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    16 May 2021, Volume 54 Issue 10
    CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS
    OsCSC11 Mediates Dry-Hot Wind/Drought-Induced Ca2+ Signal to Regulate Stamen Development in Rice
    REN ZhiJie,LI Qian,SUN YuJia,KONG DongDong,LIU LiangYu,HOU CongCong,LI LeGong
    Scientia Agricultura Sinica. 2021, 54(10):  2039-2052.  doi:10.3864/j.issn.0578-1752.2021.10.001
    Abstract ( 599 )   HTML ( 86 )   PDF (6692KB) ( 592 )   Save
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    【Objective】When rice is occasionally stressed by dry-hot wind (DHW) or drought at flowering stage, the yield greatly decreases due to the rapid water loss in developing gamete cells. During this process, calcium as a universal second messenger mediates cellular signal transduction in response to drought or other stresses. However, the underlying molecular mechanism remains elusive. This study attempts to dissect the physiological and molecular function of Oryza sativa Calcium-permeable Stress-responsive Cation channels (OsCSCs) which will provide a new strategy for studying the stress responsive mechanism to DHW in crops.【Method】Based on the methods of genetics, electrophysiology and bioinformatics, a typical receptor-like-channel named OsCSC11 was identified. The expression pattern of OsCSC11 were analyzed by qRT-PCR and GUS staining. Further, subcellular localization of OsCSC11-GFP was observed in Arabidopsis protoplasts and onion epidermal cells. Meanwhile, the oscsc11 mutants were generated by CRISPR/Cas9 gene editing tool. Finally, the phenotype and physiological functions were analyzed by cytological method. 【Result】Sequence alignment results in DUF221 family revealed that OsCSC11 includes a typical conserved domain and some unique motifs, which belongs to an independent subfamily. OsCSC11 is mainly expressed in anthers and leaves in rice. Further analysis showed that the activity of full-length protein of OsCSC11 which is in a resting state can be activated by the hypertonic solution. However, OsCSC11ΔTM1-3(TM1-3 was truncated in OsCSC11) has constitutive channel activity that specifically mediates divalent cations of calcium and magnesium. Thus, we speculated that TM1-3 is a receptor domain in CSCs/OSCAs channel for sensing DHW stresses, and the rest part of OsCSC11 (OsCSC11ΔTM1-3) generates calcium signal. OsCSC11 and OsCSC11ΔTM1-3 are both localized in the plasma membrane which may be related to the receptor function. In loss of function mutants oscsc11-1 and oscsc11-2, anthers become smaller and wrinkler compared to wild type, and pollen sterility rate reached to 60%-70% and water content dramatically reduced.【Conclusion】OsCSC11 functions in regulating anther water status and pollen development through mediating calcium influx and possibly involves in the primary sensing step under DHW stresses.

    Genome-wide Association Analysis of Wheat Grain Size Related Traits Based on SNP Markers
    ZHANG Fang,REN Yi,CAO JunMei,LI FaJi,XIA XianChun,GENG HongWei
    Scientia Agricultura Sinica. 2021, 54(10):  2053-2063.  doi:10.3864/j.issn.0578-1752.2021.10.002
    Abstract ( 524 )   HTML ( 60 )   PDF (1525KB) ( 503 )   Save
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    【Objective】Grain traits are important factors affecting wheat yield. the significant locus of controlling wheat grain traits was explored by genome-wide association analysis of wheat grain traits, which provided a theoretical reference for the study of genetic improvement of wheat grain traits. 【Method】The genome-wide association analysis (GWAS) based on mixed linear model MLM (Q+K) was performed on 121 wheat grown in Xinjiang using wheat 50 K SNP chips for 6 traits which including grain length, grain width, grain length-width ratio, grain area, grain perimeter and 1000-grain weight.【Result】Six grain traits showed wide phenotypic variation in different environments, in which the maximum coefficient of variation of 1000-grain weight was 13.91-17.79 and the heritability of each grain trait was between 0.85-0.90. The polymorphism information content PIC value was 0.09-0.38, and the minimum allele frequency MAF value was 0.05-0.50. Group structure analysis shows that the natural groups used in the experiment can be divided into 4 subgroups. GWAS results showed that a total of 592 significant association sites (P<0.001) were detected in 6 traits, of which 29 SNPs were repeatedly detected in 2 or more environments, distributed in 1A(5), 1B(2), 1D, 2A(5), 3B, 5A, 5D, 6B(4), 6D, 7B and 7D(7) chromosomes, can explain 9.3% to 22.7% of the phenotypic variation. Six markers associated with stable grain length were detected, which distributed on 1A, 2A, and 7D chromosomes to explain the phenotypic variation of 9.9%-22.7%. Two markers associated with stable grain width were detected, which distributed on 3 B and 5 D chromosomes to explain the phenotypic variation of 9.6%-12.2%. Six markers associated with stable grain length-width ratio were detected, which distributed on 2A(2), 5A, 7B, and 7D(2) chromosomes to explain the phenotypic variation of 10.1%-19.4%. Three markers associated with stable grain area were detected, which distributed on 1A, 1B and 1D chromosome to explain the phenotypic variation of 9.9%-18.2%. Six markers with stable correlation with grain perimeter were detected, which distributed on 1A(2), 2A, 6D and 7D(2) chromosomes to explain the phenotypic variation of 9.3%-22.6%. Six markers associated with stable 1000-grain weight were detected, which distributed on 1B, 2A and 6B chromosomes to explain the phenotypic variation 9.7%-12.9%. Five dominant loci of pleiotropism with were found to control wheat grain traits, which distributed 1A, 2A(2) and 7D(2) chromosomes, explaining the phenotypic variation of 9.9%-22.7%.【Conclusion】In this study, the genetic diversity of the materials was abundant, a total of 29 multi-environment stability loci were found in natural population with 2 or more environmental associated with 6 grain traits.

