中国农业科学 ›› 2021, Vol. 54 ›› Issue (3): 619-628.doi: 10.3864/j.issn.0578-1752.2021.03.015
毕蒙蒙1(),刘迪1,高歌1,祝朋芳1,2,毛洪玉1,2()
收稿日期:
2020-05-04
接受日期:
2020-07-13
出版日期:
2021-02-01
发布日期:
2021-02-01
通讯作者:
毛洪玉
作者简介:
毕蒙蒙,E-mail: 基金资助:
BI MengMeng1(),LIU Di1,GAO Ge1,ZHU PengFang1,2,MAO HongYu1,2()
Received:
2020-05-04
Accepted:
2020-07-13
Online:
2021-02-01
Published:
2021-02-01
Contact:
HongYu MAO
摘要:
【目的】菊花白色锈病是菊花重要病害之一,严重影响其观赏品质。分析CmWRKY15-1在菊花抗白色锈病中的功能及防卫机制,为利用分子育种解决菊花产业中白色锈病病害问题提供借鉴。【方法】以菊花白色锈病抗病品种‘黄莺’为试验材料,构建CmWRKY15-1过表达载体与干扰载体,通过农杆菌介导法转化菊花,分别获得超表达与沉默植株株系,通过表型观察、病情指数统计及转基因植株内源SA含量变化、SA合成关键基因及病程相关基因的表达分析,探究CmWRKY15-1在水杨酸信号途径响应菊花白色锈病的功能。【结果】经遗传转化后获得超表达植株OE-9株系与沉默植株RNAi-4株系,接种后的表型鉴定和病情指数调查中RNAi-4株系病情指数为53.67,抗性反应表现为感病,而野生型植株接菌前后表型无变化,表明CmWRKY15-1的沉默降低了‘黄莺’对白色锈病的抗性。同时OE-9株系中内源水杨酸含量在接种前后与WT相比呈上升趋势,而RNAi-4株系中水杨酸含量降低,即CmWRKY15-1的过表达与沉默分别显著上调和下调内源SA含量,对SA的积累有正向调节作用。OE-9系、RNAi-4系和WT在接菌处理前后的SA合成基因表达情况分析结果表明,在接种处理后,OE-9株系中ICS1的表达呈先增加后降低的趋势,其最高表达量为对照的3.8倍。而在RNAi-4株系中,ICS1在接种后出现降低,且一直保持较低水平。PAL在OE-9株系中的表达相较于对照增加,其最高表达量为对照的2.3倍,而在RNAi-4株系中,该基因的表达总体呈下降趋势,这与SA含量测定的变化结果基本一致,也进一步验证CmWRKY15-1通过SA介导的信号途径响应菊花白色锈病。此外,响应SA信号的病程相关基因表达分析结果显示,随着白色锈病病原菌处理时间的延长,OE-9株系中这些基因的表达除PR5外,均在40 h时出现峰值,PR5相对延后,在48 h时出现峰值。RNAi-4株系中除PR5外,其他基因的表达量显著低于WT、OE-9中的表达量。CmWRKY15-1的过表达提高了水杨酸信号途径中PR的转录水平,与此相反,CmWRKY15-1的沉默使这些基因表达下调。【结论】CmWRKY15-1对菊花抗白色锈病具有正调控作用,且通过水杨酸介导的信号途径调控抗病。
毕蒙蒙,刘迪,高歌,祝朋芳,毛洪玉. CmWRKY15-1通过水杨酸信号通路调控菊花白色锈病抗性[J]. 中国农业科学, 2021, 54(3): 619-628.
BI MengMeng,LIU Di,GAO Ge,ZHU PengFang,MAO HongYu. CmWRKY15-1 Regulates Resistance of Chrysanthemum White Rust Through Salicylic Acid Signaling Pathway[J]. Scientia Agricultura Sinica, 2021, 54(3): 619-628.
表2
定量所需引物"
引物 Primer | 序列 Sequence(5′-3′) |
---|---|
ICS1-F | TCCCTACTGAAGAGGCACGG |
ICS1-R | CCAACAGCGGGTTCACTCTC |
PAL-F | ATGGCACCGAAGCAAGTCACAC |
PAL-R | GATACCCGAGTAACCCTGGAGGAG |
NPR1-F | TGTCGAGAAGGATGGAAAGCC |
NPR1-R | GGAGGCACCCATCATCAACA |
PR1-F | CTCAACCAAAAGGAATAGTCGG |
PR1-R | CCCTGCCAGTTTACGCTGTA |
PR2-F | GGCAATGGTGGTGTTGGAAC |
PR2-R | CTTCCTCCGTCAGCAGAAGG |
PR5-F | CCAATGGAGTTTAGCCCCGT |
PR5-R | GTCCACAACTACCACGCTCA |
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