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Analysis of VvBES1-1 Involvement in Flower Bud Differentiation of Red Globe Grape Based on Red and Blue Light Regulation TANG XueShen, DANG ShiZhuo, ZHOU Juan, LI JiaHao, LI MeiHua, HU Hao, ZHANG YaHong Scientia Agricultura Sinica    2025, 58 (8): 1650-1662.   DOI: 10.3864/j.issn.0578-1752.2025.08.015 Abstract （56）   HTML （1）    PDF （5271KB）（33）       Knowledge map    Save 【Objective】 BRI1-EMS-Suppressor 1 (BES1), a key transcription factor in brassinosteroid (BR) signaling, regulates plant photomorphogenesis and photoperiodic flowering. This study aimed to investigate the role of BES1 in flower bud differentiation of Vitis vinifera Red Globe under red and blue light regulation, thereby elucidating the BR-mediated mechanisms driven by light quality and providing insights into flowering regulation in other woody fruit trees.【Method】 Bioinformatics analysis was performed to characterize the protein structure and sequence alignment of VvBES1-1. Flower buds of Red Globe grape were collected under greenhouse natural light (CK, control) and red:blue=4:1 (R4B1) light treatment. qPCR was used to analyze the spatiotemporal expression patterns and tissue specificity of VvBES1-1 during flower bud differentiation. Subcellular localization was determined via tobacco (Nicotiana benthamiana) transient transformation. Protein-protein interactions were examined using bimolecular fluorescence complementation (BiFC), and transcriptional activation activity was assessed via yeast autoactivation assays. 【Result】VvBES1-1 contains a BES1_N domain and belongs to the BES1-S-type protein family, showing the closest phylogenetic relationship with Populus trichocarpa BES1. It is expressed throughout all developmental stages of Red Globe grapevines. During R4B1 treatment, the expression level of VvBES1-1 was significantly reduced compared to the control. Its expression peaked during the development of secondary inflorescence axes. Additionally, treatment with 10 mmol·L-¹ EBR enhanced VvBES1-1 expression in Red Globe grape flower buds. Yeast autoactivation assays demonstrated that VvBES1-1 possesses self-activation activity. Subcellular localization analysis revealed that VvBES1-1 is localized in the nucleus. Overexpression of VvBES1-1 in tobacco delayed flowering time, promoted stem elongation, and increased meristem number. Downregulation of VvBES1-1 expression integrated brassinosteroid (BR) and photoperiod signaling pathways to promote flower bud differentiation in Red Globe grapes. During flower bud differentiation, VvBES1-1 expression was significantly regulated by red and blue light. Under R4B1 treatment, its expression peaked after 6 hours of 10 mmol·L-¹ EBR exposure.【Conclusion】VvBES1-1 plays a critical role in flower bud differentiation of Red Globe grapes. It integrates BR and photoperiod signals to inhibit grape flower bud differentiation. However, under red and blue light conditions, the expression of VvBES1-1 is effectively downregulated, thereby promoting flower bud differentiation. This study provides insights into the regulatory mechanisms of light and phytohormones in grape reproductive development. Table and Figures | Reference | Related Articles | Metrics

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Effect of VdF3′5′H2 Overexpression on the Accumulation of Anthocyanin Composition in Spine Grape Cells GUO AoLin, LIN JunXuan, LAI GongTi, HE LiYuan, CHE JianMei, PAN Ruo, YANG FangXue, HUANG YuJi, CHEN GuiXin, LAI ChengChun Scientia Agricultura Sinica    2025, 58 (4): 802-818.   DOI: 10.3864/j.issn.0578-1752.2025.04.014 Abstract （81）   HTML （12）    PDF （12810KB）（59）       Knowledge map    Save 【Objective】This study aims to clone the key gene VdF3′5′H2 involved in anthocyanin biosynthesis and analyze its regulatory role of anthocyanin biosynthesis and component content in spine grape (Vitis davidii Foëx) cells, thereby providing technical support and a theoretical foundation for constructing spine grape cells with high-yield anthocyanin production and enabling targeted regulation of anthocyanin biosynthesis.