Scientia Agricultura Sinica

Genome-Wide Association Study of Grain Quality Related Characteristics of Spring Wheat

YAN YongLiang,SHI XiaoLei,ZHANG JinBo,GENG HongWei,XIAO Jing,LU ZiFeng,NI ZhongFu,CONG Hua

Scientia Agricultura Sinica. 2021, 54(19): 4033-4047. doi:10.3864/j.issn.0578-1752.2021.19.001

Abstract ( 587 )

HTML ( 233 )

PDF (702KB) ( 569 )

Save

Figures and Tables | References | Related Articles | Metrics

【Objective】Identify SNPs and candidate genes that are significantly related to wheat grain quality traits, and reveal their genetic mechanism. 【Method】In this study, 298 introduced varieties (lines), Xinjiang landrace (lines) and Xinjiang bred varieties (lines) were used for an association population. Seven grain quality traits, including protein content (PRC), wet gluten content (WGC), sedimentation value (SV), starch content (STC), grain hardness (GH), flour yield (FY) and test weight (TW), were measured under five environments. Based on phenotypes of seven quality traits and genotypes of 55K SNP markers in this population, the Q+K association mixed model was used for genome-wide association analysis to obtain significantly associated SNP loci.【Result】The coefficients of variation of the seven grain quality traits of introduced varieties (lines), landraces and bred varieties (lines) under different environments were 1.3%-13.4%, 1.1%-18.6% and 1.0%-13.9%, respectively. Among them, the protein content, wet gluten content and sedimentation value of introduced varieties (lines) have the highest coefficient of variation (CV); Xinjiang bred varieties (lines) have the largest CV of starch content, grain hardness and flour yield. Whereas, for other six grain quality traits, including protein content, wet gluten content, sedimentation value, starch content, grain hardness, and the coefficients of variation of Xinjiang landraces are all between those of the introduced varieties (lines) and Xinjiang bred varieties (lines). Population structure analysis showed that 298 wheat varieties (lines) can be divided into 3 subgroups. Subgroup 1 contains 128 (43.0%) the materials mainly from landrace (lines); Subgroup 2 has 24 (8.1%) materials, mainly including introduced varieties (lines) and landraces; Subgroup 3 contains 146 (48.9%) materials, mainly introduced varieties (lines). The linkage disequilibrium analysis showed that the LD attenuation distances of the A, B and D genomes and the whole genome respectively were 10, 10, 6 and 8 Mb, according to the LD attenuation distance of the whole genome, the loci in the 8 Mb interval after the physical map were identified as a candidate loci. A total of 85 loci were simultaneously detected in two or more environments, that were significantly associated with 7 wheat grain quality traits detected by GWAS, with a contribution rate of 3.7%-10.9%. Stable SNPs associated with multiple traits were detected on chromosomes 1B, 1D, 2D, 3A, 3D, 4A, 4B, 5A, 6A, 6D, 7A and 7D. Among them, AX-109452823-AX-110545157 on chromosome 7A is related to protein content, starch content, wet gluten content, sedimentation value, flour yield and grain hardness, and was detected across four environments. Candidate genes at stable loci associated with multiple traits were searched, and 10 candidate genes that might be related to wheat grain quality were screened. Among them, TraesCS4A01G299800 (cationic amino acid transporter), TraesCS7A01G059500 (tryptophan decarboxylase), TraesCS7A01G331200, TraesCS7D01G418700 (xyloglucan endoglucosylase/hydrolase) play important roles in regulating the amino acid content in wheat grains.【Conclusion】The 85 loci were simultaneously detected in two or more environments, and 10 candidate genes related to wheat grain quality traits were predicted.

Genome-Wide Identification of VQ Gene Family in Fagopyrum tataricum and Its Expression Profiles in Response to Leaf Spot Pathogens

ZHENG FengSheng,WANG HaiHua,WU QingTao,SHEN Quan,TIAN JianHong,PENG XiXu,TANG XinKe

Scientia Agricultura Sinica. 2021, 54(19): 4048-4060. doi:10.3864/j.issn.0578-1752.2021.19.002

Abstract ( 533 )

HTML ( 224 )

PDF (3796KB) ( 937 )

Save

Figures and Tables | References | Related Articles | Metrics

【Objective】VQ gene family plays important roles in plant growth, development and responses to biotic or abiotic stress. The aim of this study is to comprehensively identify Fagopyrum tataricum L. Gaertn. VQ (FtVQ) gene family on genome-wide scale and analyze its expression profiles under challenge of leaf spot pathogens Alternaria. alternata and Nigrospora osmanthi, and treatment of defense-related hormones, such as salicylic acid (SA), jasmonic acid (JA) and ethylene (ET), thus providing a solid foundation not only for further elucidation possible roles of members of the VQ genes in defense response to leaf spot pathogens and underlying mechanisms in tartary buckwheat, but also for mining gene resources of breeding for crop disease resistance. 【Method】Based on the Hidden Markov Model profile of the conserved VQ domain (PF05678), HMMER 3.0 software was used to identify FtVQ genes from F. tataricum cv. Pinku1 genome database. Bioinformatic tools such as DNAMAN, MapInspect, MEGA, MEME, OrthoFinder and PLACE were used to analyze gene structure, chromosomal location of genes, cis-elements of gene promoters, physicochemical properties of proteins, conserved motifs of proteins, subcellular localization of proteins, and phylogenetic relationships. Quantitative real-time PCR (qPCR) was employed to analyze the expression profiles of leaf FtVQ genes of tartary buckwheat plants under infection of the pathogens and treatment of the hormones. 【Result】A total of 28 VQ genes were identified in the genomes of tartary buckwheat, with the gene size ranging from 566 to 1454 bp. The FtVQ genes contain no introns, and distribute unevenly on chromosomes 1-8. According to their physical locations on the chromosomes, the FtVQ genes were named from FtVQ1 to FtVQ28. Each of the FtVQ proteins has a highly conserved VQ motif FxxxVQx (L/F/I/V/A/Y) TG, where x represents any amino acid. Analysis of subcellular localization showed that 21 FtVQ proteins were predicted to the nucleus, and the others to the chloroplasts or cytoplasm. Based upon their amino acid sequence and presence of various conserved motifs, the FtVQ proteins were classified into five subfamilies (Ⅰ-Ⅴ). Each subfamily shared relatively conserved gene structures and protein motifs. The analysis of gene duplication revealed that F. tataricum genome had 8 pairs of paralogous pairs, all of which were segmental duplicated genes, suggesting that segmental duplication played major roles in FtVQ gene expansion. The ratio of nonsynonymous to synonymous substitutions (Ka/Ks) of paralogous pairs was less than 1, suggesting that they underwent purifying pressure during the evolution process. Prediction of cis-elements showed that pathogen-, SA-, JA-, or ET-responsive elements, such as BIHD1OS, CGTCA, ERELEA4, W-box and W-box-like sequences, were present within the promoters of all the FtVQ genes. Especially, FtVQ10, FtVQ14, FtVQ15, FtVQ22, FtVQ23 and FtVQ27 contained more elements in their promoter regions. 55% to 70% of the detectable FtVQ genes were differentially expressed genes (DEGs), and 72.7% to 85.7% of the DEGs were significantly induced on the level of transcription under the infection of leaf spot fungi A. alternata and N. osmanthi, or the treatment of SA, methyl jasmonate and ethephon. 【Conclusion】The tartary buckwheat genome contains 28 members of VQ gene family. Some FtVQ genes may be involved tartary buckwheat defense response to leaf spot pathogens.

Cloning and Function Analysis of MsMAX2 Gene in Alfalfa (Medicago sativa L.)

