ACTA AGRICULTURAE BOREALI-SINICA
 
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2018 Vol. 33, No. 4
Published: 2018-08-28

 
1 The Identification of Drought Tolerance-related Transcription Factors in Early Developing Tassel of Maize
WU Yongbo, HAO Zhuanfang, WANG Nan, SONG Jie, ZHOU Yueheng, LIU Bojuan, ZHU Hanyong, DI Hong, WANG Zhenhua, LI Xinhai
DOI: 10.7668/hbnxb.2018.04.001
To explore more drought-related transcription factors genes in early growth and development of tassels, two drought-tolerant (DT) Tie 7922, X178 and two sensitive maize inbred lines (DS) Ji 81162, Dan 340 were used. The potted planting method was used to cultivate the seedlings, and the materials were treated by artificial simulated field drought stress and normal irrigation under the same and suitable water conditions. RNA-Seq technique was used to detect the changes in transcriptional level of the gene in drought related transcription factor family in the early stage of maize ear development compared with the normal irrigation treatment. The results showed that 287 genes were differently expressed under drought stress, and 34 of them are related to drought tolerance regulation revealed by GO enrichment and annotation, including 6 commons in two drought-tolerant line, 3 commons in two sensitive line, and all with uniform expression pattern, and 1 detected in four inbreds. The results provided the basic knowledge for drought tolerant gene identifition and molecular breeding of maize.
2018 Vol. 33 (4): 1-8 [Abstract] ( 53 ) HTML (1 KB)  PDF (1110 KB)  ( 75 )
9 The Temporal Spatial Expression of Gibberell in Key Metabolic Enzyme in Upland Cotton
SHI Jianbin, WANG Ning, ZHOU Hong, XU Qinghua, QIAO Wenqing, YAN Gentu
DOI: 10.7668/hbnxb.2018.04.002
To reveal the relationship between the key enzyme gene expression of gibberellin synthesis pathway and different tissues or stages of growth, the CCRI49 cotton was selected as material, and relative quantification Real-time PCR was developed to determine the gibberellin key gene expression of different cotton organizations in different stages of development. The results showed that, in seedling, the expression level of all gibberellin synthesis pathway genes was higher in stem, except GA2ox1 gene. In full-blooming stage, the expression of GA2ox1, GA3ox1, GID1B in root, CPS1, GA20ox1, GA3ox1, GID1B in stem and GA3ox1 in leaf had an up-regulation. In boll opening stage, the expression levels of GA2ox1 and GA3ox1 were up-regulated in root, while the remaining genes were down-regulated. The expression levels of KS and GID1B in stem were down-regulated, while the other genes were up-regulated, with GA2ox1 rise in maximum. And GA2ox1 in the leaf was down-regulated and the rest were up-regulated. In full-blooming stage, the high expression of GA20ox1 gene provided the necessary active gibberellin level for the elongation and growth of the plant stem, and the GA3ox1 gene might be associated with flowering and bell. In boll opening stage, the expression of GA2ox1 gene in root and stem gradually increased to reduce the synthesis of active, gibberellic acid. The gene expression levels of GA3ox1 and GA20ox1 in leaf tissues were raised to provide the necessary hormone levels for cotton boll growth and dehydration. The results indicated that cotton plant by changing the key enzymes gene expression of gibberellin synthesis metabolic pathway to regulate plant growth and development, and the purpose gene expression volume trends exist a certain difference along with the growth and development. It provides a theoretical basis for studying the regulation mechanism of gibberellin on upland cotton, which can help accelerate the improvement of variety and germplasm innovation.
2018 Vol. 33 (4): 9-16 [Abstract] ( 34 ) HTML (1 KB)  PDF (1101 KB)  ( 20 )
17 Cloning and Expression Analysis of Arginase Gene GhARG1 cDNA from Gossypium hirsutum L.
WANG Huifei, FENG Xue, ZHANG Yiming, FENG Lifeng, ZHANG Yu, CHEN Guang, SUN Yanxiang
DOI: 10.7668/hbnxb.2018.04.003
It is of great significance to investigate the response of arginase gene to abiotic stress and its physiological function in cotton. Bioinformatics analysis showed that the ORF sequence of GhARG1 was obtained by In silico cloning and RT-PCR technique. Salt stress, PEG6000 stress and ABA, SA and MeJA treatment cotton seedlings, and GhARG1 was expressed in E. coli prokaryotic. The ORF sequence contained 1 023 bp, and encoded 340 amino acids with typical conserved domains of arginase. Quantitative RT-PCR analysis showed that the expression of GhARG1 in leaves was up-regulated under stress condition of NaCl, PEG6000, and signal molecule of ABA, SA, respectively, but down-regulated by MeJA. The coding sequence of GhARG1 was fused to prokaroytic expression vector pCold-TF, and the combinant vectors were induced by IPTG and detected by SDS-PAGE. The molecule weight of protein GhARG1 was detected to be the expected 37 ku. The activity of arginase in the recombinant strain was 6 times that of the control. The results suggested that GhARG1, which coding product has arginase activity, might be involved in the response to salinity and drought stress through ABA and SA signaling pathway in cotton, and could lay foundation for further physiological functional identification.
2018 Vol. 33 (4): 17-24 [Abstract] ( 21 ) HTML (1 KB)  PDF (1357 KB)  ( 28 )
25 Cloning and Sequence Analysis of S and ORF3 Genes of Porcine epidemic diarrhea virus in Henan Province
ZHAO Li, HAN Haoying, ZHANG Hongxin, LU Tingting, WANG Wenjing, CHEN Hongying
DOI: 10.7668/hbnxb.2018.04.004
In order to analyze the S and ORF3 genes of PEDV in Henan Province, 150 PED suspected diseases were isolated from different pig farms in Henan Province from January to December, 2015, and the sequences of S and ORF3 genes were analyzed. The result showed that the nucleotide and amino acid homologies of S genes from 15 strains with CV777 were 93.6%-93.9% and 92.2%-93.1%, respectively. And some base insertions and deletions were detected. Compared with CV777, the nucleotide homology of ORF3 gene was 97.7%-100% and the amino acid homology was 93.7%-100%. Additionally, there was no base insertion and deletion compared with the vaccine strain. The Phylogenetic tree was based on the 15 PEDV of S gene amplified and the relationship of these strains were relatively distant from other reference strains. The relationships of the 15 strains, amplified based on the ORF3 gene, were closely related to domestic isolates, American strains and Korean strains. For the entire evolutionary relationship of S and ORF3 genes, 15 Henan strains in this experiment were relatively independent and their relationships with classic strain CV777 and other vaccine strains used in China were distant.The experiment analyzed of the S and ORF3 gene sequence features in prevailing of Henan region, which provided a support of molecular epidemiology study and prevention and control of PED in Henan Province.
2018 Vol. 33 (4): 25-30 [Abstract] ( 16 ) HTML (1 KB)  PDF (995 KB)  ( 12 )
31 RT-PCR Detection of Plum pox virus and the Screening of DNA Markers Linked to PPV-resistance in Apricot
ZHANG Junhuan, SUN Haoyuan, Boris Krška, YANG Li, JIANG Fengchao, ZHANG Meiling, WANG Yuzhu
DOI: 10.7668/hbnxb.2018.04.005
In order to rapidly detect the Plum pox virus (PPV) from the introduced apricot resources, and screening PPV-resistant materials from domestic resources, we employed RT-PCR and quantitative Real-time PCR, and the rapid detection technology system of PPV was established in apricot, by comparing and verifying these two methods. The PCR specific parameters of the PCR amplification system and reaction conditions were determined.RT-PCR amplifications were performed in 20 μL reaction mixture containing 10×PCR Buffer 2 μL, 25 mmol/L Mg2+ 1.6 μL, 10 mmol/L dNTP 0.4 μL, 0.5 U/μL Taq DNA polymerase 0.2 μL, both forward and reverse primer 0.4 μL, cDNA 1 μL, and ddH2O 14 μL. Reaction parameters for RT-PCR were as follows:94℃ for 5 min; 35 cycles of 94℃ for 45 s, 55℃ for 30 s, and 72℃ for 45 s; with a final extension step at 72℃ for 10 min. In addition, F1 population crossed from two different PPV-resistant apricot parents had been constructed by Faculty of Horticulture, Mendel University. Based on these F1 individual materials, we used SSR method coupled with BSA method to screen the markers linked to PPV-resistant genes. Firstly, the polymorphic markers were screened between the PPV-resistant gene pool and susceptible one, and six SSR loci (PGS1.23, PGS1.24, PaCITA5, UDAP-414, Pchcms4 and Pchgms4) had been screened out. The polymorphism between two gene pools and between two parents was consistent. Then these six locus were further verified among the F1 progenies. As the final results, one SSR locus PGS1.23 with a high degree of linkage to PPV resistance was identified. There was a high coincidence of 78.3% between expected marker genotype and phenotype for the PGS1.23. Furthermore, this linkaged marker was used to evaluate the PPV resistance for 37 Chinese apricot germplasms, and five cultivars with PPV-resistance including Caizihuang, Yangjiyuan, Chuizhishanxing, Dazaoshu and Xifuxing were screened out. These results would provide better parents materials for PPV-resistant apricot breeding, and also lay the foundation for the further study on the candidate PPV-resistant gene.
