, and then was transformed into peanut embryonic leaflets explants by Agrobacterium EHA105 mediated transformation. The regenerated plants were then obtained by grafting and transplanting those regenerated somatic embryo seedlings.These transgenic positive plants were further screened and verified by PCR amplification, and the increased expression level of Ah-Chi was confirmedin transgenic plants by RT-PCR amplification. Cercosporidium personatum was inoculated to the detached leaves of transgenic and non-transgenic plants. Seven days later, it was found that the non-transgenic plants showed more serious leaves' browning and necrosis than transgenic plants, which indicated that the disease resistance of transgenic plants were enhanced.
2018 Vol. 33 (6): 42-48 [Abstract] ( 31 )   全文: HTML   PDF (1261 KB)  ( 40 )
49 Molecular Detection and Evaluation of Disease Resistance Genes of 39 Introduced Wheat Germplasms
DONG Na, CHEN Xiangdong, HU Tiezhu, LI Gan, ZHANG Yajuan, RU Zhengang
DOI: 10.7668/hbnxb.2018.06.007
To preliminarily clarify the composition of the resistance genes to stripe rust and powdery mildew in 39 introduced wheat germplasms, co-segregated or tightly-linked markers were used to detect stripe rust resistance genes Yr5, Yr10 and Yr18, and powdery mildew resistance gene Pm4, Pm13 and Pm21. Simultaneously, the disease resistant level of these germplasms were evaluated in the field. The results indicated that there were 6 germplasms carrying Yr18, which showed stable stripe rust resistance and the reaction type was varied from near immune to middle resistance. 21 germplasms were Yr5 linked marker S1320 positive, and 2 ones were Yr10 linked marker SC200 positive. These same marker positive germplasms showed differential disease resistant character, which might be associated with the genetic backgrounds of the carrier varieties. In assistant breeding, the detection results of these markers should be further confirmed by inoculation identification. The resistance of Pm4 to powdery mildew was almost lost, for only 1 of the 7 germplasms carrying Pm4 was of disease resistance. In all 39 germplasms, the powdery mildew resistance genes Pm13 and Pm21 could not be detected. Additionally, 2 germplasms Aoayou No.1 and bermude were of both resistance to stripe rust and powdery mildew, which could be used appropriately in disease resistance breeding.The results would be helpful for parental selection and rational utilization of germplasm resources in wheat disease resistance breeding.
2018 Vol. 33 (6): 49-55 [Abstract] ( 51 )   全文: HTML   PDF (1109 KB)  ( 25 )
56 Identification of PCD Induced by Salicylic Acid and Expression Analysis of Genes Related with the PCD in Cucumber
CHEN Chong, LIU Shuang, WANG Dandan, CHI Chunyu, ZHU Hong, JIN Xiaoxia, DING Guohua
DOI: 10.7668/hbnxb.2018.06.008
To reveal the role of chloroplast in the PCD of cucumber induced by salicylic acid (SA),10 mmol/L salicylic acid was added to the leaves of cucumber seedlings at the four-leaf period,and the samples were collected at the location where SA was added. ROS in situ detection,Trypan blue staining,PI and FDA staining,and TUNEL detection were performed. These results showed that SA could induce PCD process in cucumber. According to the results of high-throughput sequencing,15 genes annotated with chloroplast were screened out among 1 759 differentially expressed genes. These genes were expressed in cucumber leaves treated with SA was confirmed by RT-PCR. It was preliminarily identified that these genes were involved in the PCD process induced by SA in cucumber. According to the expression of qRT-PCR,the 15 genes played a role of positive regulation or negative regulation in PCD induced SA in cucumber,upon regulating characteristics five kinds of genes were raised:Probable thylakoidal processing peptidase 1 gene, 6-phosphogluconate dehydrogenase gene, Probable fructose-bisphosphate aldolase 3 gene, Alpha-glucan water dikinase 1 gene and Allene oxide cyclase 4 gene, down-regulated expression of 10 genes were:Betaine aldehyde dehydrogenase 1 gene, Calcium sensing receptor gene, Pheophorbide a oxygenase gene, Probable chlorophyll(ide) b reductase1 gene, Sigma factor binding protein 1 gene, NADPH-protochlorophyllide oxidoreductase gene, 2-succinylbenzoate-CoA ligase gene, ATP-dependent zinc metallopro-tease FtsH1 gene, Nicotinamide adenine dinucleotide transporter 1 gene and CBS domain-containing protein1 gene. It lays a foundation for further revealing the role of chloroplast in cucumber PCD induced by SA.
2018 Vol. 33 (6): 56-63 [Abstract] ( 38 )   全文: HTML   PDF (1185 KB)  ( 20 )
64 Cloning and Expression Analysis of Phenylalanine (PAL) Gene in Vitis amurensis
CHEN Meng, ZHANG Xue, ZHANG Yu, YANG Minghui, LIU Haifeng
DOI: 10.7668/hbnxb.2018.06.009
To investigate the expression of key enzymes affecting the composition of anthocyanins in the transformation stage of grape peel of Vitis amurensis. With the young leaves of Vitis amurensis as the test material, the genomic DNA of Vitis vinifera was extracted and the full length sequence of PAL (GenBank accession number:MH045991) was obtained by homologous cloning, and conducted a biological information analysis. The Q-PCR method was used to analyze the expression levels of 8 grapevine skins at different pigmentation stages. The results showed that the DNA sequence of the PAL gene was 1 763 bp in length and had a 1 671 bp open reading frame (ORF) encoding 556 amino acids. The molecular weight was 61.07 ku and the isoelectric point was 5.76, which was a stable protein. The whole polypeptide chain had a transmembrane helix structure with no signal peptide. Molecular phylogenetic tree analysis showed that the homology was higher with European and Asian grape species; The expression pattern of PAL gene in the 8 stages of the grape rind of P. villosa. The PAL gene was analyzed in the last 4 periods. With irregular expression levels, the PAL gene was hardly expressed during both the 50% color shift and 100% color shift. The regulation of PAL gene expression was regulated by many factors. It changed regularly in the early stage of growth and development of Vitis vinifera. In the last 4 periods, the expression level changes irregularly. PAL gene was in 50% color rotation and 100% color rotation. Almost no expression, which may be affected by some kind of inhibition of PAL enzyme activity factors.