【Method】Using spine grape cells as materials, the VdF3′5′H2 was cloned and a plant expression vector was constructed. This vector was subsequently transformed into spine grape cell, with positive cell lines were screened in antibiotic media and identified through fluorescence observation and PCR analysis. The phenotypic characteristics of the transgenic spine grape cells were analyzed, along with quantification of anthocyanin, flavonoid, and proanthocyanidin. Additionally, the metabolite profile of anthocyanins was detected by UPLC-MS/MS. The expression levels of genes associated with anthocyanin biosynthesis were assessed using real-time quantitative PCR (RT-qPCR), and a comprehensive analysis of gene expression and differential metabolites was carried out by the O2PLS method.【Result】The open reading frame (ORF) of VdF3′5′H2 gene is 1 527 bp, encoding a protein comprising 508 amino acid residues. The VdF3′5′H2 protein exhibits high homology with its counterparts in related plants within the same family, containing the CYP75 conserved domain, a heme-binding site, and three characteristic conserved motifs. Phylogenetic analysis revealed that F3′H and F3′5′H cluster within the same branch, implying an evolutionary relationship from F3′H to F3′5′H, with VdF3′5′H2 positioned at a more ancestral state compared to VdF3′5′H1. Furthermore, subcellular localization indicated that the VdF3′5′H2 protein resides in the cytoplasm. The contents of anthocyanins, flavonoids and proanthocyanidins in two transgenic spine grape cells overexpression VdF3′5′H2 showed a significant increase compared to the wild type control. The content ratio of delphinidin-anthocyanins in the two transgenic cells increased to 7.82% (T6) and 14.32% (T10), respectively, Petunidin-anthocyanins increaseed to 7.30% and 10.16%, respectively, while the content ratio of malvidin- anthocyanins increased to 58.08% and 42.30%, respectively. In contrast, the content ratios of the three types of anthocyanins in the wild-type cells were 1.92%, 1.48%, and 8.49%, respectively. Additionally, the content ratios of cyanidin- and paeonidin-anthocyanins in the two transgenic cells decreased significantly. Overexpression of VdF3′5′H2 led to the downregulation of PAL, CHS, CHI and F3H expression levels, while upregulating the expression levels of VdF3′5′H1, LDOX and UFGT genes. The comprehensive analysis integrating gene expression data with differential metabolites showed that the overexpression of VdF3′5′H2 modulated the transcriptional regulation of genes involved in anthocyanin biosynthesis, thereby influencing both the synthesis pathways and accumulation patterns of various types of anthocyanins.【Conclusion】The overexpression of VdF3′5′H2 gene significantly enhanced the biosynthesis and accumulation of anthocyanins in spine grape cells, by regulating gene expression and altering the metabolic flux of the anthocyanin biosynthesis pathway, there by modulating the composition and content ratio of antocyanins in spine grape cells. Table and Figures | Reference | Related Articles | Metrics

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Effect of FGF5 and FGF21 on Proliferation of Dermal Papilla Cells in Cashmere Goat WANG Niu, SHI XinRan, ZHANG WeiDong, WANG Xin Scientia Agricultura Sinica    2025, 58 (4): 819-830.   DOI: 10.3864/j.issn.0578-1752.2025.04.015 Abstract （61）   HTML （6）    PDF （6578KB）（71）       Knowledge map    Save 【Background】The hair follicle cycle in goat is divided into three phases: the growth phase (anagen), the regression phase (catagen), and the resting phase (telogen). Fibroblast growth factor 5 (FGF5) and fibroblast growth factor 21 (FGF21) are important regulators in goat hair cycle. However, the precise localization of FGF5 and FGF21 in goat skin tissue and their regulatory mechanisms remain unclear during hair follicle cycle transition. Dermal papilla cells (DPCs) located in the dermal papilla region, are essential for hair follicle development. When DPCs lose their function, hair follicle remains in the telogen, leading to hair loss.【Objective】Therefore, this study aimed to identify the expression and localization of FGF5 and FGF21 in Cashmere goat skin tissue, understand their roles in DPCs, and to analyze their regulatory mechanisms. This would enrich the knowledge of fibroblast growth factor family proteins in hair follicle cycle regulation and provide a theoretical basis for further elucidating the molecular mechanisms of Cashmere goat hair follicle cycle transition. 