2018 Vol. 33 (4): 31-38 [Abstract] ( 18 ) HTML (1 KB)  PDF (1297 KB)  ( 15 )
39 Cloning and Sequence Analysis of Yak IGF-IR Gene
LIANG Chunnian, WANG Hongbo, WU Xiaoyun, CHU Min, GUO Xian, YAN Ping
DOI: 10.7668/hbnxb.2018.04.006
The complete cDNA encoding yak insulin like growth factor 1 receptor (IGF-IR) gene was cloned with RT-PCR and homologous cloning methods. Sequence analysis showed that the entire coding region was 4 104 bp in length encoding 1 367 amino acids with a putative mass of 154.95 ku and pI of 5.71. Compared to IGF-IR gene sequence from cattle, there were 20 base changes in yak IGF-IR gene, including 2 substitutions for G-T, 2 transitions for G-A, 7 transitions for C-T, 3 transitions for A-G and 6 transitions for T-C, respectively. In addition, amino acid changed at 3 sites (G6R, T29I and S30I). The phylogenetic analysis revealed that yak IGF-IR gene had high homology (> 92%)with that of cattle, pig, human, dog, mouse and chimpamzee, which were classified into the same branch. The secondary structure predication indicated that yak IGF-IR protein might have the typical domains of the IGF-IR family, containing the furin-like cysteine rich region (FU), fibronectin type Ⅲ domain (FN3), transmembrane domain (TM), and tyrosine kinase domain (TyrKc), respectively. 3D model showed the mature fragment in yak IGF-IR had 98.68% homology with human IGF-IR. The present results provided the valuable foundation for further studies on yak IGF-IR functions.
2018 Vol. 33 (4): 39-45 [Abstract] ( 20 ) HTML (1 KB)  PDF (2091 KB)  ( 26 )
46 H-FABP Gene Polymorphism Screening and Protein Function Prediction in Gansu Black Pig
ZHANG Li, LIU Lixia, NONG Weilun, LI Huijie, ZHANG Guohua, LU Jianxiong
DOI: 10.7668/hbnxb.2018.04.007
In order to identify the genetic characteristics of the H-FABP gene, DNA pooling and direct sequencing technique were used for rapid screening of H-FABP gene polymorphism, the physicochemical properties of the encoded amino acids and the protein structure and function were predicted by bioinformatics in Gansu black pig. The result showed that 4 mutation sites were found in the region of the H-FABP gene amplification in the Gansu black pig. C734T-Intron1, T152C-Exon2, G30A-Intron2, T121G-Intron2, respectively, it was shown that the allele frequencies were significant differences before and after mutation;the CDS region of H-FABP gene in Gansu black pig was 402 bp, encoding 133 amino residues, and T152C were missense mutations, was found in CDS region. Amino acids were converted from threonine (Ile)to threonine (Thr). The H-FABP gene encoding protein of black pig was a stable hydrophilic protein, protein secondary structure and tertiary structure were mixed type, and the proportion of beta folding was the highest. The results of protein function prediction showed that there were 19 potential phosphorylation and 1 N-glycosylation sites, it might affect the increase of fat content in the H-FABP gene regulation muscle;Homology comparison and phylogenetic tree analysis showed that it had most similar genetic relationship between Artiodactyl and Gansu black pig. These provided a theoretical basis for the molecular genetic markers of the intramuscular fat content deposition in Gansu black pigs, the polymorphism screening of the H-FABP gene, it has become a hot spot of molecular research in the improvement of meat quality traits in domestic animals.
2018 Vol. 33 (4): 46-52 [Abstract] ( 20 ) HTML (1 KB)  PDF (1155 KB)  ( 15 )
53 Prokaryotic Expression of Blue Light Receptor Genes Cmwc-1 and Cmwc-2 and Detection of Expressed Proteins in Cordyceps militaris
ZHONG Xueqing, YE Dexiao, MAO Yumei, FU Mingjia, YANG Zhihai
DOI: 10.7668/hbnxb.2018.04.008
The blue light receptor protein is the factor that receives blue light signal in living organisms. To reveal the existence form of blue light receptor protein, the partial sequences of the blue light receptor genes Cmwc-1 and Cmwc-2 in Cordyceps militaris were constructed in prokaryotic expression vector pET-41a (+), and the correct expression of Cmwc-1 and Cmwc-2 was carried out in Escherichia coli. Using the expressed fusion protein as antigen to prepare antibodies, we obtained the antibodies of high titer of the C. militaris blue light receptor CmWC-1 and CmWC-2. By Western Blotting analysis, 42 ku (CmWC1-42)and 48 ku (CmWC1-48)protein for CmWC-1 were detected in mycelia and fruiting bodies of C.militaris, which in the mycelium was dominated by CmWC1-48, more than the CmWC1-42 relative quantity, while in the fruiting body was dominated by CmWC1-42, no CmWC1-48 was detected. 50 ku (CmWC2-50)protein for CmWC-2 was detected in both mycelia and fruiting bodies in C.militaris. These results suggested that CmWC-1 degraded differently in mycelia and fruiting bodies, while CmWC-2 remained stable after completing the biological functions of CmWC-1 and CmWC-2 complexes.
2018 Vol. 33 (4): 53-60 [Abstract] ( 13 ) HTML (1 KB)  PDF (1325 KB)  ( 13 )
61 Comparative Analysis of BLEC1 Gene Sequences in Different Chicken Breeds
ZHANG Xin, CHANG Guobin, ZHENG Shenghan, WANG Tongmiao, CHEN Bowen, ZHANG Yang, WANG Zhixiu, CHEN Guohong
DOI: 10.7668/hbnxb.2018.04.009
The BLEC1 gene (C-type lectin-like receptor 1)encodes a C-type lectin-like receptors that is associated with the activated antigen of the CD69 family and is closely linked to the innate immune function of poultry. In order to explore the genetic diversity of BLEC1 gene and different disease resistance of different chicken breeds. We used phenol and chloroform to extract the DNA of 26 different chicken breeds. Gallus gallus as a reference, the serial number was NC_006103.4, target sequence capture assay and MEGA 6 software was used to detect SNPs, Indels and amino acid sites in the BLEC1 gene of 26 chicken breeds. Finally, MEGA 6 software was used to construct phylogenetic trees. The results showed that 14 SNPs and 12 Indels loci were detected in 26 chicken breeds, of which 5 SNPs and 3 Indels loci were exon, 3 Indels sites on exons are widely present in chicken breeds. Indel1 was presented in all chicken breeds excepted Black Wolf Pheasant. These led to a great change in amino acids. In exons, 3 amino acids missense mutations, the rest of synonymous mutations. The neighbour joining cluster analysis was used to classify 26 chicken breeds into three groups, Anka chicken, SPF-Lai Hang chicken and Taihewugu chicken grouped together, Xiaoshan chicken alone for a categorize, the rest of the breeds were grouped together.All these suggested that BLEC1 gene was rich in genetic diversity in local chicken breeds in China and it was associated with innate immunity and disease resistance of different chicken.
