Research On The Change Of Micro Rna In The Paulownia Witches Seedlings Treated With Dimethyl Sulphonate

PaWB is a destructive plant disease caused by phytoplasma.Paulownia Witches Broom(Pa WB)causing great losses to the economic development of Paulownia industry and alsoseriously affect the enthusiasm of paulownia growers.Paulownia witches' is one of the problems needs to solve in the forestry production in China.In the past 40 years,the science researchers of paulownia witches' and planting are studied in two aspects about the disease.The science researchers have studied a lot about Pa WB of paulownia.but the molecular mechanism of Pa WB occurrence is still unclear so far.In this study,healthy seedlings,Pa WB phytoplasma-infected seedlings and Pa WB phytoplasma-infected seedlings treated with DMS of P.fortunei,P.tomentosa and P.fortunei × P.tomentosa were used to construct nine small RNA libraries.The differential expression profiles of miRNAs after phytoplasma infection had been analyzed.The main results of this study are as follows:(1)Nines RNA librarys of three kindspaulownia were sequencing,each samples have nearly 10 million raw reads sequences.After using bioinformatics software analysis,then get Valid reads(PF)11378626,(PFI)10311261,(PFI-D15)7447741,(PT)7283265,(PTI)8660319,(PTI-D15)6784031,(PTF)10376326,(PTFI)6791333,and(PTFI D15)7698718 paulownia genome sequences.(2)Nine paulownia genomic librarys have been sequenced with Illumina,made s RNA sequences alignment to the paulownia genome,systematically compares the amounts and types of each type s RNA series,then got 519 miRNAs.In P.fortunei,a total of 328 miRNAs were identified in PF,PFI and PFI-D15 libraries including 206 conserved miRNAsand 122 novel miRNAs.In P.tomentosa,a total of 451 miRNAs were identified in PT,PTI and PTI-D15 libraries including 295 known miRNAs and 156 novel miRNAs.In P.fortunei × P.tomentosa,a total of 431 miRNAs were identified in PTF,PTFI and PTFI-D15 libraries including 291 known miRNAs and 140 novel miRNAs.(3)We have indentified 27 differential expressed miRNAsin P.fortunei,P.tomentosa and P.fortunei × P.tomentosa by comparing scheme(23 conserved miRNAs and 4 novel miRNAs).And we have 56 differential expressed miRNAsin P.fortuneand P.tomentosa(46 conserved miRNAs and 10 novel miRNAs).There are different number miRNAs in fortunei × P.tomentosa with their parents plants.51 miRNAs have been indentified in fortunei × P.tomentosa and P.fortune(40 conserved miRNAs and 11 novel miRNAs).106 miRNAs have been indentified in P.tomentosa and P.fortune(85 conserved miRNAs and21 novel miRNAs).By comparing the differences between the three kinds of Paulownia mi RNA target genes function,we have found 27 miRNAs play an important role in the Pa WB infected paulownia process.(4)Degradome analysis were used to determine the target genes of differential expressed miRNAs.We have found some miRNAs with Pa WB infected.They are involved in Biosynthesis of other secondary metabolites?Metabolism of other amino acids?Cell growth and death?Immune system? Xenobiotics biodegradation and metabolismand other metabolic pathways.(5)Seven miRNAs and four target genes were selected randomly for q RT-PCR validation.The results showed that the expression patterns obtained from q RT-PCR data were obviously consistent with those obtained from Illumina sequencing analysis,indicating the reliability of the RNA-seq analysis.