Screening of host factors interacting with wheat yellow mosaic virus P2 by yeast two-hybrid system

doi:10.3969/j.issn.1004-1524.2021.03.15

Acta Agriculturae Zhejiangensis ›› 2021, Vol. 33 ›› Issue (3): 497-505.DOI: 10.3969/j.issn.1004-1524.2021.03.15

• Plant Protection • Previous Articles Next Articles

Screening of host factors interacting with wheat yellow mosaic virus P2 by yeast two-hybrid system

HAN Xiaolei¹^,²^,³, GAO Shiqi¹^,²^,³, ZHANG Fan³, YANG Jian³, LIU Peng³, JIANG Hongming¹, LI Linzhi¹^,^*()

1. Yantai Academy of Agricultural Sciences, Yantai 265500, China
2. College of Life Sciences, Yantai University, Yantai 265500, China
3. Ningbo University/State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products, Ningbo 315000, China

Received:2020-10-27 Online:2021-03-25 Published:2021-03-25
Contact: LI Linzhi

Abstract

Abstract:

Wheat yellow mosaic virus (WYMV) is classified in the genus Bymovirus, family Potyviridae. The genome is composed of two sense RNA single-stranded RNA1 and RNA2 and encodes 10 proteins, among which P2 plays an important role in virus replication. In order to identify the targets of P2 in host, co-transformation of cDNA library vector and pGBKT7-P2 was constructed, eighteen candidate host targets of P2 were screened out, which were divided into several types based on their functions, including salicylic acid signal pathway factor, E3 ubiquitin ligase, defense response, oxidoreductase activity and photosystem II assembly. Additionally, the interaction of P2 and TaTIFY or Ta14-3-3 was further verified by Y2H system. Based on the results, it could be speculated that P2 may interact with E3 ubiquitin transcription factor, salicylic acid signal transcription factor, or the factors associated with plant photosystem and chloroplasts formation. In conclusion, P2 may participate in multiple signaling pathways during viral infection, and provide an insight into molecular interaction mechanism between WYMV and the host.

Key words: wheat yellow mosaic virus, wheat, P2, cDNA library, yeast two-hybrid

CLC Number:

S435.12

HAN Xiaolei, GAO Shiqi, ZHANG Fan, YANG Jian, LIU Peng, JIANG Hongming, LI Linzhi. Screening of host factors interacting with wheat yellow mosaic virus P2 by yeast two-hybrid system[J]. Acta Agriculturae Zhejiangensis, 2021, 33(3): 497-505.

Figures/Tables 7

References 32

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编号 Number	引物名称 Primers name	引物序列 Primers sequence(5'-3')	酶切位点 RE site
1	pGADT7 forward primer	TAATACGACTCACTATAGGGCGA	SamⅠ
	pGADT7 reverse primer	CTGTGCATCGTGCACCATCT	BamHⅠ
2	pGBKT7 forward primer	GTAATACGACTCACTATAGGGCGA	NotⅠ
	pGBKT7 reverse primer	ATTCTCAGTGAAATTTTAAACATA	PstⅠ
3	Ta14-3-3 forward primer	TCCCCCGGGATGGCCTTCTTCTCCCACCAC	SamⅠ
	Ta14-3-3 reverse primer	CGCGGATCCTCACATATATATCTCCATGCC	BamHⅠ
4	WYMV-P2 forward primer	AAAGCGGCCGCATGTCAGCTGAGGAAACAGGT	NotⅠ
	WYMV-P2 reverse primer	GCACTGCAGATGGCCCGGCCTCCTGAGGAA	PstⅠ
5	TaTIFY 10A forward primer	TCCCCCGGGATGGCGGCTTCCGCGAGGCAG	SamⅠ
	TaTIFY 10A reverse primer	CGCGGATCCTTATTGGTTTGGCTTCAGGGC	BamHⅠ

