薇甘菊花芽分化前后内源激素及相关基因表达的研究
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国家林业和草原局推广项目(2019TG026号);云南省万人计划科技支撑项目(194306)


Study on Endogenous Hormones and the Expression of Related Genes before and after Flower Bud Differentiation of Mikania micrantha
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    摘要:

    以云南省瑞丽市勐秀林场扦插种植的薇甘菊为试材,采用液相色谱串联质谱(LC-MS/MS)技术对花芽未分化期和花序原基分化期花芽中的生长素(IAA)、赤霉素(GA)、脱落酸(ABA)、反式玉米素(tZ)、异戊烯腺嘌呤(IP)、1-氨基环丙烷羧酸(ACC)、茉莉酸(JA)和水杨酸(SA)含量进行定量分析,同时基于转录组基因功能注释数据对内源激素合成、代谢及信号转导途径相关基因进行表达分析,以探讨不同内源激素对薇甘菊花芽形成的调控作用,以及内源激素合成和信号转导途径相关基因调控薇甘菊花芽分化的机制,为后期通过外源激素调控薇甘菊内源激素水平的方式来控制薇甘菊的有性繁殖提供理论和技术支持。结果表明:(1)薇甘菊未分化期花芽中GA15、GA19、GA20、GA24、IAA、ABA和ETH含量低于花序原基分化期,而未分化期花芽中两种细胞分裂素tZ和IP含量显著高于花序原基分化期。(2)基于RNA-seq测序结果,在薇甘菊两个花芽分化时期共获得7 116个差异表达基因(DEGs),其中上调3 907个,下调3 209个。(3)在内源激素合成方面,参与GA15、GA19、GA20、GA24、IAA、ABA和ACC合成的大量DEGs在花序原基分化期上调表达,这与它们在薇甘菊花序原基分化期的高含量趋势相一致;参与IAA合成的YUCCA基因家族和ACC合成的ACS基因在花序原基分化期的高表达也可能参与促进薇甘菊花芽分化。(4)在植物激素转导途径中,在花序原基分化期,生长素信号转导途径通过AUX/IAA(gene-E3N88_07743)的下调表达和ARF(gene-E3N88_41119)的上调表达,乙烯信号转导途径通过ERF(gene-E3N88_41547)的上调表达,赤霉素信号转导途径通过GID1(gene-E3N88_19448)基因的上调表达,细胞分裂素信号转导途径通过B-ARR(gene-E3N88_28086)和A-RRR(gene-E3N88_40764)基因的下调表达,脱落酸途径通过AREB(gene-E3N88_18558)基因的上调表达,茉莉酸信号转导途径通过JAZ(gene-E3N88_05628)的上调表达和MYC2(gene-E3N88_32405)的下调表达来调控薇甘菊花芽分化。研究发现,高水平的GA15、GA19、GA20、GA24、IAA、ABA和ACC有利于薇甘菊的花芽分化;薇甘菊在花芽分化过程中通过改变不同种类内源激素合成、代谢基因的表达来调控激素浓度,而激素又通过信号转导途径引起下游基因的表达,进而调控薇甘菊的花芽分化。

    Abstract:

    Using Mikania micrantha cutted and planted in Mengxiu forest farm of Ruili City, Yunnan Province as test materials, we quantitatively analyzed the contents of indole-3-acetic acid (IAA), gibberellins (GAs), abscisic acid (ABA), trans-zeatin (tZ), N6-Isopentenyladenine (IP), 1-aminocyclopropanecarboxylic acid (ACC), jasmonic acid (JA) and salicylic acid (SA) in flower buds at the undifferentiated stage of flower buds and the differentiation stage of inflorescence primordium by liquid chromatography mass spectrometry (LC-MS/MS). Based on the transcriptome gene function annotation data, we also analyzed the expression of genes related to endogenous hormone synthesis, metabolism and signal transduction pathways. The aim of this study was to explore the regulatory effects of different endogenous hormones on flower bud formation of M. micrantha, and analyze the mechanism of endogenous hormone synthesis and signal transduction pathway related genes regulating flower bud differentiation of M. micrantha. It provides theoretical and technical support for controlling the sexual reproduction of M. micrantha by regulating the endogenous hormone level of M. micrantha with exogenous hormones in the later stage. The results showed that: (1) the contents of GA15, GA19, GA20, GA24, IAA, ABA and ACC in flower bud at undifferentiated stage were lower than those at inflorescence primordium differentiation stage, while the contents of tZ and IP in flower bud at undifferentiated stage were significantly higher than those at inflorescence primordium differentiation stage. (2) Based on RNA-seq results, a total of 7 116 differentially expressed genes (DEGs) were obtained at two flower bud differentiation stages of M. micrantha, of which 3 907 were up-regulated and 3 209 were down regulated. (3) In terms of endogenous hormone synthesis, a large number of DEGs involved in the synthesis of GA15, GA19, GA20, GA24, IAA, ABA and ACC were up-regulated during inflorescence primordium differentiation, which was consistent with their high content trend during inflorescence primordium differentiation of M. micrantha. The high expression of YUUCA gene family involved in IAA synthesis and ACS gene involved in ETH at inflorescence primordium differentiation stage may also be involved in promoting flower bud differentiation of M. micrantha. (4) In the plant hormone transduction pathways, flower bud differentiation of M. micrantha was regulated by down-regulation of AUX/IAA gene (gene-E3N88_07743) and up-regulation of ARF (gene-E3N88_41119) in auxin signal transduction pathway, up-regulation of ERF (gene-E3N88_41547) in ethylene signal transduction pathway, up-regulation of GID1 (gene-E3N88_19448) in gibberellin signal transduction pathway, down-regulation by B-ARR (Gene-E3N88_28086) and A-RRR (gene-E3N88_40764) in cytokinin signal transduction pathway, up-regulation of AREB (gene-E3N88_18558) in abscisic acid signal transduction pathway, up-regulation of JAZ (gene-E3N88_05628) and down-regulation of MYC2 (gene-E3N88_32405) in jasmonic acid signal transduction pathway. Therefore, high levels of GA15, GA19, GA20, GA24, IAA, ABA and ETH are conducive to flower bud differentiation of M. micrantha. The flower bud differentiation process of M. micrantha regulates the hormone concentration by changing the expression of different kinds of endogenous hormone synthesis and metabolism genes, and the hormone causes the expression of downstream genes through signal transduction pathway to regulate the flower bud differentiation of M. micrantha.

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梁 晨,泽桑梓,高燕珠,等.薇甘菊花芽分化前后内源激素及相关基因表达的研究[J].西北植物学报,2023,43(2):229-241

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  • 在线发布日期: 2023-03-09
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