Transcriptome Sequencing Of Cistanche Deserticola And Host Haloxylon Ammodendron And Preliminary Analysis On Transfer RNA

Rou Congrong(Cistanche deserticola Y.C.Ma)belongs to the genus Cistanche Orobanchaceae and is a perennial holoparasitic herbaceous plants.Cistanche deserticola is an obligate parasitic plant grown on the roots of Haloxylon ammodendron,a small arbor in Chenopodiaceae family.Cistanche deserticola is known as "desert ginseng" for its high medicinal value.Haloxylon ammodendron is an important drought sand fixing plant and plays an important role in the ecological environment in the arid area.To date,the research on Cistanche deserticola and Haloxylon ammodendron only focus on identification of its medicinal value the separation of bioactive components.Wheheas,study on its genetic material and the molecular biology research is rarely reported.The main goal of present study is to enrich genetic information of Cistanche deserticola and Haloxylon ammodendron through transcriptome sequencing and to reveal the possible RNA transfer events between them.Transcriptome sequencing was performed on Cistanche deserticola and Haloxylon ammodendron using Illumina high-throughput sequencing platform,as enriched the sequence data of these two species and laid foundation for future molecular biology research.At the same time,the putative RNA transfer event betwwn these two species was revealed by bioinformatics of transcriptome sequencing results and verification by molecular detection.The main results are as follows:1.Illumina high-throughput sequencing platform produced more than 750,000,000 raw reads and 723,000,000 clean reads.After assembly,Unigenes of 200?500 bp were 130,000,Unigenes of 500?1,000 bp were 30,000,Unigenes of 1,000?2,000 bp were 25,000,Unigenes of more than 2,000 bp were 17,000,totalling up to 222,899 Unigenes.Among these Unigenes,95,123 were from Haloxylon ammodendron,108,575 were from Haloxylon ammodendron parasitized with Cistanche deserticola,85,131 were from Cistanche deserticola,and 56,084 were from haustoria.2.? 16,860 Unigenes were assumed to transfer from Haloxylon ammodendron to Cistanche deserticola,2,712 Unigenes were putatively transferred from Cistanche deserticola to Haloxylon ammodendron by the analysis of Perl language program.The selected Unigenes have very important research value as they may also be induced by the parasitization of Cistanche deserticola.? The use of Python language program analysis in the National Center for Biotechnology Information(NCBI)database indicated that 5,016 Unigenes were assumed to transfer from Haloxylon ammodendron to Cistanche deserticola,and 454 Unigenes were putatively transferred from Cistanche deserticola to Haloxylon ammodendron.? Local blast comparison analysis of transcriptome sequences with 11 Chenopodiaceae databases and 7 Orobanchaceae databases found that 33 Unigenes were transferred from Haloxylon ammodendron to Cistanche deserticola,as they were highly homologous to Chenopodiaceae sequences,but no similar sequences were found in Orobanchaceae databases.Fifteen Unigenes were transferred from Cistanche deserticola to Haloxylon ammodendron,as they were highly homologous to Orobanchaceae sequences,but no similar sequences were found in Chenopodiaceae databases.3.PCR test with different templates and subsequent DNA sequencing indicated that 4 Unigenes were transferred from Cistanche deserticola to Haloxylon ammodendron,further supporting the mRNA transfer events between these twe species.