2018 Vol. 33 (4): 61-66 [Abstract] ( 14 ) HTML (1 KB)  PDF (967 KB)  ( 23 )
67 Cloning and Expression of Chitinase Gene AO-190 of Arthrobotrys oligospora XJ-A1 and Analysis of the Chitinase Activity of Recombinant Protein
GONG Shasha, MENG Qingling, QIAO Jun, ZHONG Wenqiang, HUANG Yunfu, ZHANG Guowu, CHEN Ying, CAI Xuepeng
DOI: 10.7668/hbnxb.2018.04.010
Nematode-trapping fungi can secrete chitinase in the process of infecting nematodes.In order to study the functions and molecular characteristics of chitinase, chitinase gene AO-190 of Arthrobo trys oligospora XJ-A1 isolate was cloned, analyzed and expressed in Escherichia coli by constructing prokaryotic expression vector pET32a-AO-190. The prokaryotic expression recombinant fusion protein was purified by Ni column and the chitinase activity was purified by chitinase ELISA kit.The results showed chitinase AO-190 gene had a length of 1 574 bp and four intron sequences. The sequence contained one open reading frame (ORF)with 1 251 bp that encoded 416 amino acids.The homology of the chitinase AO-190 gene sequence was 93.33% and the amino acid sequence was 92.55% respectively with Arthrobotrys oligospora standard strain (ATCC 24927)chitinase AO-190. There was a signal peptide, a Arginine-rich region profile, a Bipartite nuclear localization signal, a N-glycosylation site, a Protein kinase C phosphorylation site, a N-myristoylation site, a Casein kinase Ⅱ Amidation site and a Low-Complexity region.The proteins belonged to the family 18 glycoside hydrolase with prevalent conserved substrate binding domains SLGG and catalytic domains VDGVDLDLE.The major structural elements of secondary structure included random coils, alpha helix and extended strand while a typical (α/β)8 phosphorylation site, a rounded bucket structure were predicted about tertiary structure.Phylogenetic analysis showed that the phylogenetic relationship of chitinase AO-190 was more close to the chitinase (EPS43772.1)originate from Dactylellina haptotyla who could produce short handle sticky ball and the chitinase (EWC46603.1) originate from Drechslerella stenobrocha who could produce constricting rings and had a significant evolutionary distance with chitinase of insects and vertebrates.The recombinant protein was identified by SDS-PAGE analysis that showed it had a molecular weight of about 63 ku, which was consistent with prediction. Western Blot analysis indicated that the recombinant protein could specifically react with polyclonal antibody against protein crude extracts.The purified recombinant chitinase activity was 222 IU/L that was measured using by chitinase ELISA kit.
2018 Vol. 33 (4): 67-74 [Abstract] ( 14 ) HTML (1 KB)  PDF (1405 KB)  ( 20 )
75 Study on Physiological Function of Arabidopsis CPK6 in Ca2+ Signal Transduction
YU Yahui, YANG Ju, ZHANG Yuchen, ZHANG Lin
DOI: 10.7668/hbnxb.2018.04.011
The objective was to research the physiological function of CPK6 in Ca2+ signal transduction and to reveal the mechanism of plants to adapt to Ca2+ deficient environment. By homozygous identification of the CPK6 deletion mutant (cpk6), two homozygous mutants, cpk6-2 and cpk6-3 were obtained. And the growth phenotype of wild-type Arabidopsis Col-0 and cpk6 mutants in the deficiency of Ca2+, Fe2+, Mg2+, Zn2+ and Mn2+ were analyzed. The results showed that the mutants showed a growth-inhibiting phenotype relative to wild-type Col-0 in the deficiency of Ca2+. The Arabidopsis transgenic material expressing CPK6 promoter-driven β-glucuronidase were obtained. Using GUS histochemical staining, it was found that CPK6 promoter had activity in the roots and the growth point of the leaves. And the activity of CPK6 promoter in leaves was visibly enhanced under Ca2+ deficient condition. The total Ca content of Col-0 and cpk6 was determined by atomic absorption spectrometry (AAS). It was found that there was no significant difference in Ca content between Col-0 and cpk6 under control and Ca2+ deficiency conditions. The results indicated that CPK6 gene did not participate in the absorption and accumulation of Ca in Arabidopsis. Using histochemical staining and fluorescent probes methods, it was found that the accumulation of H2O2 in the leaves and roots of cpk6 was visibly higher than that in Col-0 under Ca2+ deficient condition. The results showed that CPK6 was a negative regulator of the accumulation of H2O2 induced by Ca2+ deficiency in plants.
2018 Vol. 33 (4): 75-81 [Abstract] ( 22 ) HTML (1 KB)  PDF (1116 KB)  ( 24 )
82 Cloning and Expression Analysis of GsTIFY6B Associated with Saline and Alkali Stress in Glycine soja
YAN Wenfei, CHENG Fansheng, JIANG Xinqiang, LIU Cuixia, ZHU Dan
DOI: 10.7668/hbnxb.2018.04.012
In order to determine the molecular characterization and expression patterns of TIFY family genes involved in saline and alkali stress in Glycine soja, according to previous transcriptome data, GsTIFY6B was obtained by homologous cloning using cDNA of Glycine soja G07256, with extremely tolerant ability to saline and alkali stress, as its template. The full-length CDS was 1 047 bp. GsTIFY6B protein was an unstable protein with 348 amino acids. Its molecular weight was 36.8 ku and its isoelectric point was 9.12. Phylogenetic tree analysis showed the closest homology of GsTIFY6B to the TIFY family proteins of GmTIFY6B, VrTIFY6B, CcTIFY6B and AdTIFY6A. All those proteins had the conserved domains TIFY and Jas. The transcription level of GsTIFY6B in different parts of Glycine soja as well as response to saline and alkali stress and hormone treatment was detected by Real-time fluorescence quantitative PCR. The results showed that the highest relative expression level of GsTIFY6B in roots. Under salt stress, the expression of GsTIFY6B in roots and leaves showed an up-regulation.Under alkali stress, the expression of GsTIFY6B in roots and leaves showed the first down-regulation and then up-regulation expression. It is speculated that the gene might be involved in the defense reaction of saline-alkali stress.Under the ABA treatment, GsTIFY6B showed down-regulated expression in the roots and up-regulated expression at 6, 12 h in leaves. Under MeJA stress, GsTIFY6B expression was decreased first and then increased in roots, but showed up-regulation in leaves. This study suggested that GsTIFY6B could positively respond to saline and alkali stress by participating in the ABA and MeJA signaling pathways. These results will provide a theoretical basis for the later study on the molecular mechanism of saline-alkali-tolerant response of Glycine soja.
2018 Vol. 33 (4): 82-89 [Abstract] ( 20 ) HTML (1 KB)  PDF (1269 KB)  ( 11 )
90 Cloning and Expression Analysis of CO4 Gene in the Different Light Qualities and Light Transfers of Grapevine
FAN Qi, MA Yanni, CHEN Baihong, ZUO Cunwu, MAO Juan
DOI: 10.7668/hbnxb.2018.04.013
To explore the effects of different light qualities on the expression of CO4 gene in grapevine plantlet in vitro. The full-length cDNA sequence of CO4 was cloned from Vitis riparia×V.labrusca Beta in vitro by RT-PCR. Bioinformatics analysis of the second level structure, and phylogenetic tree of protein. The result showed that the size of CO4 gene fragment was 663 bp, the open reading frame (ORF)of CO4 gene was 747 bp, including 248 amino acid coding regions. By use bioinformation analysis, CO4 of Beta was a hydrophobic, unstable protein with good lipid solubility. CO4 had a signal peptide, it can be inferred that it was a transmembrane protein. Phylogenetic tree indicated that CO4 of Beta (Vitis riparia×V.labrusca)had the highest evolutionary relationship with Eucalyptus grandis (Eucalyptus grandis Hill ex Maiden).qRT-PCR analysis showed that the CO4 expressed specifically and strongly in the treatment of red to blue light (control), which increased 5.39 fold in comparison to the control. The difference was not significant when exposed to the treatment of white to red light and red to white light. However the expression of CO4 was the lowest in the white to blue light treatment, which was 54% of the control. Fluorescence parameter analysis showed that the maximum photochemical efficiency (Fv/Fm), potential activity (Fv/Fo)and photochemical quenching coefficient (qP)were significantly higher than those of the control under red to blue light. The non photochemical quenching coefficient (NPQ)was the lowest under that treatment, but was the highest under red light treatment, it was the lowest under this treatment. The CO4 of Beta was more sensitive to light. The treatments of red to blue light could not only promote the expression of CO4, but also could improve the Beta Grapevine in vitro photosynthetic capacity. Thus, red to blue light played an important role in the process of morphogenesis of Beta Grapevine in vitro.
