外源水杨酸对水稻苗期生长与防卫相关基因表达的影响

doi:10.3969/j.issn.1004-1524.2021.10.01

浙江农业学报 ›› 2021, Vol. 33 ›› Issue (10): 1789-1796.DOI: 10.3969/j.issn.1004-1524.2021.10.01

外源水杨酸对水稻苗期生长与防卫相关基因表达的影响

刘寒¹^,²(), 戴远兴¹^,², 吕明芳², 袁正杰², 李静², 严成其², 张恒木²^,^*()

1.浙江师范大学化学与生命科学学院,浙江金华 321004
2.浙江省农业科学院病毒学与生物技术研究所,浙江杭州 310021

收稿日期:2021-04-15 出版日期:2021-10-25 发布日期:2021-11-02
通讯作者: 张恒木
作者简介:*张恒木,E-mail: zhhengmu@tsinghua.org.cn
刘寒(1994—),女,河南新乡人,硕士研究生,主要从事植物病理学研究。E-mail: 2916120823@qq.com
基金资助:
浙江省重点研发计划(2019C02018)

Effects of exogenous salicylic acid on growth and defense-related genes of rice seedlings

LIU Han¹^,²(), DAI Yuanxing¹^,², LYU Mingfang², YUAN Zhengjie², LI Jing², YAN Chengqi², ZHANG Hengmu²^,^*()

1. College of Chemistry and Life Science, Zhejiang Normal University, Jinhua 321004, China
2. Institute of Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China

Received:2021-04-15 Online:2021-10-25 Published:2021-11-02
Contact: ZHANG Hengmu

摘要/Abstract

摘要：

为探究外源水杨酸(SA)对水稻苗期生长与SA相关防卫反应的影响,用不同浓度外源SA喷施苗期日本晴水稻,利用分光光度计测定水稻苗期生长关键指标叶绿素含量,采用qRT-PCR定量分析外源SA对水稻苗期SA合成基因PAL1、SA受体基因NPR1、SA信号途径下游的转录因子WRKY45/WRKY76和防卫基因PR1a/PR1b的表达水平。结果表明,外源SA对上述水稻基因的影响不同,即低浓度SA在一定程度上促进水稻苗期叶绿素积累,并显著影响水稻苗期防卫相关基因的表达水平;高浓度SA抑制叶绿素的积累,影响水稻的正常生长。综合比较结果显示,喷施2.0 mmol·L^-1外源SA对水稻苗期防卫相关基因表达的影响最为有效,该结果为进一步探索外源SA促进水稻苗期生长并提高水稻苗期防卫能力的作用研究奠定基础。

关键词: 水稻, 水杨酸, 防卫相关基因, 叶绿素

Abstract:

To explore the effects of exogenous salicylic acid (SA) on plant growth and SA-related defense responses at the stage of rice seedlings, gradient concentrations of exogenous SA were used to spray rice seedlings of Nipponbare. Spectrophotometer was applied to measure the content of chlorophyll, a key indicator of rice seedling growth, and qRT-PCR was further used to analyze the expression levels of SA-biosynthesis gene PAL1, SA-receptor gene NPR1, and transcription factors WRKY45/WRKY76 and defense genes PR1a/PR1b at the downstream of SA signal pathway. It was found that different concentrations of exogenous SA exerted varying influences on expression patterns of tested genes, i.e. lower concentrations of exogenous SA promoted chlorophyll accumulation a certain extent and markedly affected on the expression of defense-related genes, while high concentrations of SA inhibited the accumulation of chlorophyll and induced abnormal growth at the stage of rice seedlings. Comprehensive comparison of quantitative results suggested that 2.0 mmol·L ^-1 of SA could be most effective in inducing the expression of defense-related genes by spraying rice seedlings. These results laid the foundation for further investigating roles of exogenous SA in promotion of plant growth and defense responses at the stage of rice seedlings.

Key words: rice, salicylic acid (SA), defense-related genes, chlorophyll

中图分类号:

S511.01

刘寒, 戴远兴, 吕明芳, 袁正杰, 李静, 严成其, 张恒木. 外源水杨酸对水稻苗期生长与防卫相关基因表达的影响[J]. 浙江农业学报, 2021, 33(10): 1789-1796.

LIU Han, DAI Yuanxing, LYU Mingfang, YUAN Zhengjie, LI Jing, YAN Chengqi, ZHANG Hengmu. Effects of exogenous salicylic acid on growth and defense-related genes of rice seedlings[J]. Acta Agriculturae Zhejiangensis, 2021, 33(10): 1789-1796.

图/表 6

表1 qPCR采用的引物

Table 1 Primers used in qPCR

目的基因 Target gene	上游引物(5'→3') Forward primer (5'→3')	下游引物(5'→3') Reverse primer (5'→3')	参考文献 Reference
OsNPR1	TTTCCGATGGAGGCAAGAG	GCTGTCATCCGAGCTAAGTGTT	[29]
OsPR1b	CAGCAACTGGAACAACCTTGG	TATGGACCGTGAAGGCGTGG	[25]
PAL1	CCGACCACCTGACTCACAA	ATCTCACGCTCGATGGACTT
OsPR1a	TATGCTATGCTACGTGTTTATGC	CACTAAGCAAATACGGCTGACA
OsWRKY45	TCCACGCGTGTGTACAGAAA	TGCTAGCATGTCTGCAGCTTA	[29]
OsWRKY76	CTGCCCGAATTCTAGCTTCCT	GCCCAAGGACCAACAGGTTAT
UBC	CCGTTTGTAGAGCCATAATTGCA	AGGTTGCCTGAGTCACAGTTAAGTG	[28]

图1 SA处理的水稻苗叶绿素含量和表型变化 *表示与0 h相比差异达显著水平(P<0.05)。下同。

Fig.1 Changes in chlorophyll content and phenotypes of rice seedlings caused by SA * represented significant differences at 0.05 level compared with 0 h. The same as below.

图2 SA处理水稻苗中OsPAL1的相对表达量

Fig.2 Relative expression level of OsPAL1 in SA-treated rice seedlings

图3 SA处理水稻苗中NPR1的相对表达量

Fig.3 Relative expression level of NPR1 in SA-treated rice seedlings

图4 SA处理水稻苗中OsWRKY45和OsWRKY76的相对表达量

Fig.4 Relative expression levels of OsWRKY45 and OsWRKY76 in SA-treated rice seedlings

图5 SA处理水稻苗中OsPR1a和OsPR1b的相对表达量

Fig.5 Relative expression levels of OsPR1a and OsPR1b in SA-treated rice seedlings

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外源水杨酸对水稻苗期生长与防卫相关基因表达的影响

Effects of exogenous salicylic acid on growth and defense-related genes of rice seedlings

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