MA Lin,WEN HongYu,WANG XueMin,GAO HongWen,PANG YongZhen

Scientia Agricultura Sinica. 2021, 54(19): 4061-4069. doi:10.3864/j.issn.0578-1752.2021.19.003

Abstract ( 393 )

HTML ( 213 )

PDF (4431KB) ( 1584 )

Save

Figures and Tables | References | Related Articles | Metrics

【Background】 Branching is one of the key yield components, which plays an important role in alfalfa (Medicago sativa L.) breeding. Exploring and functional characterization of key branching-related genes are of significance in accelerating breeding of alfalfa with high yield and quality. MAX2 is an important branching-related gene, which is involved in the regulation of branching in several plant species. 【Objective】 Our research on the functional characterization of MsMAX2 in alfalfa will lay a foundation for the molecular mechanism of MsMAX2 in regulating branch development in alfalfa. 【Method】 The gene sequence of MAX2 in alfalfa was isolated by using homologous cloning. Sequence characteristics and phylogenetic tree of MsMAX2 were analyzed by using bioinformatics tools including Expy Protparatam, DNAMAN, and MEGA-X. The real-time quantitative PCR (qPCR) was applied to detect the tissue-specific expression pattern of MsMAX2 in alfalfa. The subcellular localization of the MsMAX2 protein was determined by using transient expression system in tobacco. The biological function of MsMAX2 was clarified by transformation in the Arabidopsis mutant via Agrobacterium-mediated transformation. Proteins interacting with MsMAX2 were determined by using yeast two-hybrid assay. 【Result】The length of MsMAX2 CDS is 2 136 bp, encoding a protein of 711 amino acids, and it belongs to the F-box protein super-family. Phylogenetic analysis showed that the evolution of MAX2 homologs was highly similar to the differentiation of species, indicating that MsMAX2 was a functionally conserved gene. It was showed that MsMAX2 was expressed in the neck at the highest level, followed by in the leaves of seedling, the inflorescences on pollination day and the roots; the expression level of MsMAX2 was relatively low in other tissues, indicating it functions in multiple tissues. Subcellular localization assay showed that the MsMAX2 protein was localized in the nucleus. Complementation assay in Arabidopsis max2 mutant showed that the multi-branch phenotype was recovered by the ectopic expression of MsMAX2. Yeast two hybrid assay demonstrated that the interaction between MsMAX2 and hormone receptor D14 depended on the existence of strigolactones. 【Conclusion】The MsMAX2 was obtained from alfalfa and it was highly expressed in the neck and the encoding MsMAX2 protein was localized in nucleus. When the MsMAX2 was over-expressed in the Arabidopsis max2 mutant, its multi-branch phenotype was recovered, indicating that MsMAX2 regulates branch development in alfalfa plant, and its function was conserved.

Effects of Nitrogen Fertilizer and Plant Density on Carbon Metabolism, Nitrogen Metabolism and Grain Yield of Two Winter Wheat Varieties

WANG JinFeng,WANG ZhuangZhuang,GU FengXu,MOU HaiMeng,WANG Yu,DUAN JianZhao,FENG Wei,WANG YongHua,GUO TianCai

Scientia Agricultura Sinica. 2021, 54(19): 4070-4083. doi:10.3864/j.issn.0578-1752.2021.19.004

Abstract ( 516 )

HTML ( 83 )

PDF (612KB) ( 431 )

Save

Figures and Tables | References | Related Articles | Metrics

【Objective】A field study was conducted to investigate the effects of nitrogen fertilizer, plant density and its interaction on carbon metabolism, nitrogen metabolism and grain yield of winter wheat in moist clayey soil of southeastern Henan province, and to clarify the appropriate nitrogen-density treatment for winter wheat in this area, so as to provide the technical support for optimizing high yield and high efficiency cultivation of winter wheat in this area. 【Method】The treatments included two varieties (Xinhua 818 with medium tiller ability and high heading rate and Bainong 207 with high tiller ability and medium heading rate), three nitrogen rates (N0, 0; N240, 240 kg·hm^-2; N360, 360 kg·hm^-2) and three plant densities (M1, 225×10⁴ plant/hm²; M2, 375×10⁴ plant/hm²; M3, 525×10⁴ plant/hm²) during two consecutive winter wheat growing seasons from 2018 to 2020 in moist clayey soil of southeastern Henan province. The differences of carbon metabolism (soluble sugar content; SPS activity; SS activity), nitrogen metabolism (soluble protein content; NR activity; GS activity) and yield under three-factor treatments were analyzed. 【Result】Variety, nitrogen fertilizer and density and their interactions significantly affected the carbon and nitrogen metabolism of winter wheat, and nitrogen fertilizer was the main effect that affected the yield and its components of the two winter wheat varieties. The effects of nitrogen application amount and planting density on carbon and nitrogen metabolism of the two winter wheat varieties in different growth stages were different. On the whole, the advantage of nitrogen-density regulation on carbon metabolism of the two winter wheat was mainly in the late filling stage, while the advantage on nitrogen metabolism was mainly in the middle stage of filling, during which the average increase of carbon and nitrogen metabolism indexes under M2N240 was 358.28% compared with the minimum treatment. The balance of carbon and nitrogen metabolism had a greater impact on the yield formation of winter wheat varieties with different tiller heading rates, especially in the late growth stage, which was the main physiological reason that the yield of Xinhua 818 was higher than that of Bainong 207 as a whole. In the winter wheat growing season from 2018 to 2020, the yield under treatment of M2N240 was the highest. Compared with the M1N0 treatment with the lowest yield, the average yield increases in two years under M2N240 were 96.49%. 【Conclusion】Considering the effects of variety, nitrogen fertilizer, density and their interactions on the balance of carbon-nitrogen metabolism and yield of winter wheat, the advantages of nitrogen application and planting density on the carbon-nitrogen metabolism of two winter wheat varieties were mainly reflected in the middle and late stages of filling. The M2N240 combination treatment could be used as a suitable nitrogen-density cultivation mode for winter wheat in the moist clayey soil area of southeast Henan province.

Effects of Planting Density on Photosynthetic Characteristics, Yield and Stem Lodging Resistance of Soybean in Maize-Soybean Strip Intercropping System

CHENG Bin,LIU WeiGuo,WANG Li,XU Mei,QIN SiSi,LU JunJi,GAO Yang,LI ShuXian,Ali RAZA,ZHANG Yi,Irshan AHMAD,JING ShuZhong,LIU RanJin,YANG WenYu

Scientia Agricultura Sinica. 2021, 54(19): 4084-4096. doi:10.3864/j.issn.0578-1752.2021.19.005

Abstract ( 618 )

HTML ( 78 )

PDF (1593KB) ( 407 )

Save

Figures and Tables | References | Related Articles | Metrics

【Objective】The aim of this study was to reveal the light environment change law of soybean canopy under different planting densities in maize-soybean strip intercropping, and to clarify the effects of density on leaf photosynthetic characteristics, yield and stem lodging resistance of soybean, so as to provide the theoretical reference for the construction of reasonable population density of intercropped soybean in low radiation area. 【Method】In this study, soybean genotype of Chuandou-16 and maize genotype of Zhenghong-505 were used as experimental materials. The two-factor random expulsion design was adopted, among which maize-soybean strip intercropping and monocropping were the main factors, and three planting densities of soybean (PD1 = 17 plants/m², PD2 = 20 plants/m², PD3 = 25 plants/m²) were the secondary factors. Effects of planting density on light environment of canopy, photosynthetic characteristics, growth dynamics, lodging percentage and yield composition of soybean were investigated. 【Result】Two-year data showed that the growth of soybean was affected by the shading of maize and self-shade at the middle and later stages in the maize-soybean strip intercropping system. The photosynthetic active radiation (PAR) in the canopy of the plant population, leaf area index (LAI), leaf photosynthetic capacity, number of branches and yield were significantly decreased, while the degree of being affected by maize varied with soybean planting densities. In the strip intercropping, compared with PD3, the PAR in soybean population canopy of PD1 and PD2 increased by 45.4% and 24.8% respectively, the Pn of leaves increased by 46.1% and 12.3%, respectively, the Ep increased by 53.2% and 27.2%, respectively, the Bn increased by 270.4% and 140.9%, respectively, and the lodging percentage decreased by 50.3% and 19.3%, respectively. Correlation analysis showed that lodging percentage was significantly negatively correlated with the PAR, net photosynthetic rate (Pn), stem bending force (SBF), dry weight of stem / leaf ratio (S﹕L), number of branches per plant (Bn) and number of effective pods per plant (Ep), and positively correlated with plant height (PH), LAI and number of ineffective pods per plant (nEp). 【Conclusion】Therefore, in the maize-soybean strip intercropping, the appropriate planting density (20 plants/m²) was beneficial to create a better light environment of soybean population, reduce the lodging percentage, increase the accumulation of photosynthates, and thus improve the yield of soybean.

