利用酵母双杂交系统筛选与柑橘黄化脉明病毒CP互作的寄主因子

doi:10.3864/j.issn.0578-1752.2023.10.006

Abstract

Abstract:

【Objective】Citrus yellow vein clearing virus (CYVCV) is one of the viruses mostly threatening the stable development of citrus industry in China, but its infection and pathogenic mechanism in citrus is still unclear. In this study, the coat protein (CP) of CYVCV was used as a bait to screen the Eureka lemon (Citrus limon Burm. f.) cDNA library, and the function of obtained host factors in the interaction between host and virus was analyzed by bioinformatics method.【Method】The total RNA of Eureka lemon leaves was extracted by the Trizol method, and then reversely transcribed to the First-Stand cDNA with SMART method, which was used as a template for obtaining ds cDNA through Long-Distance PCR. After homogenization, the ds cDNA fragments were ligated to pGADT7 plasmid vector by recombination junctions to construct the primary cDNA library of Eureka lemon. The recombinant plasmids were transfected into Escherichia coli DH10B to obtain the E. coli cDNA library of Eureka lemon, and its quality was identified. Simultaneously, the CP sequence of CYVCV was amplified by PCR and ligated into the yeast two-hybrid (Y2H) bait vector pGBKT7, and the plasmids of pGBKT7-CP and pGADT7 were co-transfected into yeast Y2H Gold. The positive yeast clones were grown on the plate of SD/-Trp, SD/-Leu-Trp, SD/-Leu-Trp/X-α-Gal and SD/-Leu-Trp-His medium, respectively, and then the growth situation of the yeast was tested to identify the toxicity of pGBKT7-CP on yeast Y2H Gold and the self-activating effect of pGBKT7-CP on the reporter gene of yeast was analyzed. Then the Y2H Gold containing bait vector pGBKT7-CP was transformed with the primary cDNA library of Eureka lemon, the co-transformed yeasts were coated on the plate of SD/-Leu-Trp, SD/-Leu-Trp-His/X-α-Gal and SD/-Leu-Trp-His-Ade/X-α-Gal medium in turn. Finally, the blue and well grown positive clones were selected. The plasmids of positive yeast clones were extracted and sequenced. The candidate genes were preliminarily compared in the GenBank, and the interacted protein factors were annotated and the protein’s biological functions were analyzed with gene ontology (GO) pathway of Uniprot online websites. According to the results of the analysis, candidate factors that may be involved in host disease resistance or symptom development were selected, their CDS full-length sequences were amplified and constructed into the target vector pGADT7 and then were verified with pGBKT7-CP by one for one in yeast, respectively.【Result】The titer of the Eureka lemon-E. coli cDNA library was 1.02×10⁸ cfu/mL, and it demonstrated that the cDNA library measured up to the experiment standard. The bait vector of pGBKT7-CP was constructed, which had no ability to activate the reporter gene and had no virulence to the yeast. The 41 positive clones were finally selected by using the SD/-Leu-Trp-His-Ade/X-α-Gal medium. After sequence similarity comparison, removing the repetitive sequences, the vector sequences and the frameshift sequences, the 32 host factors that interacted with CYVCV CP were screened. The GO pathway annotation results indicated that these host factors were involved in several chloroplast-related biological processes, including photosynthesis, metabolic process, carbohydrate metabolic process, response to light stimulus, etc. The molecular functions of the 32 host factors were multiple, including catalytic activity, hydrolase activity, transferase activity, protein binding, DNA-binding transcription factor activity, and translation factor activity, etc. Moreover, the cell components of the 32 host factors were involved in the cytoplasm, membrane, chloroplast, thylakoid, nucleolus, and golgi apparatus, etc. Validation of one-to-one yeast two-hybrid crosses of 14 important proteins selected from candidate host factors with CP showed that CP interacted with all 14 proteins.【Conclusion】The cDNA library of Eureka lemon was constructed successfully, and 32 host factors of Eureka lemon interacted with CYVCV CP were preliminarily screened. According to the results, the function of the important proteins was analyzed. It was presumed that CYVCV CP affects photosystem stability, thylakoid structure and chlorophyll synthesis by interacting with multiple chloroplast-related proteins such as oxygen-evolving enhancer protein 2 (PsbP), chlorophyll a-b binding protein 8 (Lhca3) and ribulose bisphosphate carboxylase small subunit (RbcS), resulting in reduced photosynthesis and damaged chloroplast morphology and function. The interaction of CP with these chloroplast-associated factors was separately verified one-to-one in yeast, which will provide a theoretical basis for revealing the molecular mechanism of CYVCV CP in viral pathogenicity.