【Method】Real-time quantitative PCR (RT-qPCR) and enzyme-linked immunosorbent assay (ELISA) were used to measure the expression levels of FGF5, FGF21 and their receptor 1 (FGFR1) in the skin tissue of Cashmere goats at anagen and telogen. Single-cell transcriptome sequencing data were analyzed to determine the expression and localization of FGF5, FGF21 and FGFR1, which were validated through immunofluorescence methods. Adenovirus was explored to overexpress FGF5 and FGF21 in DPCs. The effects on DPCs proliferation were measured using MTT, EdU and flow cytometry. The expressions of FGFR1 and proliferation-related genes were detected using RT-qPCR, immunofluorescence and western blotting methods. RT-qPCR, immunofluorescence and western blotting were used to investigate the effect of FGF5 and FGF21 on the proliferation of DPCs. 【Result】FGF5, FGF21 and FGFR1 exhibited higher expression levels in anagen than that in telogen, all of which were located within DPCs. Both FGF5 and FGF21 restrained DPCs viability, which significantly reduced the number of EdU-positive cells. FGF5 decreased the percentage of DPCs in G2 and S phases, while FGF21 specifically affected the G2 phase. The expression of FGFR1 was significantly increased following the overexpression of FGF5 and FGF21. Moreover, the expressions of cell proliferation markers KI67 and PCNA were also significantly inhibited with the overexpression of FGF5 and FGF21. Additionally, the expression of β-catenin, a key component of Wnt/β-catenin signaling pathway, was reduced. Thereby the activity of transcription factors TCF3 and JUN was inhibited, which were the downstream effectors of Wnt/β-catenin signaling pathway. Thus the inhibition further led to the decreased expression of proliferation-related genes MYC and CYCLIND1. 【Conclusion】FGF5 and FGF21 played a critical role as regulatory factors in controlling the biological function of DPCs. This study provided the first evidences that FGF21, similar to FGF5, restrained the proliferation of DPCs by suppressing the Wnt/β-catenin signaling pathway. These findings contributes to our understanding of how fibroblast growth factors influence hair follicle cycle transition, particularly in the context of Cashmere goat hair growth and cashmere production. Table and Figures | Reference | Related Articles | Metrics

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Screening of Interaction Proteins with AhSAP1 in Peanut Using the Yeast Two-Hybrid System ZHU YanTing, DANG Hao, NIU SiJie, LIN JingYi, YANG Hua, YANG Qiang, ZHANG Chong, CAI TieCheng, ZHUANG WeiJian, CHEN Hua Scientia Agricultura Sinica    2024, 57 (21): 4376-4390.   DOI: 10.3864/j.issn.0578-1752.2024.21.016 Abstract （131）   HTML （11）    PDF （3959KB）（135）       Knowledge map    Save 【Objective】Seed size directly affects the yield of peanut. The key transcription factor AhSAP1, which regulates peanut seed size, was obtained by QTL mapping in the early stage, but the molecular mechanism of AHSAP1 regulating peanut seed size remains unclear. In this paper, a peanut embryo yeast two-hybrid cDNA library was constructed and AhSAP1 was used as bait to screen interacting proteins, and the spatial and temporal expression characteristics of candidate interacting protein genes were analyzed. It laid the foundation for further study on the molecular mechanism of AhSAP1 regulating peanut kernel development. 【Method】The peanut embryo Escherichia coli cDNA library was constructed and identified by SMART (switching mechanism at 5′ end of the RNA transcript) method. The decoy vector pGBKT7-AhSAP1 was constructed and its toxicity and self-activation to yeast cells were evaluated. The peanut embryo cDNA library plasmid and the bait plasmid pGBKT7-AhSAP1 were co-transformed into Y2H Gold yeast strains, and the positive colonies with good growth and blue color were screened and sequenced to obtain the candidate interacting protein gene sequences and predict the biological functions. The expression characteristics of candidate interacting protein genes in different tissues and organs of peanut, induced by exogenous plant hormones and induced by low calcium stress were determined by RNA-seq. According to the functional annotation results, the candidate factors that may be involved in plant seed development were selected, their CDS full-length sequences were amplified, and the target vector pGADT7 was constructed, and point-to-point yeast two-hybrid interaction was verified with pGBKT7-AhSAP1 co-transformed yeast cells. 