2018 Vol. 33 (4): 90-97 [Abstract] ( 22 ) HTML (1 KB)  PDF (1472 KB)  ( 13 )
98 The Development of a SYBRGreen Real-time qPCR for the Detection of Porcine Circovirus Type 2
FENG Hua, LIU Yunchao, CHEN Yumei, WEI Qiang, ZHANG Gaiping
DOI: 10.7668/hbnxb.2018.04.014
To develop a SYBRGreen Real-time quantitative PCR (qPCR)assay for PCV2 diagnose, the recombinant plasmid pMD19T-ORF2 was constructed as template based on PCV2 ORF2 (706 bp), and the specific primers were designed. The primer concentration and annealing temperature were optimized, respectively. Besides, the sensitivity, specificity, reproducibility and clinical samples detection of this method were further tested. The results showed that the detection limit for this assay was 3.0×102 copies/mL, while it was 3.0×104 copies/mL for conventional PCR. The melt curve analysis presented a single melt peak located at 77-79℃. And no cross reaction with CSFV, PPV, PRRSV and PRV was detected. The inter-assay coefficient of variation ranged from 0.29% to 0.32% and the inter-assay coefficient of variation ranged from 0.32% to 0.36%. In addition, a higher PCV2 positive rate was detected by SYBRGreen Real-time qPCR (81.5%, 31/38)compared with conventional PCR (68.4%, 26/38)in clinical samples detection. Therefore, the PCV2 SYBRGreen Real-time qPCR is more rapid, sensitive, specific and stable, which was more suitable for PCV2 clinical infection diagnose.
2018 Vol. 33 (4): 98-103 [Abstract] ( 10 ) HTML (1 KB)  PDF (1017 KB)  ( 10 )
104 Construction of Expression Vector and Expression Analysis of Tobacco ERF Gene NtRAP2-7
CHEN Qian, ZHUO Wei, LUO Jing, YANG Shangyu, LU Liming, LI Liqin
DOI: 10.7668/hbnxb.2018.04.015
In order to study the function of tobacco NtRAP2-7 gene in abiotic stress response, NtRAP2-7 was cloned from K326 by homologous cloning. And the bioinformatics software was used to analyze the physicochemical properties, spatial structure and phylogenetic analysis of the encoded protein. Sequence analysis results showed that the gene contained 1 398 bp in length which encoded 465 amino acid residues and the predicted molecular weight of this protein was 51.16 ku. Based on 3-D modeling and secondary structure analysis, the protein consisted of three β-sheet and one α-helix and contained 58 phosphorylation sites. The subcellular localization prediction showed that the protein was mainly located in the nucleus and contained a monopartite nuclear location signal sequence. Phylogenetic analysis showed that NtRAP2-7 protein had the highest homology with Nicotiana sylvestris RAP2-7 protein sequence, which was 98%.The expression pattern analysis of the gene was carried out by qRT-PCR. Tissue expression analysis showed that the gene was expressed in the roots, stems, leaves and flowers, but had the highest expression level in roots. The expression levels of NtRAP2-7 was affected by abiotic stress treatments, indicating that the NtRAP2-7 was involved in the tobacco abiotic stress response. The pBI121-NtRAP2-7 overexpression vector was successfully constructed by double enzyme digestion, which laid the foundation for further study on the function of this gene in abiotic stress.
2018 Vol. 33 (4): 104-111 [Abstract] ( 12 ) HTML (1 KB)  PDF (1274 KB)  ( 9 )
112 Genetic Analysis of Salt Tolerance Related Traits in Sorghum Recombinant Inbred Lines
LÜ Jianpeng, LI Jinwang, DUAN Xiafei, LI Yanling, LI Oujing, PEI Zhongyou
DOI: 10.7668/hbnxb.2018.04.016
Salt tolerance is an important agronomic trait of sorghum. Salt tolerance genetic analysis is important for sorghum breeding. In order to select new varieties of salt-tolerant sorghum and study the genetic rules of salt tolerance, 482 F5 recombinant inbred lines were obtained from the salt-tolerant cultivar Sanchisan and salt-sensitive cultivars Tx622B, and the germination rate, seedling height, fresh weight and dry weight were used to identify salt tolerance at seedling stage. Genetic analysis was carried out by single-generation combined quantitative trait analysis. The results showed that the sal-tolerance traits of the Sanchisan and salt-sensitive cultivars Tx622B were significantly different. The relative mean values of germination rate, seedling height, fresh weight and dry weight of the F5 generation group were all different among parents, and the tolerance to salt tolerance was different in different degree. The germination rate, seedling height, fresh weight and dry weight were analyzed. There were significant positive correlations between the two traits. The relative germination rate, relative seedling height, relative fresh weight and relative (B-5)genetic model of two pairs of major genes were found by genetic model analysis. Through genetic model analysis, it was found that salt tolerance of F5 population of recombinant inbred lines of sorghum was dominant to salt, and inheritance of salt-tolerance trait of sorghum was in line with complete dominant (B-5)genetic model of two major genes. The heritability of the major gene of germination rate was 40.94%. The heritability of major gene of seedling height was 31.22%. The heritability of major gene of fresh weight was 39.19%. The heritability of main gene of dry weight was 63.98%.The germination rate, seedling height, fresh weight and dry weight have high heritability as the main characters of salt tolerance evaluation, and have far-reaching significance for the selection and cultivation of sorghum salt-tolerant varieties.
2018 Vol. 33 (4): 112-119 [Abstract] ( 20 ) HTML (1 KB)  PDF (1030 KB)  ( 9 )
120 Genetic Diversity Analysis of Main Parents for Hybrid Soybean Based on SSR Markers
BAI Zhiyuan, ZHANG Ruijun, WU Guoping, LIAN Shichao, YANG Yuhua, WEI Baoguo
DOI: 10.7668/hbnxb.2018.04.017
To determine the genetic diversity and genetic relationship of the main parents of hybrid soybean in Shanxi Province, 64 pairs of SSR primers were used to analyze the genetic diversity of 32 hybrid soybean parents including 15 maintainer lines and 17 restorer lines. Totally 357 alleles were revealed among 32 parents, 2-12 alleles could be detected at different SSR locus, with the average of 5.578 alleles per SSR marker, of which 322 alleles were detected in the maintainer line, ranging from 2 to 9 with an average of 5.031, and the detection of 320 alleles in the restorer ranged from 2 to 12 with an average of 5. The polymorphism information content (PIC)of the total polymorphism was 0.368-0.900, with an average of 0.666, of which, the PIC variation of maintainer lines ranged from 0.370 to 0.868 with an average of 0.661, and the PIC variation of restorer lines ranged from 0.329 to 0.904 with an average of 0.630. Cluster analysis result showed that the average genetic similarity coefficient (GS)between maintainer lines was 0.637. At the genetic similarity coefficient (GS)of 0.61, the maintainer lines could be grouped into two major groups. At the similarity coefficient of 0.65, the second group could be divided into three subcategories. The average genetic similarity coefficient (GS)between restorer lines was 0.669. At the genetic similarity coefficient (GS)of 0.65, the restorer lines could be grouped into two major groups. The clustering results were related to the geographical origin of cultivars.
2018 Vol. 33 (4): 120-125 [Abstract] ( 13 ) HTML (1 KB)  PDF (940 KB)  ( 26 )
126 Expression of a Novel Xylanase Gene in Pichia pastoris
LI Qi, CHENG Lifeng, DUAN Shengwen, FENG Xiangyuan, ZHENG Ke, YANG Qi, LIU Zhiyuan, PENG Yuande
DOI: 10.7668/hbnxb.2018.04.018
To obtain a new extracellular xylanase in high yield, the xynA (GenBank number:U57819) can be expressed in different ways. A xynA was early expressed in prokaryotic microbiology. Subsequently, we cloned the partial xynA encoding mature xylanase into expression vector pPICZαA, and then transformed into Pichia pastoris X33, so eukaryotic expression system was successfully constructed to produce heat-resistant xylanase without signal peptide. 0.5% (V/V)methanol was added in batch to induce the engineering Pichia pastoris which produced extracellular xylanase. The extracellular xylanase was analyzed by the xylan-degrading activity, SDS-PAGE and Western Blot. The results showed that the XynA contained typical catalytic structure domain of glycosidic hydrolyzyme 11 (GH11), and the predicted molecular weight of mature protein was 38.6 ku, and the isoelectric point was 6.86. Xylanase activities secreted by the seven genetic-engineering Pichia pastoris X33/pPICZαA-xynA were more than 300 U/mL, and the highest xylanase activity was up to 487.2 U/mL;the molecular weights of extracellular xylanases on SDS-PAGE and Western Blot were about 45 ku by the characteristic strips, bigger than the predicted molecular weight (38.6 ku). Therefore, a novel extracellular xylanase has been effectively expressed and may lay a foundation for its separation and purification to study the property, structure and function.