编号 Number	引物名称 Primers name	引物序列 Primers sequence(5'-3')	酶切位点 RE site
1	pGADT7 forward primer	TAATACGACTCACTATAGGGCGA	SamⅠ
	pGADT7 reverse primer	CTGTGCATCGTGCACCATCT	BamHⅠ
2	pGBKT7 forward primer	GTAATACGACTCACTATAGGGCGA	NotⅠ
	pGBKT7 reverse primer	ATTCTCAGTGAAATTTTAAACATA	PstⅠ
3	Ta14-3-3 forward primer	TCCCCCGGGATGGCCTTCTTCTCCCACCAC	SamⅠ
	Ta14-3-3 reverse primer	CGCGGATCCTCACATATATATCTCCATGCC	BamHⅠ
4	WYMV-P2 forward primer	AAAGCGGCCGCATGTCAGCTGAGGAAACAGGT	NotⅠ
	WYMV-P2 reverse primer	GCACTGCAGATGGCCCGGCCTCCTGAGGAA	PstⅠ
5	TaTIFY 10A forward primer	TCCCCCGGGATGGCGGCTTCCGCGAGGCAG	SamⅠ
	TaTIFY 10A reverse primer	CGCGGATCCTTATTGGTTTGGCTTCAGGGC	BamHⅠ

编号 Number	序列名 Sequence ID	基因名 Gene name	功能 Function
1	TraesCS2A02G407700.1	TapTOM75	Aquaporin PIP-type pTOM75
2	TraesCS5A02G322500.1	TaCB36	Chlorophyll a-b binding protein 36
3	TraesCS6A02G245100.1	TaRS31	Serine/arginine-rich splicing factor RS31-like
4	TraesCS2D02G248400.2	TaOEE1	Oxygen-evolving enhancer protein 1
5	TraesCS5A02G164600.1	TaLIP	Light-induced protein
6	TraesCS3D02G055500.1	Ta14-3-3	14-3-3-like protein C
7	TraesCS2B02G195600.1	TaTIFY 10A	Protein TIFY 10A-like
8	TraesCS3D02G247300.2	TaID31	Protein IQ-DOMAIN 31-like
9	TraesCS7A02G235200.3	TaAGWD	Alpha-glucan water dikinase
10	TraesCS7D02G276300.1	TaCBP21	Chlorophyll a-b binding protein 21
11	TraesCS3D02G232300.1	TaUBA2c	UBP1-associated protein 2C-like
12	TraesCS2B02G126500.1	TaRL19-3	60S ribosomal protein L19-3-like
13	TraesCS3D02G079600.2	TaBAG6	Large proline-rich protein BAG6
14	TraesCS4A02G159700.2	TaCGS1	Cystathionine gamma-synthase 1
15	TraesCS2D02G253900.1	TaPSP	Photosystem II 10 kDa polypeptide
16	TraesCS3A02G309800.1	TaCNR8	Cell number regulator 8-like
17	TraesCS5D02G169600.2	TaRBCSC	Ribulose bisphosphate carboxylase small chain
18	TraesCS4D02G215600.2	TaBOI	BOI-related E3 ubiquitin-protein ligase 3

编号 Number	序列名 Sequence ID	基因名 Gene name	功能 Function
1	TraesCS2A02G407700.1	TapTOM75	Aquaporin PIP-type pTOM75
2	TraesCS5A02G322500.1	TaCB36	Chlorophyll a-b binding protein 36
3	TraesCS6A02G245100.1	TaRS31	Serine/arginine-rich splicing factor RS31-like
4	TraesCS2D02G248400.2	TaOEE1	Oxygen-evolving enhancer protein 1
5	TraesCS5A02G164600.1	TaLIP	Light-induced protein
6	TraesCS3D02G055500.1	Ta14-3-3	14-3-3-like protein C
7	TraesCS2B02G195600.1	TaTIFY 10A	Protein TIFY 10A-like
8	TraesCS3D02G247300.2	TaID31	Protein IQ-DOMAIN 31-like
9	TraesCS7A02G235200.3	TaAGWD	Alpha-glucan water dikinase
10	TraesCS7D02G276300.1	TaCBP21	Chlorophyll a-b binding protein 21
11	TraesCS3D02G232300.1	TaUBA2c	UBP1-associated protein 2C-like
12	TraesCS2B02G126500.1	TaRL19-3	60S ribosomal protein L19-3-like
13	TraesCS3D02G079600.2	TaBAG6	Large proline-rich protein BAG6
14	TraesCS4A02G159700.2	TaCGS1	Cystathionine gamma-synthase 1
15	TraesCS2D02G253900.1	TaPSP	Photosystem II 10 kDa polypeptide
16	TraesCS3A02G309800.1	TaCNR8	Cell number regulator 8-like
17	TraesCS5D02G169600.2	TaRBCSC	Ribulose bisphosphate carboxylase small chain
18	TraesCS4D02G215600.2	TaBOI	BOI-related E3 ubiquitin-protein ligase 3

Screening of host factors interacting with wheat yellow mosaic virus P2 by yeast two-hybrid system

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