Construction of Tomato (Solanum lycopersicum) Leaf cDNA Library Under Ralstonia solanacearum Stress and Screening and Identification of SlMYB86-like Interacting Proteins
CHEN Na1, WEN Yi-Jun2, SHAO Qin1,*
1 College of Life Science and Resources and Environment, Yichun University, Yichun 336000, China; 2 Yichun Academy of Sciences (Jiangxi Rich-Selenium Industry Research Institute), Yichun 336000, China;
Abstract：The MYB transcription factor is one of the largest transcription factor families in plants and plays a significant role in the response of plants to both biotic and abiotic stresses. Previous research has indicated that the tomato (Solanum lycopersicum) SlMYB86-like (Solyc06g071690) transcription factor can be induced by the Ralstonia solanacearum and participates in the resistance response process of bacterial wilt. Nonetheless, the regulatory mechanism remains to be elucidaded. To screen the protein that interacted with the MYB transcription factor SlMYB86-like in tomato under the infection with R. solanacearum, the tomato inbred line BY 1-2 (susceptible to bacterial wilt) was used as the material. Total RNA was extracted from tomato leaves at 0, 3, 6, 9 and 24 h post-inoculation with R. solanacearum and subsequently mixed equally to create a tomato cDNA yeast two-hybrid library. The library capacity and recombination efficiency were calculated. Meanwhile, the constructed library was utilized to explore the interaction proteins during the infection process of R. solanacearum by using SlMYB86-like as the bait protein, and the potential proteins that might be involved in the defense response to pathogen were verified for interaction in yeast. The yeast library was constructed with a capacity of 1.9×107 CFU and showed complete recombination, corresponding to a 100% recombination rate. Furthermore, the average insert size was confirmed to be over 1 000 bp. Screening of the yeast two-hybrid system identified a total of 44 potential proteins that interacted with SlMYB86-like. Further verification through yeast rotation showed that SlMYB86-like interacted with the transcription factors vascular plant one-zinc finger 1 (VOZ1), teosinte branched 1, cycloidea, and proliferating cell factors 15 (TCP15), E3 ubiquitin protein ligase ring finger protein 14 (RNF14), serine/threonine protein kinase D6 protein kinase (D6PK), ethylene-responsive transcription factor 1 (ERF1), and calcium-dependent protein kinases (CDPK). In conclusion, these results offer significant experimental evidence for further investigation into the molecular mechanisms underlying SlMYB86-like's resistance to bacterial wilt in tomato.
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