Function Of The MiR482b/c In Tomato For The Resistance To Late Blight

As a commercial crop around the world,tomato is one of the most universal planted freshproduct.Where as,its product always be severely affected by the late blight.The research about the functional mechanism between tomato and bacterial is important for prevention and control of late blight.mi RNA regulated nectwork was built which is related to the tomato disease in the interaction between tomato and bacterial,intergrate the QRT-PCR analysis,found that mi R482 play an important role in this process.In order to study function of mi R482,we bulid the stable expression plant.The first step is bioinformatic analysis.Firstly,the sequence information of mi R482 family members from the mi Rbase database was obtained and blasted them using Clustal X,mi R482b/c located on the same chromosome and had the highest sequence similarity.After that,target genes of mi R482 family members from ps RNATarget was got and built regulatory network between them,found that the target were mostly NBS-LRR family disease resistance genes,which accounted for about 3/4 of the total amount of disease resistance genes in plants.Besides this,mi R482 b and mi R482 c shared more common target genes.In view of the above analysis,study target mi R482 b and mi R482 c was determined.Further more,the mi R482b/c was cloned from Zao Fen 2 tomato and overexpression vector and CRISPR/Cas9 targetd vectors was constrated.In the transient assays,after the infection of Phytophthora infestans,from the relative lesion area statistical results,tomato leaves had lager relative lesion area which was treated using the engineering Agrobacterium tumefaciens with over-expression vector,the relative expression of mi R482 was significantly increased and the relative expression of disease resistance genes NBS-LRR decreased.This results of targeted vector is contrary to that.This results preliminarily proved that mi R482b/c can regulate the NBS-LRR family to resist the outbreak of late blight.To further study the function of mi R482 in tomatoes,the stable expression plants was built,the results is consistant to the transient assays,demonstrated the function of mi R482 in the tomato disease resistance.Finally,physiological and biochemical indexes detection of Phytophthora infestans treated transgenic plants was posessed.The cell death will be detected by trypan blue staining.The accumulation of H2O2 and O2-in the leaves will be tested by DAB and NBT staining.Thisserious of assays futher illustrate the function of mi R482 in the interaction between tomato and Phytophthora infestans.In summary,this research mainly used two technologys such as overexpression and CRISPR/Cas9 to edit mi R482,with the help of transient assays,stable trangenic plants and physiological and biochemical indexes detection to certify the function of mi R482 in tomatoes.the overexpression of mi R482 can reduce the instance to tomato late blight...