Molecular Characteristic of Squash Leaf Curl China Virus (SLCCNV) Infecting Cucurbitaceae Crops in Guangdong Province

ZHANG Li,TANG YaFei,LI ZhengGang,YU Lin,LAN GuoBing,SHE XiaoMan,HE ZiFu

Scientia Agricultura Sinica. 2021, 54(19): 4097-4109. doi:10.3864/j.issn.0578-1752.2021.19.006

Abstract ( 338 )

HTML ( 33 )

PDF (2808KB) ( 373 )

Save

Figures and Tables | References | Related Articles | Metrics

【Objective】Squash leaf curl China virus (SLCCNV) is one of the main viruses infecting Cucurbitaceae crops, which can infect pumpkin, cucurbits, wax gourd, cucumber, muskmelon and honeydew melon, and severely affect Cucurbitaceae crop production. The objective of this study is to investigate the molecular characteristics, genetic relationship and pathogenicity of SLCCNV Guangdong isolates, and to provide a theoretical basis for the prevention and control of SLCCNV.【Method】Fifty-three suspected samples infected by Begomovirus were collected from Leizhou City and Xuwen County of Zhanjiang City, Boluo County of Huizhou City, Yuancheng District of Heyuan City and Gaoming District of Foshan City, Guangdong Province. Total DNA was extracted from all suspected samples, respectively, and used as template for PCR amplification with degenerate begomovirus primers AV₄₉₄/CoPR. The full-length DNA-A sequence of each isolate was obtained by RCA amplification, gene cloning and sequencing from positive samples by PCR detection. The full-length DNA-B sequence of each isolate was obtained by PCR amplification using designed abutting primers. All full-length sequences of SLCCNV obtained from Guangdong Province were analyzed with BLAST in NCBI. The similarity was compared using MUSCLE alignment of SDT1.2 software. Phylogenetic analysis between SLCCNV Guangdong isolates and the other SLCCNV isolates was performed using MEGA7. Infectious clone of SLCCNV isolate HYNG was constructed using homologous recombination technology. The infectious clone was inoculated to the cotyledon of pumpkin plants by agro-inoculation.【Result】The PCR detection result showed that 52 out of 53 suspected samples were infected by Begomovirus. The full-length sequences of eight isolates of SLCCNV from Guangdong Province were obtained from pumpkin, cucurbits, wax gourd and white melon, respectively. The full DNA-A sequences of eight isolates ranged from 2 735 to 2 739 nt in size, contained six ORFs, which were identical to the reported SLCCNV. The full DNA-B sequences of eight Guangdong isolates of SLCCNV ranged from 2 701 to 2 721 nt in size, contained two ORFs, encoding proteins associated with viral movement BC1 and BV1, respectively. The similarity analysis of the full sequence showed that the DNA-A of eight isolates from Guangdong shared >94.9% nt identities with each other, and shared >88% nt identities with other isolates of SLCCNV in GenBank database, and the DNA-B of eight isolates from Guangdong shared >86.7% nt identities with each other, and shared >83% nt identities with isolates of SLCCNV in GenBank database. Phylogenetic analysis indicated that the eight Guangdong isolates clustered with SLCCNV isolates from Guangxi, Henan, Hainan of China, Vietnam, Thailand, Philippines, Indonesia to form one branch, and the isolates from India clustered to form the other branch. Pathogenicity results showed that the new leaves of pumpkin inoculated with infectious clones of SLCCNV displayed crimple symptoms at 15 days post-inoculation (dpi), and more severe symptoms at 30 dpi. SLCCNV could be detected by PCR from the symptomatic plants, Western blot detection further verified the above results.【Conclusion】SLCCNV was detected infecting Cucurbitaceae crops in Guangdong Province. The SLCCNV is the pathogen causing pumpkin curl leaf disease in Guangdong Province. The SLCCNV isolates from Guangdong are closely related to isolates from Guangxi, Hainan etc., belonging to the same branch, while it is more distantly related to isolates from India.

Cloning of Nicotiana benthamiana NAC062 and Its Inhibitory Effect on Potato Virus Y Infection

QU XiaoLing,JIAO YuBing,LUO JianDa,SONG LiYun,LI Ying,SHEN LilLi,YANG JinGuang,WANG FengLong

Scientia Agricultura Sinica. 2021, 54(19): 4110-4120. doi:10.3864/j.issn.0578-1752.2021.19.007

Abstract ( 243 )

HTML ( 33 )

PDF (2050KB) ( 270 )

Save

Figures and Tables | References | Related Articles | Metrics

【Objective】 Potato Y virus (PVY) is one of the most important viruses that endanger the tobacco production in China. NAC transcription factors are closely related to plant disease resistance and stress resistance. The objective of this study is to clone NbNAC062, analyze its bioinformatics and research its role in the process of PVY infection, and to provide a target for the development of tobacco antiviral agents. 【Method】 Nicotiana benthamiana was used as the material to clone NbNAC062, and MEGA, UniProt, SMART, TMHMM Server 2.0, Sol Genomics Network, PlantCARE and other technologies were used for bioinformatics analysis. Laser confocal microscope and quantitative real-time PCR (qRT-PCR) were used to clarify the localization of NbNAC062 protein and the change of NbNAC062 mRNA expression before and after PVY infection. Based on virus-induced gene silencing (VIGS) technology and over-expression technology, the pTRV::NbNAC062 silencing vector and the pEarleyGate100::RFP::NbNAC062 over-expression vector were constructed. qRT-PCR and Western blot were used to detect the changes of PVY accumulation and the expression of unfolded protein response (UPR) related gene BiP after silencing and over-expression in N. benthamiana.【Result】NbNAC062 encodes 646 amino acids, the N-terminal 28-179 aa is the NAC domain, 129-185 aa is the DNA binding region, and the C-terminal 621-643 aa is a hydrophobic transmembrane structure. Phylogenetic tree and protein sequence analysis show that N. benthamiana NbNAC062 is closely related to N. attenuata NaNAC062. The NbNAC062 promoter contains a variety of cis-acting elements related to abscisic acid, methyl jasmonate, salicylic acid and stress response. PVY infection activates NbNAC062 to transfer from cell membrane to nucleus and induces NbNAC062 up-regulation of expression. For 5 and 7 days after PVY infection, the NbNAC062 mRNA level in the treatment group was 2.52 and 1.95 times of that of the control group, respectively. For 3 days after PVY infection, the BiP mRNA expression was 2.39 times of that of the control group, and for 7 days after PVY infection, the expression of BiP was significantly lower than that of the control group, which was down-regulated by 56.77%. NbNAC062 was silenced and PVY was inoculated, compared with the control group, the expression of PVY CP mRNA was up-regulated in the silence group at 3, 5, and 7 days after inoculation, which was 2.12, 2.41, and 1.38 times of that of the control group, respectively. However, the expression of BiP mRNA was down-regulated by 28.19%, 58.11%, and 10.77%, respectively. The PVY CP protein content of the silence group was also significantly higher than that of the control group at 5 and 7 days after vaccination. NbNAC062 was over-expressed and PVY was inoculated, compared with the control group, the expression of PVY CP mRNA in the over-expression group at 24, 48, 72 hours after inoculation was down-regulated by 22.60%, 34.51%, and 36.21%, respectively, and BiP mRNA was up-regulated at 48 and 72 hours after inoculation, which was 1.56 and 1.35 times of that of the control group, respectively. The content of PVY CP in the over-expression group was also lower than that of the control group.【Conclusion】NbNAC062 belongs to the NAC class of membrane-bound transcription factors, which can be activated by PVY infection and transferred to the nucleus. It may regulate the expression of the UPR-related gene BiP to promote cell survival and inhibit early PVY infection.