【Result】The titer of peanut embryo Escherichia coli secondary cDNA library was 1.05×108 cfu/mL, the recombination rate was 98%, the average insert fragment size was more than 1 000 bp, and the library quality was high. The yeast two-hybrid decoy vector pGBKT7-AhSAP1 was successfully constructed, which had no self-activation in yeast cells and no toxicity to yeast. Sixty-eight yeast-positive clones were screened, and 60 candidate interacting proteins were obtained by sequence similarity comparison and removal of duplicating, which were mainly involved in energy production and metabolism, translation, ribosome structure and biological development, transcription, signal transduction mechanism, post-translational modification, inorganic ion transport and metabolism, chromatin structure, etc. Twelve candidate interacting proteins were selected for one-to-one yeast two-hybrid verification with AhSAP1, and 8 candidate interacting proteins were found to interact with AhSAP1. 【Conclusion】The mixed cDNA library of peanut embryo development at different stages was successfully constructed, and 60 candidate interacting proteins with AhSAP1 were screened. The candidate interacting proteins were mainly involved in energy production and metabolism, translation, ribosome structure and biogenesis, transcription, signal transduction mechanism, post-translational modification, inorganic ion transport and metabolism, chromatin structure, etc. These candidates interacting protein genes were expressed in root, stem, leaf, inflorescence, fruit needle, pericarp, seed coat and embryo, and the interaction between 8 candidate interacting proteins and AhSAP1 was confirmed. Table and Figures | Reference | Related Articles | Metrics

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A Study on the Quality Stability of Wheat Grains at Designated Locations Across Multiple Years ZHAO BoHui, ZHANG YingQuan, JING DongLin, LIU BaoHua, CHENG YuanYuan, SU YuHuan, TANG Na, ZHANG Bo, GUO BoLi, WEI YiMin Scientia Agricultura Sinica    2024, 57 (9): 1833-1844.   DOI: 10.3864/j.issn.0578-1752.2024.09.016 Abstract （255）   HTML （25）    PDF （2211KB）（151）       Knowledge map    Save 【Objective】The stable quality of wheat raw grain can keep the processing technology and its parameters basically unchanged, reduce human intervention in the processing process and the loss of process adjustment, and ensure the stability of the quality of flour products. The evaluation of variety quality stability could provide the reference for processing enterprises to purchase stable quality raw grain.【Method】In this experiment, seven wheat varieties, including Shiluan 02-1, Jimai 22, Liangxing 99, Han 6172, Yingbo 700, Luyuan 502, and Gaoyou 2018, were planted in Xingtai and Handan cities from 2013 to 2019 and selected as trial samples. The coefficient of variation was used to quantitatively characterize the quality stability of varieties over multiple years and locations. The product of test weight, crude protein in grains, wet gluten content, and dough stability time was calculated, and its coefficient of variation was used as a comprehensive statistical measure of quality stability. Pairwise analysis of varieties in the same year and location was conducted to analyze the stability of their quality indicators.【Result】The fluctuation ranges of test weight, crude protein in grains, wet gluten content, dough stability time, and the coefficient of variation of the product of quality indicators were 0.06%-5.50%, 0.01%-12.21%, 0.03%-10.02%, 0.4%-138.69%, and 0.32%-140.01%, respectively. Pairwise analysis showed that the stability of test weight from high to low was Gaoyou 2018, Luyuan 502, Han 6172, Liangxing 99, Yingbo 700, Jimai 22, and Shiluan 02-1. The stability of crude protein in grains from high to low was Gaoyou2018, Han 6172, Yingbo 700, Luyuan 502, Liangxing 99, Jimai 22, and Shiluan 02-1. The stability of wet gluten content from high to low was Gaoyou 2018, Han 6172, Luyuan 502, Yingbo 700, Liangxing 99, Shiluan 02-1, and Jimai 22. The stability of dough stability time from high to low was Yingbo 700, Jimai 22, Gaoyou 2018, Luyuan 502, Han 6172, Liangxing 99, and Shiluan 02-1. The stability of the product of the above quality indicators from high to low was Yingbo 700, Gaoyou 2018, Luyuan 502, Liangxing 99, Han 6172, Jimai 22, and Shiluan 02-1. 【Conclusion】Among the tested quality indicators, test weight showed higher stability, while dough stability time exhibited lower stability. The stability of crude protein in grains and wet gluten content fell between the two. By using the coefficient of variation of the product of four quality indicators as the evaluation criterion, Yingbo 700 demonstrated higher quality stability among the trial varieties. Table and Figures | Reference | Related Articles | Metrics