Key words: citrus yellow vein clearing virus (CYVCV), Eureka lemon, yeast two-hybrid, coat protein, host factor

BIN Yu, ZHANG Qi, WANG ChunQing, ZHAO XiaoChun, SONG Zhen, ZHOU ChangYong. Screening of the Host Factors Interacting with CP of Citrus Yellow Vein Clearing Virus by Yeast Two-Hybrid System[J].Scientia Agricultura Sinica, 2023, 56(10): 1881-1892.

Figures/Tables 8

Table 1

Fig. 1

Fig. 2

Fig. 3

Fig. 4

Table 2

Interacting candidate proteins obtained by Y2H screening of a lemon cDNA library using CYVCV CP as bait"

克隆号 Clone number	登录号 GenBank accession	蛋白注释ID Uniprot ID	寄主因子（蛋白名称缩写） Factor of host (Abbreviations of protein name)	与CP一对一酵母双杂交验证结果 Validation results for one-to-one Y2H with CP
1	XP_015383675.1	A0A067E581	可能的甲基转移酶20（PMT20）Probable methyltransferase 20	互作Interaction
2	XP_006487560.1	A0A067ESK0	转醛缩酶（TAL）Transaldolase	互作Interaction
3	XP_006488195.1	A0A067G0G4	L-抗坏血酸过氧化物酶（APX）L-ascorbate peroxidase	互作Interaction
4	XP_006494191.1	V4UB67	U11/U12小核糖核蛋白35 kDa类蛋白 U11/U12 small nuclear ribonucleoprotein 35 kDa protein-like	—
5	XP_006479566.1	V4TUF9	类真核翻译起始因子6-2（eIF6） Eukaryotic translation initiation factor 6-2-like	互作Interaction
6	NP_001306987.1	A0A067G915	WD40重复蛋白（WD40-repeat）WD40 repeat protein	互作Interaction
7	XP_006493749.1	A0A067F2B8	琥珀酸-半醛脱氢酶，线粒体亚型X2（SSADH） Succinate-semialdehyde dehydrogenase, mitochondrial isoform X2	—
8	XP_006480341.1	A0A067EMJ8	疏水蛋白LTI6B Hydrophobic protein LTI6B	—
9	XP_006469744.1	A0A067EA24	木葡聚糖内转葡糖基化酶蛋白6（XTH6） Xyloglucan endotransglucosylase/hydrolase protein 6	互作Interaction
10	XP_006466254.1	A0A067F028	Ras相关蛋白Rab11A Ras-related protein Rab11A	—
11	XP_006476755.1	A0A067FQW5	醛脱氢酶家族2成员B4，线粒体（ALDH2B4） Aldehyde dehydrogenase family 2 member B4, mitochondrial	—
12	XP_024949159.1	—	含有RAP结构域的蛋白质，叶绿体 RAP domain-containing protein, chloroplast	—
13	XP_006481758.1	A0A067FSV8	异质核糖核蛋白Q（HNRNPAQ） Heterogeneous nuclear ribonucleoprotein Q	—
14	XP_006493717.1	A0A067F2C0	类T复合蛋白1亚基 T-complex protein 1 subunit gamma-like	—
15	XP_006466732.1	A0A067GIJ6	氨基肽酶P1亚型X1 Aminopeptidase P1 isoform X1	—
16	XP_006488080.1	A0A067GCU3	类丝氨酸/苏氨酸蛋白磷酸酶6调节亚基3 Serine/threonine-protein phosphatase 6 regulatory subunit 3-like	—
17	XP_006473996.1	A0A067EWB7	可能的嘌呤渗透酶5 Probable purine permease 5	—
18	NP_001275850.1	Q8LPM6	蔗糖转运蛋白1（SUT）Sucrose transporter 1	—
19	XP_006490909.1	A0A067H1D4	转酮醇酶，叶绿体（TK）Transketolase, chloroplast	—
20	XP_006471243.2	A0A067DX06	光捕获叶绿素a-b结合蛋白3，叶绿体（Lhca3） Light-harvesting chlorophyll a-b binding protein 8, chloroplast	互作Interaction
21	XP_006477045.2	A0A067DAX7	类FCS锌指蛋白1（FLZ1）FCS-like zinc finger protein 1	互作Interaction
22	XP_006474598.1	A0A067GEE4	过氧化物还原酶Q，叶绿体（PrxQ）Peroxiredoxin Q, chloroplast	互作Interaction
23	XP_006492323.1	A0A067H7T2	通用应激蛋白PHOS34（USP PHOS34）Universal stress protein PHOS34	互作Interaction
24	XP_006490626.1	A0A067EZZ6	重金属相关异戊二烯化植物蛋白39（HIPP39） Heavy metal-associated isoprenylated plant protein 39	互作Interaction
25	XP_006482254.1	Q84LN2	核酮糖二磷酸羧化酶小亚基，叶绿体（RbcS） Ribulose bisphosphate carboxylase (RuBisCO) small subunit, chloroplast	互作Interaction
26	XP_006482548.1	A0A067G267	V型质子ATP酶亚基d2 V-type proton ATPase subunit d2	—
27	XP_006484494.1	A0A067G359	F盒/LRR重复蛋白4（FBXL4）F-box/LRR-repeat protein 4	互作Interaction
28	XP_006469469.1	A0A067D341	葡萄糖-6-磷酸1-脱氢酶，类叶绿体（G6PD） Glucose-6-phosphate 1-dehydrogenase, chloroplast-like	—
29	XP_006485001.1	A0A067DPL0	光系统II放氧强化蛋白2，叶绿体（PsbP） Photosystem II oxygen-evolving enhancer protein 2, chloroplast	互作Interaction
30	XP_006476226.1	—	未知蛋白Uncharacterized protein	—
31	XP_006491226.1	A0A067HEJ5	通用植物调节因子1亚型X2 Common plant regulatory factor 1 isoform X2	—
32	XP_006478862.1	A0A067E694	类溶质载体家族35成员F2 Solute carrier family 35 member F2-like	—

