| Empoasca vitis is one of the main pests in Camellia sinensis,which seriously affect the quality of tea,and Beauveria bassiana and Metarhizium anisopliae were the main pathogenic fungi for biological control,the paper screened strong virulence of B.bassiana and M.anisopliae strain Ma1775 of BLK against E.vitis.Their biological characteristics and the compatibility with botanical pesticides and the biological controlling experiments with them in forest on E.vitis were studied.Based on these,the transcriptome sequencing analysis of Ma1775 strain was studied.It may serve as a theoretical and application basis for biological control on E.vitis.1.The virulence of 8 strains M.anisopliae and 5 strains B.bassiana against E.vitis were determined by spraying.The results showed that various strains of M.anisopliae had certain virulence against E.vitis after inoculation with(1±0.5)×107spore·mL-1 for 12 days.Among these strains,BLK and Ma1775 exhibited the most effective virulence against E.vitis,with corrected mortality of(86.92±1.96)%and(76.92±1.72)%,the cadaver rate of(84.67±0.82)%and(76.67±1.55)%,the median lethal time(LT50)of 6.35 d and 6.76 d,The LC50 of BLK and Ma1775 strain to E.vitis on the 6th day were 9.539×106 spore·mL-1 and 4.156×107 spore·mL-1.A time-dose-mortality model was employed to estimate parameters of time and dose,which showed that BLK and Ma1775 had a high lethal effect on E.vitis during 3-5 d after treatment,so they has been proved to be a great potential application in biological control of E.vitis.2.The research on biological characteristics of the BLK and Ma1775,the results showed that dextrose was the best carbon source for the most spore germination rate of two strains,peptone or beef extraction was the most ideal nitrogen for BLK strain and peptone was the best nitrogen for Ma1775 strain among 5 kinds of carbon and nitrogen source.The optimum temperature for the spore germination of two strains was 25? to 28?,while the optimal humidity was more than 90%.PPDA medium was the most suitable for their mycelia growth and conidia production among 5 kinds of commonly used medium,and then was PDA medium.3.The spore germination rate of BLK and Ma1775 were studied after compositing stabilizer with 7 kinds of botanical pesticides,the results showed that the spore germination rate of two strains was inhibited variably by botanical pesticides,BLK and Ma1775 strains respectively with 1.5%Pyrethrum EC and 3%Avermectin EC compatibility were better.Using the method of scattering powders,evaluated the effect of B.bassiana and M.anisopliae powders and their mixtures on E.vitis.Results showed that:after scattering powders 15 days,the control effects of BLK with Pyrethrum EC were best,which the decrease rate and control effect of Empoasca vitis were respectively(76.68±0.96)%and(73.90±1.04)%significantly above others.They can be promoted in production application.4.Differential transcriptome response of M.anisopliae Ma1775 strain on different medium(SDB medium and E.vitis powder medium)with different period have been analyzed.Results showed that four samples matched sequencing requirements,four compared-groups had diverse differences expression genes(DEGs).The GO function and pathway significant enrichment analysis of DEGs showed that only CK-MG VS Treat-MG in "biological process" category,had 2 terms significantly enriched and 1 term in"molecular function" category,and had varying amounts of significant concentration pathway in four compared-groups.Quantitative four samples with RPKM screened out 175 DEGs in common.These DEGs' cluster analysis were done that there were 11 genes which the strain of Ma1775 strain gene expression on insect powder medium was higher than that on SDB medium,with 6 fungal pathogenicity related genes. |
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