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Characteristics of Good Taste and High Yield Type Japonica Rice in the Lower Reaches of the Yangtze River ZHU TianCi, MA TianFeng, KE Jian, ZHU TieZhong, HE HaiBing, YOU CuiCui, WU ChenYang, WANG GuanJun, WU LiQuan Scientia Agricultura Sinica    2024, 57 (4): 820-830.   DOI: 10.3864/j.issn.0578-1752.2024.04.015 Abstract （221）   HTML （18）    PDF （1590KB）（72）       Knowledge map    Save 【Objective】In order to explore the common agronomic and physiological characteristics of good taste and high yield type japonica rice in the lower reaches of the Yangtze River, the study could provide theoretical basis and technical support for realizing synergistic improvement of rice yield and quality in this region. 【Method】The cultivar screening tests were conducted in 2018 and 2022 using 14 and 13 conventional japonica rice cultivars, respectively, to systematically compare the yield and composition, taste quality and textural characteristics, as well as agronomic and physiological indicators such as biomass, stem non-structural carbohydrates (NSC), and leaf area duration (LAD) among the different cultivars.【Result】The taste value and yield varied across the different cultivars, so through clustering by taste value and yield, they can be further divided into three types: medium taste and low yield (ML), medium taste and high yield (MH), good taste and high yield (GH). The average taste value and yield of GH in two years were 68.5 and 10.2 t·hm-2, respectively, which were 6.8% higher in taste value and 14.6% higher in yield than MH and ML. In terms of performance for yield and quality traits, GH showed the highest grain filling percentage and taste scores compared to MH and ML. In terms of the dry matter accumulation, GH increased the proportion of dry matter accumulation at the early stage of grain filling stage, and showed the highest dry matter accumulation from heading stage to 15 days after heading stage. Further analysis of the causes of dry matter accumulation at this stage revealed that GH significantly increased the NSC remobilization rate while steadily increasing LAD. Correlation analyses showed highly significant positive correlations between grain filling percentage and taste, dry matter accumulation and LAD from heading stage to 15 days after heading stage, and NSC remobilization rate.【Conclusion】 While maintaining a high LAD from heading stage to 15 days after heading stage (100.4 m2·m-2·d), further increasing NSC remobilization rate during grain filling stage (79.9%) to promote the initiation of grain filling, increasing the dry matter accumulation from heading stage to 15 days after heading stage (3.6 t·hm-2), then improving the grain filling percentage (95.4%), and taste (9.6), which are the common characteristics of good taste and high yield type japonica rice in this region. In addition, the development of water and fertilizer management techniques targeting the enhancement of NSC remobilization at the grain filling stage and NSC accumulation at heading stage is expected to further exploit the yield and taste quality potential of the above good taste and high yield type japonica rice. Table and Figures | Reference | Related Articles | Metrics

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Comparison of Heat Tolerance of Maize Hybrids and Their Parental Inbreds with Different Genotypes XU TianJun, LÜ TianFang, LI ZiHao, ZHANG Yong, LIU HongWei, LIU YueE, CAI WanTao, ZHANG RuYang, SONG Wei, XING JinFeng, ZHAO JiuRan, WANG RongHuan Scientia Agricultura Sinica    2024, 57 (2): 403-415.   DOI: 10.3864/j.issn.0578-1752.2024.02.014 Abstract （298）   HTML （22）    PDF （4552KB）（279）       Knowledge map    Save 【Objective】 In recent years, the adverse weather of high temperature and heat damage in the Huang-Huai-Hai maize region of China occurred frequently, which has become an important adverse factor threatening maize production. Study and clarify the effects of high temperature stress on male and female ear characteristics and yield of maize can provide useful guidance for the cultivation and selection of high temperature tolerant maize varieties. 