The Effects Of Transcription And MiRNA About PaWB20 Of Paulownia Witches' Broom Phytoplasma In Populus Trichocarpa

Phytoplasma is a prokaryote without cell wall.It can cause more than 1000 plant diseases in98 families in the world.However,the pathogenic mechanism of phytoplasma is not clear.At present,the research on the pathogenic mechanism of phytoplasma effector protein to plants has become a research hot spot.In order to explore the pathogenic mechanism of phytoplasma,our team have completed the sequencing and assembly the genome of PaWB-phytoplasma completed to drew the map,and used the bioinformatic softwares to predict and analyze the whole genome,and found 73 effect proteins including PaWB20(Tengu like),which has the 100% homology ratio with the TENGU from Onion Yellow strain M(OY-M).Previous experimental studies showed that Tengu transgenic tobacco and Arabidopsis can make the plants produce arbuscular and hydrolyze them into several small peptide segments in vivo,but the molecular mechanism of arbuscular is not clear.In order to further understand the pathogenic mechanism of the effector protein of Paulownia witches' broom(PaWB)and accelerate the research progress of the interaction between Paulownia and PaWB-phytoplasma,we used wild-type Populus trichocarpa and PaWB20 transgenic Populus trichocarpa as experimental materials,and used Illumina Nova SEQ technology platform to sequence and analyze the transcriptome and small RNA,and obtained the corresponding transcriptome and miRNA data.The main results are as follows:1.The transgenic PaWB20 positive plants of Populus trichocarpa were obtained.Agrobacterium mediated method was used to transform the effect protein PaWB20 of Paulownia into Populus trichocarpa.The resistant seedlings were detected by PCR and the results were positive.Compared with the wild-type Populus trichocarpa,the phenotype of the transgenic Populus trichocarpa showed a significant increase in branching.2.The transcriptome expression profile of PaWB20 gene positive transgenic plants of Populus trichocarpa was constructed.High throughput sequencing of Illumina yielded 54.27 Gb of clean data.The clean data of all samples reached 7.63 Gb,the content of G+C was about 45%,the percentage of Q30 base was over 93.22%,and the comparison rate of reads from all samples to genome was between 92.28% and 96.28%.A total of 41933 transcripts and 873 new genes were identified in the two libraries.There were 6248 differential transcripts,2981 of which were up-regulated and 3267 down-regulated.After screening and identification,13 differential expressed transcripts related to branches were identified.3.The miRNA expression profile of the transgenic plants of Populus trichocarpa was constructed.A total of 123.34 M raw reads and 70.31 M clean reads were obtained by high-throughput sequencing of Illumina,and the Q30 in each library was over 96.84%,indicating that the sequencing data was of good quality.521 miRNAs were identified in the two s RNA libraries,including 369 known miRNAs,152 new miRNAs,and 8061 predicted miRNA target genes.106 miRNAs were differentially expressed,of which 60 were up-regulated and 46 were down-regulated.Four differentially expressed miRNAs related to branches were identified.4.Four branch related miRNAs and their target genes were identified.These miRNAs are related to plant-pathogen interaction and plant hormone signal transduction.