2018 Vol. 33 (4): 126-130 [Abstract] ( 15 ) HTML (1 KB)  PDF (1100 KB)  ( 19 )
131 Effects of Calcium Fertilizer and Plastic Film Mulching Cultivation on Dry Matter Production,Maturity Performance,Yield Components and Quality of Peanut in Red Soil under Ca Deficiency
WANG Jianguo, ZHANG Hao, LI Lin, LIU Dengwang, WAN Shubo, WANG Fei, LU Shan, GUO Feng
DOI: 10.7668/hbnxb.2018.04.019
In order to solve the problem of unfruitful peanut pods and to explore the effect of calcium application and film mulching on the dry matter production, maturity performance, yield and quality of peanut under Ca deficiency in red soil of Southern China, large seed variety Xianghua 2008 and the typical red soil under calcium deficiency were used as material for the soil column experiment. Three calcium fertilizer gradients and two kinds of cultivation methods (open field and plastic film cultivation)were used in this experiment including no calcium (Ca0), medium calcium fertilizer (Ca375)and high calcium fertilizer (Ca750). The net photosynthetic rate, chlorophyll content, maturity performance, dry matter, yield composition and quality of peanut were studied. The results showed that film mulching cultivation increased the dry matter of vegetative organs and genitals. While calcium application promoted genital growth better and increased harvest index, but reduced the ratio of root to shoot. Compared to Ca0-OF, the genital dry matter of Ca750-OF and Ca750-PF increased 74.0%, 94.3%, respectively. The net photosynthetic rate of the leaves was increased gradually at seedling phase and flowering and pedicel forming phase, but it decreased in the late growth period by calcium fertilizer application. The SPAD of open field (OF)at seedling phase was Ca750 > Ca375 > Ca0. The SPAD of calcium treatment higher than that of the no calcium application treatment (Ca0)under the plastic film cultivation in the whole growth period except pod setting phase. The yield per plant of peanut at maturity presented a quadratic equation curve relation with SPAD value (y=-0.020x2+1.034x-1.930, R2=0.308**), and was significantly correlated. The maturity performance of no calcium application treatment was greedy late, and the plant could not be into normal senescence. With the calcium application increased, maturity performance of peanut became obvious, and leaf color turned green to yellow, so the peanut was normal senescence, mature and had higher yield. The number of total pods and full pods per plant, the kernel rate to pods, plumpness, fat content and the ratio of oleic acid to linoleic acid increased, and the number of rotten, empty pods and pods per kilogram reduced, thus pod yields increased and quality was improved by calcium application and film mulching cultivation.
2018 Vol. 33 (4): 131-138 [Abstract] ( 17 ) HTML (1 KB)  PDF (1230 KB)  ( 21 )
139 Effects of Short-day Photoperiod Inducement after Early Flowering on Physiological Parameters in Adzuki Bean Leaves
DONG Weixin, YIN Baozhong, ZHANG Yanli, ZHANG Yuechen
DOI: 10.7668/hbnxb.2018.04.020
In order to clarify the effects of short-day photoperiod inducement after early flowering on physiological parameters in adzuki bean, medium maturing variety Jihong 9218 and late maturing variety Tangshanhongxiaodou were treated by different short-day photoperiod at Early Flowering-Early Podding (EF-EP), Early Podding-Early Seed (EP-ES)and Early Flowering-Early Seed (EF-ES)three stages, to investigate the effects of short-day photoperiod inducement on physiological parameters in adzuki bean leaves. Short-day photoperiod inducement decreased the chlorophyll content of two adzuki bean varieties, net photosynthetic rate also showed a decreasing trend at the same time.Which leads to decreased the maximum photochemical quantum yield (Fv/Fm);Soluble sugar and soluble protein content of two adzuki bean varieties were decreased by short-day photoperiod inducement after early flowering, and 12 h short-day photoperiod inducement had a greater influence at the EF-ES stage, and then leading to slower carbon and nitrogen metabolism, meanwhile, sucrose synthetase (SS)activity showed a decreasing trend, it was thus clear that the rate of carbon metabolism was related to SS activity, but the activities of sucrose phosphate synthase (SPS)in two varieties were different, sucrose phosphate synthase (SPS)activity of Jihong 9218 increased at three stages compared with CK while Tangshanhongxiaodou decreased, which might be related to the ripening of varieties;glutamic-oxalacetic transaminase (GOT)and glutamic-pyruvic transaminase (GPT)activity were not affected by photoperiod. The induction effect of short-day induction effect after anthesis was weaker than that before anthesis, but it also had a certain control effect, physiological parameter in adzuki bean could be regulated by short-day photoperiod inducement after early flowering, thereby affecting the growth and development of adzuki bean.
2018 Vol. 33 (4): 139-146 [Abstract] ( 16 ) HTML (1 KB)  PDF (1501 KB)  ( 19 )
147 Effects of Gibberellin Presoaking on Maize Germination under Chill
XU Gaoping, LIU Xiufeng, YUAN Wenya, WANG Pu, LOU Chenjun, YANG Zhaoshun
DOI: 10.7668/hbnxb.2018.04.021
The objective of this experiment is to study the effects of chill and drought stresses on seed germination characteristics, and the effects of stresses resistance by presoaking with gibberellin (GA3)under different concentrations. An experiment was conducted in order to investigate the effect of chill stress and chill-drought double stresses on gemination and seedling growth, the difference of the effects of presoaking with GA3 was also analyzed. Two varieties of Zhengdan 958 (ZD958 with good cold resistence)and CB15098 (CB15098 with poor cold resistence)were presoaked in gibberellin solution with three concentrations (10, 50, 100 mg/L)before sowing. The result showed that the effect of gibberellin presoaking on dry and fresh weight of shoot exhibited the opposite effect under two kinds of stress:the promoting effect under chill stress and the inhibition effect under double stresses. ZD958 had better chill resistance in compared with CB15098, and shoot elongation of ZD958 was promoted by presoaking with GA3 under both chill and chill-drought double stresses. GA3 presoaking increased germinability of CB15098 under both stresses as well as its shoot length under double stresses, contrary to its decreased plant length under chill stress. The responses of germination, fresh and dry weight and length of shoot of ZD958 was significantly higher than CB15098. Lower concentration of GA3 presoaking was recommended for maize variety under chill and drought stresses. High concentration of gibberellin solution was recommended for seed soaking under single chill stress, and low concentration of gibberellin solution for seed soaking was better for maize under chill and drought stress.
2018 Vol. 33 (4): 147-152 [Abstract] ( 19 ) HTML (1 KB)  PDF (1044 KB)  ( 15 )
153 Effect of Transplanting Period on Growth and Yield of Rice at Different Growth Stages
WANG Chuankai, NIU Jianhang
DOI: 10.7668/hbnxb.2018.04.022
In order to explore the effects of transplanting periods on growth and yield of rice and optimize the suitable transplanting date and rice varieties, the dynamic change laws of agronomic traits, carbonitride and yield factors of rice were analyzed using Yangliangyou 6, Shenliangyou 5814, Gangyou 1237, Yixiangyou 1577 andⅡyou 1259 as the experiment materials. Two transplanting periods for each of the five varieties were set up:4 leaf stage and 6 leaf stage. The results showed that the plant height, LAI and dry matter accumulation in vegetative growth stage, as well as the spike number per square meter and theoretical yield at mature stage were lower in 6 leaf transplanting treatments than in 4 leaf transplanting treatments. For the 4 leaf transplanting treatment, the plant height of Gangyou 1237, Yixiangyou 1577 and Ⅱyou 1259 were higher than that of other varieties at 90 days after planting and heading stage, and the LAI of Gangyou 1237 and Yixiangyou 1577 were also higher at 75 d and 90 d after planting, which quickly decreased substantially at heading stage, 40.0% and 32.7% lower, respectively, compared with that at 90 d after planting.At 90 d after planting, the fresh of Yixiangyou 1577 transplanted at 4 leaf stage was extremely significantly higher than those of other varieties, and its dry weights was significantly higher than other varieties.In the growth stage, the soluble sugar content of Gangyou 1237 transplanted at 4 leaf stage was the highest, followed by Yixiangyou 1577. The nitrogen content, mature spike number per square meter, seed setting rate and theoretical yield of Yixiangyou 1577 transplanted at 4 leaf stage were higher than those of other varieties, and its theoretical yield was 32.2% higher than that of Yixiangyou 1577 (6 leaf), followed by Gangyou 1237 (4 leaf). The plant height, dry weight and soluble sugar were positively correlated with mature spike number per square meter, grain number per spike, 1000-grain weight and theoretical yield at 90 d after planting and heading stage. LAI at heading stage was negatively correlated with yield factors, and its correlation coefficient with theoretical yield was-0.779, reaching the extremely significant level. In summary, the plant height, LAI, dry fresh quality, carbon nitrogen compound and yield index of Yixiangyou 1577 and Gangyou 1237 transplanted at 4 leaf stage were better at 90 d after planting and heading stage. To distribute photosynthetic products effectively and increase the grain yield, the LAI decreased greatly at heading stage. Therefore, Yixiangyou 1577 and Gangyou 1237 should be transplanted at 4 leaf stage so as to achieve the high yield of rice.
