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‘红地球’葡萄花芽分化过程中的转录组分析
引用本文:刘鑫,张亚红,袁苗,党仕卓,周娟. ‘红地球’葡萄花芽分化过程中的转录组分析[J]. 中国农业科学, 2022, 55(20): 4020-4035. DOI: 10.3864/j.issn.0578-1752.2022.20.013
作者姓名:刘鑫  张亚红  袁苗  党仕卓  周娟
作者单位:1宁夏大学农学院,银川 7500212宁夏大学食品与葡萄酒学院,银川 750021
基金项目:宁夏回族自治区重点研发计划(2021BEF02016);宁夏大学研究生创新项目(GIP2020-09)
摘    要:【目的】 葡萄是我国重要的果树树种,花芽分化直接影响葡萄的质量和数量。对‘红地球’葡萄花芽分化过程中的花芽进行比较分析,探索‘红地球’葡萄花芽分化机制,挖掘关键基因,为了解‘红地球’葡萄花芽分化提供理论基础。【方法】 对‘红地球’葡萄花芽分化过程中4个发育阶段:S1(未分化期)、S2(花原始体发育期)、S3(花序主轴发育期)和S4(花序二级轴发育期)的芽进行形态学观察和植物激素测定,并进行转录组测序分析及验证。【结果】 ‘红地球’葡萄花芽分化过程中共发现13 729个差异基因,其中S1-S2、S2-S3、S3-S4和S1-S4分别有4 158、2 050、3 425和7 652个差异基因。在S1-S4差异基因的富集调控网络中发现差异基因在激素介导的信号通路、脱落酸代谢过程、对酸性化学物质的反应和植物细胞壁组织或生物发生等通路富集。在激素介导的信号通路中发现大量与生长素、赤霉素和脱落酸等相关基因,测定表明,生长素在S2时期含量最高,而在S3和S4时期含量最低;赤霉素含量在花芽分化过程中不断降低,在S4时期为S1时期的80%;脱落酸含量在S1和S4时期较高,而在S2时期最低。此外,S1-S4差异基因来自转录因子家族(MYB、ERF、bHLH和MADS-box等),表明这些转录因子家族基因参与了‘红地球’葡萄花芽分化。对差异表达的13个MADS-box家族基因进一步分析表明,MADS8AGL65AGL15AGL12MADS2在花芽分化进程中表达上调,而AGL30LeMADSFBP24AGL14MADS3表达下调。对这些MADS-box基因进行qRT-PCR验证,基因表达趋势与转录组数据一致且相关系数较高,表明数据分析结果可靠。【结论】 ‘红地球’葡萄花芽分化是一个复杂的生物过程,其中,植物激素介导的信号通路以及MADS-box家族基因在花芽分化中发挥重要作用。研究结果提供了一个关于转录因子、基因和激素的信息,有助于揭开这一复杂的发育过程,并为‘红地球’葡萄花芽分化综合模型的建立提供理论基础。

关 键 词:‘红地球’葡萄  花芽分化  转录组测序  植物激素  MADs-box  
收稿时间:2022-01-19

Transcriptome Analysis During Flower Bud Differentiation of Red Globe Grape
LIU Xin,ZHANG YaHong,YUAN Miao,DANG ShiZhuo,ZHOU Juan. Transcriptome Analysis During Flower Bud Differentiation of Red Globe Grape[J]. Scientia Agricultura Sinica, 2022, 55(20): 4020-4035. DOI: 10.3864/j.issn.0578-1752.2022.20.013
Authors:LIU Xin  ZHANG YaHong  YUAN Miao  DANG ShiZhuo  ZHOU Juan
Affiliation:1College of Agriculture, Ningxia University, Yinchuan 7500212School of Food & Wine, Ningxia University, Yinchuan 750021
Abstract:【Objective】 Grape is an important fruit tree species of China, and the flower bud differentiation directly affects the quality and quantity of this grape. In this study, Red Globe grape developmental stages during flower bud differentiation under natural growth conditions were compared and analyzed, the mechanism of flower bud differentiation of this grape variant was evaluated, and the essential genes were mined, so as to provide a theoretical basis for understanding the flower bud differentiation of this grape species. 【Method】During flower bud differentiation of the Red Globe grape, the morphological observation, phytohormone determination, and transcriptome sequencing analysis were performed on the buds of four developmental stages, including S1 (undifferentiated), S2 (flower anlagen development), S3 (formation of the main cob of inflorescence), and S4 (formation of the second cob of inflorescence). 【Result】A total of 13 729 differentially expressed genes were determined during the flower bud differentiation process of the ‘Red Globe’ grape, which included 4 158, 2 050, 3 425, and 7 652 genes in S1-S2, S2-S3, S3-S4, and S1-S4, respectively. In the enrichment-regulation network of the S1-S4 differential genes, those differential genes were found to be enriched in the hormone-mediated signaling pathways, abscisic acid metabolism, acid chemical reactions, plant cell wall tissues, or biogenesis. Several genes related to auxin, gibberellin, and abscisic acid were detected in the hormone-mediated signaling pathway. The results revealed that the content of auxin was the highest in S2 and the lowest in S3 and S4. The gibberellin content decreased continuously during flower bud differentiation, 80% of that at S4 in S1; the abscisic acid content was higher in S1 and S4 and the lowest in S2. In addition, the S1-S4 differential genes belonged to the transcription factor families (MYB, ERF, bHLH, and MADS-box), indicating that these family genes were involved in the flower bud differentiation of the Red Globe grape. The further analysis of the 13 differentially expressed MADS-box genes revealed upregulated expressions of MADS8, AGL65, AGL15, AGL12, and MADS2 during flower bud differentiation. In contrast, the expressions of AGL30, LeMADS, FBP24, AGL14, and MADS3 were downregulated. These MADS-box genes were verified via quantitative reverse transcription polymerase chain reaction, and the expression trend was found to be consistent with the corresponding transcriptome. 【Conclusion】The flower bud differentiation of the Red Globe grape was a complex biological process. The plant hormone-mediated signaling pathways and the MADS-box family genes played essential roles in flower bud differentiation. These results provided information about transcription factors, genes, and hormones to help understand this complex developmental process and provided a theoretical basis for establishing a comprehensive model for flower bud differentiation in the Red Globe grape.
Keywords:Red Globe grapes  flower bud differentiation  transcriptome sequencing  plant hormone  MADs-box  
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