【Method】 The variety of Zhengdan958 (Zheng58×Chang7-2), Xianyu335 (PH6WC×PH4CV), Jingnongke728 (JingMC01×Jing2416), MC812 (JingB547×Jing2416), and their parents were used as the test materials. High temperature stress before and after flowering (from V12 stage to 7 d after silking) were conducted. The effects of high temperature stress on the growth and development of male and female panicles, ASI, pollen activity, yield and yield components of different genotypes of maize hybrids and their parents were studied. 【Result】 High temperature stress before and after anthesis significantly reduced the ear length, rows per ear and grains per row of the tested maize varieties and their parents, and then resulted significant decrease in yield. Compared with the control, the grain number per spike of Zhengdan958, Xianyu335, Jingnongke728 and MC812 decreased by 22.28%, 47.69%, 6.13% and 8.11% respectively under high temperature stress, resulting yield decrease of 9.50%, 50.61%, 3.17% and 5.00% respectively. Among the parental materials, the decrease of rows per panicle, grains per row and yield of Jing2416 under high temperature treatment was the smallest and not significant, while the decrease of PH6WC was the largest. Under high temperature stress, the total number of tassel branches, the length of tassel, the total amount of loose pollen and pollen activity decreased significantly, the silking period of loose pollen was prolonged, and the duration of loose pollen was shortened. Among them, Jingnongke728 had the smallest decline, followed by MC812, showing good heat resistance, while Zhengdan958 had the largest decline in the length of tassel, but the amount of pollen was the largest due to the large number of tassel branches. Xianyu335 has fewer male panicle branches, a large decrease in male panicle length, the least amount of pollen and low activity. Among the parental materials, Jing2416 had a large amount of total loose pollen and strong pollen vitality under high temperature treatment, with the smallest decline, only 4.50 and 3.98 percentage points. Compared with the control, the interval of loose pollen silking (ASI) was prolonged by 1.6d under high temperature stress. The decrease in male spike length is manifested as Zhengdan958>Xianyu335>MC812>Jingnongke728. Zhengdan958 had the largest decrease in male spike length, but had more branches and the largest pollen yield; Xianyu335 has fewer branches of male spikes, a significant decrease in male spike length, the least pollen quantity, and the lowest activity; Jing 2416 has a large amount of loose pollen and strong pollen vitality, with the smallest decrease (only 4.50% and 3.98%). 【Conclusion】 High temperature stress before and after anthesis has a significant impact on the grain yield, male and female ear development process, pollen activity and filament microstructure of the tested maize varieties. Under high temperature stress at this stage, the decline of yield and pollen activity of Jingnongke728 and MC812 is significantly less than Xianyu335, showing higher single ear yield and heat tolerance. By comparing the heat resistance of the parental inbred lines of the tested maize hybrids, it was found that the heat resistance of the paternal inbred lines was better than that of the maternal inbred lines. The male panicle branch and length of the parent material Jing2416 decreased slightly, the amount of pollen was large, the pollen activity was high, the filaments were hairy, the ability to capture pollen was strong, the single panicle yield was high, and the heat resistance was the best. Therefore, in the planting area with frequent high temperature and heat damage, selecting maize varieties such as Jingnongke728 can achieve stable and high yield; and during the maize breeding process, we should pay more attention to the utilizing of the higher temperature resistant inbred such an Jing2416 and then combination higher temperature maize varieties. Table and Figures | Reference | Related Articles | Metrics