    Creation of New Maize Variety with Fragrant Rice Like Flavor by Editing BADH2-1 and BADH2-2 Using CRISPR/Cas9
    ZHANG Xiang,SHI YaXing,LU BaiShan,WU Ying,LIU Ya,WANG YuanDong,YANG JinXiao,ZHAO JiuRan
    Scientia Agricultura Sinica. 2021, 54(10):  2064-2072.  doi:10.3864/j.issn.0578-1752.2021.10.003
    Abstract ( 500 )   HTML ( 40 )   PDF (3919KB) ( 490 )   Save
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    【Objective】Fragrance is an important trait for quality improvement of crops. The 2-acetyl-1-pyrroline (2-AP) is the major component of the aroma flavor. BADH2 controls fragrance in plants, and its null or weak alleles lead to 2-AP accumulation. In this study, the fragrance related genes were modified in JING724, a maize elite inbred line invented by Beijing Academy of Agriculture and Forestry Sciences, using CRISPR/Cas9 to improve its trait of fragrance. 【Method】To find BADH gene family of target species, OsBADH2 protein sequence was used to search against protein databases of Arabidopsis, rice and maize with the Ensembl online BLAST tool. All BADH family members were verified by protein domain information in UniProt database. Furthermore, phylogenetic analysis conducted in MEGA software was used to search for maize BADH2 homologs as gene-editing targets. Based on principles of CRISPR/Cas9, we designed highly specific sgRNA to target the candidate genes. The CRISPR/Cas9 vector containing this sgRNA was introduced into the maize variety JING724 by Agrobacterium-mediated transformation. We obtained transgenic maize plants with PMI resistance. Sanger sequencing was used to confirm the CRISPR/Cas9-mediated mutations. Finally, we used gas chromatography-mass spectrometry (GC-MS) to measure the 2-AP content in T1 seeds of the gene-editing lines. 【Result】Phylogenetic analysis showed that there were two BADH2 homologs in the maize gnome, subsequently they were named ZmBADH2-1 and ZmBADH2-2. ZmBADH2-1 is located in chromosome 4, whereas ZmBADH2-2 was in chromosome 1. Both genes have 15 exons and 14 introns. The 4th exon of ZmBADH2-1 shares high nucleotide identity with that of ZmBADH2-2. A specific target sequence, which is located in the 4th exons of both genes, was designed and introduced into a CRISPR/Cas9 vector. Using this vector, 10 gene-editing lines were acquired after maize transformation. PCR amplification and sanger sequencing revealed that, in each of the 10 gene-editing lines, different type of insertions or deletions were introduced into the target sites of both ZmBADH2 genes successfully. Genotypes of mutations included biallelic and multi-allelic mutations. Mess spectra analysis showed that Zmbadh2-1/Zmbadh2-2 double mutants had 2-AP, which is the same substance of flavor with that in fragrant rice. Using GC-MS, we found that 2-AP contents in grains gathered from four randomly selected T0 gene-editing lines were 438.29, 404.63, 348.65 and 161.82 μg·kg-1, respectively. On the contrary, no 2-AP was detected in JING724 wild type. 【Conclusion】With site specific mutations introduced into ZmBADH2-1 and ZmBADH2-2 simultaneously using CRISPR/Cas9, new maize variety with fragrant rice like flavor was created successfully.

    TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY·AGRICULTURE INFORMATION TECHNOLOGY
    Research Progress of Soil Microbial Mechanisms in Mediating Plant Salt Resistance
    KONG YaLi,ZHU ChunQuan,CAO XiaoChuang,ZHU LianFeng,JIN QianYu,HONG XiaoZhi,ZHANG JunHua
    Scientia Agricultura Sinica. 2021, 54(10):  2073-2083.  doi:10.3864/j.issn.0578-1752.2021.10.004
    Abstract ( 712 )   HTML ( 66 )   PDF (464KB) ( 728 )   Save
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    Soil salinization has seriously hindered the sustainable agricultural production. Remediation of salt affected areas with efficient, low cost and adaptable method is a challenging goal for scientists. Soil microorganisms play important roles in regulating rhizosphere environment of plants, enhancing plant development and productivity. Adaptation of plants to stress driven by soil microbes has been attracted extensive attention. The identification and exploitation of soil microorganisms that interact with plants in alleviating salt stress provides a new strategy for the improvement of saline area, as well as new approaches to discover mechanisms involved in stress tolerance. Knowledge of the underlying physiological mechanisms by which diverse microbes mediate stress tolerance, is critical to the effective use of these microbes to assure sustained agricultural production. This paper reviewed the recent studies about the mechanisms of soil microorganisms mediated in plant salt stress tolerance from the aspects of plant nutrient absorption, osmosis balance, hormone levels and antioxidant function. The beneficial effects and lack of current researches related to soil microorganism in the regulation of plant salt tolerance were evaluated, and the directions of the future research were also proposed. At present, improving nutrient and water uptake efficiency to maintain plant ion homeostasis under salt stress, increasing auxin synthesis and reducing ethylene release to regulate plant hormone levels seem to be promising target processes for soil biota-improved plant salt tolerance. However, limited success has been obtained in application of microorganism to large-scale agricultural production, due to the competition of introduced single microbial strains with native soil microbial communities which resulted in many bacterial strains has little colonization efficiency. The researches related to microbial mediated plant salinity tolerance should break through the single microbial inoculation, further clarify the mechanism of plant-microbial interaction at the community level, and solve the key problems of microbial utilization in agricultural production.

    Remote Sensing Inversion of Nitrogen Content in Apple Canopy Based on Shadow Removal in UAV Multi-Spectral Remote Sensing Images
    LI MeiXuan,ZHU XiCun,BAI XueYuan,PENG YuFeng,TIAN ZhongYu,JIANG YuanMao
    Scientia Agricultura Sinica. 2021, 54(10):  2084-2094.  doi:10.3864/j.issn.0578-1752.2021.10.005
    Abstract ( 427 )   HTML ( 37 )   PDF (1373KB) ( 292 )   Save
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    【Objective】The shadows in UAV multi-spectral remote sensing images were removed to improve the accuracy of the nitrogen inversion model for apple canopy. 【Method】Using the UAV multi-spectral images collected in June 2019 at the apple orchard of Qixia city in Shandong province, as the experimental area, normalized shaded vegetation index (NSVI) and normalized canopy shadow index (NDCSI) were respectively used to remove shadow and to extract the spectral information of the canopy in non shadow area. The correlation analysis method was used to analyze the correlation between the spectral data, including the data obtained based on the original spectral images and the images after removing the shadow based on NSVI and NDCSI, and the measured leaf nitrogen content data, respectively, and then the sensitive wavelength of nitrogen content were screened and spectral parameters were constructed. Partial least squares (PLS) and support vector machine (SVM) methods were used to build the inversion model of nitrogen content and to carry out the precision inspection in the fruit tree canopy. 【Result】The results showed that the green band and red band were sensitive bands for the inversion of nitrogen content in fruit tree canopy based on UAV multi-spectral images. The spectral information of fruit tree canopy was weakened by shadow, and the spectral difference of canopy multispectral bands before and after shadow removal was significant, especially in red-edge band and near-infrared band. The accuracy of nitrogen inversion model based on two shadow indexes after shadow removal was improved, and the optimal model was the support vector machine nitrogen content inversion model based on NDCSI, the modeling set of this model R2 and RPD was 0.774 and 1.828, the validation set R2 and RPD were 0.723 and 1.819 respectively. 【Conclusion】NDCSI could effectively remove the shadow in the multi-spectral fruit tree canopy image of the UAV to improve remote sensing inversion accuracy of nitrogen content in apple canopy, so as to provide a useful reference for precise nitrogen management in orchard.