2018 Vol. 33 (6): 64-71 [Abstract] ( 33 )   全文: HTML   PDF (1620 KB)  ( 23 )
72 Cloning, Molecular Characteristics of ACBP Gene and Its Expression Profile Analysis in Binglangjiang Buffalo
SHENG Dan, LIU Qin, TENG Xiaohong, ZHANG Yongyun, JI Li, MIAO Yongwang
DOI: 10.7668/hbnxb.2018.06.010
The coding product of ACBP gene is an important transport protein for long chain acyl coenzyme A esters in cells. Recently, it is found that this gene is closely related to lactation traits in common dairy cows, but it is rarely reported in water buffalo. The purpose of this study is to explore the sequence and gene expression characteristics of buffalo ACBP gene in order to reveal its function. The coding region of ACBP gene of Binglangjiang buffalo was cloned by RT-PCR method, and the gene was then subjected to analysis of functional bioinformatics and gene expression profiling. The sequence of complete coding region of ACBP gene in Binglangjiang buffalo was 264 bp in length which encoded a protein composed of 87 amino acid residues. This protein had a ACBP functional domain and contained 4 phosphorylated sites. It had no signal peptide and transmembrane region and was a hydrophilic protein located in the cell. The homology analysis of ACBP protein showed that Binglangjiang buffalo and the species of bovine subfamily were clustered together, which revealed the functional similarity of their ACBP gene.In the 13 tissues assayed, buffalo ACBP gene was highly expressed in the rumen, brain, heart during lactating stage and this gene was highly expressed in the rumen, heart, spleen during non-lactating stage. In addtition, ACBP gene was expressed in mammary gland, small intestine and liver during lactating stage, however it was not expressed in those tissues during non-lactating stage.The ACBP of buffalo and other species had similar amino acid composition and structural characteristics, which might participate in the transport of long chain acyl coenzyme A esters and plays an important role in the synthesis of milk fat.
2018 Vol. 33 (6): 72-79 [Abstract] ( 22 )   全文: HTML   PDF (1146 KB)  ( 28 )
80 Construction of Expression Vector and Expression Analysis of Tobacco Protein Kinase Gene NtCIPK3
CHEN Qian, YANG Shangyu, ZHUO Wei, LI Jiahao, PENG Shuang, WANG Jing, LI Liqin
DOI: 10.7668/hbnxb.2018.06.011
In order to study the function of CIPK3 in abiotic stress response,a CIPK family gene NtCIPK3 was cloned from common tobacco K326 by homologous cloning. The gene contained a 1 272 bp ORF encoding 423 amino acid. Protein sequence analysis indicated that the protein contained a transmembrane domain and belonged to a hydrophilic membrane protein. Protein alignment analysis revealed that NtCIPK3 contained a highly conserved N-terminal kinase region,a junction region,and a C-terminal regulatory region,with the highest homology to wild tobacco CIPK3,reaching 99%.Subcellular localization predictions showed that the protein was mainly located in the nucleus and had a bipartite typing nuclear localization signal sequence. The expression characteristics of this gene were analyzed by qRT-PCR. Analysis of tissue expression revealed that the gene was expressed in the roots,stems,leaves and flowers of tobacco and had tissue-specific expression and the expression level in leaves was significantly higher than that in other tissues,and it was speculated that it might function mainly in leaves.Meanwhile,NtCIPK3 gene was induced by low potassium,high salt,drought,ABA,H2O2,and low temperature treatment. It is speculated that NtCIPK3 gene played an important role in tobacco abiotic stress response. And successfully constructed the pBI121-NtCIPK3 overexpression vector,which laid the foundation for the future research of the gene under abiotic stress.
2018 Vol. 33 (6): 80-87 [Abstract] ( 35 )   全文: HTML   PDF (1223 KB)  ( 17 )
88 Analysis of the Relationship Between NtCIPK Family Gene Expression and Potassium Content in Tobacco
YANG Shangyu, ZHUO Wei, CHEN Qian, JIANG Yao, TONG Zhu, LI Liqin, REN Xueliang, LU Liming
DOI: 10.7668/hbnxb.2018.06.012
To explore the relationship between tobacco NtCIPK family gene expression and the potassium content of tobacco, and to provide reference for the molecular mechanism of potassium absorption in tobacco. For common tobacco varieties K326, Guiyan 5 and Yun 87 three different potassium concentrations (6.0, 1.0,0.2 mmol/L) were cultured in solution and their dry weight and potassium content were measured. At the same time, 11 genes of the NtCIPK family were analyzed for different potassium concentrations using qRT-PCR. Induced expression pattern. With the increased of potassium concentration in the culture medium, the dry weight and potassium content of the tested plants showed an increasing trend, and there were significant differences between treatments. Among them, Yun 87 is treated with different potassium concentrations (1.0, 0.2 mmol/L). Dry weight and potassium content were the highest. The expression of 11 genes in the NtCIPK family was induced by the change of potassium concentration, and showed up-regulated expression with the decreased of K+ concentration; only the expression of NtCIPK9 in K326 and NtCIPK24 in Guiyan 5 showed a downward trend.The results of path analysis showed that the expression of NtCIPK3 and NtCIPK5 was significantly positively correlated with the potassium content of K326 and Guiyan 5, respectively, at the concentration of 0.2 mmol/L K+, while NtCIPK23 was treated at the concentration of 6.0 mmol/L K+. The expression had a significant negative correlation with the potassium content per plant of Yun 87. The potassium content of tobacco was closely related to the expression of NtCIPK gene, and the enhanced expression of NtCIPK gene contributes to the absorption and accumulation of potassium by tobacco.
2018 Vol. 33 (6): 88-94 [Abstract] ( 28 )   全文: HTML   PDF (1113 KB)  ( 11 )
95 Prediction of miR-181a-5p Target Gene and Its Expression Analysis in Goose Follicle Granular Layer
MO Yuanliang, DENG Yan, WANG Jiwen
DOI: 10.7668/hbnxb.2018.06.013
To lay a foundation for further explore of the possible function and mechanism of miR-181a-5p in geese granulosa cell. Based on various network database, we predicted the potential targets and associated signaling pathways of miR-181a-5p through bioinformatics methods. Then the expression of miR-181a-5p and its targets in different stages granular layer were detected by qPCR.The results showed that miR-181a-5p was highly conserved in vertebrates. The GO and KEGG analysis revealed that miR-181a-5p mainly involved in the regulation of cell proliferation, differentiation and apoptosis. Most of target genes were mainly enriched in FOXO, MAPK, and PI3K-Akt signaling pathways. According to the interaction network of target genes, 10 core target genes (VEGFA, MAPK1, FOS, KRAS, HRAS, SIRT1, BCL2, ESR1, PTEN, CDKN1A) were screened out and most of these targets were associated with cell proliferation and apoptosis. The levels of miR-181a-5p was the highest in 2-4 mm follicles,but declined sharply in the 4-6 mm and showed an upward trend in the 4-6 mm, 8-10 mm, and F5 follicles.miR-181a-5p might regulate the SIRT1, BCL2, CDKN1B, PTEN, etc, then participate in FOXO, MAPK, PI3K-Akt signaling pathway to regulate the proliferation and apoptosis of granulosa cells.