Cloning and Analysis of P-glycoprotein Gene and Its Transcriptional Response to Insecticide in Chilo suppressalis

MENG XiangKun,WU ZhaoLu,YANG XueMei,GUAN DaoJie,WANG JianJun

Scientia Agricultura Sinica. 2021, 54(19): 4121-4131. doi:10.3864/j.issn.0578-1752.2021.19.008

Abstract ( 328 )

HTML ( 39 )

PDF (4406KB) ( 279 )

Save

Figures and Tables | References | Related Articles | Metrics

【Objective】The CsPgp was cloned from Chilo suppressalis, and the molecular characteristics and expression profiles of CsPgp were analyzed. Transcriptional responses as well as the potential transcriptional regulation mechanism of CsPgp to two common used insecticides (chlorantraniliprole and abamectin) were also studied.【Method】The full length of CsPgp was cloned from C. suppressalis using the gene cloning technology. The molecular characteristics and the transcription factor binding sites in 5′ transcriptional regulatory region of CsPgp were analyzed employing the bioinformatics technologies. Expression profiles of CsPgp in different stages and tissues of C. suppressalis, and the transcriptional responses of CsPgp to different doses of chlorantraniliprole and abamectin treatment were determined using the real-time quantitative PCR.【Result】The full length of CsPgp cDNA is 4 584 bp and consists of 23 exons. The encoding protein has 1 259 amino acids containing two transmembrane regions and two nucleotide binding domains and the typical structural features of ABC transporter family such as the Walker A, Walker B and D, H, P, Q-Loop which have important function in substrate transfer. CsPgp was mainly expressed in larval stage of C. suppressalis, especially in the 3rd and 4th instar larvae, while CsPgp showed low expression levels in the pupal and adult stages. Analysis of the tissue expressions showed that CsPgp was predominately expressed in the foregut and midgut, and had very low expression levels in other tissues including hindgut, fat body and Malpighian tubule. No significant change of CsPgp expression was found in the 3rd instar larvae of C. suppressalis after treated with LC₃₀ and LC₇₀ of chlorantraniliprole for 12 and 24 h, respectively, when compared with the control groups. However, the expressions of CsPgp were significantly up-regulated in larvae after treated with LC₃₀ of chlorantraniliprole for 36 h, while the expressions of CsPgp were significantly down-regulated in larvae after treated with LC₇₀ of chlorantraniliprole for 36 h. In the 0.05 mg·L^-1 of abamectin treatment, CsPgp was remarkably down-regulated at 12 h post-treatment, while the expressions of CsPgp were not significantly changed at 24 and 36 h post-treatment, respectively. However, CsPgp was significantly induced in larvae after treated with 0.15 mg·L^-1 of abamectin for 24 and 36 h, respectively. Sequence analysis of the 5′ transcriptional regulatory region of CsPgp showed that multiple transcription factor binding sites were predicted in the 5′ transcriptional regulatory region of CsPgp, including five potential CncC binding sites.【Conclusion】CsPgp was highly expressed in the midgut of C. suppressalis and could be induced by chlorantraniliprole and abamectin, which indicated that CsPgp might involve in the detoxification metabolism of chlorantraniliprole and abamectin in C. suppressalis. Multiple CncC binding sites were found in the 5′ transcriptional regulatory region of CsPgp which might have important regulatory effects on the expression of CsPgp. It was speculated that CsPgp might be regulated by transcription factor CncC and participated in the detoxification metabolism of chlorantraniliprole or abamectin when C. suppressalis was exposed to chlorantraniliprole or abamectin.

Evolution Characteristics and Influencing Factors on Inherent Soil Productivity Across Dryland

LI GuanMo,ZHANG WenJu,QU XiaoLin,QIAO Lei,HUANG YaPing,XU Hu,XU MingGang

Scientia Agricultura Sinica. 2021, 54(19): 4132-4142. doi:10.3864/j.issn.0578-1752.2021.19.009

Abstract ( 342 )

HTML ( 53 )

PDF (1634KB) ( 263 )

Save

Figures and Tables | References | Related Articles | Metrics

【Objective】Inherent soil productivity of cropland is an important index of its productivity potential. The main objective of this study was to explore the evolution patterns and to identify the main influencing factors on the contribution of inherent soil productivity (CISP), which could provide the guidance for sustainable utilization of cultivated land to improve of CISP. 【Method】Based on the national long-term field monitoring network in China, the monitoring dataset were grouped according to regional distributions, monitoring years, soil types, and soil physical and chemical properties to explore the spatial-temporal evolution patterns and importance factors on the CISP for wheat/maize season.【Result】The median values of CISP for wheat and maize were 48.9% and 53.4% across main producing regions. The CISP for maize in Northeast and Northwest China were 60.8% and 57.0%. In southwest China, the CISP for the wheat and maize were both the lowest, with the median values of 35.8% and 21.3%. During the past 30 years, the CISP of China’s cropland showed an increasing trend. The CISP in the 2010s significantly were increased by 15 percentage points compared with the value in the 1980s. The results of random forest model showed that soil type and soil pH were the main factors influencing the CISP at national scale. As for wheat on regional scale, soil organic matter (SOM) was the most important factor in the North China, and available phosphorus (AP) was the most important factor in the Southwest China and the middle and lower reaches of Yangtze River. However, as for maize, regardless of soil type and area, available potassium (AK) and soil pH were mainly important influencing factors in the North China and the lower reaches of Yangtze River. Soil organic matter (SOM) was the most important factor in the Northeast and Southwest China. 【Conclusion】The CISP in China is increasing on the whole with obvious differences among regions. On the national scale, soil type and pH were main factors casing variation of CISP. The soil type, SOM, AP and pH were mainly important factors casing variation of CISP on regional scale.

Difference in Nitrogen Responses and Nitrogen Efficiency of Different Paddy Soils in Southern and Northern China Under the Same Climatic Condition

HUANG QiuHong,LIU ZhiLei,LI PengFei,CHE JunJie,YU CaiLian,PENG XianLong

Scientia Agricultura Sinica. 2021, 54(19): 4143-4154. doi:10.3864/j.issn.0578-1752.2021.19.010

Abstract ( 299 )

HTML ( 46 )

PDF (504KB) ( 287 )