    PLANT PROTECTION
    Effects of Proteins Encoded by “C4 ORFs” of Cotton Leaf Curl Multan Virus on Viral Pathogenicity
    ZHENG XinShi,SHANG PengXiang,LI JingYuan,DING XinLun,WU ZuJian,ZHANG Jie
    Scientia Agricultura Sinica. 2021, 54(10):  2095-2104.  doi:10.3864/j.issn.0578-1752.2021.10.006
    Abstract ( 260 )   HTML ( 25 )   PDF (3457KB) ( 444 )   Save
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    【Objective】Cotton leaf curl Multan virus (CLCuMuV) was found to encode three proteins with different sizes near the open reading frame (ORF) of C4, and the size of C4 protein encoded by each isolate of CLCuMuV registered in NCBI database was not the same. The objective of this study is to analyze the effect of proteins encoded by three hypothesized “C4 ORFs” on viral pathogenicity, and to determine the exact location of C4 ORF of CLCuMuV.【Method】According to the conserved protein of geminiviruses, three ORFs with different sizes were found near C4 ORF of CLCuMuV, which were labeled as C4-L (567 nt), C4-M (546 nt) and C4-S (303 nt). The three ORFs were constructed into PVX heterologous expression vector by homologous recombination, respectively. Using an agroinoculation method, the effect of PVX mediated expression of the three proteins on the symptoms of Nicotiana benthamiana were analyzed. Using a homologous recombination method, the infectious clones of CLCuMuV wild type and C4-L or C4-S deletion mutant were constructed, and then together with infectious clone of the associated DNA-β, they were inoculated into N. benthamiana respectively by agroinoculation. The effect of C4-L or C4-S deletion on CLCuMuV pathogenicity was analyzed. Meanwhile, Southern blot and Western blot were used to analyze the accumulation of viral genome and the expression level of viral C4 protein. At the same time, the subcellular localization of C4-L and C4-S proteins in the epidermal cells of N. benthamiana leaves was analyzed by a Gateway system vector.【Result】In the PVX heterologous expression assay, the results showed that the leaf curling and petiole elongation were observed in PVX-C4-L and PVX-C4-S inoculated N. benthamiana plants, and the symptoms of PVX-C4-S inoculated plants were more severe, while those of PVX-C4-M inoculated plants were mild or almost asymptomatic. The expression of C4 protein was also detected by Western blot in PVX-C4-L and PVX-C4-S inoculated plants. The results revealed that the protein encoded by C4-S ORF had the greatest effect on the pathogenicity of PVX heterologous virus. In the infectious clone assay, the results showed that the wild type (CLCuMuV) and C4-L mutant (CLCuMuV-ΔL) inoculated N. benthamiana plants had leaf wrinkled, petiole and stem distorted, while those inoculated with C4-S mutant (CLCuMuV-ΔS) and control (Mock) did not show any symptoms. Furthermore, the height of the CLCuMuV and CLCuMuV-ΔL inoculated plants was significantly shorter than that of CLCuMuV-ΔS and Mock inoculated plants. At the same time, the accumulation of viral genome was detected by Southern blot in both CLCuMuV and CLCuMuV-ΔL inoculated plants, and the expression of C4 protein was also detected by Western blot. The results revealed that the protein encoded by C4-S ORF played a key role in the process of CLCuMuV induced symptoms. The subcellular localization showed that YFP-C4-L was mainly located in the chloroplast of epidermal cells of N. benthamiana leaves, while YFP-C4-S was located in the cell membrane or the cytoplasmic periphery, and formed a point like aggregate structure. 【Conclusion】The protein encoded by C4-S ORF is essential for the infection of CLCuMuV, while the proteins encoded by C4-L and C4-M ORF are not necessary.

    Gene Cloning, Expression Pattern and Molecular Characterization of Chitin Deacetylase 2 in Cydia pomonella
    SHI GuoLiang,WU Qiang,YANG NianWan,HUANG Cong,LIU WanXue,QIAN WanQiang,WAN FangHao
    Scientia Agricultura Sinica. 2021, 54(10):  2105-2117.  doi:10.3864/j.issn.0578-1752.2021.10.007
    Abstract ( 258 )   HTML ( 35 )   PDF (2146KB) ( 254 )   Save
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    【Objective】In order to provide a scientific basis for the selection of novel target for pest control, molecular characterization and expression patterns of chitin deacetylase 2 genes (CpCDA2) of the codling moth (Cydia pomonella) were studied. 【Method】The cDNA sequences of chitin deacetylase 2 genes were identified from the transcriptome data of C. pomonella by bioinformatic methods, and the conserved domain was analyzed, the homologous sequences from other insects were selected to construct phylogenetic tree by using neighbor-joining (NJ) method of MEGA7 software. The protein properties including molecular weight (Mw), theoretical isoelectric point (pI), glycosylation site and hydrophilic/hydrophobic properties were analyzed by online software. RT-qPCR was performed to detect the relative expression levels of CpCDA2a and CpCDA2b in different tissues and developmental stages. To explore the functions of these genes in the process of larval molting, the ecdysone (20E) was injected to the 5th instar larvae, and the expression dynamics of them were detected by RT-qPCR.【Result】The full length cDNA sequences of CpCDA2a and CpCDA2b, alternative splicing variants of C. pomonella CDA2, were identified. The amino acid analysis showed that both variants of CpCDA2 possess the signal peptide, and the open reading frames contain three conserved domains, including chitin binding domain (ChBD), low-density lipoprotein receptor class A domain (LDLa) and chitin deacetylase catalytic domain (CDA). The multi-sequence alignment analysis revealed that the CpCDA2a and CpCDA2b are similar to other insect, and the phylogenetic tree showed that the sequences of insect in the same order are clustered into a clade with high confidence. Three-dimensional structure simulation and comparison revealed that the ChBD functional domains of the two splicing variants are different in structure. Physical and chemical properties analysis showed that there are also differences in hydrophilic/hydrophobic properties and glycosylation site of the two splicing variants. Developmental expression patterns showed that the CpCDA2a expression was high in larval stage, CpCDA2b in early and middle stages of pupae, and the expression of CpCDA2a and CpCDA2b was significantly up-regulated before and after larval molting. Tissue-specific expression analysis indicated that the expression of CpCDA2a and CpCDA2b was the highest in integument, and the expression of CpCDA2b in head was also higher. Expression dynamics of the larvae after injection of ecdysone (20E) indicated that both CpCDA2a and CpCDA2b were up-regulated, but with very different levels.【Conclusion】The CpCDA2 has two alternatively spliced variants, namely CpCDA2a and CpCDA2b, which plays a key role in the development of C. pomonella. While, according to the comprehensive analysis of molecular characterization, developmental stages and tissue expression profiles, and expression trends after ecdysone injection, it is deduced that the differences in protein structure and physical and chemical properties of CpCDA2a and CpCDA2b caused by alternative splicing, resulted in the differentiation of function between them.