2018 Vol. 33 (6): 95-102 [Abstract] ( 27 )   全文: HTML   PDF (1241 KB)  ( 26 )
103 Establishment of Mammary Epithelial Cell Line Transfected with HSP72 and Its Expression Identification
YU Wenhui, ZHAN Xizhen, FENG Yanni, CAO Rongfeng, JIANG Zhongling, LI Huatao, TIAN Wenru
DOI: 10.7668/hbnxb.2018.06.014
The objective of present study was to establish stably expressing HSP72 mammary epithelial cell (bMECs) line with expression vector of HSP72 slow virus constructed. The primers for PCR amplification purpose were designed and synthesized according to the template of bovine HSP72 gene sequence and was connected to plasmid virus. The bMECs were infected with the slow virus obtained after packaging. The cells with purpose gene or no-load virus were sorted out with puromycin and nuclear localization were determined by Hochest33342 method. The transfection efficiency was examined under an inverted fluorescence microscope. The positively expressed cell lines were verified by immunohistochemical technique and Western Blot method. The results showed that the titer of the lentivirus carrying HSP72 gene was about 1.0×109 TU/mL, the optimum screening concentration of purinamycin was 2 μg/mL, the bovine HSP72 lentivirus infected cells expressed HSP72, and the infected cow mammary epithelial cells could see green fluorescence under the inverted fluorescence microscope, and the transfection efficiency could be over 90%. The results showed that normal cells and empty carriers infected cells, HSP72 showed low expression, transfected with HSP72 target gene carrier cells, HSP72 high expression and dark brown color, and Western Blot assay proved that the expression of HSP72 in milk gland epithelial cells transfected by lentivirus empty carrier was slightly lower than that of normal transfected cells. There was no statistical difference, but the expression of HSP72 transfected by lentivirus carrier with HSP72 gene was highly expressed, and the difference was very significant (P<0.01). Compared with the mammary epithelial cells transfected with the lentivirus empty carrier, the HSP72 of the cells transfected with the lentivirus carrier with HSP72 gene showed high expression, and the difference was very significant (P<0.01).In conclusion the bovine mammary epithelial cell line, which stably expressing HSP72 protein, was successfully established. It laid solid foundation for further studying HSP72 function in inflammatory response of bovine mammary epithelial cells.
2018 Vol. 33 (6): 103-107 [Abstract] ( 19 )   全文: HTML   PDF (1002 KB)  ( 16 )
108 Polymorphisms of MyoG Gene and Analysis of Relevance with Growth Performances in Yak
CHAI Zhixin, XIN Jinwei, WANG Hui, GUO Lin, ZHONG Jincheng, ZHANG Chengfu, JI Qiumei
DOI: 10.7668/hbnxb.2018.06.015
The aims of this study were to compare the characteristics of maiwa yak in Sichuan, leiwuqi yak, shenzhayak, pali yak, sibu yak in Tibet, select the MyoG gene to detection of SNPs, to reveal the association between different genotypes and growth traits. This study provided a reference for the effective evaluation of the growth performance to the main yak population in Tibet. The DNA pool technology and direct sequencing were adopted to study in the genetic variation of the candidate genes. GLM statistical model of SPSS 17.0 software was used to analyze the association with different genotypes corresponding to the SNPs MyoG with growth traits, such as body height, body length, chest circumference, cannon circumference and body weight. The results showed that the growth indexes of shenzha yak were significantly lower than maiwa yak, leiwuqi yak and pali yak, but the traits of leiwuqi yak were better. Different ecological environments and climatic conditions directed affect grassland, vegetation types and their growth conditions, which were affect the feed intake and growth of yak. Four SNPs were found in MyoG gene (g.757 T>C,g.662 G>A,g. 539 A>G and g. 2216 A>G). There were three genotypes. The fitness test showed that MyoG gene on polymorphic loci in Hardy-Weinberg equilibrium state. The polymorphism of g.757 T>C,g.662 G>A and g. 539 A>G was higher in shenzha yak and pali yak, and lower in other groups. The different significance test indicated that the g.757 T>C,g.662 G>A and g. 539 A>G of the MyoG gene were significantly related to the body height(P<0.05). There were the major effect genes influencing the body height of the yak or loci might be closely linked to the major effect gene and they could be regarded as the genetic markers for the assistant selection.
2018 Vol. 33 (6): 108-115 [Abstract] ( 28 )   全文: HTML   PDF (1027 KB)  ( 15 )
116 Allelic Variation of Starch Synthesis Related Genes and Population Structure in Japonica Rice Cultivars
ZHANG Shanlei, YUE Hongliang, ZHAO Chunfang, CHEN Tao, ZHANG Yadong, ZHOU Lihui, ZHAO Ling, LIANG Wenhua, WANG Cailin
DOI: 10.7668/hbnxb.2018.06.016
To evaluate the allelic variation of starch synthesis related genes in Japonica rice cultivars, their population structure and genetic relationship, 34 molecular markers of starch synthesis related genes were used to detect the genetic diversity of 181 Japonica rice cultivars and their population structure and genetic relationship were analyzed combine with 185 SSR/Indel markers. A total of 74 alleles were detected by 34 molecular markers of starch synthesis related genes, ranging from 2 to 4, with an average value of 2.176 alleles per marker. Genetic diversity index ranged from 0.011 to 0.494 with an average value of 0.148. The polymorphism information content (PIC) ranged from 0.011 to 0.399 with an average value of 0.128. Six moderately polymorphic loci (0.25 < PIC < 0.50) and zero highly polymorphic locus (PIC>0.50) were detected, indicating that the genetic diversity of starch synthesis related genes in Japonica rice cultivars was not enough at present. Population structure analysis and UPGMA cluster analysis based on Nei's genetic distance divided the populations into three main groups (POP1-POP3) and one mixed group (Mixed). Most of the cultivars derived from the neighboring regions were clustered into one large group. It could provide a basis for genetic improvement of eating quality traits of rice, and lay a foundation for the subsequent association analysis of starch synthesis-related genes and starch quality traits.
