| Drought,salinity,high temperature,low temperature,and various diseases and pests are the serious constraint to the yield and quality of crops.In order to avoid biotic and abiotic stresses,plants have developed elaborate mechanisms to perceive external signals and adjust metabolic pathways by modulating the expression of genes.Responses of plants to adversity stress are a complex process involving multiple genes,multiple signal transduction pathways and multiple gene expression products,which includes four stages: the awareness of plants to adversity stress signal,signal transmission,signal recognition and related gene expression(Pei et al.2012).Researches show that Lectin Receptor-like Protein Kinases(LecRLKs)is a kind of important protein family that contribute to mediating the perception and transduction of external environment signals,regulating the expression of downstream resistant genes,and triggering multiple physiological and biochemical reactions to minimize and eliminate hazards(Vaid et al.2013).As a kind of candidate gene related to resistance,a number of successful applications of LecRLKs in molecular breeding have been reported.Their members are huge in higher plants and they are believed to play indispensable roles in the growth and development process of plants.In this study,we cloned a lectin receptor-like protein kinase gene TaLecRLK1 gene which from transcriptional profiling of wheat under the drought stress treatment.The genes can be induced by drought stress.Through a series of experiments,including the analysis of expression characteristics,subcellular localization,validation of protein interactions,and resistance identification of transgenic Arabidopsis,we got a preliminary understanding of the expression features of TaLecRLK1 and the signal transduction pathways that it may be involved in.Through the above experiments,we drew the following conclusion:(1)The cloning and expression feature analysis of TaLecRLK1: In this article,we cloned TaLecRLK1 gene.TaLecRLK1 was located in the cytoplasm,cell nucleus and the cell membrane by analyzing for subcellular localization.We analyzed the characteristics of gene expression through real-time PCR technology.The results showed that TaLecRLK1 were induced to expression under the stresses of drought,high salt,low temperature,and hormone like ABA,SA,BR that associated with biological stress.The results indicated that TaLecRLK1 gene may participate in a variety of signal transduction pathways.(2)The resistance identification of TaLecRLK1 in transgenic Arabidopsis: We constructed a plant expression vector and translated the vector into Col-0 Arabidopsis through the method of inflorescence infection mediated by Agrobacterium.The seedlings of transgenic Arabidopsis were treated with different concentration of ABA and drought stresses.We analyzed the resistance of transgenic Arabidopsis through seeds germination rate and root length experiments.The results show that the TaLecRLK1 gene in Arabidopsis is sensitive to ABA.Under the ABA treatment,the germination of transgenic Arabidopsis seeds was restrained.The primary results indicated that stress-tolerance of TaLecRLK1 gene may depends on ABA signal transduction pathways.(3)The interaction of TaLecRLK1 and TaARP1 in wheat: In this study we found that in the process of TaLecRLK1 improve wheat resistance,another protein TaARP1 also participated in this process.We verified that Ta LecRLK1 and Ta ARP1 interact with each other through BiFC and GST-pull down assay.This result suggested that TaLecRLK1 and TaARP1 co-regulate a signaling pathway or the function of TaLecRLK1 gene can be performed depend on the interaction with other proteins. |
PDF Full Text Request