    Cloning, Preparation of Antibody and Response Induced by 20-Hydroxyecdysone of Target of Rapamycin in Apolygus lucorum
    TAN YongAn,ZHAO XuDong,JIANG YiPing,ZHAO Jing,XIAO LiuBin,HAO DeJun
    Scientia Agricultura Sinica. 2021, 54(10):  2118-2131.  doi:10.3864/j.issn.0578-1752.2021.10.008
    Abstract ( 217 )   HTML ( 21 )   PDF (1775KB) ( 348 )   Save
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    【Objective】The objective of this study is to clone the target of rapamycin gene (TOR) of Apolygus lucorum (AlTOR), analyze the expression profile of AlTOR as well as its expression patterns after injection of 20-hydroxyecdysone (20E) and its inhibitor U73122, obtain the recombinant protein and antibody against AlTOR protein, and to lay the foundation for the further study of AlTOR protein function.【Method】The full-length of AlTOR was cloned by RACE method. Using qRT-PCR technique, the expression pattern of AlTOR was determined at different developmental stages and different tissues in A. lucorum, as well as its expression patterns after injection of 20E and U73122. To construct its prokaryotic expression pCzn1-AlTOR, T vector containing the target gene was digested with the restriction enzyme EcoR I and Xho I and subcloned. The recombinant plasmid containing target gene was specifically expressed after induction by IPTG. The target recombinant protein has over expressed and has been purified by using Ni-NTA agarose. The polyclonal antibody was obtained by rabbit immunity, cell fusion and ascites preparation, and Western blot was used to detect the specificity of antibody.【Result】The open reading frame of AlTOR encodes 435 amino acids. AlTOR protein has the typical domains including the SIN1 domain, highly conserved CRIM domain and PH domain. qRT-PCR results showed that AlTOR was expressed in 1-day-old to 16-day-old of A. lucorum, in which the expression level was the highest at 1-day-old. AlTOR was expressed in all tissues of A. lucorum female adults, and the fat body had the highest expression level. 20E could induce the expression of AlTOR at different day-old nymphs, and could also induce the up-regulated expression of AlTOR in the head, wing, ovary and fat body of the female adults, which was increased by 200.00%, 118.89%, 20.53% and 60.82%, respectively, while U73122 significantly suppressed AlTOR expression in the midgut and ovary. The vector containing AlTOR was digested and linked to the vector pCzn1. The recombinant plasmid pCzn1-AlTOR expressed the target recombinant protein of 36 kD after induction by IPTG. The 36 kD target protein from the strain containing AlTOR was obtained by the Ni-NTA agarose and used as the antigen, and one cell line polyclonal antibody could specifically bind to the AlTOR recombinant protein and the total protein of the 3rd instar nymph of A. lucorum, indicating that the prepared AlTOR polyclonal antibody has good specificity.【Conclusion】The expression of AlTOR shows the developmental stage- and tissue-specificity. After the induction of 20E and its inhibitor, AlTOR shows the opposite responses. The obtained polyclonal antibody against AlTOR recombinant protein is highly specific.

    SOIL & FERTILIZER·WATER-SAVING IRRIGATION·AGROECOLOGY & ENVIRONMENT
    Spatial Variations and Relationships of Topsoil Fertility Indices of Drylands in the Typical Black Soil Region of Northeast China
    YIN SiJia,LI Hui,XU ZhiQiang,PEI JiuBo,DAI JiGuang,LIU YuWei,LI AiMeng,YU YaXi,LIU Wei,WANG JingKuan
    Scientia Agricultura Sinica. 2021, 54(10):  2132-2141.  doi:10.3864/j.issn.0578-1752.2021.10.009
    Abstract ( 413 )   HTML ( 28 )   PDF (747KB) ( 272 )   Save
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    【Objective】In order to clarify the latitude characteristics and relationships among soil fertility indices in the typical black soil region of northeast China, clarify the problems of fertility control in the heart of black land and provide strong support for the quality control of black land cultivated land and grain yield stable and increase.【Method】Soil samples were collected following by latitude gradient in the study area after the autumn harvest in 2018. The soil fertility indices, such as soil organic matter, pH, total phosphorus, total potassium, available phosphorus, available potassium, alkali-hydrolyzable nitrogen, bulk density, soil water content, soil temperature the ratio of macro- and micro-aggregates, soil water content and soil temperature, were measured to investigate these indices’ latitudinal variation characteristics and their relationships by means of principal component analysis, redundancy analysis and correlation analysis. 【Result】The soil fertility indices of typical black soil region in Northeast China showed obvious latitudinal variation characteristics. In addition to the weak variation of bulk density, other fertility indices showed medium variation. Soil organic matter was the core influencing index in this region, which was increasing with the increase of latitude, and had a significant positive correlation with the ratio of macro- and micro-aggregates, alkali-hydrolyzable nitrogen, available phosphorus, available potassium, total potassium, total phosphorus and pH. Nevertheless, it was negatively correlated with soil water content, soil temperature and bulk density. The soil fertility indices with latitude between 46.07 ° and 49.18 ° were greatly affected by the change of soil bulk density, while the indices with latitude between 42.99 ° and 45.59 ° were combined action with soil bulk density, organic matter content and pH.【Conclusion】In the latitude range of 46.07 ° to 49.18 °, the soil fertility indices could be regulated by improving soil bulk density, while in the latitude range of 42.99 ° to 45.59 °, the soil nutrient should be comprehensively regulated by soil testing and formulated fertilization. For the whole study area, the regulation of soil organic matte was still an important content of soil fertility.