2018 Vol. 33 (6): 116-122 [Abstract] ( 21 )   全文: HTML   PDF (1059 KB)  ( 20 )
123 Effects of Weak Light Stress on Physiological and Biochemical Characteristics and Relative Gene Expression of Yinhong Grape
HE Jingwen, LI Chen, QIU Tian, WU Yanyan, SHEN Zili, WU Yueyan
DOI: 10.7668/hbnxb.2018.06.017
The stress response and its physiological and relative gene characteristics of grape under the weak light conditions remains unclearly due to the lacking of research. The plant cell membrane system and metabolic process is also affected by low-light environment. As studies on response to weak light stress could provide for prevention of weak light adversity. Low light stress on one year Yinhong grape as the tested material with pot experiment, which was used to examine their growth changes, protective enzymes, osmotic adjusting materials, gene expression of MYC2 and TRX used Q-PCR under the different levels. Ranging from shading intensity of 0 to 85%(0, 25%, 40%, 70%, 85%).The results showed as follows:the growth and enzyme activities indicated that they all good at shading of T1, while the plant growth was gradually constrained with the enhanced weak light stress, SOD, POD, CAT, soluble protein activity, MYC2 and TRX gene expression increased firstly and then decreased gradually. At the same time, the activity of PRO and MDA was positively related with the degree of shade. The physiological indexes under higher intensity of shading (85%) were decreased significantly, which indicating that severe stress could cause irreversible damage by mediating itself and not adapt the weak light stress by regulating the cell protective enzymes and synthesizing the related genes. The conservation, the breeding of weak light varieties of grape was also discussed on the basis of the research results, it was also believed that more genes related to the weak light response need to be verified in the future to confirm whether they were induced by weak light.
2018 Vol. 33 (6): 123-129 [Abstract] ( 26 )   全文: HTML   PDF (1293 KB)  ( 28 )
130 Effects of Exogenous Calcium on the Development and Photosynthetic Characteristics of Peanut in Saline-alkali Soil and Normal Soil
TIAN Jiaming, ZHANG Zhimeng, DAI Liangxiang, ZHANG Guanchu, CI Dunwei, DING Hong, YANG Jishun, SHI Xiaolong, SHI Shubing
DOI: 10.7668/hbnxb.2018.06.018
In order to explore the effect of exogenous calcium on peanut planting in saline-alkali soil,Huayu 25 had been used as experimental material to study the effects of exogenous calcium on the agronomic traits and photosynthetic characteristics of saline-alkali soil and non-saline-alkali soil. The results showed that the main stem height,lateral branch length,leaf area index,SPAD value and net photosynthetic rate of peanut in saline-alkali soil were severely inhibited compared with non-saline-alkali soil. The application of exogenous calcium to non-saline-alkali soil increased the peak of leaf area index (LAI) of peanut and increased the net photosynthetic rate (Pn),SPAD value and leaf area index (LAI) of peanut leaves after pod setting phase,which could delay the senescence of peanut.The application of exogenous calcium to saline-alkali soil not only delayed the senescence of peanut after pod setting phase,but also reduced the degree of leaf SPAD value and the net photosynthetic rate increased by 55% before pod setting phase. When the amount of exogenous calcium was 52.2 kg/ha,the effect was not obvious,only to promote the growth of peanut plant height,with the increased of exogenous calcium application,the biological accumulation and its maximum growth velocity increased significantly,thereby increasing peanut kernel percent and yield. Under the experimental conditions,when the amount of exogenous calcium was 156.6 kg/ha,the yield of peanut increased by 43%. The results of this study would provide a theoretical basis for the rational use of calcium fertilizer in the practice of peanut production in saline-alkali soil.
2018 Vol. 33 (6): 130-136 [Abstract] ( 35 )   全文: HTML   PDF (1092 KB)  ( 51 )
137 Effect of Some Plant Growth Regulators Used by Stages Treatments on Endosperm Cell and Starch Granule in Maize Endosperm Development
LIU Xiaoqing, ZHAN Yanting, WANG Yue, WANG Ping, YIN Shikai, HAN Caixia, DONG Xuehui
DOI: 10.7668/hbnxb.2018.06.019
In order to study the effect of exogenous plant growth regulators on maize grain filling, maize inbred line B73 was used by some approaches such as cell counting and starch granule particle analysis to analyze the process of endosperm development of kernels. The main work focused on the development of grain filling, endosperm cells and starch granules after ethephon treatment in jointing stage and in combination with some plant regulators in silking stage. It was found that the grain filling rate in DAP 20 dropped by almost 17.7% in the ethephon treated plant, the 100-kernel weight in the ethephon treated plant decreased by 11.3% in contrast to the control plant in DAP 44. The endosperm cell number in the ethephon treated plant decreased significantly 22.9% compared with the control plant in DAP 12 and the starch granule number with the endosperm area of IV, V and VI in ethephon treated plant dropped notably. The starch granule diameter distribution in the ethephon treated plant (Treatment B) changed significantly in different stages after pollination compared with the control plant. The endosperm cell number and starch granule distribution showed nearly no difference in ethylene treatment in combination with abscisic acid treatment (Treatment E) in silking stage and in treatment B. Compared with the treatment B, the 100-kernel weight, the grain filling rate, the starch granule diameter distribution and the starch granule number per endosperm were all improved in some degree after ethylene treatment and with the compounding of 1-MCP (Treatment C) or 6-BA(Treatment D). The 100-kernel weight and the starch granule number per endosperm in treatment D were consistent with the control plant. This experiment provided a foundation for further research on exogenous plant growth regulators used in maize.