Table 2

Fig. 5

Fig. 6

References 34

[1]	ZHOU Y, CHEN H M, CAO M J, WANG X F, JIN X, LIU K H, ZHOU C Y. Occurrence, distribution, and molecular characterization of citrus yellow vein clearing virus in China. Plant Disease, 2017, 101(1): 137-143. doi: 10.1094/PDIS-05-16-0679-RE pmid: 30682317
[2]	CHEN H M, LI Z A, WANG X F, ZHOU Y, TANG K Z, ZHOU C Y, ZHAO X Y, YUE J Q. First report of citrus yellow vein clearing virus on lemon in Yunnan, China. Plant Disease, 2014, 98(12): 1747.
[3]	REHMAN A U, LI Z R, YANG Z K, WAQAS M, WANG G P, XU W X, LI F, HONG N. The coat protein of citrus yellow vein clearing virus interacts with viral movement proteins and serves as an RNA silencing suppressor. Viruses, 2019, 11(4): 329. doi: 10.3390/v11040329
[4]	BIN Y, XU J, DUAN Y, MA Z, ZHANG Q, WANG C, SU Y, JIANG Q, SONG Z, ZHOU C. The titer of citrus yellow vein clearing virus is positively associated with the severity of symptoms in infected citrus seedlings. Plant Disease, 2022, 106(3): 828-834. doi: 10.1094/PDIS-02-21-0232-RE
[5]	宾羽, 宋震, 崔甜甜, 周常勇. 柑橘黄化脉明病毒突变体及其构建方法: ZL201910939750.8[P]. (2019-12-31) [2023-03-02].
	BIN Y, SONG Z, CUI T T, ZHOU C Y. Citrus yellow vein clearing virus mutant and its construction method: ZL201910939750.8[P]. (2019-12-31) [2023-03-02]. (in Chinese)
[6]	LOCONSOLE G, ONELGE N, POTERE O, GIAMPETRUZZI A, BOZAN O, SATAR S, DE STRADIS A, SAVINO V, YOKOMI R K, SAPONARI M. Identification and characterization of citrus yellow vein clearing virus, a putative new member of the genus Mandarivirus. Phytopathology, 2012, 102(12): 1168-1175. doi: 10.1094/PHYTO-06-12-0140-R
[7]	SONG Z, KURTH E G, PEREMYSLOV V V, ZHOU C Y, DOLJA V V. Molecular characterization of a citrus yellow vein clearing virus strain from China. Archives of Virology, 2015, 160(7): 1811-1813. doi: 10.1007/s00705-015-2423-1 pmid: 25913691
[8]	MA X F, HONG N, MOFFETT P, ZHOU Y J, WANG G P. Functional analysis of apple stem pitting virus coat protein variants. Virology Journal, 2019, 16(1): 20. doi: 10.1186/s12985-019-1126-8 pmid: 30736799
[9]	李帅, 蒋西子, 梁伟芳, 陈思涵, 张享享, 左登攀, 胡亚会, 江彤. 利用酵母双杂交系统筛选与草莓镶脉病毒P6蛋白互作的森林草莓寄主因子. 中国农业科学, 2017, 50(18): 3519-3528. doi: 10.3864/j.issn.0578-1752.2017.18.008. doi: 10.3864/j.issn.0578-1752.2017.18.008
	LI S, JIANG X Z, LIANG W F, CHEN S H, ZHANG X X, ZUO D P, HU Y H, JIANG T. Screening of the host factors of woodland strawberry interacting with P6 of strawberry vein banding virus by yeast two-hybrid system. Scientia Agricultura Sinica, 2017, 50(18): 3519-3528. doi: 10.3864/j.issn.0578-1752.2017.18.008. (in Chinese) doi: 10.3864/j.issn.0578-1752.2017.18.008
[10]	ZHAO J, ZHANG X, HONG Y, LIU Y. Chloroplast in plant-virus interaction. Frontiers in Microbiology, 2016, 7: 1565. pmid: 27757106
[11]	CUI T, BIN Y, YAN J, MEI P, LI Z, ZHOU C, SONG Z. Development of infectious cDNA clones of citrus yellow vein clearing virus using a novel and rapid strategy. Phytopathology, 2018, 108(10): 1212-1218. doi: 10.1094/PHYTO-02-18-0029-R pmid: 29726761
