Identification And Expression Analysis Of BURP Gene Family In Wheat

BURP protein family is ubiquitous in plants and unique to the plant kingdom.It is involved in a variety of biological processes such as seed and anther development,and plays an important role in plant response to stress.Although the systematic analysis of the BURP gene family and related functional genes have been studied in many plant species,such as rice and corn,it has been no report in wheat.Therefore,the wheat BURP family members of protein structure,evolutionary relationships,abiotic stress and tissue specific expression were analysised in this study,we found that TaBURP14B is specific expression in anther tapetum,and reduced expression under male condition.Using RANi interference technology to silence the TaBURP14B homologous gene OsBURP4 genes in rice,found that it is male sterile.So,TaBURP14B is likely to be an important gene involved in the regulation signal pathway of light-temperature sensitive male sterility in wheat,which also provides a new way to further analyze the molecular mechanism of light-temperature sensitive male sterility in wheat.1.54 TaBURPs gene family members of wheat were identified by bioinformatics analysis.The TaBURP gene family members of wheat had different structures,and the gene location showed that all chromosomes except chromosome 2A and 4B were distributed.Evolutionary analysis showed that TaBURP gene family could be divided into 5 subfamilies in wheat.Structural analysis shows that there are differences between subfamilies beyond the structural domain,but there are similarities among the members of each subfamily.2.The expression of wheat BURP gene family members were analyzed under drought,salt,cold and ABA stress using RT-PCR.The results showed that nearly half of the gene family members have at least one kind of stress response.Among these genes,TaBURP4B-1,TaBURP5A,TaBURP6D,TaBURP7A and TaBURP20 is response to drought,salt,cold and ABA stress.These results showed the wheat BURP gene families involved in ABA-medicated abiotic stress signal regulation pathway.3.Using public microarray data and transcriptome data genes,the tissue specificity of TaBURP gene family revealed that TaBURP11A/B/D,TaBURP14B/D and TaBURP15B/D were specifically expressed in anther.RT-PCR analysis showed that TaBURP8D,TaBURP1IA/B/D,TaBURP14B/D,TaBURP15B and TaBURP25 were highly expressed in anthers,and these genes may be involved in the development of wheat stamens.4.RT-PCR was used to analyze the expression patterns of TaBURP8D,TaBURP11A/B/D,TaBURP 14B/D,TaBURP 15B and TaBURP25 in stamens of BS366 and JING411 in fertile and sterile environments during the continuous pollen development of microspore mother cell stage,meiotic stage and microspore stage.The results showed that TaBURP14B expression in sterile environment was nearly 30 times less than that in fertile environment,while the expression in sterile and fertile environment of common wheat was basically unchanged.Furthermore,in situ hybridization analysis showed that TaBURP14B was specifically expressed in wheat anther tapetal layer and significantly decreased in sterile line BS366.These results showed TaBURP14B may be involved in the light-temperature sensitive male sterility signaling pathway in wheat.5.In order to further confirm whether TaBURP 14B is involved in the male sterility process of BS366,the expression vector of RNAi-OsBURP4(TaBURP 14B homologous gene)was constructed and transformed into rice,and the results showed that pollen abortion was carried out in transgenic rice.The above results indicated that TaBURP 14B may play an important role in the development of wheat stamens.