    Effects of Long-Term Application of Organic Fertilizer on Carbon and Nitrogen Accumulation and Distribution of Sweetpotato in Fluvo- Aquic Soil Area
    ZHAO Peng,LIU Ming,JIN Rong,CHEN XiaoGuang,ZHANG AiJun,TANG ZhongHou,WEI Meng
    Scientia Agricultura Sinica. 2021, 54(10):  2142-2153.  doi:10.3864/j.issn.0578-1752.2021.10.010
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    【Objective】The accumulation and distribution of carbon and nitrogen in sweet potato is the key factor of yield formation, the response relationship between carbon and nitrogen accumulation and distribution in sweetpotato under long-term organic fertilizer addition was studied to provide scientific basis for high-yield and high-efficiency cultivation of sweetpotato in fluvo-aquic soil region. 【Method】Based on the 40-year long-term locating test of fluvo-aquic soil (Xuzhou), the treatments of no fertilizer (CK), nitrogen, phosphorus and potassium fertilizer (NPK), organic fertilizer (M), organic fertilizer + nitrogen, phosphorus and potassium fertilizer (MNPK) were selected as the research objects,soil properties, aboveground / underground biomass and carbon and nitrogen contents of main functional organs of sweet potato under different fertilization measures were measured and analyzed, the effects of different fertilization treatments on the content of carbon and nitrogen in sweet potato and the distribution ratio of carbon and nitrogen in various functional organs, as well as the changes of the ratio of carbon and nitrogen (C/N) in the aboveground and underground parts of sweet potato under different fertilization measures were expounded. The relationship between the distribution of carbon and nitrogen in sweet potato and soil properties was analyzed by principal component analysis.【Result】Compared with single application of organic fertilizer (M) or chemical fertilizer (NPK), long-term application of organic fertilizer combined with nitrogen, phosphorus and potassium fertilizer (MNPK) significantly increased the biomass and dry matter quality of sweet potato root tuber (P<0.05). At the same time, the contents of total nitrogen and available potassium in soil were significantly increased (P<0.01). The correlation analysis between soil properties and C/N fixation and C/N ratio of sweet potato showed that the C/N fixation of sweet potato organs was significantly positively correlated with soil organic carbon (SOC) and total nitrogen (TN) (P<0.01), and significantly positively correlated with soil available potassium (AK) (P<0.05). However, the content of soil available phosphorus (AP) was not correlated with the content of carbon and nitrogen in sweet potato leaves, but positively correlated with the nitrogen fixation of root tuber (P<0.01), with a correlation coefficient of 0.839. The C/N ratio of leaves of sweet potato was positively correlated with soil EC (P<0.01), and the C/N ratio of petioles and stems of sweet potato was negatively correlated with soil EC (P<0.01) . The C/N ratio of underground tubers of sweet potato was negatively correlated with soil available phosphorus (P<0.01); principal component analysis (PCA) was used to analyze the distribution ratio of carbon and nitrogen fixation in aboveground and underground parts and the C/N ratio of aboveground and underground parts. The results showed that the first two axes explained 66.6% of the variation, and the contribution rate of the first principal component axis was 42.8%. The order of CK and NPK treatments on axis 1 was higher, which indicated that the soil pH and EC values were higher under both treatments, and the explanations for the distribution of carbon and nitrogen in the aboveground part and C/N in the underground part of sweet potato were higher.【Conclusion】It can be seen that organic material addition can reasonably allocate C/N of each organ, improve the distribution ratio of carbon and nitrogen in the storage root part of sweetpotato, and promote the formation of sweetpotato yield.

    HORTICULTURE
    Research Advances of Applying Virus-Induced Gene Silencing in Vegetables
    LI Jie,LUO JiangHong,YANG Ping
    Scientia Agricultura Sinica. 2021, 54(10):  2154-2166.  doi:10.3864/j.issn.0578-1752.2021.10.011
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    Recently, the virus-induced gene silencing (VIGS) as a reverse genetics tool is used for gene function analysis. Due to its advantages of simple construction, low cost and short cycle, VIGS technology has been extensively and deeply studied in the field of functional genomics. VIGS technology, as a fast, effective, high-throughout new technology, has played an important role in research of vegetable functional genes in plant development processes, disease resistance, stress resistance, biosynthesis and metabolic regulation. Herein, it is of great significance to excavate new genes and identify the function of disease resistance, stress resistance genes, crop improvement and molecular breeding by using VIGS technology. Many VIGS systems with virus as vector have been successfully established in vegetable crops, but they still have some shortcomings. With the in-depth exploration of the mechanism of VIGS and the continuous development of virus vectors, VIGS has been applied to a wider range of vegetable crops. This paper reviewed the current status and research progresses of gene function of eggplant, melons and leafy vegetables based on VIGS technology in recent years, and the mechanism of VIGS technology, the application of virus vector and the progress of VIGS technology was briefly analyzed. Meanwhile, the advantages and disadvantages of VIGS technology, RNA interference (RNAi) and current CRISP/CAS9 technology were compared and analyzed. It focused on the application of VIGS technology in vegetable fruit development and disease resistance, and the latest progresses of VIGS technology in vegetable crop metabolic regulation, hormone regulation, biotic and abiotic stress responses were summarized. The cases of studying target genes function and silencing phenotypes of solanaceous, melon, leafy and legume vegetables by VIGS were listed. Finally, the problems and deficiencies of VIGS technology in studying gene function of vegetable crops were summarized, such as lack of suitable VIGS vector, lack of effective virus vector infection method, difficulty in systematic silencing in some tissues, low silencing efficiency, inherent limitations of VIGS, etc. At the same time, the future research directions of VIGS technology in the development of virus vectors with higher specificity and stability, selection of efficient gene fragments, and establishment of virus vectors suitable for more host range were proposed. The application foreground of gene function analysis, improvement, molecular breeding of vegetable crops and production not carrying exogenous gene of vegetable varieties by VIGS technique was prospected. This review would provide a guidance and give ideas for future studies on the growth and development of vegetable crops, secondary metabolism and adversity stress related gene function research and breakthrough in the key factors restricting VIGS technique.

    Fine Mapping and Candidate Gene Analysis of Seed Coat Color Gene CmSC1 in Melon
    MA Jian, LI CongCong, HUANG YaTing, XIE YuLi, CHENG LingLing, WANG JianShe
    Scientia Agricultura Sinica. 2021, 54(10):  2167-2178.  doi:10.3864/j.issn.0578-1752.2021.10.012
    Abstract ( 422 )   HTML ( 40 )   PDF (1247KB) ( 422 )   Save
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    【Objective】In this study, the genetic analysis and fine mapping of melon seed coat color gene were performed, and the candidate gene was isolated and the molecular marker developed, so as to provide a foundation for gene function and rational utilization. 【Method】The inbred line HP22 with white seed coat was crossed with the inbred line B8 and B150 with yellow seed coat, and the offspring population was obtained, respectively. The seed coat colors of offspring plants were investigated by visual inspection. The locus of gene was fine mapped through map-based cloning strategy, and the candidate target gene in the mapping region was determined by gene sequencing and function analysis of gene coding region. 【Result】The white seed coat was dominant to yellow, which was controlled by a single dominant gene locus, named CmSC1, showed delayed genetic characteristic. Using 368 yellow seed coat individuals from F2 population, the target gene CmSC1 was delimited to a 95 kb interval flanked by markers S27 and S28 on chromosome 5 that contained twelve annotated open reading frames (ORFs). Among of them, the MELO3C014406 gene encoded a bHLH transcription factor protein which was homologous to the Arabidopsis AtTT8 gene. Sequence analysis of MELO3C014406 gene in the yellow seed coat inbred lines indicated that two types of sequence variation were identified. The first type was an A insertion at the 47th base pairs position downstream of ATG, and the other was a 14 bp deletion at the 260th base pair position downstream of ATG, which caused premature termination and produced truncated proteins, respectively. Furthermore, the sequencing analysis of 65 yellow seed coat accessions were detected by specific molecular marker YS, and the results showed that they all contained one mutation type of two variation types. These results suggested that the MELO3C014406 gene was an ideal candidate gene for CmSC1 controlling seed coat color.【Conclusion】In this study, the CmSC1 gene regulating seed coat color was finally mapped to a 95 kb interval, and the MELO3C014406 gene was presumed to be the candidate gene. In addition, a specific marker YS was developed based on the 14 bp deletion of MELO3C014406 gene.