[12]	QIU Y, ZHANG Y, WANG C, LEI R, WU Y, LI X, ZHU S. Cucumber mosaic virus coat protein induces the development of chlorotic symptoms through interacting with the chloroplast ferredoxin I protein. Scientific Reports, 2018, 8(1): 1205. doi: 10.1038/s41598-018-19525-5 pmid: 29352213
[13]	马丹丹. 柑橘黄化脉明病毒在尤力克柠檬中的时空分布及细胞结构变化研究[D]. 重庆: 西南大学, 2016.
	MA D D. Spatial and temporal distribution of citrus yellow vein clearing virus and cell structural changes in infected Euneka lemon[D]. Chongqing: Southwest University, 2016. (in Chinese)
[14]	邓雨青. 柑橘黄化脉明病毒诱导尤力克柠檬细胞程序性死亡研究[D]. 重庆: 西南大学, 2017.
	DENG Y Q. Study on programmed cell death in Euneka lemon induced by citrus yellow vein clearing virus[D]. Chongqing: Southwest University, 2017. (in Chinese)
[15]	MATHIOUDAKIS M M, RODRIGUEZ-MORENO L, SEMPERE R N, ARANDA M A, LIVIERATOS I. Multifaceted capsid proteins: Multiple interactions suggest multiple roles for pepino mosaic virus capsid protein. Molecular Plant-Microbe Interactions, 2014, 27(12): 1356-1369. doi: 10.1094/MPMI-07-14-0195-R pmid: 25162316
[16]	BHATTACHARYYA D, CHAKRABORTY S. Chloroplast: The Trojan horse in plant-virus interaction. Molecular Plant Pathology, 2018, 19(2): 504-518. doi: 10.1111/mpp.12533 pmid: 28056496
[17]	QIAO Y, LI H F, WONG S M, FAN Z F. Plastocyanin transit peptide interacts with potato virus X coat protein, while silencing of plastocyanin reduces coat protein accumulation in chloroplasts and symptom severity in host plants. Molecular Plant-Microbe Interactions, 2009, 22(12): 1523-1534. doi: 10.1094/MPMI-22-12-1523 pmid: 19888818
[18]	SUN X, LI Y, SHI M, ZHANG N, WU G, LI T, QING L, ZHOU C. In vitro binding and bimolecular fluorescence complementation assays suggest an interaction between tomato mosaic virus coat protein and tobacco chloroplast ferredoxin I. Archives of Virology, 2013, 158(12): 2611-2615. doi: 10.1007/s00705-013-1778-4
[19]	GRUBER A V, FEIZ L. Rubisco assembly in the chloroplast. Frontiers in Molecular Biosciences, 2018, 5: 24. doi: 10.3389/fmolb.2018.00024 pmid: 29594130
[20]	KAWAKAMI N, WATANABE A. Translatable mRNAs for chloroplast- targeted proteins in detached radish cotyledons during senescence in darkness. Plant and Cell Physiology, 1993, 34(5): 697-704. doi: 10.1093/oxfordjournals.pcp.a078472
[21]	ROY H. Rubisco assembly:A research memoir//BISWAL B, KRUPINSKA K, BISWAL U C. Plastid Development in Leaves during Growth and Senescence. Dordrecht: Springer Netherlands, 2013: 117-129.
[22]	ATKINSON N, LEITAO N, ORR D J, MEYER M T, CARMO-SILVA E, GRIFFITHS H, SMITH A M, MCCORMICK A J. Rubisco small subunits from the unicellular green alga Chlamydomonas complement Rubisco-deficient mutants of Arabidopsis. New Phytologist, 2017, 214(2): 655-667. doi: 10.1111/nph.2017.214.issue-2