2018 Vol. 33 (6): 137-144 [Abstract] ( 35 )   全文: HTML   PDF (1404 KB)  ( 43 )
145 Study on High Yielding Effects of Row and Plant Spacing of Mechanized Sowing in Sesame
WU Yin, GAO Tongmei, WANG Dongyong, LI Feng, TIAN Yuan, ZENG Yanjuan, WEI Shuangling
DOI: 10.7668/hbnxb.2018.06.020
In order to explore the high yield effect of different seeding methods and clarify row and plant spacing of mechanized on high yielding in sesame. Field experiments were conducted to study functional-leaves photosynthetic characteristics, agronomic traits, yield and its components and grain quality of sesame under different row and plant spacing configuration with Zhengzhi 98N09 and Zhengtaizhi 1 in experiment base of Henan Pingyu in 2014. The results showed that the SPAD values and net photosynthesis rate in sesame functional-leaves were different under different row and plant spacing configuration. The net photosynthetic rate decreased with plant spacing reduced, effects of row and plant spacing on agronomic traits, yield and its components was great, while effects on quality of sesame was little. Among them, plant spacing had a greater impact on plant height, height of the first capsule, length of no capsule tip and length of main fruit axis, while row spacing had a smaller impact. The number of capsules per plant decreased with the decrease of plant spacing. The variation regularity of seed number per capsule with different varieties, but also showed the change rule with plant spacing reduced. 1000-grain weight was also decreased with plant spacing, and the 1000-grain quality of equal row spacing was higher than that of wide and narrow row spacing. The effect of row spacing on quality was not significant. The yield of sesame seed was the highest under the condition of row spacing was no more than 11.11 cm(the density was less than 225 000 seedlings/ha), under the condition of plant spacing allocation, not only the chlorophyll content of sesame leaves was higher, the photosynthetic assimilation ability of sesame leaves was stronger, but also the yield components were more coordinated, and the agronomic characters were better, which could be used as a reference for production.
2018 Vol. 33 (6): 145-151 [Abstract] ( 21 )   全文: HTML   PDF (1000 KB)  ( 37 )
152 Association Analysis of Verticillium Wilt-resistance in Upland Cotton Germplasm Population Based on SSR Markers
ZHANG Sujun, TANG Liyuan, LI Xinghe, WANG Haitao, LIU Cunjing, ZHANG Xiangyun, ZHANG Jianhong
DOI: 10.7668/hbnxb.2018.06.021
The molecular markers related with Verticillium wilt-resistance were explored by association analysis, which could provide candidate markers for disease resistance for use in marker-assisted breeding (MAS). In 214 upland cotton materials, 237 SSR markers were assayed for polymorphism, and cluster analysis was performed by PowerMarker v3.25, population structure was analyzed by Structure 2.2, an association analysis between SSR markers and Verticillium wilt-resistance traits were performed using the Tassel 2.1 GLM(general linear model) program. The results showed that the 237 SSR markers included 695 alleles belong to 280 loci,with an average of 2.479 0 alleles per SSR marker(range, 2-6). Structure analysis showed that all the accessions could be divided into 2 subgroups. Of the SSR loci, 27 were significantly(P<0.01) associated with Verticilium wilt-resistance under GLM program in different years, among which 2 marker loci (BNL3442 and BNL1064) were found to be consistent associated with Verticillium wilt-resistance in 3 years, and the ratio of explanation on the phenotype variation of above markers was 12.10% and 8.02% respectively. These stable SSR markers in different years are likely to be closely linked to disease resistance genes and are expected to be used in the screening of cotton Verticillium wilt-resistance material and resistance gene mining.
2018 Vol. 33 (6): 152-159 [Abstract] ( 36 )   全文: HTML   PDF (1200 KB)  ( 27 )
160 GhWRKY22 Isolation and Function Analysis in Cotton Resistance to Verticillium wilt
LEI Yu, ZHANG Zhennan, HU Guang, LIU Jianfen, TANG Ye, ZHANG Ning, SI Huaijun, WU Jiahe
DOI: 10.7668/hbnxb.2018.06.022
The development of plant disease-resistant cultivars by disease-resistant gene mining becomes a most available measure. The bioinformation,virus induced gene silencing,qPCR and fungus inoculation were employed to analyze cotton WRKY resistance to Verticillium dahliae. Eight cotton orthologous WRKY genes corresponding to 8 Arabidopsis defense-related WRKYs was selected by phylogenetic tree analysis. And their expression was induced by V. dahliae infection,indicating that they participate in cotton defense to this fungus. Among these genes,six GhWRKYs showed up-regulation expression, and expression levels of GhWRKY70 and GhWRKY48 exhibited up-or down-regulation over time. Five cotton WRKY genes including showed two expression peaks at 24,72 h after pathogen inoculation. GhWRKY22 gene was chosen for further charactering its function in cotton resistance to V. dahliae infestation.GhWRKY22 expressed predominantly in stem,and its expression level was obviously increased upon pathogen infestation and responded positively to the defense signaling molecules such as salicylic acid and jasmonic acid hormones. Knockdown of GhWRKY22 gene increased susceptibility of plants to V. dahliae infection. Additionally,the expression level of defense marker genes,including PR1,PR3,PR4,PR5,PAL and PDF1.2,significantly decreased in GhWRKY22-silenced plants compared to the control plants,which suggesed that GhWRKY22 participates in cotton defense against Verticillium wilt through SA and JA signaling pathways. Taken together,these results showed that 8 GhWRKYs were identified and characterized in regulation of resistance to V. dahliae. Among them,GhWRKY22 was a positive regulation component in cotton defense,which could be regarded as a candidate gene to breed plant defense cultivars.
2018 Vol. 33 (6): 160-168 [Abstract] ( 16 )   全文: HTML   PDF (1186 KB)  ( 26 )
169 The Damage and Identification of Bradysia odoriphaga on Chuanxiong
DU Lixin, SONG Jian, JIA Haimin, CAO Weiping, LI Yaofa, ZHANG Xiao
DOI: 10.7668/hbnxb.2018.06.023
In order to determine the occurrence and species of root maggot pests in cultivated Chuanxiong fields in Anguo City, Hebei Province, the larvae of root maggot were collected and reared in simulated field habitats. The species of root maggots of Chuanxiong in Anguo City were identified by using traditional morphological classification technology and modern molecular biological classification technology. The damage characteristics and degree of root maggots of Chuanxiong in Anguo City were systematically investigated by five-point sampling method. The results showed that the damage rate of root maggots was 4%-82%, the death rate of seedlings was 22%, and the average number of insects per plant was 37.76. The root maggots of Chuanxiong mainly damaged the root and stem of Chuanxiong through larvae, resulting in yellowing of the upper leaves of Chuanxiong, and the whole plant died when the roots were eaten completely. By comparing the traditional morphological characteristics of the adults, it was found that the morphological characteristics of the root maggot of Chuanxiong in Anguo were basically the same as those of the Bradysia odoriphaga, and the results of molecular marker identification showed that the sequence of mtDNA-COI gene of the root maggot of Chuanxiong in Anguo was 99% consistent with that of the B. odoriphaga. Root maggots were B. odoriphaga larvae. The results of this study showed that the root maggot species harmed and seriously affected the yield and quality of Chuanxiong in Anguo City of Hebei Province in recent years were B. odoriphaga, and the damage characteristics and degree of the root maggot of Chuanxiong were clarified, which would provide a scientific basis for the green control of the root maggot of Chuanxiong in the future.