Save

Figures and Tables | References | Related Articles | Metrics

【Objective】Soil is one of the factors affecting crop yield and nitrogen fertilizer uptake or utilization. In order to provide suggestions for high yield and high quality of rice producing, we clarify the effect of paddy soil in northern and southern China on rice growth and nitrogen efficiency.【Method】In 2018 and 2019, the pot experiments were conducted in Harbin, Heilongjiang Province. The experimental soils were black paddy soil from Heilongjiang and gleyed paddy soil from Jiangsu. Three nitrogen fertilization levels were set for each soil, including no nitrogen application (N0), 0.87 g N/pot (N1, equivalent to 150 kg N·hm^-2), and 1.74 g N/pot (N2, equivalent to 300 kg N·hm^-2). Tiller numbers, SPAD value, yields and earing rates of rice, as well as nitrogen mineralization amount and nitrogen utilization efficiency of two soils, were determined. 【Result】Rice tiller numbers on black paddy soil increased with the increase of nitrogen fertilizer application at early growth stage, however, which on gleyed paddy soil was responded to nitrogen application after the elongation stage. Soil type has a significant effect on rice tiller number. In 2018, rice tiller numbers on gleyed paddy soil was 4.41%-43.04% higher than that on black paddy soil without nitrogen application, while tiller numbers was 8.25%-12.98% lower than that on the black paddy soil after nitrogen application. In 2019, the most of tiller numbers on black paddy soil was 4.41%-46.53% higher than that on gleyed paddy soil. In 2018, the leaf SPAD value and the earbearing tiller percentage of rice showed significant differences between two soil types. The leaf SPAD value on gleyed paddy soil was 19.28%-21.19% higher than that on black paddy soil, and also, earbearing tiller percentage of rice on gleyed paddy soil was 23.89%-40.53% higher than that on black paddy soil, but no significant difference between two soil types was observed in leaf SPAD value and earbearing tiller rate in 2019. Water-logged incubation over 28 days showed that two types of soils had the same inorganic nitrogen content. Initial nitrogen mineralization rates in gleyed paddy soil was higher than that in black paddy soil, while nitrogen mineralization rate in gleyed paddy soil at later stage was lower than that in black paddy soil. A higher nitrogen mineralization potential was observed in black paddy soil, indicating the greater mineralization capacity. The nitrogen agronomic efficiency (AE_N) of black paddy soil was higher in comparison with gleyed paddy soil, while the partial factor Productivity of applied N (PFP_N ) showed the opposite trend. A higher Y₀/Nr (Y₀is the yield of rice field without nitrogen fertilizer application, and Nr is the amount of nitrogen fertilizer application) was found in gleyed paddy soil, suggesting a better coordination between soil nitrogen supply and application. The nitrogen recovery efficiency (RE_N) and nitrogen physiological efficiency (PE_N) of black paddy soil was remarkably higher in comparison with gleyed paddy soil in 2018, but the two soil types had no significant difference in the RE_N and PE_N in 2019.【Conclusion】Soil difference was not the decisive factor of nitrogen efficiency difference which was observed between southern and northern paddy fields in China, but rather the results of the combined effects of factors such as climate, crop variety, soil type, etc. Compared with black paddy soil, the gleyed paddy soil should decrease base and tiller nitrogen fertilizer, and increase the panicle nitrogen fertilizer to maintain sufficient nitrogen supply in the later stages and obtain high rice yield.

Effects of Straw Interlayer with Different Thickness on Saline-Alkali Soil Temperature, Water Content, and Sunflower Yield in Hetao Irrigation Area

WANG GuoLi,CHANG FangDi,ZHANG HongYuan,LU Chuang,SONG JiaShen,WANG Jing,PANG HuanCheng,LI YuYi

Scientia Agricultura Sinica. 2021, 54(19): 4155-4168. doi:10.3864/j.issn.0578-1752.2021.19.011

Abstract ( 277 )

HTML ( 32 )

PDF (588KB) ( 276 )

Save

Figures and Tables | References | Related Articles | Metrics

【Objective】The effects of different thickness of straw interlayer on soil temperature, water content dynamic change and crop yield of sunflower field in saline-alkali land were studied to provide a basis for selecting reasonable thickness straw interlayer measures suitable for the growth of sunflower in Hetao irrigation area. 【Method】 From 2015 to 2017, 4 straw interlayers of different thicknesses were set up in the typical saline-alkali farmland in the Hetao area of Inner Mongolia, namely CK (no straw interlayers), S3 (straw interlayers with a thickness of 3 cm), S5 (straw interlayers with a thickness of 5 cm), and S7 (straw interlayer with a thickness of 7 cm), to study the effects of different thickness of straw interlayer on the dynamic changes of soil temperature, water content, and sunflower yield during the growth period of sunflower under. 【Result】The straw interlayer treatment (S3, S5 and S7) significantly increased the soil temperature of 0-40 cm soil layer during the whole growth period of sunflower. Among them, compared with CK treatment, the soil temperature in sunflower seedling stage significantly increased by 0.7℃, 0.6℃, and 0.5℃, respectively, from 2015 to 2017 (P<0.05), and the increase in temperature gradually decreased with the increase of straw burying time. There was a significant difference between straw interlayer treatments during flowering period, among which S5 and S7 treatments increased by 0.4℃ and 0.6℃, respectively, compared with CK treatment in average for 3 years (P<0.05); 40-50 cm soil layer straw interlayer treatment showed a trend of increasing temperature in seedling and bud stage of sunflower, and showed a cooling trend in the later growth period. Under different treatments, the soil temperature during the whole growth period of sunflower decreased as the soil layer deepens, and there was a very significant positive correlation between soil temperature and atmospheric temperature. The distribution range of R² value in three years was 0.628-0.735. Straw interlayer treatments enhanced the sensitivity of soil temperature to atmospheric temperature, and the response of soil temperature to atmospheric temperature decreases with the increase of straw burying time. The interaction between different straw interlayer treatments and different irrigation periods also had significant effects on soil water content (P<0.05). Straw interlayer treatment could reduce the average soil water content of the 0-40 cm soil layer before irrigation and after harvest. Among them, the S7 treatment had the largest reduction, and the three-year average decreased by 7.9% and 5.4% (P<0.05), compared with the CK treatment; after irrigation, the average soil water content of S3, S5 and S7 treatments increased by 2.3%, 3.4%, and 3.6%, respectively, compared with CK treatment for 3 years (P<0.05). The straw interlayer treatment could promote the growth of sunflower and increase the yield of sunflower, and improve irrigation water productivity and water use efficiency, among which 5 and 7 cm thick straw interlayer treatments increased the most and had the largest increase, but there was no significant difference between the two treatments (P>0.05). 【Conclusion】Straw interlayers of different thicknesses could increase the soil temperature in the 0-40 cm soil layer during the growth period of sunflower, and the increase of temperature decreased with the increase of the straw burying time. The difference between different treatments was significant during the flowering stage. In addition, the straw interlayer treatment could improve the average soil water content of 0-40 cm soil after irrigation, and provide a suitable growth environment for sunflower. Considering soil temperature and crop water use efficiency, straw separation with thickness of 5 cm is the most suitable for promotion and application in Hetao irrigation area of Inner Mongolia.

Cloning and Identification of γ-Glutamyl Transpeptidase AcGGT Gene from Onion (Allium cepa)

XU HuanHuan,LI Yi,GAO Wei,WANG YongQin,LIU LeCheng

Scientia Agricultura Sinica. 2021, 54(19): 4169-4178. doi:10.3864/j.issn.0578-1752.2021.19.012

Abstract ( 329 )

HTML ( 35 )

PDF (2533KB) ( 755 )