[23]	SUZUKI Y, MAKINO A. Availability of Rubisco small subunit up-regulates the transcript levels of large subunit for stoichiometric assembly of its holoenzyme in rice. Plant Physiology, 2012, 160(1): 533-540. doi: 10.1104/pp.112.201459 pmid: 22811433
[24]	LIN L, LUO Z P, YAN F, LU Y W, ZHENG H Y, CHEN J P. Interaction between potyvirus P3 and ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO) of host plants. Virus Genes, 2011, 43(1): 90-92. doi: 10.1007/s11262-011-0596-6 pmid: 21400205
[25]	ZHAO J P, LIU Q, ZHANG H L, JIA Q, HONG Y G, LIU Y L. The Rubisco small subunit is involved in tobamovirus movement and Tm-2²-mediated extreme resistance. Plant Physiology, 2013, 161(1): 374-383. doi: 10.1104/pp.112.209213
[26]	BARBER J, NIELD J. Organization of transmembrane helices in photosystem II: Comparison of plants and cyanobacteria. Philosophical Transactions of the Royal Society of London, Series B, Biological Sciences, 2002, 357(1426): 1329-1335.
[27]	IFUKU K, YAMAMOTO Y, ONO T, ISHIHARA S, SATO F. PsbP protein, but not PsbQ protein, is essential for the regulation and stabilization of photosystem II in higher plants. Plant Physiology, 2005, 139(3): 1175-1184. pmid: 16244145
[28]	IFUKU K, ISHIHARA S, SHIMAMOTO R, IDO K, SATO F. Structure, function, and evolution of the PsbP protein family in higher plants. Photosynthesis Research, 2008, 98(1/3): 427-437. doi: 10.1007/s11120-008-9359-1
[29]	IDO K, IFUKU K, YAMAMOTO Y, ISHIHARA S, MURAKAMI A, TAKABE K, MIYAKE C, SATO F. Knockdown of the PsbP protein does not prevent assembly of the dimeric PSII core complex but impairs accumulation of photosystem II supercomplexes in tobacco. Biochimica et Biophysica Acta-Bioenergetics, 2009, 1787(7): 873-881. doi: 10.1016/j.bbabio.2009.03.004
[30]	IFUKU K. The PsbP and PsbQ family proteins in the photosynthetic machinery of chloroplasts. Plant Physiology and Biochemistry, 2014, 81: 108-114. doi: 10.1016/j.plaphy.2014.01.001 pmid: 24477118
[31]	IFUKU K, NOGUCHI T. Structural coupling of extrinsic proteins with the oxygen-evolving center in photosystem II. Frontiers in Plant Science, 2016, 7: 84. doi: 10.3389/fpls.2016.00084 pmid: 26904056
[32]	BALASUBRAMANIAM M, KIM B S, HUTCHENS-WILLIAMS H M, LOESCH-FRIES L S. The photosystem II oxygen-evolving complex protein PsbP interacts with the coat protein of alfalfa mosaic virus and inhibits virus replication. Molecular Plant-Microbe Interactions, 2014, 27(10): 1107-1118. doi: 10.1094/MPMI-02-14-0035-R pmid: 24940990
[33]	KONG L, WU J, LU L, XU Y, ZHOU X. Interaction between rice stripe virus disease-specific protein and host PsbP enhances virus symptoms. Molecular Plant, 2014, 7(4): 691-708. doi: 10.1093/mp/sst158 pmid: 24214893
[34]	BIN Y, ZHANG Q, SU Y, WANG C, JIANG Q, SONG Z, ZHOU C. Transcriptome analysis of Citrus limon infected with citrus yellow vein clearing virus. BMC Genomics, 2023, 24(1): 65. doi: 10.1186/s12864-023-09151-5