2018 Vol. 33 (4): 153-159 [Abstract] ( 33 ) HTML (1 KB)  PDF (1483 KB)  ( 29 )
160 Effects of Subsoiling During Fallow Period and Sowing Date on Dry Matter Accumulation and Yield of Wheat in Dryland
GAO Peifang, JIN Yonggui, SUN Min, LIANG Yanfei
DOI: 10.7668/hbnxb.2018.04.023
To determine the appropriate sowing date of high-yielding cultivation in Yuncheng Area of Shanxi Province, the experiment was conducted in the wheat growing season of 2013-2015 under two tillage subsoiling (SS)and traditional tillage (CK), and September 20 (early sowing date, D1), October 1 (conventional sowing date, D2), October 10 (late sowing date, D3)three sowing date study the effects of SS on the water changes of soil and contribution of dry matter accumulation and remobilization to yield and the adjusting effects of sowing dates. The results showed that the soil water storage was increased under SS in fallow period at different growth stages. Adoption of SS obviously improved dry matter weight, dry matter accumulation and at different growth stages, yield. In fallow period, the soil water storage was the highest under D2 treatment between winter to booting stage and the lowest under D1 treatment; the soil water storage was the lowest after anthesis under D2 treatment. D2 treatment could significantly increase the dry matter accumulation and accumulation rate in each stage. The yields were significantly increased by 5%-10% under D2 treatment under the condition of subsoiling during the fallow period. Under the control conditions, the D2 treatment significantly increased the yield by 9% to 11%. The dry matter accumulation in different organs at anthesis stage showed stem > leaf > ear. The formation of yield was more closely related to the dry matter accumulation after anthesis. The leaf and stems + sheaths dry matter accumulation showed a linearly relationship with the yield and it was positively correlated. The accumulation of dry matter in the glume + spike was parabolic with yield. The dry matter accumulation of ears was significantly parabolic correlated with the yield. In short, subsoiling during the fallow period plus the sowing date of October 1 was beneficial to promote that storage of soil water moisture and increasing dry matter accumulation and translocation, thereby increasing the yield of wheat.
2018 Vol. 33 (4): 160-166 [Abstract] ( 25 ) HTML (1 KB)  PDF (1081 KB)  ( 23 )
167 Effects of Planting Method and Density on the Dry Matter Accumulation, Distribution,Root Yield,and Quality of Fresh Edible Sweet Potatoes
DUAN Wenxue, WANG Jinli, ZHANG Haiyan, XIE Beitao, WANG Baoqing, ZHANG Liming
DOI: 10.7668/hbnxb.2018.04.024
In order to determine suitable planting method and density for fresh edible sweet potato planting in barren hilly in the central region of Shandong Province, L9 and J26 were used as test varieties under field conditions. Each variety was planted vertically and horizontally, with planting densities of 37 500 (D1), 52 500 (D2), and 67 500 plants/ha (D3). We studied the effects of different planting methods and densities on the dry matter accumulation, distribution, yield, and quality of fresh sweet potatoes. An increase in density increased the dry matter accumulation in the middle and late growth stages of the two varieties and significantly decreased the ratio of the dry matter distribution of the branch leaves of J26 planted vertically and horizontally. D2 treatment decreased the ratio of the dry matter distribution of the branch stems of L9 planted vertically and horizontally in the late growth stage and significantly increased the ratio of the dry matter distribution of the tuber roots of the two varieties planted vertically and horizontally in the middle growth stage. The D2 and D3 treatments significantly increased the ratio of the dry matter distribution of the tuberous roots of J26 planted vertically and horizontally in the late growth stage. For the two cultivars at the same transplanting method, D2 and D3 treatments significantly improved the soluble sugar content and starch content in tuberous roots and D2 treatment significantly increased the tuberous root yield of the two cultivars. Compared with vertical planting, horizontal planting significantly increased the dry matter accumulation in the early growth stage under D2 treatment and in late growth stage of J26 under D2 and D3 treatments and in the middle growth stage of L9 at the same density. Compared with vertical planting, horizontal planting increased the soluble sugar content of the tuberous roots of J26 under D1 treatment and that of L9 with different densities. Horizontal planting was also benificial for increasing tuberous root weight per plant and tuberous root yield of the two cultivars. With regard to yield and quality, horizontal planting + D2 treatment is the optimal combination under the test conditions.
2018 Vol. 33 (4): 167-174 [Abstract] ( 14 ) HTML (1 KB)  PDF (1022 KB)  ( 14 )
175 Effects of Nitrogen Fertilization on Dry Matter Accumulation and Yield Components of Following Wheat Which Planted after Maize Peanut Intercropping
MENG Weiwei, ZHANG Zheng, XU Jie, NAN Zhenwu, LIU Lingyan, GAO Huaxin, GUO Feng, WAN Shubo
DOI: 10.7668/hbnxb.2018.04.025
In order to study the suitable, stable and efficient nitrogen fertilizer management for following wheat after maize peanut intercropping, the effects of nitrogen fertilization on yield and yield structure, accumulation and distribution of dry matter and nitrogen use efficiency of following wheat after maize peanut intercropping were studied. Jimai 22 was used as experimental material. With traditional nitrogen application (225.0 kg/ha)as control, different nitrogen and nitrogen reduction treatments were set up respectively. The results indicated that there were no significant differences in the dry matter accumulation, total nitrogen accumulation, pre-anthesis dry matter translocation amount, post-anthesis dry matter accumulation amount and grain yield among the treatment increased the amount of nitrogen fertilizer, two increased nitrogen treatments, decreased 10% treatment and the control. The dry matter accumulation after jointing, total nitrogen accumulation, pre-anthesis dry matter translocation amount, post-anthesis dry matter accumulation amount and grain yield were significantly reduced by 30% nitrogen reduction. The study also found that the spike number per hectare decreased with the decreased of nitrogen fertilizer in the range of increase or decrease 30%. There was no significant difference in the number of hectare per hectare treated with nitrogen reduction 10%, which was significantly higher than that of nitrogen reduction over 20%. Nitrogen application had no significant effect on grain number per ear and thousand kernel weights. The nitrogen partial factor productivity (NPFP)of following wheat after maize peanut intercropping was significantly improved by reduction of nitrogen. Under the conditions of this experiment, there was no significant difference on dry matter accumulation and yield of wheat, but the nitrogen partial productivity was sharply increased when the nitrogen fertilization was reduced from 225.0 kg/ha to 202.5 kg/ha. That the amount of nitrogen fertilizer reduced from 225.0 kg/ha to 202.5 kg/ha was beneficial to the efficient and safe production of following wheat after maize peanut intercropping.
2018 Vol. 33 (4): 175-180 [Abstract] ( 16 ) HTML (1 KB)  PDF (960 KB)  ( 21 )
181 Research on Increasing and Maintaining Yield from Reducing N Application Rate and Enhancing Planting Density by Using Multiple Spike Cultivar of Huacheng 3366
ZHANG Xiangqian, LIU Liangbai, QIAO Yuqiang, CAO Chengfu, CHEN Huan, WANG Pingxin, GUO Ran, LIU Fei, DU Shizhou, ZHAO Zhu, LI Wei
DOI: 10.7668/hbnxb.2018.04.026
In order to reveal the feasibility of increasing and maintaining yield with green agriculture development by using multiple spike cultivar to enhance density based on decreasing nitrogen application rate, we studied the effects of the interaction of four nitrogen fertilization levels and three planting density on root traits, photosynthesis, grain quality and yield of wheat. Under the same nitrogen level, the length, surface area, average diameter of root were insignificantly affected by planting density, while root volume and root tips were significantly affected by planting density, the root length, root surface area, root volume and root tips of N1 (traditional N application amount)were significant higher than that of N3 (70% N1)under the same planting density. Under the same planting density, the flag leaf area was not significantly reduced at booting stage when decreasing 15% nitrogen (N2, 85% traditional N application amount)application amount, while was significantly reduced by decreasing 30% N application amount. Chlorophyll content was not significantly affected by planting density, the chlorophyll content of N1 was significantly higher than that of N3 under the density of D1 (180×104 plants/ha)and D3 (360×104 plants/ha). Under the same nitrogen level, to increase planting density would led to photosynthetic rate, stomatal conductance and transpiration rate increased and intercellular CO2 concentration decreased, when compared to N3, N1 significantly increased photosynthetic rate and stomatal conductance by 20.03%, 18.44%, 17.36% and 24.11%, 20.40%, 19.76% under D1, D2 (270×104 plants/ha), D3. The grain protein, starch, wet gluten and sedimentation value was insignificantly affected by density, the difference in protein content and sedimentation value between N1 and N3 was significant, while in starch and wet gluten content were in significant. Under the same nitrogen level, to increase the density of 180×104 plants/ha basic seedlings could significantly enhance the population dry matter weight of wheat at each stages, the population dry matter weight was significantly decreased when N application amount reduced from N1 to N3, the biological yield of N2D3 and N3D3 were higher than or equal to N1D1 and N1D2, and the biological yield of N3D3 was higher than or equal to N2D1 and N2D2. Compared to D1, under N1, N2, N3, D3 increased the yield by 11.83%, 11.42%, 14.03% and 5.52%, 10.31%, 10.88% respectively in 2016 and 2017, the yield of N3D2 and N3D3 were higher than or equal to N1D1. All the findings showed that when to reduce 15%-30% N and increase the density of 90×104-180×104 plants/ha basic seedlings could lead to biological and economic yield higher than or equal to that of the treatment without N reduction.
