苹果炭疽病抗性miRNA的筛选

doi:10.11924/j.issn.1000-6850.casb2020-0519

中国农学通报 ›› 2021, Vol. 37 ›› Issue (7): 106-111.doi: 10.11924/j.issn.1000-6850.casb2020-0519

所属专题：农业生态

苹果炭疽病抗性miRNA的筛选

张亚楠(), 陈新慧, 张杰()

农业应用新技术北京市重点实验室/植物生产国家级实验教学示范中心/北京农学院植物科学技术学院,北京 102206

收稿日期:2020-09-30 修回日期:2020-12-14 出版日期:2021-03-05 发布日期:2021-03-17
通讯作者: 张杰
作者简介:张亚楠,女,1992年出生,北京人,硕士研究生在读,主要研究方向为果树种质资源利用与创新。通信地址：102206 北京市昌平区回龙观镇北农路7号,E-mail： yananzhang2019@gmail.com。
基金资助:
国家重点研发计划课题“苹果果园间作物趋避效应研究与趋避植物选择技术”(2016YFD0201116)

Screening of Malus miRNA: Mediated Resistance to Colletotrichum gloeosporioides

Zhang Yanan(), Chen Xinhui, Zhang Jie()

Beijing Key Laboratory of New Technology in Agricultural Application/ National Demonstration Center for Experimental Plant Production Education/ Plant Science and Technology College, Beijing University of Agriculture, Beijing 102206

Received:2020-09-30 Revised:2020-12-14 Online:2021-03-05 Published:2021-03-17
Contact: Zhang Jie

摘要/Abstract

摘要：

为筛选炭疽病菌侵染苹果属植物后相关miRNA的响应情况,以抗病品种‘八棱脆’海棠、感病品种‘嘎啦’苹果2个为试验材料,分别用胶胞炭疽孢子悬浮液侵染组培苗,4天后发病,提取植物总RNA并反转miRNA,用qRT-PCR分析2种植物中抗病相关miRNA相对表达量的差异,并对其靶基因进行预测。在抗性品种中miR390a和miR396b/c/f表达量下调,而在感病品种中表达量上调,且其靶基因均有LRR受体样丝氨酸/苏氨酸蛋白,miR482b靶基因为抗性蛋白。miR390a、miR482b和miR396b/c/f可能在苹果炭疽病的侵染过程中起重要作用。

关键词: 苹果, 炭疽病, 侵染, miRNA, 抗病

Abstract:

To screen the response of related miRNAs after Colletotrichum gloeosporioides infection on genus Malus, the disease resistant variety Malus robusta and the susceptible variety ‘Gala’ were taken as test materials to cultivate the seedlings respectively by using the C. gloeosporioides spore suspension infection group. Two varieties were infected after 4 days. Total RNAs of plants were extracted, and miRNAs were reversed. On this basis, qRT-PCR was used to analyze the difference of two varieties in relative expression of disease resistance related miRNAs and predict the target genes. The expression levels of miR390a and miR396b/c/f were down-regulated in resistant varieties, while they were up-regulated in susceptible varieties. Moreover, their target genes had LRR receptor like serine/ threonine protein, and miR482b target genes were resistance protein. miR390a, miR482b and miR396b/c/f might play an important role in the infection of apple anthracnose.

Key words: Malus× domestica;, anthracnose, infection, miRNA, disease resistance

中图分类号:

S661.1

张亚楠, 陈新慧, 张杰. 苹果炭疽病抗性miRNA的筛选[J]. 中国农学通报, 2021, 37(7): 106-111.

Zhang Yanan, Chen Xinhui, Zhang Jie. Screening of Malus miRNA: Mediated Resistance to Colletotrichum gloeosporioides[J]. Chinese Agricultural Science Bulletin, 2021, 37(7): 106-111.

