Screening of Malus miRNA: Mediated Resistance to Colletotrichum gloeosporioides

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

Abstract

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

CLC Number:

S661.1

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.

Figures/Tables 6

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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