    Expression Characteristics and Function of FveD27 in Woodland Strawberry
    SUN HongYing,WANG Yan,LI WeiJia,ZHU TianShu,JIANG Ying,XU Yan,WU QingYue,ZHANG ZhiHong
    Scientia Agricultura Sinica. 2021, 54(10):  2179-2191.  doi:10.3864/j.issn.0578-1752.2021.10.013
    Abstract ( 361 )   HTML ( 26 )   PDF (3396KB) ( 305 )   Save
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    【Objective】It is known that the new plant hormone strigolactone is a key factor regulating plant branching development, but the role of strigolactone in the growth and development of strawberry plants is unclear. In this study, the expression characteristics and function of the key gene DWARF27 (D27) for strigolactone biosynthesis in woodland strawberry (Fragaria vesca) was revealed, and the role of FveD27 in strawberry branching growth and development was explored, which would lay a theoretical foundation for studying strawberry plant architecture. 【Method】The coding sequence region of strigolactone synthesis key gene FveD27 was cloned by RT-PCR from woodland strawberry. The phylogenetic relationship between FveD27 and D27 in apple, Arabidopsis and other species was analyzed by using MEGA 6.0. A fusion vector of FveD27 and GFP was constructed to inject tobacco leaves for analyzing the subcellular location of FveD27. The expression level of FveD27 in different organs of woodland strawberry was analyzed by qRT-PCR technology. The FveD27 promoter and GUS fusion expression vector was constructed and transformed into woodland strawberry by Agrobacterium-mediated method, and the expression characteristic of FveD27 was further analyzed by GUS staining. The overexpression vector of FveD27 was constructed and the FveD27 overexpression strawberry lines were obtained by Agrobacterium-mediated transformation with leaf disks of woodland strawberry as explants. 【Result】The coding sequence region of FveD27 with the length of 789 bp was cloned from woodland strawberry. The subcellular localization in tobacco showed that FveD27 was located in the chloroplast. The expression levels of FveD27 in woodland strawberry organs from high to low were followed by young leaves, shoot tips, petiole, mature leaves and roots. The FveD27 promoter sequence with a length of 1670 bp was cloned, and transgenic plants with pFveD27::GUS gene showed that the young leaves and buds had strong GUS activity, the mature leaves and petioles had weak GUS activity, while the roots had little GUS activity. The result of FveD27 expression revealed by GUS analysis was consistent with the result by qRT-PCR. The FveD27 overexpression vector was constructed and the transgenic woodland strawberry lines overexpressing FveD27 were obtained with Agrobacterium-mediated transformation. Overexpressing FveD27 in woodland strawberry inhibited significantly the formation of branch crowns and increased the number of inflorescences. 【Conclusion】FveD27 had the functions of regulating the development of branch crowns and the number of inflorescences in woodland strawberry, and this study provided new ideas for regulating the number of branch crowns and yield of strawberry.

    FOOD SCIENCE AND ENGINEERING
    Research Progress on Mechanisms of Apoptosis to Postmortem Tenderization in Muscle
    HUANG Feng,WEI QiChao,LI Xia,LIU ChunMei,ZHANG ChunHui
    Scientia Agricultura Sinica. 2021, 54(10):  2192-2202.  doi:10.3864/j.issn.0578-1752.2021.10.014
    Abstract ( 373 )   HTML ( 30 )   PDF (1940KB) ( 276 )   Save
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    Tenderness has been considered as one of the most important eating quality characteristics of meat, while inconsistent changes of tenderness in postmortem (PM) muscles can significantly reduce the purchasing intention. Therefore, investigating mechanism of postmortem muscle tenderization is becoming more and more important in the past decade. Since apoptosis definition was highlighted in the PM tenderization process, it has been widely concerned. Shortly after slaughter, the reactive oxygen species (ROS) was significantly accumulated, and ATP (adenosine triphosphate) was gradually exhausted within muscle fibers, which inevitably led to skeletal cell death. Both PM cell death and meat tenderization refer to the activation of muscle endogenous enzymes by a series of regulatory factors, followed by the degradation of muscle structural proteins. The two biochemical processes are considered to be highly related. PM muscle cells die mainly through apoptosis. Besides, in the early stage of PM, when a small amount of ROS is produced, the cells start their defense system by autophagy and ATP therefore gradually deplete muscle cells in the later stage, which may change from apoptosis to necrosis. It has been documented that mitochondrial pathway is crucial for the apoptosis activation in PM muscles. The release of apoptotic factors from mitochondria is the master node of the intracellular death cascade reaction. The opening status of mitochondrial outer membrane directly determines the way, in which the muscle fiber dies. In this paper, the release mechanism of apoptotic factors induced by PM mitochondrial damage was discussed from the perspectives of mitochondrial membrane permeability and cristae remodeling, and the regulation of mitochondrial damage on PM muscle tenderization was discussed. Moreover the underlying mechanism behind it was also analyzed to reveal the effect of mitochondria on muscle pH regulation through energy metabolism, the release of apoptotic factors and regulation of apoptosis enzyme activity. At the same time, the interaction between mitochondria and endoplasmic reticulum were discussed, focusing on Ca2+ signal transduction and cell apoptosis process. The interaction between mitochondria and lysosome was further investigated, by highlighting the stability of lysosomal membrane and the subsequently released cathepsin to activate Bax and Bid to accelerate mitochondrial membrane permeability. Caspases were activated and involved in the limited degradation of some myofibrils in the early stage of PM tenderization, followed by the inactivation resulting from the decreasing ATP or other factors with the extended PM time. Therefore, caspases maybe only involve the early stage of tenderization. This review could provide a theoretical reference for the perfection of PM muscle tenderization investigations.