Save

Figures and Tables | References | Related Articles | Metrics

【Objective】Alliin metabolized by Allium plants had important pharmaceutical value. γ-glutamyl transpeptidase was a key enzyme in the deglutamylation step of the process of alliin synthesis. Studying the function of γ-glutamyl transpeptidase gene in onion can reveal the role of γ-glutamyl transpeptidase in alliin synthesis pathway, providing theoretical basis for alliin synthesis in vitro and laying a foundation for further study on alliin synthesis mechanism. 【Method】Using onion as material, the primers were designed according to onion RNA-seq database, and the gene, γ-glutamyl transpeptidase, was cloned from onion by RT-PCR and analyzed by bioinformatics. The CAMV 35S-AcGGT-GFP vector was used to bombard onion inner epidermis cells with gold powder plasmid microcarrier by particle bombardment technology, and the subcellular localization of AcGGT was determined by fusion green fluorescent expression protein. The Saccharomyces cerevisiae expression vector with AcGGT was constructed, transforming and inducing the expression of AcGGT, and using the method of transforming glutamyl-p-nitroaniline to p-nitroaniline by γ-glutamyl transpeptidase to determine the glutamyl transpeptidase activity of the total protein of Saccharomyces cerevisiae transferred into AcGGT. Real time quantitative PCR was used to analyze the differential expression pattern of the gene in onion tissues. The activity of endogenous transpeptidase in onion tissues was determined by the method of γ-glutamyl transpeptidase catalyzing the production of p-nitroaniline from p-nitroaniline. 【Result】AcGGT was cloned and its length was 1 869 bp. Bioinformatics analysis showed that AcGGT encoded 622 amino acids, protein domain prediction showed that it had glutamyl transpeptidase domain, secondary structure was mainly α - helix, transmembrane region analysis suggested that GGT protein had transmembrane region, amino acid multiple alignment results showed that GGT in plants had certain conservation, evolutionary analysis showed that AcGGT was related to garlic AsGGT2, and the relationship is closest. The fluorescence signal of CaMV 35S-AcGGT-GFP fusion protein was located in the vacuole, indicating that the protein encoded by CaMV 35S-AcGGT-GFP was located in the vacuole. The results of glutamyl transpeptidase activity assay showed that the glutamyl transpeptidase activity of yeast transformed with AcGGT was significantly higher than the control, indicating that the protein encoded by AcGGT had transpeptidase activity. The results of differential expression analysis of AcGGT showed that the expression of AcGGT was mainly in leaf sheath, bulb and leaf sheath followed by. The activity of glutamyl transpeptidase in different tissues, the highest activity in leaf, followed by leaf sheath. Correlation analysis showed that there was no significant correlation between the activity of transglutaminase and the expression of AcGGT. 【Conclusion】The enzymatic reaction data of exogenous AcGGT expression were obtained. The deglutination of alliin synthesis pathway preceded S-oxygenation; there was no significant correlation between AcGGT expression and endogenous transglutaminase activity in onion, suggesting that there may be multiple transglutaminase genes in onion.

Metabolomics Comparative Study on Fruits of Edible Seed Watermelon, Egusi and Common Watermelon

YUAN PingLi,HE Nan,ZHAO ShengJie,LU XuQiang,ZHU HongJu,DIAO WeiNan,GONG ChengSheng,MUHAMMAD Jawad Umer,LIU WenGe

Scientia Agricultura Sinica. 2021, 54(19): 4179-4195. doi:10.3864/j.issn.0578-1752.2021.19.013

Abstract ( 402 )

HTML ( 38 )

PDF (2718KB) ( 410 )

Save

Figures and Tables | References | Related Articles | Metrics

【Objective】Watermelon is a very popular summer fruit. Citrullus mucosospermus (egusi), Citrullus lanatus var. megalospermus (edible seed watermelon) and Citrullus lanatus var. vulgaris (common dessert watermelon) are the three main cultivation types. The appearance and taste of their fruits are quite different, but there is no detailed report on the comprehensive fruit metabolome. In order to clarity the influence of selection on metabolomics in watermelon domestication and improvement processes, the metabolic profiling of three types of watermelon fruits was analyzed. 【Method】 In this study, 5 egusi watermelon, 5 edible seed watermelon and 6 common watermelon accessions were used for metabolomics profiling analysis by widely targeted metabolic profiling method based on liquid chromatography tandem mass spectrometry (LC-MS/MS). The software SIMCA-P, MetaboAnalyst 5.0, and Origin were used to analyze the metabolomics data. 【Result】A total of 323 metabolites were detected, including 51 amino acids and their derivatives, 21 nucleotides and their derivatives, 14 carbohydrates, 32 organic acids, 52 lipids, 36 flavonoids, 32 hydroxycinnamoyl derivatives, etc. PCA and cluster analysis showed that the difference of metabolomics profile between egusi watermelon and edible seed watermelon was small, and the metabolomics profile of edible seed watermelon was between that of egusi watermelon and common watermelon. The major metabolites contributing to the classification were sucrose, citric acid, L-glutamic acid, L-threonine, cucurbitacin, vanillic acid glycoside isomers, lipids, and so on. Heatmap analysis showed that cucurbitacin and their derivatives were the unique metabolites in egusi watermelon. The contents of chlorogenic acid, LysoPE, etc. in edible seed watermelon were higher than those in egusi and common watermelon. The contents of carbohydrate, arginine, ferulic acid and C18-2 and C18-3 unsaturated fatty acids in common watermelon were higher than those in egusi and edible seed watermelon. There were 156 differentially accumulated metabolites, which were identified by comparison analysis. The main divergent metabolites between edible seed watermelon and egusi watermelon were p-coumaraldehyde, ferulic acid, cinnamic acid, sucrose, cucurbitacin D O-glucoside, cucurbitacin E isomer, vitexin, coniferyl alcohol, and so on. The main divergent metabolites between edible seed watermelon and common watermelon were lipids, flavonoids, organic acids, sugars, and others. With the development of crop evolution, the content of nutritious metabolites increased, whereas the content of resistance related metabolites decreased. 【Conclusion】The watermelon fruit metabolomics was reported for the first time based on widely targeted metabolic profiling method. The metabolomics difference between edible seed watermelon and egusi watermelon was smaller than that of edible seed watermelon and common watermelon, and the metabolic profile of edible seed watermelon lay between that of egusi watermelon and common watermelon. In addition to phenotypic and genomic differences, the differential metabolites could also be used as biomarkers to distinguish different varieties. The metabolomics data provided the new insight for complementary understanding watermelon germplasm resources and for metabolomics-based genetic improvement of functional watermelon.

Specific Length Amplified Fragment (SFLA) Sequencing Mapping Construction and QTL Analysis of Fruit Related Traits in Muskmelon

WANG Ling,CAI Yi,WANG GuiChao,WANG Di,SHENG YunYan

Scientia Agricultura Sinica. 2021, 54(19): 4196-4206. doi:10.3864/j.issn.0578-1752.2021.19.014

Abstract ( 293 )

HTML ( 38 )

PDF (1331KB) ( 214 )

Save

Figures and Tables | References | Related Articles | Metrics

【Objective】 Through the QTL mapping and candidate gene analysis of muskmelon (Cucumis melo L.) fruit-related traits, so as to provide a theoretical basis for muskmelon quality breeding, gene mapping and functional verification. 【Method】 The F₁ plant was made cross by 1244 as female parent with thin-skinned melon and M5 with thick-skinned melon as male parent, combined with SFLA sequencing technology to develop molecular tags and construct high-density genetic maps. Based on F_2:3 phenotype data, Mapqtl analysis method was used to detect QTL location. 【Result】 Totally 380 446 Mreads (83.12 Gb) data were obtained, the average Q30 and GC content was 93.59% and 36.87%, respectively; totally 112 844 SLAF tags and 3 274 879 SNPs were obtained; a genetic map contained 12 linkage groups with a total of 10 596 markers were constructed. The genetic map spanned 1 383.88 cM with the average distance was 0.13 cM between markers, and totally 99.92% developed markers were mapped. The gene controling fruit sutures (fst) of melon was located at the end of chromosome 11, between Marker 1993423 (62.18) and Marker 1998820 (63.05), covering 0.72 Mb of the genome, and containing 33 candidate genes. The gene for pericarp pattern (fpp) was located at chromosome 2 between Marker 459584 (90.91) and Marker 459446 (90.91) which covered the genome of 0.08 Mb and contained 5 candidate genes, among them MELO3C026292 (1-deoxy-D-xylulose-5-phosphate reductoisomerase) might be the candidate gene for pericarp pattern. At the same time, QTL locus of pericarp color (fpc) was detected which located between Marker 1229174 (7.14) and Marker 1229973 (7.14) on the chromosome 7, with the contribution rate was 9.9%. One QTL locus of fruit shape (fs) fs9.1 was detected between Marker 1705671 (76.19) and Marker 1705915 (79.23) on chromosome 9 with a contribution rate of 7.6%. Six QTL loci related to single fruit weight (sfw1.1, sfw2.1, sfw2.2, sfw6.1, sfw7.1, sfw10.1) were detected on chromosomes 1, 2, 6, 7, and 10 with the contribution rate were between 3.1% to 17%, and the LOD value were between 3.0 to 5.6. 【Conclusion】The fruit sutures and pericarp pattern genes were located on the chromosomes11 and 2, and then 33 and 5 candidate genes were obtained, respectively; One QTL for pericarp color, one QTL for fruit shape and 6 QTLs for single fruit weight were also detected.