Metrics

Viewed

Full text

Abstract

Cited

Shared

Discussed

Comments

Recommended 0

No Suggested Reading articles found!

引物名称Primer name	引物序列Primer sequence (5′-3′)
CP-Nde I-F	TGATCTCAGAGGAGGACCTGCATATGAGCTTCGACTACACTCACCC
CP-BamH I-R	TGCGGCCGCTGCAGGTCGACGGATCCTTAGATGTTGAAAGGGGTC
pGADT7-F	TAATACGACTCACTATAGGGCGAGCGCCGCCATG
pGADT7-R	TGCGGCCGCTGCAGGTCGACGGATCCTTAGATGTTGAAAGGGGTC
1-F	CCCGGGTGGGCATCGATACGGGATCCATGCAAGATGGGACAGTTCG
1-R	TATCTACGATTCATCTGCAGCTCGAGTCAACTAGAGGTTTGGTTAG
2-F	CCCGGGTGGGCATCGATACGGGATCCATGGCGACCATTTCGAAGCTTAC
2-R	TATCTACGATTCATCTGCAGCTCGAGTTACAGACTAACCAGCTTGAG
3-F	CCCGGGTGGGCATCGATACGGGATCCATGACGAAGAATTACCCCAC
3-R	TATCTACGATTCATCTGCAGCTCGAGTTAGGCTTCAGCAAATCCTAG
5-F	ACGACGTACCAGATTACGCTCATATGGCAACCAGACTTATGTTTGAG
5-R	ATCTGCAGCTCGAGCTCGATGGATCCTTAGACATAGCTGTCAATTAAAG
6-F	ACGACGTACCAGATTACGCTCATATGGAAAACTCAAGCCAAGAATC
6-R	ATCTGCAGCTCGAGCTCGATGGATCCTCAAACTTTCAAAAGTTGCATTTTG
9-F	ACGACGTACCAGATTACGCTCATATGGCTAAGTTTGATGATCTTTATC
9-R	ATCTGCAGCTCGAGCTCGATGGATCCCTAGTGGCGGTGGTGCACGC
20-F	ACGACGTACCAGATTACGCTCATATGGCAACCCAAGCTTTGGTTTC
20-R	TATCTACGATTCATCTGCAGCTCGAGTCAGTGAAACTTGAGGCTAG
21-F	ACGACGTACCAGATTACGCTCATATGGATTCTGGGGGTTCAAAGAG
21-R	ATCTGCAGCTCGAGCTCGATGGATCCTTAAGCAGCTGCAACAGTGC
22-F	ACGACGTACCAGATTACGCTCATATGGCTGCTTCACTGAATCTCCC
22-R	ATCTGCAGCTCGAGCTCGATGGATCCTCAACTGCTTTGAAGAAACTTC
23-F	ACGACGTACCAGATTACGCTCATATGAACACGAACGAGCGCAGAGTTG
23-R	ATCTGCAGCTCGAGCTCGATGGATCCTCAATTCTCCTCTGGATGCTTC
24-F	ACGACGTACCAGATTACGCTCATATGAAGAAAGCAGTGTTGAAACTTG
24-R	ATCTGCAGCTCGAGCTCGATGGATCCTCAGCAAATAACACAAGCATTAG
25-F	ACGACGTACCAGATTACGCTCATATGGCTTCCTCAATGATCTCATC
25-R	ATCTGCAGCTCGAGCTCGATGGATCCCTAGACGCCTGGTGGCTTGG
27-F	ACGACGTACCAGATTACGCTCATATGGGTTCCCTTTGCATAAACGAAG
27-R	ATCTGCAGCTCGAGCTCGATGGATCCTCAAAGTAACACATCTGGCTC
29-F	ACGACGTACCAGATTACGCTCATATGGCTTCAACTCAATGCTTCTTG
29-R	ATCTGCAGCTCGAGCTCGATGGATCCTCAAGCAACACTGAAAGAA

Screening of the Host Factors Interacting with CP of Citrus Yellow Vein Clearing Virus by Yeast Two-Hybrid System