2018 Vol. 33 (4): 181-188 [Abstract] ( 13 ) HTML (1 KB)  PDF (954 KB)  ( 21 )
189 Effects of Different Annual Tillage Treatments on the Accumulation and Translocation of Nitrogen and Dry Matter of Summer Maize in Shajiang Black Soil
TONG Xingxing, SUN Wu, LI Songjian, ZHANG Hongsheng, LIU Shutang, JIANG Wen
DOI: 10.7668/hbnxb.2018.04.027
In order to explore a most suitable tillage to attain high and stable yield in summer maize in Shajiang Black soil, continuous annual tillage experiments were started from 2010 with seven different tillage treatments of winter wheat-summer maize annual tillage (rotary tillage-no tillage (CK);no tillage-no tillage;subsoiling tillage-no tillage;deep tillage-no tillage;rotary tillage-alternate years rotary tillage;subsoiling tillage-alternate years subsoiling tillage;deep tillage-alternate years deep tillage)in summer maize and winter wheat rotation system of Huanghuai Region, China. Sampling was done in 2012 and 2013. Rotary tillage-alternate years rotary tillage attained the highest aboveground nitrogen accumulation amount at maturity stage, with the increased by 14.57% (2012)and 30.95% (2013)compared with CK, and there was no significant difference between other tillage treatments and CK.For both two years, the nitrogen accumulation in shoot during kernel filling was lowest in control, followed by the treatments of no tillage-no tillage and the deep tillage-no tillage. The dry matter was lowest in CK, which was significantly lower than those of rotary tillage-alternate year rotary tillage treatment, subsoiling tillage-no tillage (2012), subsoiling tillage-alternate years subsoiling tillage (2013). The difference between CK and other tillage treatments was not significant, while in 2012, subsoiling tillage-no tillage attained highest yield, significantly higher than those of deep tillage-no tillage and deep tillage-alternate years deep tillage. Therefore, the alternate seasons of wheat and maize or years rotary tillage and subsoiling could be suggested to increase nitrogen accumulation, especially grain filling stage, and improve grain nitrogen concentration, dry matter production.
2018 Vol. 33 (4): 189-195 [Abstract] ( 12 ) HTML (1 KB)  PDF (925 KB)  ( 7 )
196 Analysis of Bacterial Diversity in Cultivated Soils in Middle and Southern Hebei Province Based on High-throughput Sequencing
ZHANG Xiang, SONG Shuishan, HUANG Yali, JIA Zhenhua, HUANG Yuanyuan, SONG Cong
DOI: 10.7668/hbnxb.2018.04.028
To investigate the difference of bacterial diversity in greenhouse soil and field soil, 14 samples of greenhouse and field soil in middle and Southern Area of Hebei Province were sequenced by using Miseq high-throughput sequencing technology. And the species composition information and the relative abundance information of two sets of samples were obtained from the sequencing result. The results showed that in the greenhouse (M1)and field (N1)soil samples, the composition of the dominant microflora was basically the same. The dominant phylum of all samples were Proteobacteria, Actinobacteria, Acidobacteria, Bacteroidetes, Chloroflexi, Firmicutes and Gemmatimonadetes. The relative abundance of Actinobacteria in the field was more than it in the greenhouse, but the relative abundance of Firmicutes in the field was less than it in the greenhouse. At the class level, more than 15 dominant species were found in all samples, such as Alphaproteobacteria, Acidobacteria, Gammaproteobacteria, Actinobacteria, Deltaproteobacteria, Betaproteobacteria, Gemmatimonadetes, Cytophagia and so on. Among the dominant species, the most abundant relative abundance in greenhouse was Cytophagia, Bacilli and Flavobacteriia, while the relative abundance was significantly reduced by Acidobacteria and β-Proteobacteria. At the genus level, Arthrobacter, Sphingomonas, Devosia, Bacillus, Xanthomonas and Lysobacter were six dominant genera. Arthrobacter was more in the soil of field, while Bacillus, Xanthomonas and Lysobacter were more in the greenhouse. In addition, some of the genera that had nitrogen fixation and degradation and repair capabilities are relatively more abundant in the field. This should be caused by the different planting environments in fields and greenhouses. The research results have certain value for improving arable land, increasing yield and disease resistance.
2018 Vol. 33 (4): 196-203 [Abstract] ( 15 ) HTML (1 KB)  PDF (1321 KB)  ( 11 )
204 Effect of Enhanced Ammonium Nutrition on Morphological Characteristics and Dry Matter Accumulation of Cotton Seedlings
MA Tongtong, CHEN Li, WAN Hualong, LI Jin, LIU Liantao, SUN Hongchun, ZHANG Yongjiang, BAI Zhiying, LI Cundong
DOI: 10.7668/hbnxb.2018.04.029
In order to provide references for nitrogen fertilization in field production, the effects of different NH4+/NO3- ratio of nitrogen nutrition (NH4+-N/NO3--N were 0/100, 25/75, 50/50, 75/25, 100/0)on morphological characteristics and dry matter accumulation, hydroponic experiments were conducted using Nongda 601 as material. The results showed that the plant height, stem diameter, leaf area and main root length of cotton increased gradually with the growth stages changed. In addition to the main root length showed a continuous elongation, the plant height, stem diameter, leaf area and dry weights of cotton increased firstly and then decreased with the increasing of NH4+ concentration in the ammonium nutrient solution. The plant height of cotton seedlings was the highest at the NH4+/NO3- ratio of 50/50 and 25/75 respectively 7, 14, 21 d after enhanced ammonium nutrient solution treatment;both stem diameter and leaf area were the largest at the NH4+/NO3- ratio of 50/50 after enhanced ammonium nutrition treatment 7, 14, 21 d, indicating that the concentration of NH4+ more than 50% could inhibit significantly the growth of cotton seedlings. In terms of dry matter, total dry weight, stem dry weight and leaf dry weight were the highest at the NH4+/NO3- ratio of 75/25, while there was no significant difference in root dry weight and root to shoot ratio, indicating that the effect of enhanced ammonium nutrition on the dry matter weight was positive mainly in the aboveground, while the effect in the underground was not obvious. Moreover, the ratio of root to shoot was the highest, the main root length was the longest when supplied with only NH4+, indicating that single NH4+ could promote root growth. All results showed that ammonium and nitrate nitrogen had different mechanisms for regulating growth and development in cotton underground and aboveground organs, mixed nitrogen was more favorable to the aboveground growth of cotton seedlings.