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Characteristics of Endophytic Microbial Community Structures in Stems Between Hylocereus undatus and H. polyrhizus ZHOU XinYan, CHEN SiYu, WEI YuFei, ZHU Yu, FENG JunQian, DING DianCao, LU GuiFeng, YANG ShangDong Scientia Agricultura Sinica    2024, 57 (2): 416-428.   DOI: 10.3864/j.issn.0578-1752.2024.02.015 Abstract （190）   HTML （11）    PDF （4336KB）（2901）       Knowledge map    Save 【Objective】 The differences of the endophytic microbial community structures between Hylocereus undatus and H. polyrhizus were analyzed, so as so to explore the correlation between the pulp color of H. polyrhizus and the endophytic microbial community composition and their functional microorganisms. 【Method】 Based on high-throughput sequencing technology, the diversity and richness of endophytic microorganisms (bacteria and fungi) in stems between H. undatus and H. polyrhizus were analyzed. Meanwhile, based on LEfSe analysis, the differences of endophytic microorganisms in stems between H. undatus and H. polyrhizus were also investigated.【Result】 The significant differences of the endophytic microbial community structures were found in stems between H. undatus and H. polyrhizus. Meanwhile, the numbers of specific bacterial and fungal OTUs in stems of H. polyrhizus were all higher than those of H. undatus. At the phylum level, the relative abundance ratio of Ascomycota in stems of H. polyrhizus was 1.15 times higher than that of H. undatus. At the genus level, Streptomyces and Penicillium were the highest abundant dominant bacterial and fungal genera in stems of H. polyrhizus, which were 1.24 and 4.27 times higher than those of H. undatus, respectively. In addition, some bacterial genera, such as Lechevalieria, Glycomyces, unclassified_f__Enterobacteriaceae, Actinomadura, and some fungal gerera, such as Talaromyces, unclassified_f__Serendipitaceae, unclassified_c__GS13, unclassified_o__Atractiellales, unclassified_o__Auriculariales were enriched in stems of H. polyrhizus. LEfSe analysis also showed that Promicromonospora and Xylomyces were significant enriched in stems of H. polyrhizus. 【Conclusion】 All above results suggested that the formation of pigment was closely related to the compositions of endophytic microbial community in stems of H. polyrhizus. The bacterial genera, such as Streptomyces, unclassified_f__Enterobacteriaceae, Promicromonospora, and the fungal phylum and genera, such as Ascomycota, Penicillium, Talaromyces and Xylomyces, were all the potential microorganisms in relating to pigment synthesis and metabolic accumulation in stems of H. polyrhizus. Table and Figures | Reference | Related Articles | Metrics

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Changes of Endogenous Hormones and Polyamines During Ovule Development of Stenospermocarpic Seedless Grape ZHU PeiPei, QIN HaoXiang, ZHANG JianXia Scientia Agricultura Sinica    2023, 56 (23): 4789-4800.   DOI: 10.3864/j.issn.0578-1752.2023.23.018 Abstract （201）   HTML （11）    PDF （536KB）（1438）       Knowledge map    Save 【Objective】By exploring the effects of endogenous hormones and polyamines on embryo development during the ovule development of seedless grape, this study provided a theoretical basis for promoting embryo development by spraying exogenous hormones before anthesis and adding exogenous hormones into the medium in vitro culture of ovule.【Method】In this study, the European grape (Vitis vinifera L.) variety Jingxiu and its F1 generation stenospermocarpic seedless variety Qinxiu were used as test materials. The content of endogenous hormones and polyamines in ovule of fruit at different developmental stages were compared by high performance liquid chromatography mass spectrometry (HPLCMS) and ultra performance liquid chromatography (UPLC).【Result】The content of IAA, ZT, ZR, and iPAS reached their highest values at 39 DAF (days after flowering) for Jingxiu and 42 DAF for Qinxiu, and the highest values of the former were higher than those of the latter (more than 1.5 times). For the content of JA and SA, Jingxiu started to rise sharply at 36 DAF (2 000 ng∙g-1 and 6 500 ng∙g-1, respectively), and reached the highest values at 39 DAF (6 500 ng∙g-1 and 10 000 ng∙g-1, respectively), and then, which were declining and remaining at a high level at 42-45 DAF, while Qinxiu was declining and remaining at a low level from 36 DAF (3 500 ng∙g-1 and 3 000 ng∙g-1, respectively). The ACC content of Jingxiu was almost 0 ng∙g-1 at 39 DAF, then increased rapidly and reached its highest value (1 200 ng∙g-1) at 45 DAF, while Qinxiu reached its highest value (900 ng∙g-1) at 39 DAF, then declined to 0 ng∙g-1 at 42 DAF, and remained at a very low level. The ABA content of Jingxiu was almost 0 ng∙g-1 from 36 to 45 DAF, while that of Qinxiu rose sharply from 0 ng∙g-1 at 39 DAF to the highest value (900 ng∙g-1) at 42 DAF. The trends of Put (putrescine), Spm (spermine), and Spd (spermidine) were similar for the two varieties, but Jingxiu was consistently higher than Qinxiu. The highest values of Put and Spd were 42 DAF for Jingxiu and 39 DAF for Qinxiu, except for the highest values of Spm, which were all found at 42 DAF. 