RichHTML

PDF

Abstract

Cite this article

share this article

Figures/Tables 8

References 34

Related Articles 15

Metrics

Comments

Recommended 0

[1]	ZHENG YaQin, LIU XueQing, WU SiWen, TANG XiaoYan, YANG DanNi, WANG YongKang, AHMAD Aftab, KHAN Afrsyab, WANG ChengGang, CHEN GuoHu. Cloning and Expression of BcDET2 Gene and Functional of Its Regulatory Effect on Bolting and Flowering in Wucai (Brassica campestris L.) [J]. Scientia Agricultura Sinica, 2025, 58(5): 991-1003.
[2]	QI MengNan, ZHAO DingLing, ZHANG XueYan, ZHANG YuJie, WANG RongNa, LIU BingQiang, YAN Long, ZHANG Jie, WANG DongMei. Identification of GmSZFP-Interacting Proteins and Functional Analysis of GmERF7 in Soybean Resistance to SMV Infection [J]. Scientia Agricultura Sinica, 2025, 58(14): 2739-2750.
[3]	LI ChuXin, SONG ChenHu, ZHOU JinHuan, LI JiaXin, WANG XinLiang, TIAN XuBin, SONG Zhen. Research on Prevention and Control Technology of Citrus Yellow Vein Clearing Virus Based on VIGS [J]. Scientia Agricultura Sinica, 2024, 57(22): 4473-4482.
[4]	ZHU YanTing, DANG Hao, NIU SiJie, LIN JingYi, YANG Hua, YANG Qiang, ZHANG Chong, CAI TieCheng, ZHUANG WeiJian, CHEN Hua. Screening of Interaction Proteins with AhSAP1 in Peanut Using the Yeast Two-Hybrid System [J]. Scientia Agricultura Sinica, 2024, 57(21): 4376-4390.
[5]	HE Yong, FAN XiaoZhu, CHEN XinYue, DUAN ShuJing, HU TingTing, XIE RuXue, WANG YuQing, CHEN Jing. Screening and Verification of Pepper Host Factors Interacting with the 126 kDa Protein of Pepper Mild Mottle Virus by Yeast Two-Hybrid System [J]. Scientia Agricultura Sinica, 2024, 57(15): 2986-2996.
[6]	LI TianCong,ZHU Hang,WEI Ning,LONG Feng,WU JianYing,ZHANG Yan,DONG JinGao,SHEN Shen,HAO ZhiMin. The Expression Pattern and Interaction Analysis of the Homologues of Splicing Factor SC35 in Setosphaeria turcica [J]. Scientia Agricultura Sinica, 2021, 54(4): 733-743.
[7]	SUN YuChen,JIA RuiPu,FAN KuoHai,SUN Na,SUN YaoGui,SUN PanPan,LI HongQuan,YIN Wei. Detection of Interaction Between Porcine Type I Complement Receptor and C3b Active Fragment in Vitro [J]. Scientia Agricultura Sinica, 2021, 54(19): 4243-4254.