2018 Vol. 33 (6): 169-173 [Abstract] ( 25 )   全文: HTML   PDF (1129 KB)  ( 22 )
174 Effects of Nitrogen Fertilizer Operation on Dry Matter Production and Yield of Maize Cultivars with Contrasting Nitrogen Efficiency
LI Qiang, KONG Fanlei, YUAN Jichao
DOI: 10.7668/hbnxb.2018.06.024
Two maize cultivars, nitrogen (N) efficient cultivar Zhenghong 311 (ZH311) and nitrogen inefficient cultivar Xianyu 508 (XY508) were used in a two years field experiments in 2015 to 2016. One N-free treatment and four N fertilizer operations were set up under the N application rate of 225 kg/ha (100% basal fertilizer, 75% basal fertilizer+25% topdressing, 50% basal fertilizer+50% topdressing, and 25% basal fertilizer+75% topdressing) to investigate the effects of N fertilizer operation on dry matter accumulation characteristics and yield, and the differences in response to N fertilizer operation on dry matter accumulation, distribution, translocation, and yield of maize cultivars with contrasting nitrogen efficiency. There were important effects of cultivars and N fertilizer operations on dry matter accumulation, distribution, translocation, and yield and its component. The stronger root uptake ability and higher proportion of photosynthetic organs in N-efficient cultivar ZH311 significantly increased its dry matter production ability, meanwhile the lower pre-silking dry matter translocation and translocation efficiency effectively delayed the senescence of photosynthetic organs and kept higher post-silking dry matter productivity resulted in a significantly higher yield than N-inefficient cultivar XY508. Increasing topdressing ratio significantly improved post-silking dry matter productivity and yield, while the promotion for N-inefficient cultivar XY508 was obviously higher than N-efficient cultivar ZH311. The yield advantage of ZH311 over XY508 increased first and then decreased with the increasing of topdressing ratio, reaching the maximum of 1.48 t/ha at 61.35% basal fertilizer+38.65% topdressing. In summary, a balanced basal-to-topdressing N operation is conductive to N-efficient cultivar ZH311 to maintain higher pre-silking dry matter translocation and post-silking dry matter productivity to full play yield advantages, whereas higher topdressing ratio can significantly improve post-silking dry matter productivity of N-inefficient cultivar XY508 to make up for the lack of yield.
2018 Vol. 33 (6): 174-182 [Abstract] ( 23 )   全文: HTML   PDF (1053 KB)  ( 27 )
183 Effect of NPK Fertilizers on Nutrient Uptake and Dry Matter Accumulation and Yield Components of Red Kidney Beans
JIN Fansheng, HAN Yanlong, LI Jie, LI Haijin, LI Xiaoping
DOI: 10.7668/hbnxb.2018.06.025
Study on effect of NPK fertilizers on nutrient uptake and dry-matter accumulation and yield components of red kidney beans, which could provide the theoretical basis for rational fertilization and efficient production of red kidney bean. This experiment was based on the Yingguohong red kidney bean as materials, field fertilizer experiment with nitrogen (N), phosphorus (P) and potassium (K) nutrient deficiency, collecting plant samples at full growth stage, measuring nutrient content, dry matter accumulation and yield constitute factor data. The results showed that the nitrogen, phosphorus and potassium fertilizer had significant effect on the yield of red kidney beans. In the yield component factor, 100 grain weight had the greatest influence on the yield formation. Nutrient limiting factors affecting yield was N > P > K. It could be seen from the unit content of N, P and K, N fertilizer had the greatest influence on the leaves, P fertilizer had the greatest influence on grain, K fertilizer had the greatest influence on the stem and pod skin.There was a clear reflux phenomenon in the mature period of N fertilizer. The accumulation amounts of red kidney bean plants during the whole growth period of N, P and K increase, the accumulation amount reached 161.15, 38.27, 126.70 kg/ha, the ratio was 4.21:1.00:3.31. The apex of N accumulation occured from early flowering period to full bloom stage, P and K appear from full bloom stage to pod bearing stage. The vegetative organ was the distribution center of dry matter accumulation on front stage of growth, dry matter accounted for 72.22% of the total. The reproduction organ was the distribution center of dry matter. Accumulation on later stage of growth, dry matter accounts for 70.26% of the total. The accumulation rate of dry matter was in the shape of a parabolic line on all growth period, from full bloom stage to pod bearing stage reached its peak. The accumulation of single plant was 0.952 g/d. The peak of the accumulating rate of each organ was different, root and leaf in early flowering stage, stem and pods in the full bloom stage, pod and grain in the pod bearing stage. The correlation coefficient between nitrogen accumulation and drymatter accumulation was 0.95 and 0.96 on pod bearing stage and mature stage, the performance was extremely remarkable. The correlation coefficient between phosphorus accumulation and drymatter accumulation was 0.93 on pod bearing stage, the performance was extremely remarkable. The correlation between potassium accumulation and drymatter accumulation was not remarkable. The correlation coefficient between pod number and drymatter accumulation of red kidney bean on early flowering stage was 0.95, the performance was extremely remarkable. The correlation coefficient between 100-grain weight and dry matter accumulation on pod bearing stage was 0.94, the performance was extremely remarkable. The correlation coefficient between yield and dry matter accumulation on full-bloom stage was 0.86, and the performance was remarkable. On pod bearing stage was 0.98, and the performance was extremely remarkable. The dry matter accumulation in the reproductive growth stage was the key factor affecting the yield results.
