大豆孢囊线虫侵染对乙烯合成及信号传导基因表达调控的研究

doi:10.13560/j.cnki.biotech.bull.1985.2021-1414

摘要/Abstract

摘要：

乙烯是一种重要的植物激素,其合成和信号传导相关基因参与调控植物生长发育及防御反应。为阐明大豆孢囊线虫侵染和定殖对寄主植物乙烯合成及信号传导途径的影响,本研究以8个抗病品种和2个感病品种为材料,利用荧光定量PCR方法对比接种与未接种孢囊线虫大豆根中乙烯合成基因GmACS和信号传导基因GmEIN的表达水平。实验结果表明,与感病品种相比,抗病品种中乙烯的生物合成受到显著调控,其中PI 548316中GmACS显著上调,而在PI 437654和PI 88788中GmACS显著下调。同时,GmEIN在4个抗病品种中轻微上调,但在其它抗病品种和2个感病品种中均未受到显著调控。研究表明GmACS和GmEIN 的表达受到孢囊线虫侵染与定殖调控,其中被显著调控的GmACS基因可以作为提高大豆抗病育种的候选基因。

关键词: 大豆, 大豆孢囊线虫, GmACS基因, GmEIN基因, 基因表达分析

Abstract:

Ethylene is an important plant hormone. Its synthesis and signal transduction related genes are involved in the regulation of plant growth,development and defense response. In order to clarify the effects of soybean cyst nematode infection and colonization on ethylene biosynthesis and signal transduction pathway,the expressions of ethylene synthesis gene GmACS and signal transduction gene GmEIN in soybean roots inoculated with and without cyst nematode were compared by fluorescence quantitative PCR. The results showed that compared with susceptible varieties,the ethylene biosynthesis in resistant varieties was significantly regulated,and in which 6 GmACSs genes were significantly induced in PI 548316,while significantly inhibited in PI 437654 and PI 88788. Meanwhile,several GmEINs was slightly up-regulated in 4 resistant varieties,but not significantly regulated in other resistant varieties and two susceptible varieties. The results show that ethylene synthesis in some soybean varieties are significantly regulated by soybean cyst nematode infection and colonization,and the regulated genes can be used as candidates for improving soybean disease resistance breeding.

Key words: soybean, Heterodera glycines, GmACS, GmEIN, gene expression pattern

郭宾会, 宋丽. 大豆孢囊线虫侵染对乙烯合成及信号传导基因表达调控的研究[J]. 生物技术通报, 2022, 38(8): 150-158.

GUO Bin-hui, SONG Li. Transcription of Ethylene Biosynthesis and Signaling Associated Genes in Response to Heterodera glycine Infection[J]. Biotechnology Bulletin, 2022, 38(8): 150-158.

图/表 6

表1 乙烯相关基因及其RT-qPCR引物序列

Table 1 Ethylene related genes and their RT-qPCR primer sequences

基因名称Gene name	基因 ID Gene ID	正向引物Forward primer	反向引物Reverse primer
GmACS#009	Glyma.07G065700	ATGTGAGCGTTGCTGAAGTG	AAGCCAGGGTACCCCATATC
GmACS#018	Glyma.16G032200	TAAGGACATGGGGTTTCCTG	TCCGAAACTCGACATTTTCC
GmACS#014	Glyma.09G255000	CTTGAAGGGTTTGCTGAAGG	CTGACCCTGGTGACACATTG
GmACS#021	Glyma.18G238100	TCATGGATTGCCAGAGTTCA	CCCCACTCATCAAAATACGG
GmACS#007	Glyma.05G211700	GGGTGGTTTAGGGTGTGCTA	GGCACCATGATCTCCTTGTT
GmACS#011	Glyma.08G018000	CTGGCTCTTCTTTCCATTGC	CGAAGGTTCGAATTCTTTGC
GmACS#015	Glyma.11G021500	GCGACGAAATATACGCTGGT	ATAAGGTCGCGGTCACATTC
GmACS#005	Glyma.05G108900	CGTATCGGGGCTATTGAAAA	GAACGTGTTCGAATGCAAAA
GmACS#020	Glyma.17G158100	TACCTCCTCTCTGCCATGCT	CCGAAACGAGCATTTTCTGT
GmEIN#001	Glyma.02G274600	CAAAGCCCTCCACCTTACAA	TTGGCAATATCAGGGGACAT
GmEIN#005	Glyma.13G076800	TTGGGATGGAAAGAGTGAGG	CCTCTTCCGAATGAAATCCA
GmEIN#006	Glyma.13G076700	CAAAATTTGCAAGCTCGTGA	TGCCAAGTCCTCCTCTTGAT
GmEIN#008	Glyma.14G041500	TATGATGTTGATGGGGCTGA	CATTCTCTCCATCCCAAGG
GmEIN#010	Glyma.20G051500	CAGAGAAGGGAAAGCCAGTG	GTTATGGCAGCAGGACCATT
GmEIN#007	Glyma.13G145100	GGCATCATTGATCTGGCTTT	GGAGGTGAGCCTGACTTCTG
GmACT11	Glyma.18G290800	ATCTTGACTGAGCGTGGTTATTCC	GCTGGTCCTGGCTGTCTCC

图1 GmACS和GmEIN基因家族的组织表达模式 A：GmACS基因家族,B：GmEIN基因家族

Fig. 1 Tissue expression patterns of GmACS and GmEIN gene families A：GmACS gene family. B：GmEIN gene family

图2 SCN（race 3）侵染大豆Magellan品种后根组织染色 A：接种后2 d;B：接种后4 d;C：接种后6 d;D和E：接种后8 d。比例尺：A-D：0.5 mm,E：1 mm

Fig.2 Root staining after SCN（race 3）infecting soybean Magellan variety A：2 d after inoculation;B：4 d after inoculation;C：6 d after inoculation;D and E：8 d after inoculation. Scal bar：A-D：0.5 mm, E：1 mm

图3 GmEIN基因在SCN侵染10 d后与未侵染对照的相对表达水平

Fig.3 Relative expressions of GmEIN genes in SCN infected root 10 d after inoculation compared with uninfected control

图4 GmACS#007、GmACS#011和GmACS#015在SCN侵染10 d后与未侵染对照的相对表达水平

Fig.4 Relative expressions of GmACS#007,GmACS#011and GmACS#015 in SCN infected root 10 d after inoculation compared with uninfected control

图5 6个GmACS基因在SCN侵染10 d后与未侵染对照的相对表达水平

Fig.5 Relative expressions of 6 GmACS genes in SCN infected root 10 d after inoculation compared with uninfected control

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