[8]	MA ZhiMin,XU JianJian,DUAN Yu,WANG ChunQing,SU Yue,ZHANG Qi,BIN Yu,ZHOU ChangYong,SONG Zhen. Establishment of RT-RPA for Citrus Yellow Vein Clearing Virus (CYVCV) Detection [J]. Scientia Agricultura Sinica, 2021, 54(15): 3241-3249.
[9]	YANG HongKai,YANG JingWen,SHEN JianGuo,CAI Wei,GAO FangLuan. Multi-Gene-Based PCR Detection and Identification of Chilli veinal mottle virus [J]. Scientia Agricultura Sinica, 2020, 53(16): 3412-3420.
[10]	ZHANG HuiYuan,LIU YongWei,YANG JunFeng,ZHANG ShuangXi,YU TaiFei,CHEN Jun,CHEN Ming,ZHOU YongBin,MA YouZhi,XU ZhaoShi,FU JinDong. Identification and Analysis of Salt Tolerance of Wheat Transcription Factor TaWRKY33 Protein [J]. Scientia Agricultura Sinica, 2018, 51(24): 4591-4602.
[11]	XU HaiFeng,YANG GuanXian,ZHANG Jing,ZOU Qi,WANG YiCheng,QU ChangZhi,JIANG ShengHui,WANG Nan,CHEN XueSen. Molecular Mechanism of Apple MdWRKY18 and MdWRKY40 Participating in Salt Stress [J]. Scientia Agricultura Sinica, 2018, 51(23): 4514-4521.
[12]	WANG YuKui,BAI XiaoJing,LIAN XiaoPing,ZHANG HeCui,LUO ShaoLan,PU Min,ZUO TongHong,LIU QianYing,ZHU LiQuan. Cloning and Expression Analysis of BoSPx in Brassica oleracea [J]. Scientia Agricultura Sinica, 2018, 51(22): 4328-4338.
[13]	ZHAO QingQing, LI JunPing, LIANG LiBin, HUANG ShanYu, ZHOU ChenChen, ZHAO YuHui, WANG Qian, ZHOU Yuan, JIANG Li, CHEN HuaLan, LI ChengJun. Interaction between Influenza Virus PA Protein and Host Protein PCBP1 [J]. Scientia Agricultura Sinica, 2018, 51(17): 3389-3396.
[14]	LI Shuai, JIANG XiZi, LIANG WeiFang, CHEN SiHan, ZHANG XiangXiang, ZUO DengPan, HU YaHui, JIANG Tong. Screening of the Host Factors of Woodland Strawberry Interacting with P6 of Strawberry vein banding virus by Yeast Two-Hybrid System [J]. Scientia Agricultura Sinica, 2017, 50(18): 3519-3528.
[15]	RU JingNa, YU TaiFei, CHEN Jun, CHEN Ming, ZHOU YongBin, MA YouZhi, XU ZhaoShi, MIN DongHong. Response of Wheat Zinc-Finger Transcription Factor TaDi19A to Cold and Its Screening of Interacting Proteins [J]. Scientia Agricultura Sinica, 2017, 50(13): 2411-2422.