2018 Vol. 33 (4): 204-209 [Abstract] ( 18 ) HTML (1 KB)  PDF (996 KB)  ( 12 )
210 Identification and Basic Application Research of Two Abiogenous Azotobacter
WANG Huiqiao, CHEN Weifeng, ZHUGE Yuping, HU Guoqing, YU Haiyan, FU Naixin, DONG Yuanjie
DOI: 10.7668/hbnxb.2018.04.030
In order to identify two strains of azotobacter isolated from coastal saline soil, explore the environmental adaptability of two strains and inoculated effect by pot experiment in wheat. Morphological observation, physiological and biochemical determination and 16S rDNA sequence identification were carried out. We also studied environmental adaptability of bacteria by the analysis of IAA and effect of wheat seedlings was evaluated by pot experiment in wheat.The characteristic of morphology and 16S rDNA sequence analysis revealed that strain 1 was classified as Variovorax paradoxus. And strain 2 was identified as Azospirillum zeae. The nitrogenase activity of strain 1 and strain 2 were 4.71 and 2.84 μmol/ (h·mL). The analysis of IAA indicated that both of two strains had strong ability to keep themselves under acid or alkalinity. The two strains had strong ability of IAA under 5% concentration of NaCl concentration. The strain 1 utilized mannitol most and the IAA amount of strain 2 was significantly higher under glucose. The results of strains secreted IAA under exogenous NH4+ showed that low concentration of NH4+ stimulate the strain 1 to secrete IAA.When the concentration of NH4+ was over 0.5 mmol/L, IAA was inhibited in different degrees. The curve of strain 2 showed a M curve. Pot experiments showed that strain 1 and strain 2 significantly increased height and dry weight of wheat shoots, promoted the absorption of nitrogen, phosphorus and potassium in wheat, and improved the content of available nutrients in soil.The soil urease activity of two strains increased by 16.00% and 48.15%, and the strain 2 had the best effect. The two strains have high nitrogenase activity and strong adaptability. The treatments of two strains have good effect on the growth of wheat. The results can be used for the cultivation of new low yield saline alkali soil and provide technical support for optimizing the fertilization structure of medium and high yield saline alkali farmland.
2018 Vol. 33 (4): 210-216 [Abstract] ( 22 ) HTML (1 KB)  PDF (1023 KB)  ( 8 )
217 The Study of Water Regulated Deficit on Yield Component and Water Production Function of Pepper with Mulched Drip Irrigation
WANG Shijie, ZHANG Hengjia, BA Yuchun, WANG Yucai, HUANG Caixia, XUE Daoxin, LI Fuqiang
DOI: 10.7668/hbnxb.2018.04.031
In order to study the effects of water deficit in different growth periods on growth, yield and water use efficiency of pepper with mulched drip irrigation, mild water deficit (65%-75% Field capacity, FC)and moderate water deficit (55%-65% FC)were composed during seedling period and blossom and fruit period, respectively, mild water deficit was composed during full fruit period and later fruit period, respectively, and full irrigation (75%-85% FC)during the whole pepper growth period as the control (CK). Meanwhile, water production function of pepper was modeled and solved by Jensen model. The results showed that the maximum pepper fresh fruit yield was found in the fully irrigated pepper (CK)with fruit yield of 36 203.90 kg/ha. The mild and moderate water deficit during seedling period and mild water deficit during later fruit period had no significant influence (P> 0.05)on total fresh fruit yield of pepper compared with CK, but the other water deficit treatments had significantly smaller total fresh fruit yield by 10.45% to 13.32% than CK. The mild water deficit at the full fruit period and mild water deficit at later fruit period could significantly (P<0.05)decreased pepper fresh fruit moisture content by 5.75, 5.83 percentage points during the stage with water deficit compared to CK, respectively. Pepper irrigation water use efficiency (IWUE)and WUE were similar in each water treatment, the WUE of the water treatments with moderate water deficit during seedling period and mild water deficit during later fruit period were significantly greater than CK by 11.63% and 9.41%, respectively. The water sensitive index at blossom and fruit period was the maximum of 0.517 during the different pepper growth period in Jensen water production function. Therefore, the blossom and fruit period was the critical period of water requirement of pepper, the water deficit in this growth period would lead to serious reduction of pepper yield. In order to get a higher yield, full irrigation (soil moisture was controlled at 75% to 85% of field capacity)should be conducted at this growth period.
2018 Vol. 33 (4): 217-225 [Abstract] ( 14 ) HTML (1 KB)  PDF (975 KB)  ( 12 )
226 Effects of Different Tillage Ways on Seedling Growth of Maize and Soil Physical Properties in Mollisol Region
GU Siyu, ZHU Yuwei, GUO Xingjun, CAI Yuetong, WU Shuai, ZHANG Zehui, ZHANG Huihui
DOI: 10.7668/hbnxb.2018.04.032
In order to solve the problem of low soil depth and it causes the damage to the physical properties of the plough layer in Northeast China. The effect of treatment including four tillage methods, Deep mold-board tillage (DN), No-tillage (NN), Rotary tillage (RN)and Subsoiling tillage (SN)on the soil moisture, soil temperature, soil compaction, and corn growth were studied to investigate the effect of soil physical properties on the root growth and thus the corn growth. Soil compaction increaseded sharply from soil (0-22.5 cm).In 2016, subsoiling reduced soil compaction treatment NN increased the soil compaction significantly with a maximum of 314.6 kPa respectively, SN soil compaction decreased the maximum value of 220.8 kPa. In 2017, NN increased, soil compaction maximum value was 489.3 kPa, SN decreased, soil compaction maximum value was 197.3 kPa.Compared with the treatment DN, in 2016, compared with DN, the water content of the plough layer (0-20 cm)was DN > RN > NN > SN, and the storage capacity of SN was 0.085 percentage points lower than DN. In 2017, NN increased the water storage of plough layer (0-20 cm)by 0.036 percentage points, SN was 0.066 percentage points lower than DN. Soil temperature of treatment NN dropped by 0.7℃ in 2016 and 1.5℃ in 2017. Compared with treatment DN, corn plant features of treatment SN, root length, root volume, root surface area and root dry weight showed an increasing trend by 4.54%, 19.21%, 18.57%, 14.12% respectively. Compared with DN, the root length and root dry weight of SN increased by 0.58 cm and 0.40 g in 2017.The 2016 data linear analysis showed that, maize seedling root length showed a negative linear relationship with the moisture, dry matter weight showed a negative linear relationship with the compactness, but showed the positive linear relationship with the soil temperature, the compactness had a strong linear correlation with dry matter formation in maize seedling stage. In summary, SN can reduce soil compaction, promote the growth of maize root, favorable yield formation, NN can improve the compaction and reduce the temperature, which is not conducive to the accumulation of dry matter and the formation of yield in the seedling stage of maize.
2018 Vol. 33 (4): 226-231 [Abstract] ( 27 ) HTML (1 KB)  PDF (1003 KB)  ( 19 )
232 Effects of Coupling Water and Fertilizer on Root Morphology and Yield of Summer Maize
SUI Kaiqiang, ZHAO Longgang, LIU Shutang, HUANGFU Chenghui, LIN Shaowen, CHEN Jingpei
DOI: 10.7668/hbnxb.2018.04.033
In order to study the effect of coupling water and fertilizer on the root morphology and yield of summer maize, using the "3414" optimal regression design scheme, the changes of root morphology and yield components of each treatment were analyzed, and the correlation analysis of yield and the prediction of model construction. The results showed that the total root length, total root surface area and total root volume of W3N2C2 were the largest, which were 2.89, 2.41 and 2.68 times than that of W0N0C0.The number of ear rows, row grains, ear length, 100-seed weight of W3N2C2 were 1.31, 1.19, 1.22, 1.62 times of W0N0C0.The correlation between yield factors and yield was significant. The biggest influence on maize yield was the amount of irrigation, the next was the amount of carbon and the minimum was amount of nitrogen.W3N2C2 increased the output of W0N0C0 by 72.39%.When nitrogen and organic fertilizer were at N2 and C2 level, irrigation could increase maize yield by 19.78%-48.46%. When irrigation amount and organic manure were at W2 and C2 level, nitrogen application could increase maize yield by 6.59%-15.68%. When irrigation amount and nitrogen fertilizer were at W2 and N2 level, applying organic fertilizer could increase yield of maize by 4.99%-9.67%. Under low water conditions, increasing organic manure and nitrogen fertilizer could enhance soil water holding capacity and water and nutrient utilization rate, so as to reduce water and fertilizer waste and improve maize yield. The research showed that the proper proportion of water and fertilizer under the condition of water shortage played an important role in improving crop yield. In the design range of the experiment, the yield equation was Y=6 723.94+1 589.98X1+470.79X2+182.50X3-194.23X12-74.70X22-78.66X32+42.12X1X2+98.28X1X3+89.39X2X3, the relative water content was 70%, the amount of nitrogen applied was 225 kg/ha, the amount of carbon was 2 700 kg/ha, the maximum yield was 12 393.69 kg/ha.
2018 Vol. 33 (4): 232-238 [Abstract] ( 31 ) HTML (1 KB)  PDF (1097 KB)  ( 18 )
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