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| Construction of Yeast Two-Hybrid Library and Screening the Host Targets of the Effector SDE34 from Candidatus Liberibacter asiaticus |
| HU Yan-An1, ZENG Li-Hua1, LI Zhu1, LI Feng-Yao1, HUANG Yu-Lin1, LI Rui-Min1,2, HUANG Gui-Yan1,2,* |
1 College of Life Science, Gannan Normal University, Ganzhou 341000, China;
2 National Navel Orange Engineering Research Center, Ganzhou 341000, China |
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Abstract Citrus Huanglongbing (HLB) is one of the most devastating diseases in citrus industry, mainly caused by phloem-limited Candidatus Liberibacter asiaticus (CLas). SDE34 (CLIBASIA_04055) is one of the core sec-dependent effectors (SDEs) from CLas. In order to screen the host target of SDE34 during CLas infection, a yeast two-hybrid (Y2H) cDNA library was constructed using CLas infected Newhall navel oranges (Citrus sinensis) leaves and screened with SDE34 as a bait. The titer of the yeast two-hybrid library was 2.3×108 CFU/mL, and the length of the insert fragments ranges from 300 to 2 000 bp. Three citrus proteins interacting with SDE34 were identified, among which the screening frequency of heavy metal-associated isoprenylated plant protein (HIPP7) was 83.33%. Followed bimolecular fluorescence complementation (BiFC) assay confirmed that HIPP7 interacted with SDE34 in plant cells. Subcellular localization showed that HIPP7-GFP fusion protein localized in cytoplasm and nucleus in plant cells. And the expression level of HIPP7 was very significantly down-regulated (P<0.001) after CLas infection indicated by real-time quantitative PCR assay. HIPP7 homologous genes function as susceptibility factors in various plant-pathogen interaction systems, suggesting that SDE34 might promote CLas infection by targeting the citrus susceptibility gene HIPP7. This study provides a theoretical basis for analysis of the pathogenesis of CLas and citrus disease resistance breeding.
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Received: 06 January 2023
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Corresponding Authors:
*huangguiyan@gnnu.edu.cn
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