2018 Vol. 33 (6): 183-192 [Abstract] ( 25 )   全文: HTML   PDF (963 KB)  ( 30 )
193 Effects of Spraying Selenium on Selenium Content, Conversion Rate of Organic Selenium and Quality of Foxtail Millet in Different Stages
MU Tingting, ZHANG Fuyao, LI Zhihua, LIU Zhang, TIAN Gang
DOI: 10.7668/hbnxb.2018.06.026
Exogenous selenium was sprayed at different growth stages of the foxtail millet to study the change rule of quality and grain selenium content, to determine the optimum spraying selenium period for foxtail millet, so as to provide a theoretical basis for production of selenium-rich foxtail. The varieties Changnong 35, Jigu 20 and Jingu 50 were used as test materials. The field experiment was designed by random block group, setting spray water for control (CK), foliar spraying Na2SeO3 67.84 g/ha at seedling stage, heading stage and filling stage to study on influence of quality, grain selenium content, accumulation and translocation of selenium in foxtail millet. The result showed that different growth period of spraying selenium treatment all could improve quality of foxtail millet, foxtail millet in filling stage spraying selenium on millet quality traits to improve was the best. The lysine content of Jingu 50, Jigu 20 and Changnong 35 increased by 0.03,0.05,0.02 percentage points respectively, folic acid content was 2.4%, 7.5%, 5.5% higher than that of control, respectively.The content of crude protein and crude fat in selenium spraying treatment reached extremely significant level with the control during grain filling stage(P<0.01).Spraying time impact on grain selenium content in filling stage > heading stage > seedling stage, foxtail millet grain selenium content was increased by average of 8.0-9.9 times compared to the CK within the scope of the safety of the intake of selenium. Filling stage was to improve the millet organic selenium conversion rate and the utilization ratio of the grain selenium crucial period.The organic selenium conversion rates of Jingu 50, Jigu 20 and Changnong 35 treated by spraying selenium at grain filling stage were 13, 13 and 10 percentage points higher than those of the control, respectively, and there was a significant difference between them (P<0.05), compared with the control, the selenium harvest index increased by 5.32, 5.82 and 2.70 percentage points after selenium treatment in the filling stage. The application of selenium in filling period could improve the quality of grain, increase the grain selenium content, the conversion of organic selenium and the utilization of selenium. Filling stage was an optimal period of foxtail millet foliar spraying exogenous selenium.
2018 Vol. 33 (6): 193-198 [Abstract] ( 28 )   全文: HTML   PDF (1035 KB)  ( 37 )
199 Study on the Activities of Rhizosphere Soil Enzyme in Different Fertilization Modes During Maize Growth
LIU Lei, WANG Mengliang, WANG Junhong, GAO Xiaoyin
DOI: 10.7668/hbnxb.2018.06.027
In order to reveal the changes of soil enzyme activities under different fertilization treatments and fertilization methods, field experiments were conducted to study the effects of four different microbial fertilizers on soil enzyme activities of three main rhizosphere during maize growth periods. The results showed that the activity of soil enzymes was similar in maize growth period. The activity of sucrase and urease reached the peak at jointing stage, and catalase activity was the highest at maturity stage. Under the lateral application and spread application methods, the mixed application of several microbial fertilizers increased the activity of catalase, while increasing application of Bacillus amylolyticus microbial fertilizer quality helped the activity of invertase and urease to be increased, and the activities of sucrase and urease were increased by 6.3%-9.0% and 4.0%-10.5%, respectively, while the activities of sucrase and urease were increased by 20.7%-68.0% and 18.6%-37.4%, respectively. Compared with the control, the activity of urease were increased by 21.8%-30.4% and 4.6%-14.4%, respectively at different stages. The fertilization methods indicated that the application of strip fertilizer could increase the comprehensive utilization rate of fertilizers and facilitated the full absorption of crops. The research results reflected that the combined application of inorganic fertilizer and microbial fertilizer could improve the rhizosphere soil of maize, increase soil enzyme activity and soil fertility, which enhanced the sustainable productivity of soil. Therefore, the research results could provide a theoretical basis for rational utilization of microbial fertilizers on maize.
2018 Vol. 33 (6): 199-204 [Abstract] ( 22 )   全文: HTML   PDF (1129 KB)  ( 34 )
205 Effects of Different Fertilization Regimes on Maize Field Nematode Community in Fluvo-aquic Soil in North China
KONG Yun, ZHANG Ting, LI Gang, YANG Dianlin, ZHAO Jianning, ZHANG Guilong, WANG Lili, XIU Weiming
DOI: 10.7668/hbnxb.2018.06.028
In order to better understand the influence of different fertilization regimes on nematodes community of maize field in fluvo-aquic soil, this study was conducted in the Wuqing experimental station, Agro-Environmental Protection Institute, Ministry of Agriculture. The field experiment included six treatments, i.e., contrast (A0), organic fertilizer (A1), reduce 50% chemical fertilizer plus organic fertilizer (A2), normal chemical fertilizer plus organic fertilizer treatment (A3), increase 50% chemical fertilizer plus organic fertilizer (A4), chemical fertilizer (A5). The nematodes community composition and ecological structure were analyzed. The results showed a total of 18 nematode genera were found in all the treatments, in which A0, A1, A2, A3, A4 and A5 had 16, 17, 18, 16, 15 and 14 genera, respectively.Helicotylenchus was dominant genus. The chemical fertilizer treatment (A5) reduced the species and the total amount of nematodes, and also the number of each nematode nutrient group, which was not beneficial to the stable of soil nematode community structure. In the combination of chemical fertilizer and organic fertilizer treatments (A2-A4), total amount of nematodes, number of bacterivores and fungivores had a downward trend with the increasing of the amount of chemical fertilizer, while the number of bivorous and omnivores-predatores increased first and then decreased. The combination of excessive chemical fertilizer and organic fertilizer reduced the evenness distribution of nematode species, made the species diversity to decrease, which was not conducive to the stability of the nematode community structure. In conclusion, the treatment of reduced 50% chemical fertilizer plus organic fertilizer (A2) could improve the soil nutrient, reduce the degree of soil disturbance, stabilize the food network, and be more conducive to the stability of the nematode community. It also reduced environmental pollution and accorded with the state's policy of controlling fertilizer application.
