| The green leafhopper is the main pest in tea production,which affects the quality and yield of tea.In severe cases,it can cause a loss of up to 33% of tea production.In recent years,the genetic characteristics of cytoplasmic incompatibility(CI)that may be caused by the symbiotic bacterium Wolbachia have attracted more and more attention.In this study,we combined the research on the biological control of tea tree pests,based on Wolbachia,using PCR、RT-PCR detection and FISH technology to identify and distribute Wolbachia in the green leafhopper;and then treat the small green leafhoppers with different concentrations of tetracycline solution.The symbiotic bacteria Wolbachia in the green leafhoppers established a de-symbiotic population without Wolbachia;finally,the possible mechanism of the symbiotic bacteria Wolbachia on the small green leafhoppers was explored and analyzed from the perspective of biology and genetics.The main results are as follows:1.Using the universal primers of the Wolbachia wsp gene to identify the symbiotic bacteria Wolbachia in four populations of different geographical locations,the Wolbachia-containing bands were detected except for Pingtang.By sequencing its sequence,it was found that the wsp sequence size of the Fenggang sample was 565 bp,and the sample size of Jiu’an and Meitan was the same567 bp;from the constructed evolutionary tree,it can be seen that the population of Fenggang is closely related to the Eupristina verticillate;The populations of Meitan and Jiu’an are highly similar to Aedes albopictus.2.Quantitative detection of samples from different tissues of the small green leafhopper by RTPCR technology and FISH found that Wolbachia was detected in all four populations except for the Pingtang population,which was consistent with the detection results of PCR;the detection of different parts of the tissues showed that the main focus In the abdomen,then the chest,and finally the head;the detection of samples of different genders found that the Wolbachia in females was higher than that in males,and then as the age increased,the amount of Wolbachia in the green leafhoppers also increased gradually.3.Using different concentrations of tetracycline solution,and applying the hair and systemic methods to eliminate the symbiotic bacteria Wolbachia in the green leafhopper,the results show that the method and effect of applying hair to the symbiotic bacteria Wolbachia in the green leafhopper is the most effective.Preferably,another 5 mg/m L tetracycline solution can get a stable small green leafhopper from the symbiotic population on the 25 th day,and has little effect on the population’s fecundity.4.Through the study of the influence of the symbiotic bacterium Wolbachia on the reproductive development of the green leafhopper,the lifespan,reproduction parameters and development period of the de-symbiotic population and the natural population of the green leafhopper were compared,and it was found that after the symbiotic bacterium Wolbachia was eliminated,The reproductive development parameters of the green leafhopper increased,indicating that the existence of Wolbachia is beneficial to the control of the green leafhopper population,but the amount of Wolbachia in its body is too low to achieve the effect of natural population control.5.Research on the cytoplasmic incompatibility and feminization mechanism of Wolbachia from the perspective of biology and genetics.Using the pair-by-pair combination of the desymbiotic population and the natural population,it is found through the study of the number of offspring produced and the male and female individuals,The influence of the symbiotic bacteria Wolbachia on the population of the green leafhoppers is consistent with the characteristics of cytoplasmic incompatibility and feminization. |
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