miR-286 inhibitor related to reproductive development of bactrocera dorsalis and application of miR-286 inhibitor

A technology of mir-286 and B. citrifolia, which is applied in the field of pest biological control and genetic engineering, can solve the problems of reduced hatching rate and unclear function, and achieves the reduction of single female egg production, reduction of fecundity, and total egg production. volume reduction effect

Inactive Publication Date: 2020-09-25
HUAZHONG AGRI UNIV
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  • Abstract
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  • Application Information

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Problems solved by technology

[0005] In a previous study, miR-286 was found to be highly expressed in root-knot nematode eggs (Subramanian et al2016); miR-286a and miR-286b were involved in maintaining viral replication and antiviral responses during persistent dengue virus infection in mosquitoes. (Avila-Bonilla et al 2017); depletion of miR-286b in Aedes albopictus resulted in reduced hatchability, suggesting a role in embryogenesis (Puthiyakunnon et al 2013), whereas its role in reproductive development function is unclear

Method used

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  • miR-286 inhibitor related to reproductive development of bactrocera dorsalis and application of miR-286 inhibitor
  • miR-286 inhibitor related to reproductive development of bactrocera dorsalis and application of miR-286 inhibitor
  • miR-286 inhibitor related to reproductive development of bactrocera dorsalis and application of miR-286 inhibitor

Examples

Experimental program
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Effect test

Embodiment 1

[0019] Example 1: High-throughput small RNA sequencing to obtain the nucleotide sequence of microRNA—miR-286 related to the reproduction and development of Bactrocera dorsalis

[0020] RNA was extracted from the ovaries and fat bodies of newly emerged Bacteralis dorsalis female adults with different developmental days. At least 50 female adults were sampled for each sample, with three biological replicates. RNA extraction was performed using TRIZOL Total RNA Extraction Reagent (TaKaRa). Use 2 μg of purified RNA to prepare the small RNA libraries of the ovaries and fat bodies of the first eclosion 1, 5, 9, and 13 days through the TruSeq Small RNA Sample Prep Kits (Illumina, San Diego, CA) kit, and then send them to LC-BIO company sequencing. Afterwards, the constructed library was sequenced using Illumina Hiseq2500. Finally, the differentially expressed miRNA-miR-286 in the ovary and fat body of Bacteralis dorsalis female adults at different developmental stages was selected...

Embodiment 2

[0021] Example 2: Sequence analysis of miR-286 related to reproductive development of Bactrocera dorsalis

[0022] The sequence of miR-286 was analyzed, and the folding structure showed that miR-286 had a typical hairpin structure ( figure 1 ), the mature sequence of miR-286 is located on one arm of the precursor. Through the sequence similarity of Blast alignment at NCBI, and the multiple alignment of miR-286 precursor sequences in Bacteralis dorsalis, Drosophila melanogaster and Drosophila melanogaster with MEGA6.0 software, the mature sequence of miR-286 was found Identical in these species, the B. dorsalis miR-286 precursor is highly similar to other insect miR-286 precursor sequences ( figure 2 ), these results show that miR-286 has a certain evolutionary conservation in insects.

Embodiment 3

[0023] Example 3: Synthesis of miR-286 specific inhibitors

[0024] Taking the measured mature body sequence of miR-286 as a reference, Suzhou Gemma Gene Co., Ltd. was entrusted to synthesize its inhibitor. The inhibitor sequence is: 5'-AGCACGAGUGUUCGGUCUAGUCA-3' (SEQ ID NO.2), which was named For Antagomir-286.

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Abstract

The invention discloses a miR-286 inhibitor related to reproductive development of bactrocera dorsalis and application of the miR-286 inhibitor. miR-286 is found to be related to development of ovaries and egg yield of female bactrocera dorsalis for the first time, the expression of miR-286 can be reduced by using the specific miR-286 inhibitor, compared with a control group, the ovary form of female adults of the bactrocera dorsalis with the miR-286 inhibitor shows obvious development delay, the ovary diameter is also inhibited, and the single female egg yield and the total egg yield are significantly reduced. Therefore, the expression of the miR-286 in the bactrocera dorsalis can be inhibited to affect normal reproductive development of the bactrocera dorsalis and reduce fertility, thereby facilitating green prevention and control of the bactrocera dorsalis.

Description

technical field [0001] The invention belongs to the fields of pest biological control and genetic engineering, and specifically relates to the application of microRNA-miR-286 related to the reproductive development of Bactrocera dorsalis and its inhibitor in the control of Bactrocera dorsalis. Background technique [0002] Bactrocera dorsalis, also known as oriental fruit fly, commonly known as fruit maggots, belongs to Diptera, Tephritidae, and Bactrocera. It is a destructive fruit and vegetable pest in my country and seriously harms more than 250 kinds of fruits and vegetables including citrus and mango. vegetable. At present, Bactrocera dorsalis is mainly distributed in India, the Philippines, Thailand, the United States, Australia and other countries abroad. It is mainly distributed in Taiwan Province, Hong Kong, Guangdong, Yunnan and other places in China. It can harm the fruits of various fruit trees, resulting in reduced or even extinct production. . The pest mainly ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113A01N57/16A01P23/00A01P7/04
CPCA01N57/16C12N15/113C12N2310/141
Inventor 郑薇薇张芮张宏宇张圣锋
Owner HUAZHONG AGRI UNIV
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