2018 Vol. 33 (6): 205-211 [Abstract] ( 25 )   全文: HTML   PDF (1056 KB)  ( 29 )
212 Effect of Regulating Water and Fertilizer on Maize Growth under Different Tillage Depth
SUI Kaiqiang, FU Liya, HAN Wei, LIN Shaowen, LIU Shutang, HUANGFU Chenghui
DOI: 10.7668/hbnxb.2018.06.029
In order to study the effects of water and fertilizer ratio on maize nutrient and yield under different tillage depth,16 experimental treatments were designed to analyze the nitrogen,phosphorus,potassium distribution and yield components of maize organs. The results showed that under the same water and fertilizer conditions,the yield of 30 cm of tillage depth was higher than the depth of cultivation of 20 cm, the maximum yield of treatment was A1M2W3,the maximum yield was 9 950 kg/ha,which increased 46.32% than that of A1M1W0.Nitrogen accumulation in various organs of maize,the order of quantity was grain > leaf > stalk > root. Nitrogen content in stalk and root was the highest in treatment A1M2W3, and nitrogen accumulation in grain and leaf was the highest in treatment A1M2W2. The order of phosphorus accumulation in each organ was:grain > leaf > stalk > root. The order of potassium accumulation in each organ was stem > grain > stalk > root. The highest potassium content in each organ was A1M2W3. When the tillage depth was the same as that of organic fertilizer(A1M2),the yield of each treatment increased with the increased of water content.The percentage of nitrogen content in grain to the whole corn plant nitrogen content was 49.57%,49.29%,45.21%,43.82%.The amount of phosphorus was 43.61%,36.69%,43.31%,40.88% and the amount of potassium was 26.05%,29.27%,28.65%,27.79%.Under different tillage depth,proper water and fertilizer ratio had a significant effect on maize yield. Under the control of water and fertilizer level,the tillage depth 30 cm increased the biological yield of maize and the accumulation of nitrogen,phosphorus and potassium in all organs of maize compared with the depth of 20 cm. At the tillage depth of 30 cm and the moisture value of 70%,the absorption and accumulation of nitrogen,phosphorus and potassium in maize were the largest when 7 500 kg/ha was applied to organic fertilizer. Deep rotation can change the soil structure,promote the absorption of water and nutrients by crops,promote the growth of maize roots,enhance the ability of roots to absorb water and nutrients,and improve the absorption and accumulation of nutrients to promote the growth and development of maize.
2018 Vol. 33 (6): 212-218 [Abstract] ( 24 )   全文: HTML   PDF (1157 KB)  ( 27 )
219 Effect of Different Irrigation Mode on Yield Formation Characteristics of Winter Wheat
LÜ Lihua, WANG Xueqing, HUANG Jinan, ZHANG Jingting, DONG Zhiqiang, JIA Xiuling
DOI: 10.7668/hbnxb.2018.06.030
The purposes of this study were to confirm yield formation characteristics of the integrated mode of micro-irrigation under limited water condition. A field experiment was carried out with split plot design (main plot of irrigation mode and subplot of sowing date) in Gaocheng of Hebei Province from 2015 to 2017 year to research the effects of two types of integrated irrigation modes on the growth,yield formation and water consumption characteristics of winter wheat under two sowing date conditions. The two irrigation methods were micro sprinkler irrigation(Micro-irrigation) and border irrigation,with two sowing date of Oct.10 and Oct.15. The results showed that wheat yield formation characteristics of micro-irrigation were difference in two conditions,one was normal precipitation of adequate soil water storage before sowing (2015-2016),and the other was dry precipitation of insufficient soil water storage before sowing (2016-2017). The year of adequate soil water storage before sowing and normal precipitation,growth index and yield traits of micro-irrigation mode were similar or slightly below than the border irrigation under suitable sowing date (Oct. 10),while it had more obvious advantage for micro-irrigation mode under delayed sowing (Oct. 15). Compared with border irrigation,when sowing date was postponed wheat grow faster,and LAI,leaf SPAD value of later stage and grain filling rate were significantly higher for micro-irrigation mode,so its grain weight was increased by 5.2%-8.8%,and grain yield was increased by 3.4%-3.5%. Compared with border irrigation,soil water consumption was higher,total water consumption was decreased significantly by 3.8%-4.5%,and WUE was increased significantly by 7.3%-8.5% for micro-irrigation. The water consumption of late sowing wheat was lower than that of suitable sowing wheat,decreased by 3.8%-4.5%,but the yield and WUE were obviously higher than that of suitable sowing wheat. The year of insufficient soil storage before sowing and dry precipitation,the total irrigation amount was low for micro-irrigation,so its advantages of water and fertilizer integration could not be played,so for micro-irrigation,its LAI,leaf SPAD value and grain filling rate in the late growth stage was lower than that of border irrigation under two sowing date condition,and finally resulting in lower yield,and grain yield was decreased by 6.6%-13.0%. So the year of adequate soil water storage before sowing and normal precipitation year,the wheat was recommended micro-irrigation under late sowing condition. When this technical measure was adopted,irrigation amount of 57.7 mm was saved,and grain yield and WUE were increased by 4.1% and 16.7% respectively,and the contribution to yield was similar for two technical of irrigation methods and postponed sowing date.
2018 Vol. 33 (6): 219-226 [Abstract] ( 33 )   全文: HTML   PDF (1110 KB)  ( 29 )
227 Effects of Bio-bacteria-fertilizer Combined with Biochar on Soil Properties and Dissolved Soil N and C UV Spectral Characteristics in Tobacco Growing Soil
LI Ying, LI Bin, LIU Dongyang, JIANG Guiying, SHEN Fengmin, LIU Shiliang
DOI: 10.7668/hbnxb.2018.06.031
A field experiment was conducted to study the effect of compound-organic-bio-bacteria-fertilizer or/and biochar on soil properties and dissolved soil N and C in the tobacco-growing soil.Seven treatments were set as:①no organic fertilizer (CK),②biochar(T1),③Compound-organic-bio-bacteria-fertilizer (COBBF, T2),④Bio-bacteria-biochar-fertilizer(T3),⑤50% COBBF+50% biochar(T4)⑥60% COBBF+40% biochar(T5),⑦40% COBBF+60% biochar(T6). The result showed the soil nutrient was significantly improved under treatments with organic fertilizer application.The ammonium nitrogen content under T2 treatment was significantly promoted after tobacco transplanted 60 d compared with other treatments.The nitrate nitrogen content under T5 treatment was improved in the whole growth stage. The soil available potassium content under T2 and the soil available, phosphorus under T5 treatments were increased after tobacco transplanted 60 d compared with other treatments.The DOC content and DOC/SOC were obviously promoted under T1 and T3 treatments compared with other treatments, the highest DOC/SOC ratio as 8.31 mg/g occurred under T3 treatment. The DON content under T2 treatment peaked as 57.09 mg/kg after tobacco transplanted 90 d, while which reached as 97.98 mg/kg under T5 treatment during tobacco transplanted 60 d; Aromaticity index (AI) was reduced under T3,T4,T5 and T6 treatments. The E2/E3 was increased during tobacco transplanted 90 d. In conclusion, in central area of Henan Province, it is suggested that the COBBF only and 60% COBBF+40% biochar application were the optimal practice to improve soil N content;while biochar only and Bio-bacteria-biochar-fertilizer application could improve the soil dissolved C.
2018 Vol. 33 (6): 227-234 [Abstract] ( 34 )   全文: HTML   PDF (1307 KB)  ( 32 )
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