Effects of Steaming-Retrogradation Pretreatment on Physicochemical Properties and in Vitro Starch Digestibility of the Roasted Highland Barley Flour

WANG YuLin,LEI Lin,XIONG WenWen,YE FaYin,ZHAO GuoHua

Scientia Agricultura Sinica. 2021, 54(19): 4207-4217. doi:10.3864/j.issn.0578-1752.2021.19.015

Abstract ( 295 )

HTML ( 40 )

PDF (1913KB) ( 317 )

Save

Figures and Tables | References | Related Articles | Metrics

【Objective】 This study was to explore the effects of steaming-retrogradation pretreatment on the nutritional profiles, physicochemical properties, and in vitro starch digestion of the roasted highland barley flour, with the aim to enrich the processing methods of highland barley.【Method】The effects of steaming-retrogradation pretreatment at different time (0, 6, 12, 18, and 24 h, respectively) on roasted highland barley flour was studied. The microstructure of the roasted highland barley flour was observed by scanning electron microscope and microscope. The effect of steaming-retrogradation pretreatment on short-range molecular order structure, relative crystallinity, and pasting properties of the roasted highland barley flour was determined by Fourier transform infrared spectroscopy, laser confocal microscopic Raman spectroscopy, X-ray diffraction, and rapid viscosity analysis. The effects of steaming-retrogradation pretreatment on the changes of starch digestibility in roasted highland barley flour were determined via In vitro digestion. 【Result】 Compared with the raw highland barley flour, the steaming-retrogradation pretreatment reduced the contents of starch, protein, fat, and β-glucan in roasted highland barley flour. However, steaming without retrogradation pretreatment (0-Roasted) increased the content of water-insoluble dietary fiber, water-soluble dietary fiber, and total dietary fiber in the roasted highland barley flour. The steaming-retrogradation pretreatment destroyed the starch granules in roasted highland barley flour, presenting disappearance of polarization cross. Meanwhile, the short-range molecular order structure was damaged by steaming-retrogradation pretreatment in roasted highland barley flour. Compared with the raw highland barley flour, the relatively crystallinity was reduced by steaming-retrogradation pretreatment in roasted highland barley. However, the relative crystallinity was increased with the increased time of retrogradation in roasted highland barley. The A-type diffraction pattern was changed into the V-type in roasted highland barley flour, indicating the formation of starch-lipid or starch-protein-lipid complexes after heat processing. The steaming-retrogradation pretreatment decreased the L* value while increased a* and b* values in roasted highland barley flour. Compared with the raw highland barley flour, the steaming-retrogradation pretreatment destroyed the pasting peak and caused a lower final viscosity, and increased the oil holding capacity while decreased the water holding capacity of roasted highland barley flour. Compared with the raw highland barley flour, the heat processing could increase the in vitro starch digestibility. Retrogradation at 6-24 h could decrease the content of rapidly digestible starch by 6%-16% and significantly increased the content of slowly digestible starch in roasted highland barley flour. 【Conclusion】 The steaming-retrogradation pretreatment could change nutritional profiles and physicochemical properties of roasted highland barley flour. Retrogradation at 6 h could decrease the in vitro starch digestion of the roasted highland barley flour, which was helpful in maintaining blood glucose homeostasis and could be developed as one of potential foods for diabetics and borderline diabetics.

Evaluation of Feeding Value for Whole Broussonetia papyrifera Silage in Diet of Wuchuan Black Beef Cattle

CHEN GuangJi,XIONG XianQin,HE RunXia,TIAN Xiong,SHEN YingLong,ZOU XiaoMin,YANG Hong,SHANG YiShun,ZHAO MingKun,LI XiaoDong,LI ShiGe,ZHANG Rong,SHU JianHong

Scientia Agricultura Sinica. 2021, 54(19): 4218-4228. doi:10.3864/j.issn.0578-1752.2021.19.016

Abstract ( 310 )

HTML ( 34 )

PDF (642KB) ( 237 )

Save

Figures and Tables | References | Related Articles | Metrics

【Objective】 This study aimed to explore the feeding value of whole Broussonetia papyrifera silage (WBPS) on Wuchuan black beef cattle, so as to provide a reference for rational utilization of Broussonetia papyrifera as feed.【Method】 A completely randomized trial design was used, and fifty head of black beef cattle with similar weight and age ((108.06±14.51) kg, 9 months) were randomly assigned to five dietary treatments: (A, B, C, D, and E). Each treatment had 1 replicate, with 10 steers in each replicate. Each treatment group was fed diets with the same concentrate to forage ratio () for 288 days, and the ratio of A, B, C, D, and E was 0, 17%, 41%, 66% and 83% WBPS, respectively. The dry matter intake (DMI), daily gain (ADG) and feed to weight ratio (DMI/ADG) were measured at the beginning, the middle stage (175th day and 220th day ) and the end of the experiment (288th day). The body size, rumen fermentation parameters and carcass quality were measured at the end of the experiment. 【Result】 ADG of C and D were higher than those of other groups at 0-175th day, 175-220th day, 220-288 day and the whole period (P<0.05), while DMI/ADG value was lower (P<0.05). In addition, the dietary factors affected the ADG time gradient of each group: with the prolongation of the experimental period, the ADG of A and B decreased significantly, while that of C, D and E increased firstly and then decreased. The increment of body height and oblique length of D was higher than that of other groups, and the second was group C (P<0.05). The ruminal acetic, propionic and total volatile fatty acids of A and B were higher than those of other groups, but the rumen microbial protein production of D was the highest, which was 5.27 times higher than that of A. The trial factor had no significant effect on slaughter rate and net meat rate of Wuchuan black beef cattle (P>0.05), while increasing the proportion of WBPS in the diet decreased the carcass fat percentage and muscle shear force (P<0.05). There was no significant difference in the composition and content of amino acids in muscle of Wuchuan black beef cattle (P>0.05), but WBPS decreased the content of saturated fatty acids and increased the content of polyunsaturated fatty acids (P<0.05). 【Conclusion】 WBPS replacing whole plant corn silage as a dietary component of Wuchuan beef black cattle could improve daily gain, reduce feed to weight ratio, increase rumen microbial protein production, reduce carcass fat rate, and positively change muscle fatty acid composition.

