Study On Cloning And Expression Analysis Of SERK Gene Family In Taraxacum Antungense Kitag.

Somatic embryogenesis receptor-like kinases(SERKs)are involved in multiple plant signaling pathways and are associated with apomixis,biotic stress,and abiotic stress.The amino acid sequences of SERK protein family are highly homologous,but its function is specific.The molecular mechanism is still not clear.The SERK genes are important genes related apomixs,and there is no report of SERK gene in dandelion.It is also unknownwhich pathway SERK genes may participate in dandelions.Therefore,this study mainly cloned SERK family genes from apomixis dandelion,determined the identity of SERK family genes throμgh bioinformatics methods,and analyzed the expression of SERK family genes in apomixis dandelion tissue by real-time quantitative PCR method,and according to salt Treatment of SERK gene expression under the treatment of powdery mildew,thereby speculating that SERK family genes may be involved in signal pathways in dandelion.In this study,a Da SERK3 B overexpression vector was constructed and genetic transformation was performed to provide the material basis for functional verification of SERK family genes in dandelion.The results are as follows:1.For the first time,SERK genes were cloned from the genus Taraxacum,and four genes were cloned.The ORFs were 1884bp,1923bp,1842bp,and 1872bp respectively.They were named Da SERK1,Da SERK2,Da SERK3A,and Da SERK3B by amino acid sequence comparison.Compared with the previous SERK family gene evolution analysis,the SERK family genes reported in recent years and the genes cloned in this study have been added,and 49 SERK genes from different species have been synthesized to further study the phylogenetic relationships of SERK genes.2.In the tissue-specific analysis,the expression levels of Da SERK1 and Da SERK2 in flowers were higher than those in other tissues,and the differences were extremely significant.The expression levels of Da SERK3A and Da SERK3B in roots were significantly higher than those in other tissues.3.Subcellular localization and domain analysis showed that Da SERK3B was expressed on the cell membrane,which was the same as the location of the SERK gene in other plants previously studied,and further confirmed the identity of Da SERK3B.4.Construction of 35S::Da SERK3B::GFP overexpression vector,and genetic transformation,positive transgenic plants were identified by PCR.The relative expression level of Da SERK3B gene in positive plants was analyzed by real-time quantitative PCR.The expression level of Da SERK3B in transgenic plants was significantly higher than that in wild-type and negative controls,and overexpression of Da SERK3B transgenic plants was successfully obtained.5.The SERK family genes in dandelion responded to biotic stress and abiotic stress.Under powdery mildew treatment,powdery mildew infection increased,Da SERK1 and Da SERK2 expression increased,while Da SERK3A and Da SERK3B expression decreased.