    SPECIAL FOCUS: YOUNG RUMINANT NUTRITION RESEARCH
    Improving the Health Level of Young Animals to Promote the Quality of Livestock Products
    TU Yan,DIAO QiYu
    Scientia Agricultura Sinica. 2021, 54(10):  2203-2205.  doi:10.3864/j.issn.0578-1752.2021.10.015
    Abstract ( 215 )   HTML ( 22 )   PDF (277KB) ( 198 )   Save
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    Effects of High-Fat Diet on Energy Metabolism and Slaughter Performance of Early-Weaning Lambs
    LI WenJuan,TAO Hui,ZHANG NaiFeng,MA Tao,DIAO QiYu
    Scientia Agricultura Sinica. 2021, 54(10):  2206-2216.  doi:10.3864/j.issn.0578-1752.2021.10.016
    Abstract ( 341 )   HTML ( 25 )   PDF (486KB) ( 232 )   Save
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    【Objective】Fat is an important nutrient element in animal diet, and it is also the main energy supply material, so it plays an important role in animal production. The purpose of this study was to investigate the effects of high fat diet on growth performance, energy metabolism and slaughter performance of early weaned Hu lambs before and after weaning, so as to provide theoretical basis and technical support for healthy breeding of early weaned lambs. 【Method】Thirty pairs of healthy Hu sheep twin lambs with similar birth age, similar weight were randomly divided into two groups, including high fat diet group (HF: 26.89% and 5.07%) and normal fat diet group (NF: 15.15% and 2.80%). The lambs of the two groups were fed with milk replacer and pellet with different fat levels from 7 to 60 days old and weaned milk powder at 60 days old, and then which were fed with the same pellet from 60 to 120 days. Nine pairs of twin lambs were randomly selected at the age of 50-60 and 110-120 days according to the average body weight, and the digestion and metabolism trails were conducted by the method of total feces and urine collection to evaluate the energy metabolism of lambs fed diets with different fat levels before and after weaning. According to the average body weight of lambs, nine pairs of twin lambs were randomly slaughtered at the age of 60 and 120 days to test slaughter performance, organ indexes, and gastrointestinal development. 【Result】Before weaning, the total dry matter intake (DMI), total energy intake (GE), fecal energy (FE), urinary energy (UE), apparent digestibility of total energy, total energy metabolic rate (ME/GE), empty body weight (EBW), slaughter rate, GR value of lambs in the two groups were significantly higher than those in the control group .There was no significant difference in the proportion of stomach and intestines to pre slaughter live weight (P>0.05); The body weight, DE, ME, DE/ME, HCW, head weight, heart weight, hoof weight, the proportion of hoof weight to live weight before slaughter, omasum weight and small intestine weight of 60 day old lambs in HF group were higher than those in NF group (0.05<P<0.1), and the eye muscle area and abomasum weight were significantly higher than those in NF group (P<0.05). After weaning milk powder, all lambs were fed the same pellet to 120 days of age. DMI of lambs fed high fat diet at 61-120 stage and BW, LBW, EBW, HCW, skin + gross weight, heart weight, hoof weight and rumen weight of lambs at 120 days of age were significantly higher than those in NF group (P<0.05), and spleen and kidney weight were also higher than those in NF group (0.05<P<0.1); the feeding high fat diet before weaning did not affect energy metabolism, other organ index and gastrointestinal development of lambs after weaning (P>0.05). 【Conclusion】Feeding high fat diet before weaning could improve the body weight, digestible energy and metabolizable energy, carcass weight and eye muscle area of lambs. Feeding high fat diet during lactation significantly increased feed intake, body weight, live weight before slaughter and carcass weight of lambs after weaning. In conclusion, the increasing dietary fat content before weaning had a positive effect on energy metabolism and slaughter performance of Hu sheep twin male lambs before and after weaning.

    The Effects of Early Weaning and NDF Levels of Finishing Diets on Growth Performance, Nutrient Digestion and Metabolism of Hu Lambs
    HUANG WenQin,LÜ XiaoKang,ZHUANG YiMin,CUI Kai,WANG ShiQing,DIAO QiYu,ZHANG NaiFeng
    Scientia Agricultura Sinica. 2021, 54(10):  2217-2228.  doi:10.3864/j.issn.0578-1752.2021.10.017
    Abstract ( 274 )   HTML ( 21 )   PDF (486KB) ( 187 )   Save
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    【Objective】This study was conducted to explore the effects of early weaning and NDF levels of finishing diets on the growth performance, and nutrient digestion and metabolism of Hu sheep lambs, and to filter out the best combination of feeding methods by calculating the superposition effect between the experimental factors. 【Method】One hundred and twenty Hu lambs with similar body weight [(8.26±2.14) kg] and (20±2 ) days of age were randomly divided into four groups with six replicates in each group and five lambs in each replicate. NDF levels of early finishing diets were 33%(low) and 38%(high), NDF levels of late finishing diets were 28% (low) and 38% (high). Treatments included followings: early weaning (EW) + feeding low NDF diet in early and late finishing stage (EW-LL); early weaning + feeding low NDF diet in early finishing stage and high NDF diet in late finishing stage (EW-LH); early weaning + feeding high NDF diet in early and late finishing stage (EW-HH); ewe reared (ER) + feeding high NDF diet in early and late finishing stage (ER-HH). Early weaned lambs gradually transitioned from breastfeeding to feeding milk replacer at the age of 20 days and supplemented with starter feed. The fattening period was from 61 to 180 days of age. The trial lasted 160 days. The body weight and feed intake of lambs were measured at different stages, and digestibility tests were carried out at 2, 4 and 6 months of age.【Result】There was no significant difference in body weight and average daily gain among four groups (P>0.05) except the average daily gain of lambs from 46 to 65 days of age, which was higher in ER group compared with EW group (P<0.05). The dry matter intake of early-weaned lambs from 21 to 65 days old was significantly higher than that of ewe reared lambs (P<0.05), In the early weaned groups, the pellet intake of lambs fed low-NDF diets during the finishing stage were significantly higher than the other two groups at the age of 121-180 days (P<0.05). The nutrient digestibility, energy and nitrogen utilization of early weaned lambs were lower than those in the ER group at 55 to 65 days of age. But it was inverse at fattening period, the dry matter and organic matter digestibility of lambs fed low NDF diet was higher than those fed high NDF diet in finishing stage (P<0.05). EW-LL group had the highest ADG increase scope by superposition effect analysis. The superimposing effect of the two feeding methods of early weaning and low-NDF diet during the early finishing stage was -50.57%.【Conclusion】According to the whole fattening cycle, early weaning and feeding milk replacer could promote the growth performance of lambs. The fattening effect of the low NDF level diet in the early finishing stage was worse than the high NDF level diet. When lambs have the same diet in the late finishing stage, the weight gain of lambs fed with low NDF diet was better than that of lambs fed with high NDF diet in the early finishing stage. By the comprehensive analysis of the growth performance, digestion and metabolism of lambs, the priority of the combination of feed methods was EW-LL>EW-LH=EW-HH>ER-HH. The combination of early weaning + feeding low NDF diet in early and later finishing stage was recommended in this study.