图/表 6

参考文献 25

[1]	周吉生, 周中磊. 苹果炭疽病及其防治技术[J]. 山西果树, 2019(4):76-77.
[2]	王卓. 转录组测序数据与DNA序列结构的几种差异分析方法[D]. 哈尔滨:哈尔滨工业大学, 2019.
[3]	都贝贝. 苹果轮纹病抗性相关miRNA的筛选[D]. 南京:南京农业大学, 2013.
[4]	徐涛, 张富春. 植物miRNA抗胁迫机理研究进展[J]. 生物技术通报, 2008,5:5-9.
[5]	杨丽娟, 李世访, 卢美光. miRNA在植物病原调控方面的研究进展[J]. 生物技术通报, 2020,36(1):101-109.
[6]	Navarro L, Dunoyer P, Jay F, et al. A plant miRNA contributes to antibacterial resistance by repressing auxin signaling[J]. Science, 2006,312:436-439. doi: 10.1126/science.1126088 URL pmid: 16627744
[7]	Dugas D V, Barel B. Sucrose induction of Arabidopsis miR398 represses two Cu/Zn superoxide dismutases[J]. Plant Mol Biol, 2008,67:403-417.
[8]	Jagadeeswaran G, Saini A, Sunkar R. Biotic and abioticstress down-egulate miR398 expression in Arabidopsis[J]. Planta, 2009,229(4):1009-1014.
[9]	Deng Y, Wang J, Tung J, et al. A role for small RNA in regulating innate immunity during plant growth[J]. PLoS Pathogens, 2018,14:e1006756. URL pmid: 29293695
[10]	Li F, Pignatta D, Bendix C, et al. MicroRNA regulation of plant innate immune receptors[J]. Proceedings of the National Academy of Sciences of the United States of America, 2012,109:1790-1795.
[11]	Su Y, Li H G, Wang Y, et al. Poplar miR472a targeting NBS-LRRs is involved in effective defence against the necrotrophic fungus Cytospora chrysosperma[J]. Journal of Experimental Botany, 2018,69:5519-5530.
[12]	贺晓丽. miR482b/c在番茄抗晚疫病中的功能研究[D]. 大连:大连理工大学, 2018.
[13]	Natarajan B, Kalsi H S, Godbole P, et al. MiRNA160 is associated with local defense and systemic acquired resistance against Phytophthora infestans infection in potato[J]. Journal of Experimental Botany, 2018,69(8):2023-2036.
[14]	Li Yan, Cao Xiao-Long, Zhu Yong, et al. Osa-miR398b boosts H₂O₂ production and rice blast disease-resistance via multiple superoxide dismutases[J]. New Phytologist, 2019,222:1522.
[15]	Zhou S X, Zhu Y, Wang L F, et al. Osa-miR1873 fine-tunes rice immunity against Magnaporthe oryzae and yield traits[J]. J Integr Plant Biol, 2020,8.
[16]	Chisholm S T, Coaker G, Day B, et al. Hostmicrobe interactions: Shaping the evolution of the plant immune response[J]. Cell, 2006,124:803-814.
[17]	Weaver L M, Swiderski M R, Li Y, et al. The Arabidopsis thaliana TIR-NB-LRR R-protein, RPP1A; protein localization and constitutive activation of defence by truncated alleles in tobacco and Arabidopsis[J]. Plant Journal, 2010,47(6):829-840.
[18]	Whisson S C. A translocation signal for delivery of oomycete effect or proteins into host plant cells[J]. Nature, 2007,450:115-118.
[19]	Fabro G. Multiple candidate effectors from the oomycete pathogen Hyaloperonospora arabidopsidis suppress host plant immunity[J]. PloS Pathog, 2011,7:e1002348.
[20]	Sessa Guido . Molecular Plant Immunity (Sessa/ Molecular Plant Immunity)[M]. The Roles of Salicylic Acid and Jasmonic Acid in Plant Immunity, 2012: 55-79.
[21]	Ma Chao, Lu You, Bai Songlin, et al. Cloning and Characterization of miRNAs and Their Targets, Including a Novel miRNA-Targeted NBS-LRR Protein Class Gene in Apple (Golden Delicious)[J]. Molecular Plant, 2014,7(1):218-230.
[22]	Zhang Y, Zhang Q, Hao L, et al. A novel miRNA negatively regulates resistance to Glomerella leaf spot by suppressing expression of an NBS gene in apple[J]. Horticulture Research, 2019,6(1):93.
[23]	Su Yanyan, Li Hui-Guang, Wang Yonglin, et al. Poplar miR472a targeting NBS-LRRs is involved in effective defence against the necrotrophic fungus Cytospora chrysosperma[J]. Journal of Experimental Botany, 2018,69(22):5519-5530.
[24]	Li Jianying, Hull J Joe, Liang Sijia, et al. Genome-Wide Analysis of Cotton miRNAs During Whitefly Infestation Offers New Insights into Plant-Herbivore Interaction[J] . Int J Mol Sci, 2019,20. URL pmid: 31861214
[25]	陈磊. 番茄miR1916和miR396在植物响应胁迫中的作用机制[D]. 大连:大连理工大学, 2018.

