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CN-121991990-A - Function and application of virus-derived tyrosine phosphatase in male testis development of plutella xylostella

CN121991990ACN 121991990 ACN121991990 ACN 121991990ACN-121991990-A

Abstract

The invention discloses a function and application of virus source tyrosine phosphatase in the development of male testis of plutella xylostella, namely an application of a protein tyrosine phosphatase gene CvBV _22-9 coded by plutella xylostella plate cocoon bee virus in inducing apoptosis of sperm cells of plutella xylostella. The CvBV _22-9 gene is screened and identified as a main factor for inducing the apoptosis of the testis, and is continuously and highly expressed in the spermary tissue after parasitism and has tissue specificity. After CvBV _22-9 is knocked down, the apoptosis rate of the testis is obviously reduced, and about 90.80% of the apoptosis of the testis can be induced by using recombinant baculovirus to overexpress the gene. Low concentrations of recombinant baculovirus also disrupt the structure of the testis and lead to reduced oviposition in mating females. The invention successfully constructs a transgenic drosophila model, realizes specific expression CvBV _22-9 in drosophila testis by using a UAS-Gal4 system, and induces 97.30% of testis apoptosis.

Inventors

  • WANG ZHIZHI
  • CHEN XUEXIN
  • GAO HONGSHUAI
  • Guo Mujuan
  • WU KUN

Assignees

  • 浙江大学

Dates

Publication Date
20260508
Application Date
20260112

Claims (10)

  1. 1. An application of a protein tyrosine phosphatase gene CvBV _22-9 coded by plutella xylostella plate cocoon bee virus in inducing apoptosis of plutella xylostella larva spermary cells.
  2. 2. The use according to claim 1, wherein the nucleotide sequence of the CvBV _22-9 gene comprises the sequence shown as SEQ ID No. 3 or a functional fragment thereof.
  3. 3. The use according to claim 1, wherein the CvBV _22-9 gene encodes an amino acid sequence as shown in SEQ ID No. 4 or a functional fragment thereof.
  4. 4. The use according to claim 1, wherein said use is effected by expression of said CvBV _22-9 gene.
  5. 5. The use according to claim 4, wherein said expression is effected by infection with a baculovirus expression vector.
  6. 6. The method of claim 5, wherein the recombinant baculovirus NPV-CvBV _22-9 is synthesized and used to infect plutella xylostella larvae to induce apoptosis of male plutella xylostella testis cells.
  7. 7. The application of a plutella xylostella cocoon bee virus coded protein tyrosine phosphatase gene CvBV-22-9 in preparing a preparation for biological pest control is characterized in that the preparation realizes population control by inducing male sterility of pests.
  8. 8. The use of claim 7, wherein the pest comprises a lepidopteran or dipteran pest.
  9. 9. The use according to claim 8, wherein the lepidopteran pest is plutella xylostella and the dipteran pest is drosophila.
  10. 10. A model construction method for apoptosis induction function in insect testis is characterized by comprising the following steps of constructing CvBV _22-9 gene coding sequence in a vector for drosophila melanogaster transgene, and obtaining transgenic drosophila melanogaster through embryo microinjection; crossing the transgenic drosophila with a testis-specific driving strain by using a UAS-Gal4 system to obtain a drosophila strain specifically expressing CvBV _22-9 in the testis.

Description

Function and application of virus-derived tyrosine phosphatase in male testis development of plutella xylostella Technical Field The invention relates to the technical field of biological control of agricultural pests, provides a research system for systematically verifying the phenomenon of insect testis castration in a multi-dimensional manner, and particularly relates to a function of a protein tyrosine phosphatase gene CvBV _22-9 coded by plutella xylostella cocoon bee virus (CvBV) in inducing apoptosis of plutella xylostella larva testis cells and application of the gene in pest control. Background The plutella xylostella (Plutella xylostella) is a major lepidoptera pest which damages cruciferous vegetables worldwide, has strong reproductive capacity and short generation period, and is extremely easy to generate drug resistance to chemical pesticides. At present, the control of plutella xylostella still depends on chemical pesticides seriously, which causes a series of problems such as environmental pollution, pesticide residues, rampant pests and the like. Therefore, developing an efficient, green, novel prevention and control strategy has become an urgent need and a significant challenge in this field. Biological control by using parasitic wasps is an important component of green control. Parasitic bees parasitic the host and then influence the physiological life processes of the host plutella xylostella, in particular the immunosuppression and the growth and development. The phenomenon of parasitic castration (PARASITIC CASTRATION), namely a strategy that parasitic bees destroy the reproductive organs of hosts and steal the reproduction energy of the parasitic bees to be beneficial to the development of offspring, is a key mechanism for the parasitic bees to successfully parasitic. The results of the studies on how fan and the like show that the plutella xylostella plate cocoon bee Cotesiave stalis (Haliday) carrying Bracovirus PDV or the Diadegma semiclausum Hell en parasitic of the semi-closed curved tail plate moth with Ichvovirus PDV has obvious inhibition effect on the spermatogenesis and the formation process of the plutella xylostella with different insect ages (what fan, bai Sufen, li Xin, and the like, the influence of two parasitic bees on the spermary of the plutella xylostella with different insect ages [ J ]. Insect knowledge, 2010 (3): 7). The plutella xylostella wasps (Cotesia vestalis) is a dominant natural enemy of plutella xylostella, and when the plutella xylostella wasps spawn spawns, domesticated endogenous viruses carried by the plutella xylostella wasps virus (Cotesia vestalis bracvirus, cvBV) are injected into a host together, cvBV enters the host, and then the genome of the plutella xylostella wasps is integrated into the host genome and expresses a large number of toxic genes in the host, wherein the protein tyrosine phosphatase (Protein Tyrosine Phosphatase, PTP) gene family is the family with the largest number of members and plays an important role in the processes of regulating cell signal transduction, proliferation, apoptosis and the like. However, the specific function of the vast majority of CvBV-PTP genes, particularly in regulating host reproduction, has not been known in the prior art. Earlier studies have found that the parasite of plutella xylostella coco can cause atrophy of the spermary nest of its host plutella xylostella, cvBV is an important parasitic factor （Bai SF, Cai DZ, Li X, Chen XX. Parasitic castration of Plutella xylostella larvae induced by polydnaviruses and venom of Cotesia vestalis and Diadegma semiclausum. Arch Insect Biochem Physiol. 2009 Jan;70(1):30-43. doi: 10.1002/arch.20279. PMID: 18949808）, causing this phenomenon, however its inherent molecular mechanism is not yet known. And Yang Yanyan research shows that injection of CvBV at a certain concentration can prolong the development time of plutella xylostella larvae (Yang Yanyan. Influence of plutella xylostella larva growth and endocrine activity by plutella xylostella larva parasitism [ D ]. University of Zhejiang, 2010.). The invention aims to define the key regulatory genes of parasitic wasps causing parasitic castration, reveal CvBV a molecular mechanism of causing lepidoptera host testis castration, and develop pest control technology based on male sterility. Disclosure of Invention The invention confirms that the parasitic wasps cause male castration by inducing the apoptosis of the diamondback moth spermary through the change of the size of the spermary, TUNEL apoptosis staining and other aspects. Further, the invention screens a key toxic gene CvBV _22-9 for inducing apoptosis of the spermary of plutella xylostella, utilizes various technologies such as virus particle infection, RNA interference and the like to identify the important regulation and control effect on the castration of the spermary, and verifies that the function of the carrier has conservation in model insect Dro