Mining And Bioinformatics Analysis Of Genes Related To Male Sterility In Armeniaca Sibirica

Armeniaca sibirica is an excellent ecological and economic tree species in the arid and semi-arid areas of northern China.However,there are still many problems in the development of the Armeniaca sibirica industry,such as low and unstable yield,early flowering and susceptibility to late frost damage,decline in economic traits,and poor genetic quality.Among them,the normal development of stamens is an important factor affecting the production of Armeniaca sibirica.Based on cytological observation,transcriptome sequencing and TA cloning,Armeniaca sibirica clones were used as experimental materials to study the critical period and causes of male sterility in Armeniaca sibirica,and screen the relevant genes of male sterility at the same time in this paper.The paper aimed to further increase the understanding and knowledge of the mechanism of male sterility in Armeniaca sibirica,and at the same time provide a theoretical basis for the use of genetic engineering technology to create high and stable yield varieties.The key regulatory genes that affected the male sterility of Armeniaca sibirica were screened simultaneously.The summarized results were as follows:(1)Compared with fertile flowers(FFs),the inner pollensac wall of male sterile flowers(MSFs)in Armeniaca sibirica showed no signs of degeneration in microspore stage by the observation of morphological characteristics and microstructure.When the pollen matured,the tapetum and middle layer cells were degraded insufficiently and the development of anthers stopped.After flowering,it appeared shriveled and wilted,and the pollensac couldn‘t be dehisced normally.Therefore,it was determined that the microspore stage was the critical period of anther abortion,and the pollen mature period was the post-abortion period.The analysis of relevant physiological indicators showed that during the critical period of anther abortion,the content of soluble sugar,starch and soluble protein of MSFs in Armeniaca sibirica were significantly lower than that of FFs.In the post-abortion period,the content of soluble sugar and soluble proteinof MSFs was significantly lower than that of FFs.However,the starch content was significantly higher than that of FFs.(2)Through the comparative analysis the transcriptome data between MFS and FFs of Armeniaca sibirica during microspore stage,a total of 4108 differentially expressed genes were obtained,of which 1899 were up-regulated in MSFs and 2209 was down-regulated.A total of 5 genes related to male sterility in Armeniaca sibirica were discovered which included AsMS1(Cluster-10585.0)、AsMS2(Cluster-6671.20011)、AsSUS1(Cluster-6671.6372)、As PTRE1(Cluster-6671.3208)and AsAMY1(Cluster-2681.0)by the combination of BLAST comparative analysis,function annotation,GO function enrichment analysis and KEGG pathway enrichment analysis.q RT-PCR verification results showed that the accuracy of transcriptome sequencing results was high,and could be used to analyze the dynamic changes of genes related to male sterility.(3)AsMS1、AsMS2、AsSUS1、AsPTRE1 and AsAMY1 related to male sterility in Armeniaca sibirica were selected for TA cloning and sequence analysis.The results showed that AsMS1 and the MS1 gene in Arabidopsis thaliana clustered together,and belonged to the PHD-finger family(zinc finger protein).AsMS2 and AsSUS1 were grouped together separately,and belonged to the FAR(fatty acyl-Co A reductase)and SUS(sucrose synthase)subfamily respectively.As PTRE1 had the highest similarity with PTRE1 in Arabidopsis thaliana,and clustered into one category which belonged to the P131＿Prot＿N superfamily(Proteasome regulator).AsAMY1 and the AMY1 gene in Arabidopsis thaliana clustered together,and belonged to AMY subfamily(α-amylase).