引物名称	引物序列(5′-3′)
miR171i	cggTGAGCCGAACCAATATCACTC
miR160a	aTGCCTGGCTCCCTGTATGC
miR398b	TGTGTTCTCAGGTCGCCCC
miR398a	gTGTGTTCTCAGGTCACCCCTT
miR393a	cgTCCAAAGGGATCGCATTGATCT
miR172d	ggcgAGAATCTTGATGATGCTGCAT
miR396c	cggcTTCCACAGCTTTCTTGAACTT
miR396f	cgTTCCACGGCTTTCTTGAACTG
miR396b	cgggTTCCACAGCTTTCTTGAACTG
miR390a	AAGCTCAGGAGGGATAGCGC
un3729	GGAGGACATGACACAAAACCGAGC
miR482b	aggTCTTTCCTATCCCTCCCATTCC
miR156ad	ggcgTGACAGAAGAAAGTGAGCAC
miR156t	cggTTGACAGAAGAGAGAGAGCAC
miR156p	gcgCTGACAGAAGATAGAGAGCAC
miR156a	cggTGACAGAAGAGAGTGAGCAC
miR156ab	ggcgTTGACAGAAGATAGAGAGCAC
miR156aa	gcggTGACAGAAGATAGAGAGCAC

引物名称	引物序列(5′-3′)
miR171i	cggTGAGCCGAACCAATATCACTC
miR160a	aTGCCTGGCTCCCTGTATGC
miR398b	TGTGTTCTCAGGTCGCCCC
miR398a	gTGTGTTCTCAGGTCACCCCTT
miR393a	cgTCCAAAGGGATCGCATTGATCT
miR172d	ggcgAGAATCTTGATGATGCTGCAT
miR396c	cggcTTCCACAGCTTTCTTGAACTT
miR396f	cgTTCCACGGCTTTCTTGAACTG
miR396b	cgggTTCCACAGCTTTCTTGAACTG
miR390a	AAGCTCAGGAGGGATAGCGC
un3729	GGAGGACATGACACAAAACCGAGC
miR482b	aggTCTTTCCTATCCCTCCCATTCC
miR156ad	ggcgTGACAGAAGAAAGTGAGCAC
miR156t	cggTTGACAGAAGAGAGAGAGCAC
miR156p	gcgCTGACAGAAGATAGAGAGCAC
miR156a	cggTGACAGAAGAGAGTGAGCAC
miR156ab	ggcgTTGACAGAAGATAGAGAGCAC
miR156aa	gcggTGACAGAAGATAGAGAGCAC

miRNA	靶基因	靶基因注释
miR390a	MDP0000128521	可能富含亮氨酸重复受体样蛋白激酶At1g35710
	MDP0000307313	谷氨酸受体2.7
	MDP0000144734	LRR受体样丝氨酸/苏氨酸蛋白激酶GSO1
	MDP0000322059	可能是LRR受体样丝氨酸/苏氨酸蛋白激酶At1g14390
miR482b	MDP0000160232	假定抗病蛋白RGA3
	MDP0000715013	假定抗病蛋白RGA4
	MDP0000733294	假定抗病蛋白RGA4
	MDP0000195139	假定抗病蛋白RGA4
miR396c/b/f	MDP0000261112	AtGRF8（生长调节因子8）;转录激活
	MDP0000320460	可能是LRR受体样丝氨酸/苏氨酸蛋白激酶MRH1
	MDP0000243533	AtGRF5（生长调节因子5）;转录激活
	MDP0000317856	TMV抗性蛋白

miRNA	靶基因	靶基因注释
miR390a	MDP0000128521	可能富含亮氨酸重复受体样蛋白激酶At1g35710
	MDP0000307313	谷氨酸受体2.7
	MDP0000144734	LRR受体样丝氨酸/苏氨酸蛋白激酶GSO1
	MDP0000322059	可能是LRR受体样丝氨酸/苏氨酸蛋白激酶At1g14390
miR482b	MDP0000160232	假定抗病蛋白RGA3
	MDP0000715013	假定抗病蛋白RGA4
	MDP0000733294	假定抗病蛋白RGA4
	MDP0000195139	假定抗病蛋白RGA4
miR396c/b/f	MDP0000261112	AtGRF8（生长调节因子8）;转录激活
	MDP0000320460	可能是LRR受体样丝氨酸/苏氨酸蛋白激酶MRH1
	MDP0000243533	AtGRF5（生长调节因子5）;转录激活
	MDP0000317856	TMV抗性蛋白

苹果炭疽病抗性miRNA的筛选

Screening of Malus miRNA: Mediated Resistance to Colletotrichum gloeosporioides

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