Transcriptome Sequencing And Analysis Of Zoysia Japonica Under The Mechanical Wounding Treatment

Using high-throughput sequencing technologies, the transcriptome of zoysia japonica wounded leaves was analyzed in order to research the molecular mechanism of mechanical wounding. Through the research, the main results were as followed:1.According to the PLD gene expression pattern and the results of Hydrogen peroxide content and cell membrane permeability, the stress treatment at Oh(T7),2h(T8) and 6h(T9) have the significant differences under the stress responsing trends. We confirm this moment as the condition of sequence and verification test.2.Through the sequence,15.9Gb data were obtained. After the assembled,74,385 unigenes were acquired using the Trinity software. In the T7vsT8 group,we screened 3,299 DEGs, including 2,374 up-regulated and 925 down-regulated genes. In the T7vsT9 group,3,317 DEGs were screened, including 2,433 up-and 884 down-regulated genes.3.Totally,38,084 unigenes were annotated, including 14,632 unigenes annotated in COG database, 27,857 unigenes in GO database and 7,201 unigenes in KEGG database. Among the annotated unigenes, they were classified into some important signal transduction and redox reaction mechanism pathway referring to replication, recombination and repair of gene and oxidative phosphorization and so on.4. Through analyzing the difference of gene function classification, we focused on screening of the genes related to hydrogen peroxide metabolic pathways. According to the result of screening, we speculated that the peroxidase(POD) may be more important in plants responsing to mechanical stress of antioxidant enzymes. We screened 13 unigenes that were annotated to POD. According to the different expression patterns, we speculated the POD isomers may have three functions:one is the POD isomers (2/70/56/59/62) Related to generate hydrogen peroxide; another is the POD 19 whose function is scavenging hydrogen peroxide; the third is POD 1, which may be complicit in the production and scavernging of hydrogen peroxide.5. By real-time PCR, we successfully verified the five genes expressed at T7, T8 and T9 (genes ID are c45867.graph_cO, c54560.graph_cO、 c38735.graph_cO、 c4611O.graph_cO and c49440.graph_cO) including three up-regulated genes and two down-regulated genes.