Isolation Of Laccase AcLac35 In Kiwifruit And Analysis Its Role In Defense To Bacterial Canker

Kiwifruit is an important fruit in China,especially rich in vitamin C.Kiwifruit bacterial canker of kiwifruit is cultivated mainly in the production process of the devastating bacterial disease,major damage leaves and branches,kiwifruit its pathogen Pseudomonas syringae pv.actinidiae(Psa).Pseudomonas syringae is one of the top ten plant pathogens.In the model species Arabidopsis thaliana,it was found that Pseudomonas syringae mainly harmed its leaves,and a large number of studies showed that the pathogens invaded the cells from the epiphytes through the stomata,and if the bacteria were transferred from the leaves,the damage to the branches was ultimately endangered..In plant cell walls,especially in secondary walls,lignin is a major component of its resistance to disease.Recent studies have found that plant laccase is involved in the polymerization of lignin monomers.The plant laccase gene is involved in the main pathway of lignin synthesis,as a major component of the cell wall,plays an important role in disease resistance and phenotype.To further investigate the role of kiwifruit laccase gene in lignin synthesis and anti-ulcer disease,this study by genome,transcriptome,structural analysis,gene cloning,tissue-specific expression,subcellular localization and tobacco leaves Transient expression to interpret the role of kiwifruit laccase gene in lignin synthesis and disease resistance.The main findings are as follows:1.By analyzing the genomic data of kiwifruit,combined with gene mapping,evolution analysis and gene structure analysis,a total of 39 kiwifruit laccase gene AcLacl-AcLac39 were obtained.2.Psa was inoculated into different varieties of kiwifruit leaves,and samples were taken after 24 hours of treatment.The transcriptome of the sample was sequenced and analyzed,and the transcriptome data was verified by qPCR,and finally the AcLac35 gene with different expression was screened out.3.The total lignin content of the kiwifruit samples with different symptoms was determined.The results showed that the total lignin content of the plants changed when the kiwifruit was infected with Psa,and the change was related to the disease resistance of the cultivars.4.Gene cloning analysis showed that AcLac35 gene had high tissue expression,and the expression level increased after inoculation treatment.At the same time,AcLac35 was identified as cell wall protein by transient subcellular localization analysis of tobacco.5.By transiently expressing the injected tobacco,the change of total lignin content in the 7d tobacco was observed,and the total lignin content of the recombinant vector Agrobacterium was increased,which confirmed that AcLac35 was involved in the synthesis of lignin.