【Conclusion】 The lower content of growth promoting substances (IAA, CTK, GA3, ACC, JA, SA, Put, Spd, and Spm) and the higher content of growth inhibiting substances (ABA), and lower ratios of (IAA+GA3)/ABA, (IAA+ZT+GA3)/ABA, Spm/PAs, (Spd+Spm)/PAs, and (Spd+Spm)/Put in the ovules of the seedless variety Qinxiu compared with the seeded variety Jingxiu might be one of the main reasons for seedless grape embryo abortion. Therefore, in the process of seedless grape embryo rescue, the embryo abortion could be inhibited by spraying before flowering or adding to media a certain concentration of growth promoting substances. Table and Figures | Reference | Related Articles | Metrics

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The Modification of Gene Editing Vector for Efficient GFPuv Fluorescence Screening and Its Application in Potato Genetic Transformation DU JingYa, CHEN KaiYuan, PU Jin, ZHOU HuiYing, ZHU GuangTao, ZHANG ChunZhi, DU Hui Scientia Agricultura Sinica    2023, 56 (11): 2223-2236.   DOI: 10.3864/j.issn.0578-1752.2023.11.015 Abstract （382）   HTML （36）    PDF （3831KB）（2193）       Knowledge map    Save 【Objective】The improvement and innovation of screening markers contributes to the development of transgenic technology, among which the visual screening markers are widely modified for better effect. Recent studies revealed that an enhanced Yellow Green Fluorescent like Protein (eYGFPuv (GFPuv)) obtained by mutation can emit strong and stable green fluorescence under 365 nm UV light irradiation and be easily observed. Constructing the gene editing vector with GFPuv fluorescence screening marker and carrying out experiment application and verifications in potato genetic transformation will provide technical support for the screening of positive transgenic plants in potato transformation, and lay the foundation for using genome editing technology to create potato male sterile lines in the future. 【Method】By using homologous recombination, the GFPuv expression framework and gene editing element Cas9_sgRNA were successively recombined into pCAMBIA2300 vector, and then with this new designed vector the Agrobacterium-mediated transient expression assay was conducted in tobacco plants. Six editing vectors with potato anther development conservative genes were constructed using this modified vector. The A. rhizogenes strains Ar qual and MSU440 harbouring these vectors were transformed into the potato stem segments respectively, and then the A. rhizogenes-induced hairy roots with green fluorescence were observed and counted under the portable UV lamp. The transformation efficiency and editing efficiency of these vectors were analyzed using hairy root transformation system in two different potato genotypes. In the end, the modified vectors were applied to produce transformed potato plants with modifications on target genes. 【Result】A novel gene editing vector pCAMBIA2300MGFPuv-sgRNACas harbouring a GFPuv fluorescence marker was successfully constructed, and the transient transformation in tobacco plants confirmed that the GFPuv expression framework was expressed successfully. The hairy roots with green fluorescence were screened after the transformation with two kinds of A. rhizogenes, and an additional supplement of kanamycin (Kan) significantly increased the proportion of positive fluorescent roots. Although the transformation rates of the two strains were not significantly different, the hairy roots of MSU440 formed faster. Furthermore, the transformation rates and editing rates of editing vectors for six potato anther development conservative genes in two different potato genotypes were the same, but the editing rates of six target sites differed significantly. Potato genetic transformation using the modified vector confirmed that GFPuv fluorescence could be used for the screening of transgenic callus and plants in potato. 【Conclusion】The hairy root transformation system mediated by A. rhizogenes is an essential approach to verifying the efficiency of gene editing, and GFPuv fluorescence can be used in the screening of transgenic plants in potato transformation. Table and Figures | Reference | Related Articles | Metrics

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