‘仓方早生’桃及其早熟芽变不同发育时期果实的转录组分析

doi:10.3864/j.issn.0578-1752.2023.05.012

中国农业科学 ›› 2023, Vol. 56 ›› Issue (5): 964-980.doi: 10.3864/j.issn.0578-1752.2023.05.012

‘仓方早生’桃及其早熟芽变不同发育时期果实的转录组分析

彭佳伟¹(), 张叶³(), 寇单单⁴, 杨丽¹, 刘晓飞¹, 张学英¹(), 陈海江¹(), 田义²()

¹ 河北农业大学园艺学院，河北保定 071000
² 河北农业大学山区研究所/河北省山区农业技术创新中心/国家北方山区农业工程技术研究中心，河北保定 071001
³ 河北省保定市农业农村局，河北保定 071000
⁴ 河北省盐山县望树镇中学，河北沧州 061300

收稿日期:2022-04-27 接受日期:2022-07-21 出版日期:2023-03-01 发布日期:2023-03-13
通信作者: 张学英，E-mail：yyzxy@hebau.edu.cn。陈海江，E-mail：chenhaijiang2001@163.com。田义，E-mail：tianyi@hebau.edu.cn
联系方式: 彭佳伟，E-mail：1358345303@qq.com。张叶，E-mail：20528036@qq.com。彭佳伟和张叶为同等贡献作者。
基金资助:
财政部和农业农村部：国家现代农业产业技术体系(CARS-30-2-03); 热杂果现代种业科技创新团队(21326310D)

Transcriptome Analysis of Peach Fruits at Different Developmental Stages in Peach Kurakato Wase and Early-Ripening Mutant

PENG JiaWei¹(), ZHANG Ye³(), KOU DanDan⁴, YANG Li¹, LIU XiaoFei¹, ZHANG XueYing¹(), CHEN HaiJiang¹(), TIAN Yi²()

¹ Horticultural Department, Agricultural University of Hebei, Baoding 071000, Hebei
² Mountainous Areas Research Institute, Hebei Agricultural University/Technology Innovation Center for Agriculture in Mountainous Areas of Hebei Province/National Engineering Research Center for Agriculture in Northern Mountainous Areas, Baoding 071001, Hebei
³ Baoding Municipal Bureau of Agriculture and Rural Affairs, Baoding 071000, Hebei
⁴ Wangshu Town Middle School of Yanshan County, Hebei Province, Cangzhou 061300, Hebei

Received:2022-04-27 Accepted:2022-07-21 Published:2023-03-01 Online:2023-03-13

摘要/Abstract

摘要：

【目的】通过对桃品种‘仓方早生’及其早熟芽变不同发育时期的果实进行转录组分析，挖掘参与调控桃果实成熟的关键因子，为深入研究桃果实成熟调控机理提供理论依据。【方法】以桃品种‘仓方早生’及其早熟芽变为试材，每个品种分别选择长势一致的样品树5株，分别于花后30 d（对应‘仓方早生’c1、早熟芽变y1）、45 d（对应c2、y2）、59 d（对应c3、y3）、71 d（对应c4、y4）及89 d（对应c5）对不同发育时期的桃去皮果肉进行取样和转录组测序，并利用实时荧光定量PCR（qRT-PCR）对筛选的差异表达基因进行定量验证；利用GO和KEGG对‘仓方早生’及其早熟芽变的差异表达基因进行分析；基于差异表达基因构建加权基因共表达网络分析（weighted gene co- expression network analysis，WGCNA），从中鉴定出与果实成熟密切相关的枢纽模块和枢纽基因。【结果】将处于果实相同发育时期的转录组数据进行比较，得到y1与c1、y2与c2、y3与c4和y4与c5四组对比数据，共筛选出差异表达基因4 395个，其中上调表达基因2 212个，下调表达基因2 183个。其中包括10个乙烯、11个脱落酸和18个生长素合成及其信号转导途径基因，并构建了10个IAA蛋白与预测互作ARF蛋白间的相互作用网络。由GO分类统计结果可知，差异表达基因在生物过程板块主要集中于细胞过程、代谢过程和单体过程；在细胞组分板块主要聚集于膜和细胞组分；在分子功能板块主要富集于结合蛋白和催化活性等方面。‘仓方早生’及其早熟芽变果实的差异基因主要集中在y3与c4和y4与c5对比组中，这些差异基因大多被富集到分子功能中结合活力、氧化还原酶活性等方面。对差异表达基因进行KEGG通路分析表明，在果实生长发育成熟过程中伴随着多种次生代谢产物的变化，如倍半萜和三萜生物合成、类黄酮生物合成、类胡萝卜素的生物合成和α-亚麻酸代谢等。同时，本研究发现生长素信号转导途径在不同时间节点均有富集，这意味着植物激素信号转导通路对果实成熟具有极为重要的作用。【结论】在‘仓方早生’及其早熟芽变不同发育时期果实的差异表达基因中，大量激素信号转导途径基因特别是生长素信号途径基因发生了富集，这些基因可能在调控果实发育中具有重要作用，可对这些候选的IAA和ARF功能及其如何通过相互作用调控果实成熟的机制进行进一步的解析。

