Screening And Identification Of Potential Selective Autophagy Receptors In The Process Of Puccinia Triticina

Wheat is one of the most significant food crops in the world.Wheat leaf rust disease caused by Puccinia triticina is widely distributed in the world,causing huge economic losses to the world.By revealing the physiological mechanism of wheat supplying nutrients to leaf rust,and then blocking the nutrient supply,the disease resistance can be achieved.This is a new way to explore starvation-mediated resistance(SMER)resources with both broad spectrum and persistent characteristics.Wheat can degrade its own nutrients through selective autophagy by specifically identifying autophagy receptors on the autophagosome membrane autophagy-related protein 8(TaATG8).This is a key link in wheat’s continuous supply of nutrients to leaf rust,but little is known about its degradation mechanism.In this project,the compatible leaf rust-wheat interaction as the research system,screening and identification of selective autophagy receptors in the nutrition supply process of puccinia triticina Through this project,the key points of wheat nutrition supply to the haustorium were screened to enrich the resources of SMER.It provides a basis for further searching for degradants and revealing the mechanism of selective autophagy degradation.It also provides a reference for the study on the pathogenic mechanism of other fungi such as Powdery Mildew and Magnaporthe oryzae.The research results are as follows:1.The TaATG8 subtypes that mediate the degradation of macromolecules are determined:the TaATG8 gene family is divided into 10 subtypes.The phylogenetic tree divides them into 3 subfamilies,and one of the representative genes TaATG8a/h/j of these 3 subfamilies are respectively selected for real-time quantitative detection.The results revealed that the expression level of TaATG8a/h showed an upregulated trend in the affinity combination composed of wheat variety TC and leaf rust race 260,and we used these two subtypes to study in subsequent experiments;2.Screening of receptor proteins that interact with TaATG8:construct pCM 1307-TaATG8a/h overexpression vector and transiently transform tobacco,dark induce leaf senescence and starvation,use Anti-HA purification magnetic beads to purify leaf protein samples,perform mass spectrometry after Western-Blot detection.In the final screening,10 proteins were obtained as potential interacting proteins of TaATG8a/h;3.Verify the interaction between TaATG8h and its receptor protein:mass spectrometry analysis obtains the potential interaction protein TaORM,using_Y2H,BiFC and Co-IP technology to verify their interaction,the results show that TaATG8h and TaORM interaction occurs.