    Effects of Cinnamaldehyde on Growth Performance,Health Status, Rumen Fermentation and Microflora of Dairy Calves
    YANG YunYan,WANG QiYan,PENG DiWei,PAN YiFan,GAO XiaoMei,XUAN ZeYi,CHEN ShaoMei,ZOU CaiXia,CAO YanHong,LIN Bo
    Scientia Agricultura Sinica. 2021, 54(10):  2229-2238.  doi:10.3864/j.issn.0578-1752.2021.10.018
    Abstract ( 321 )   HTML ( 24 )   PDF (467KB) ( 319 )   Save
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    【Objective】this study was conducted to research effects of addition of cinnamaldehyde in calf starter on growth performance, health status, rumen fermentation and microflora of calves, so as to provide theoretical basis for applying cinnamaldehyde in calf cultivation. 【Method】Twenty-four healthy Holstein male calves at the age of around 15 days, with similar body weight, were selected, and were divided into control group and experiment group, with three replicates in each group and four calve in each replicate. The control group were fed basic starter, while starter of experimental group was added with 0.3% encapsulated cinnamaldehyde which contained 15% effective cinnamaldehyde. The weaning period contained 6 pre-trail period, and 27 days of formal trail period, both groups were fed same amount of milk, the starter and orts grass were given at libtium, and the ruminal fluid was taken by stomach tube at the end of weaning period. After weaned, the animals were kept for another 51 days with same feeding management mode, the ruminal fluid was taken at the end of the experiment. The feed intake, growth performance, body measurement, fence score and ruminal volatile fatty acids concentrations were measured during the two phase of the experiment, and the ruminal bacterial community composition was measured based on 16SrDNA gene high through put sequencing. 【Result】The results showed that: (1) In terms of feed intake and growth performance, there were no difference in average daily gain, dry matter intake and feed efficiency between experiment and control group at pre-weaning, post-weaning stage and whole experiment period (P>0.05), there were no differences in body measurements parameters between experiment and control group as well (P>0.05); 2) In terms of calf health status, the diarrhea rate of the experimental group was lower than that of the control group, but was not significant (P>0.05). 3) For the rumen fermentation parameters, the concentration of volatile fatty acids of rumen fluid in the experimental group on weaning day was tend to lower than that in the control group (P>0.05). At the end of the experiment, the concentration of volatile fatty acids, including acetate, propionate, butyrate and valerate of experimental group was significantly higher, while acetate/propionate ratio lower than that of control group (P<0.01); 4) At the end of experiment, cinnamaldehyde supplementation increased ruminal bacteria Shannon diversity index, decreased Simpson diversity index; addition of cinnamaldehyde increased abundance of Firmicutes and Actinobacteria at phylum and prevotella at genus level (P<0.01). 【Conclusion】The results showed that addition of 0.3% encapsulated cinnamaldehyde which contained 15% effective cinnamaldehyde had no influence on growth performance, but decreased diarrhea rate, increased ruminal volatile fatty acids concentration and decreased acetate/propionate ratio, increased ruminal bacterial community diversity, and abundance of Firmicutes and Prevotella, indicated addition of high amount of cinnamaldehyde had no adverse effects on calf growth performance, but changed ruminal bacterial community composition and ruminal fermentation mode.

    Effects of High-Concentrate Diet Supplemented with Grape Seed Proanthocyanidins on Rumen fermentation, Inflammatory and Antioxidant Indicators of Rumen and Serum in Lambs
    HAO XiaoYan,MU ChunTang,QIAO Dong,ZHANG XuanZi,YANG WenJun,ZHAO JunXing,ZHANG ChunXiang,ZHANG JianXin
    Scientia Agricultura Sinica. 2021, 54(10):  2239-2248.  doi:10.3864/j.issn.0578-1752.2021.10.019
    Abstract ( 398 )   HTML ( 30 )   PDF (497KB) ( 271 )   Save
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    【Objective】This study was conducted to investigate the effects of dietary grape seed proanthocyanidins (GSPs) supplementation on growth performance, rumen fermentation, rumen and serum inflammatory factors and antioxidative activity in lambs under the condition of a high-concentrate diet, so as to provide a support for applying GSPs on ruminant. 【Method】A total of forty-eight 1/2 Dorper × 1/2 thin-tailed Han ram lambs with similar body weight (BW = 22.75±1.20 kg, mean ± SD) were randomly divided into four groups, with twelve lambs each group. Lambs were fed a 30:70 forage:concentrate diet with 0 (control), 10 (10GSPs), 20 (20GSPs), and 40 mg·kg-1 BW (40GSPs) GSPs supplemented, respectively. The feeding experiment lasted for 60 days before sampling, with the first 15 days for diet transition and adaptation. On the first day of the formal experiment period, the BW before morning feeding was weighed as the initial BW. At the end of the trial period, blood samples were collected from the jugular vein, and the serum was separated for the determination of antioxidant and inflammatory indicators and lipopolysaccharide content. At the same time, six lambs in each group were randomly selected to collect rumen fluid by using an oral stomach tube at 1, 3, 4, 6, 8 and 12 hours after feeding, respectively, and the rumen pH were measured immediately. The fluid samples at 3 hours after feeding were used to measured fermentation parameters and lipopolysaccharide content. The other 6 lambs in each group were slaughtered, and the samples of rumen tissue were collected to determine the antioxidation index and inflammatory factors. 【Result】The final weight of lambs in 10GSPs and 20GSPs groups were significant greater than that in control group (P<0.05), with no difference between 40GSPs and 10GSPs or 20GSPs (P>0.05). Lambs in 10GSPs and 20GSPs groups also had higher average daily gain and average daily feed intake than control and 40GSPs (P<0.05), with no significant difference between control and 40GSPs (P>0.05). The dietary GSPs supplementation had a certain regulatory effect on rumen pH. The rumen pH increased linearly with the supplementation of GSPs (P<0.05) at 3, 8 and 12 h after feeding, and tended to increased linearly at 4 h (P=0.057). The concentrations of acetate, butyrate and total volatile fatty acid in rumen fluid tended to decrease after GSPS was added (P<0.1), but there was no significant effect on propionate, isovalerate, valerate, and the ratio of acetic acid to propionic acid (P>0.05). After GSPs was supplemented, the serum lipopolysaccharide concentration was significantly lower than that of the control group (P<0.05), but which did not affect the lipopolysaccharide concentration in the rumen fluid. The activity of GSH-Px in rumen tissue of 20GSPs and 40GSPs groups was significantly higher than that of the control group and 10GSPs group (P<0.05) , however, the content of MDA was significantly lower. The serum SOD activity of 20GSPs and 40GSPs group was significantly higher than that of the control group, and the GSH-Px activity was higher than control and 10GSPs group (P<0.05). There was no significant effect was observed in rumen inflammatory factors when GSPs was supplemented, but there was a trend of decreasing IL-6 and IL-10 (P<0.1). The level of TNF-α in 20GSPs and 40GSPs group were significantly lower than that in the control group and 10 GSPs group (P<0.05). The level of IL-10 in 40GSPs group was significantly lower than control group, with no significant difference between 40GSPs and 10GSPs or 20GSPs (P>0.05). 【Conclusion】Supplementing appropriate amount of GSPs to high-concentrate diet could improve the rumen pH, and the antioxidant capacity of serum and rumen tissue of lambs, suggesting that appropriate amount of GSPs had the potential protective effect on lamb health. The optimal feeding dose was 20 mg·kg-1 BW under the condition of this experiment.