Transcriptome Analysis And Development Of SSR Molecular Markers For Bidens Bipinnatal.under Cadmium Stress

Bidens bipinnata L.is a typical Cd superrich plant,which has a strong ability of Cd enrichment and is also an important medicinal plant with a long history of medicinal use and many efficacy.However,due to the difference of living environment,there are many morphological variations.It is difficult to distinguish the different species of nepenthes from each other,which seriously affects the drug safety of several main medicinal nepenthes.Gene expression differences between cadmium induced and underground root tissue transcriptome under normal conditions were compared by transcriptome sequencing.At the same time,SSR markers were developed based on transcriptome and data of transcriptome,so as to distinguish different plants of nepenthes,which is of guiding significance to solve the problem that nepenthes is difficult to distinguish.In this study,IlluminaHiSeq TM 2000 high-throughput sequencing platform was used to sequence the transcriptome of the root tissues of c.diffusa under cadmium stress.Trinity software was used to assemble the transcriptome data of oncogene,and a total of 120 and 122 unigenes were obtained.With the GO(Gene Ontology)?KO(Kyoto Encyclopedia of Genes and Genomes Ontology)? KOG(clusters of euKaryotic Orthologous Groups)? NR(RefSeq non-redundant proteins)?NT(Nucleotide Sequence Database)?PFAM(Protein Family Assembly)and Swiss ProtSeven databases were functionally annotated,and the GO enrichment analysis and KEGG enrichment analysis of differentially expressed genes were performed.The results showed that in the GO enrichment analysis,the differentially expressed genes were mainly concentrated in the three pathways of REDOX process,cell wall and catalytic activity.In the KEGG enrichment analysis,the differentially expressed genes were concentrated in protein processing and phenylpropane biosynthesis in the endoplasmic reticulum.At the same time,using transcriptome results and MISA software,SSR molecular markers were developed.A total of 17 140 repeating primitives containingSSR were found.Primerswere designed by Primer3 and 50 pairs of primers were randomly selected for PCR amplification.All the 46 pairs of primers could amplify100~300 bp target bands,indicating that the primers were well designed.Among them,28 pairs had clear bands,good polymorphism and good repeatability.From the perspective of the potential of polymorphism,these SSR repeat sites obtained in this study are highly available.Experimental results of this study in ghost needle grass functional genes SSR markers laid a foundation for further development,this kind of tag the establishment of the ghost needle grass functional genes of medicinal plants in accelerating the development and utilization of resources,make the needle grass molecular marker type is rich,identification of species,comparative genomics,auxiliary selection and implement specific traits such as genetic mapping model is of great significance.