Correlation Analysis of Inosine Monophosphate Specific Deposition Related LNC_003828-gga-miR-107-3P-MINPP1 in Jingyuan Chicken Muscle Tissue

YU BaoJun,DENG ZhanZhao,XIN GuoSheng,CAI ZhengYun,GU YaLing,ZHANG Juan

Scientia Agricultura Sinica. 2021, 54(19): 4229-4242. doi:10.3864/j.issn.0578-1752.2021.19.017

Abstract ( 337 )

HTML ( 35 )

PDF (4456KB) ( 262 )

Save

Figures and Tables | References | Related Articles | Metrics

【Objective】The aim of this study was to explore the regulatory role of key regulatory factors in the process of inosine monophosphate deposition in the muscle tissue of Jingyuan chickens, and to use lncRNA-miRNA-mRNA association analysis to identify LNC_003828, gga-miR-107-3p and MINPP1 related to inosine monophosphate specific deposition, so as to provide a theoretical basis for molecular-assisted breeding to improve chicken muscle quality.【Method】 The inosine monophosphate content of the breast and leg muscles of 15 Jingyuan chickens was determined, and three samples of the breast muscles with high inosine monophosphate content and the leg muscles with low inosine monophosphate content were screened to extract total RNA. The cDNA library was constructed after passing the quality test, and PCR amplification test was carried out. Then, the cDNA library quality was evaluated by using Agilent 2100, which was sent the library to the Illumina-Hiseq platform for transcriptome sequencing. Using bioinformatics methods, the differentially expressed MINPP1, gga-miR-107-3p and LNC_003828 in different parts of the muscle tissue of Jingyuan chicken were screened out, and GO annotation and protein interaction network was used to analyze the function of MINPP1. The qRT-PCR method was used to detect the expression of LNC_003828, gga-miR-107-3p and MINPP1 in the breast and leg muscles of Jingyuan chickens, and the correlation between them and the content of inosine monophosphate was analyzed. 【Result】 R², the correlation of gene expression levels between sequenced samples, was greater than 0.9, that is, gene expression between experimental samples could be used for subsequent differential gene analysis. Three differentially expressed genes, including MINPP1, PKM, and ALDH9A1, were detected in the glycolysis/gluconeogenesis pathway involving in the synthesis and metabolism of inosine monophosphate. Interaction analysis found that there were 17 miRNAs (9 up-regulated, 8 down-regulated), 44 mRNAs (16 up-regulated, 28 down-regulated), and 155 lncRNAs (68 up-regulated, 87 down-regulated) in the lncRNA-miRNA-mRNA network diagram, of which the target gene of the core node gga-miR-107-3p interaction was MINPP1, and the target lncRNA was LNC_003828. GO enrichment analysis found that the MINPP1 gene had functions such as phosphatase activity and bisphosphoglycerate phosphatase activity; the MINPP1 gene in the protein interaction network were all interact with PGAM1 and ENO1, which were involved in glycolysis/gluconeogenesis and amino acid biosynthesis pathways BPGM genes. The results of qRT-PCR showed that the relative expression of LNC_003828 and gga-miR-107-3p in breast muscle of Jingyuan chicken was lower than that of leg muscle, but the difference was not significant; the relative expression of MINPP1 in breast muscle was significantly lower than that of leg muscle (P<0.05). The expression of gga-miR-107-3p in the breast and leg muscle tissues of Jingyuan chicken was positively correlated with the expression of LNC_003828 and negatively correlated with the expression of MINPP1. The expression of LNC_003828 and gga-miR-107-3p in breast and leg muscle tissues were positively correlated with inosine monophosphate content, and the difference was not significant; the expression of breast muscle MINPP1 was negatively correlated with inosine monophosphate content and the expression of leg muscle MINPP1. The amount was significantly negatively correlated with the content of inosine monophosphate (P<0.05). In summary, it was speculated that gga-miR-107-3p in the muscle tissue of Jingyuan chicken was used as a core regulator to adsorb LNC_003828, which affected the MINPP1 gene to regulate the specific deposition of muscle inosine monophosphate, thereby improving meat quality. 【Conclusion】 LNC_003828, gga-miR-107-3p, and MINPP1 were selected as candidate regulatory factors affecting the specific deposition of inosine monophosphate.

Detection of Interaction Between Porcine Type I Complement Receptor and C3b Active Fragment in Vitro

SUN YuChen,JIA RuiPu,FAN KuoHai,SUN Na,SUN YaoGui,SUN PanPan,LI HongQuan,YIN Wei

Scientia Agricultura Sinica. 2021, 54(19): 4243-4254. doi:10.3864/j.issn.0578-1752.2021.19.018

Abstract ( 322 )

HTML ( 35 )

PDF (984KB) ( 466 )

Save

Figures and Tables | References | Related Articles | Metrics

【Objective】In order to provide scientific data for elucidating the molecular mechanism of porcine erythrocyte immune adhesion function, it was investigated whether CR1-like (Complement receptor 1-like, CR1-like) of porcine erythrocyte could bind to the C3b or not.【Method】In this study, the recombinant plasmids of CR1-like_(3-6) and CR1-like_(8-11) functional domain fragments were constructed first, which were used to establish a yeast two-hybrid detection system. The bait plasmid (recombinant pGBKT7-CR1-like) and capture plasmid (recombinant pGADT7-C3b) were co-transformed into Y2HGold yeast cells. The single deficient SD/-Leu, SD/-Trp and double-deficient SD/-Leu/-Trp (DDO) media were used to strictly screen the co-transformed yeast cells. Then, according to the expression of report factor, the growth of transformants were identified on the double-deficient medium SD/-Leu/-Trp/X-α-Gal (DDO/X) or SD/-Leu/-Trp/X-α-Gal/Aba (DDO/X/A) combined with the color change phenomenon of the colony to comprehensively determine whether CR1-like active fragments and complement C3b bind to each other in yeast cells or not. The CR1-like-C3b binding complex in yeast cells was then separated by immunoprecipitation, and the specificity of the complex was identified by Western blot. 【Result】The co-transformed yeast clones showed normal growth on SD/-Leu, SD/-Trp, DDO and DDO/X, DDO/X/A media with blue color colonies, and this indicated that positive yeast colonies were successfully obtained. The results of PCR reverse identification showed that the co-transformed yeast contained the target genes CR1-like_(3-6) and CR1-like_(8-11). The C3b gene fragment appeared after the plasmid was digested, indicating that the recombinant plasmid pGBKT7-CR1-like and pGADT7-C3b were successfully co-transformed into yeast cells. In the immunoprecipitation test, the tag antibody c-Myc of the pGBKT7 vector was used to precipitate the fusion protein in yeast cells. Western blot detection with c-Myc as the primary antibody revealed that the fusion protein transformed pGBKT7-CR1-like_(3-6) and pGBKT7-CR1-like_(8-11) separately showed a specific band at 50 kDa; the yeast fusion protein co-transformed with pGBKT7-CR1-like_(3-6) + pGADT7-C3b and pGBKT7-CR1-like_(8-11) + pGADT7-C3b showed a specific band at 83 kDa; when the HA monoclonal antibody was used as the primary antibody for Western blot detection, no specific bands appeared in the pGBKT7-CR1-like_(3-6) and pGBKT7-CR1-like_(8-11) fusion proteins, and only the yeast fusion protein co-transformed in lane 3 and 4 showed a specific band at 83 kD. It showed that there was a complex of CR1-like and C3b in Y2HGold yeast cells. Using CR1-like monoclonal antibody to precipitate the fusion protein in yeast cells, Western blot detection with CR1-like as the primary antibody revealed that the fusion protein transformed with pGBKT7-CR1-like_(3-6) and pGBKT7-CR1-like_(8-11) separately showed a specific band at 50 kD; the yeast fusion protein co-transformed with pGBKT7-CR1-like_(3-6) + pGADT7-C3b and pGBKT7-CR1-like_(8-11) + pGADT7-C3b showed a specific band at 83 kD; when the C3 monoclonal antibody was used as the primary antibody for Western blot detection, no specific bands appeared in the pGBKT7-CR1-like_(3-6) and pGBKT7-CR1-like_(8-11) fusion proteins, lanes 3 and 4 showed that only the co-transformed yeast fusion protein had a specific band at 83 kD. This indicated that there was a biologically active CR1-like and C3b binding complex in Y2HGold yeast cells. The bait plasmid expression products CR1-like_(3-6), CR1-like_(8-11) fragments and capture plasmid expression products C3b fragment could be combined in yeast cells.【Conclusion】In summary, the recognition ligand for porcine erythrocyte CR1-like to exert immune adhesion function was C3b, which provided an important data basis for the further analysis of the molecular structure of CR1-like functional domain.

Table of Content