Study On Dormancy Release Mechanism Of Taxus Mairei (Lemee Et Levl.) S. Y. Hu Ex Liu Seeds

Objective To study the biological characteristics of the seeds of Taxus mairei(Lemee et Levl.)S.Y.Hu ex Liu,the changes of seeds morphology and physiological and biochemical characteristics before and after dormancy release.To select the best method of RNA extraction for transcriptome sequencing.The seeds of Taxus mairei(Lemee et Levl.)S.Y.Hu ex Liu were sequenced and excavated with dormancy and germination related genes.To explore the seed dormancy release mechanism of Taxus mairei(Lemee et Levl.)S.Y.Hu ex Liu.Methods ①The phenotypic traits,seed weight,seed clarity,water content,water absorption,seed vigor and germination rate of Taxus mairei(Lemee et Levl.)S.Y.Hu ex Liu were determined to study the biological characteristics.②The morphology and physiological and biochemical characteristics of the seeds of Taxus mairei(Lemee et Levl.)S.Y.Hu ex Liu were studied before and after dormancy release,including water content,soluble sugar content,starch content,protein content,crude fat content and amylase activity.③The total RNA was extracted from stems,leafs and seeds of Taxus mairei(Lemee et Levl.)S.Y.Hu ex Liu by Trizol method,CTAB method,hot phenol method,modified hot phenol method and kit method to select the best method of RNA extraction for transcriptome sequencing.④The sequences of Taxus mairei(Lemee et Levl.)S.Y.Hu ex Liu were sorted by Illumina HiseqTM2000 sequencing platform,and the sequencing results were analyzed to select the differentially expressed genes related to dormancy release.Results ①The seed weight of Taxus mairei(Lemee et Levl.)S.Y.Hu ex Liu was 63.90 g,the seed clarity was 99.42%,the water content was 10.94%,the viability was 88%,but the germination rate was 0.②After low temperature stratification treatment,water content increased significantly,starch and crude fat decreased,the soluble sugar increased obviously and the protein content increased but the change was not significant.③Trizol method,CTAB method and hot phenol method are not suitable for the extraction of total RNA from Taxus mairei(Lemee et Levl.)S.Y.Hu ex Liu,modified hot phenol method and kit method are applicable,but the kit method is more suitable for RNA extraction of Taxus mairei(Lemee et Levl.)S.Y.Hu ex Liu seeds.④A total of 49.90 Gb of valid data and 355.62 M of high quality Reads were obtained through the Illumina HiseqTM2000 sequencing platform,resulting in 64,291 unigenes,total length of 33,769,996 bp.There are 31,450 unigenes in the NR,COG,GO and KEGG database to get annotations.36 genes of material metabolism,57 genes of transcription factors,63 genes of hormone-regulated genes and 50 methionine metabolism related genes were screened out related to seed dormancy release.Conclusion Comprehensive analysis of morphological and physiological and biochemical changes and transcriptome sequencing results,excavate the relevant genes of Taxus mairei(Lemee et Levl.)S.Y.Hu ex Liu seeds dormancy release.Revealing the mechanism of Taxus mairei(Lemee et Levl.)S.Y.Hu ex Liu seeds dormancy release from the physiological level and the molecular biology level.