关键词: 桃, 果实成熟, 转录组分析, 加权基因共表达网络分析

Abstract:

【Objective】 In this study, transcriptome analyses were carried out on the fruits of Kurakato Wase peach and its early-ripening mutant at different developmental stages. The key factors involved in fruit ripening regulation were explored, so as to provide a theoretical basis for further study on the regulation mechanism of fruit ripening. 【Method】 The flesh of Kurakato Wase peach and its early-ripening mutant was sampled at 30 d, 45 d, 59 d, 71 d, and 89 d after anthesis, and transcriptome analyses were performed on the above samples. The candidate differentially expressed genes (DEGs) were verified by quantitative real-time PCR (qRT-PCR). The biological function of DEGs were analyzed through GO function and KEGG pathway. The weighted gene co-expression network analysis (WGCNA) was constructed to identify the hub modules and hub genes closely related to fruit ripening. 【Result】 Four comparison groups including y1 vs c1, y2 vs c2, y3 vs c4 and y4 vs c5 were obtained based on fruit development stages. A tatal of 4 395 DEGs were identified with 2 212 up- and 2 183 down-regulated genes. There were 10, 11 and 18 candidate genes involved in ethylene, abscisic acid and auxin synthesis and signal transduction, respectively. The interaction networks between 10 IAA proteins and their predictive interacting proteins ARF were constructed. GO function revealed that the DEGs were mainly enriched in cellular processes, metabolic processes and monomeric processes in the biological process category; in cell component category, DEGs were mainly enriched in membranes and cellular components; in molecular function category, DEGs were mainly enriched in binding protein and catalytic activity. There were more DEGs in comparison groups y3 vs c4 and y4 vs c5, and these DEGs mainly enriched in molecular functions, such as binding and catalytic activity. The KEGG pathway analysis showed that a variety of secondary metabolites changed during fruit development and ripening, such as sesquiterpene and triterpenoid biosynthesis, flavonoid biosynthesis, carotenoid biosynthesis, and α-linolenic acid metabolism. In addition, auxin signal transduction pathway was found to be enriched at different time nodes. 【Conclusion】 Among DEGs, a large number of hormone signal transduction pathway genes, especially auxin signal pathway genes, were enriched, and these genes might play an extremely important role during fruit ripening. The functions of candidate genes IAA and ARF and the molecular regulation of fruit ripening would be further elucidated in the future studies.

Key words: peach, fruit ripening, RNA-seq, WGCNA

彭佳伟, 张叶, 寇单单, 杨丽, 刘晓飞, 张学英, 陈海江, 田义. ‘仓方早生’桃及其早熟芽变不同发育时期果实的转录组分析[J]. 中国农业科学, 2023, 56(5): 964-980.

PENG JiaWei, ZHANG Ye, KOU DanDan, YANG Li, LIU XiaoFei, ZHANG XueYing, CHEN HaiJiang, TIAN Yi. Transcriptome Analysis of Peach Fruits at Different Developmental Stages in Peach Kurakato Wase and Early-Ripening Mutant[J]. Scientia Agricultura Sinica, 2023, 56(5): 964-980.

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‘仓方早生’桃及其早熟芽变不同发育时期果实的转录组分析

Transcriptome Analysis of Peach Fruits at Different Developmental Stages in Peach Kurakato Wase and Early-Ripening Mutant

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