Transgenic overexpression of UDP glycosyltransferase gene UGT41A3 induces resistance to nucleopolyhedrovirus in Bombyx mori.

IF 2.7 3区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

Transgenic Research Pub Date : 2024-12-30 DOI:10.1007/s11248-024-00422-9

Chunying Gong, Junwen Ai, Yong Liu, Xingjian He, Hong Xue, Chaohua Jia, Zhuohua Chen, Hanfu Xu, Rongpeng Liu, Yong Yang

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Abstract

Bombyx mori nuclear polyhedrosis, caused by B. mori nucleopolyhedrovirus (BmNPV), threatens sericulture seriously. To explore strategies for controlling it, the UDP glycosyltransferase gene UGT41A3 (BmUGT41A3) was targeted. UGT is involved in exogenous substances detoxification and endogenous biomass regulation in insects. Early embryos of the BmNPV-sensitive variety 'HYB' were used to obtain the transgenic line HYB-UGT41A3, overexpressing BmUGT41A3 under the IE1 promoter. qPCR results revealed that, compared with the wild-type control 'HYB', BmUGT41A3 was upregulated during the individual developmental stages of HYB-UGT41A3 from silkworm eggs to fifth-instar larvae; peak expression was observed in the third-instar larvae, which presented the most significantly upregulated expression. Individual-tissues qPCR results revealed that BmUGT41A3 expression was highest in the hemocytes of HYB-UGT41A3, followed by the midgut, whereas expression in HYB was very low. Gradient feeding of BmNPV on HYB-UGT41A3 and control 'HYB' larvae on the first day of the second-instar stage. The results revealed that the LC₅₀ of HYB-UGT41A3 reached 4.040 × 10⁷ particles/mL, which was 20-fold greater than that of HYB. The decrease in the BmNPV load was more significant in HYB-UGT41A3 than in HYB at 48 h after viral inoculation. These results indicate BmUGT41A3 overexpression inhibits BmNPV proliferation and improve resistance to BmNPV in B. mori.

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UDP糖基转移酶基因UGT41A3的转基因过表达诱导家蚕对核多角体病毒的抗性

家蚕核型多角体病是由家蚕核型多角体病毒（BmNPV）引起的，严重威胁着桑蚕养殖业。为探索其调控策略，以UDP糖基转移酶基因UGT41A3 （BmUGT41A3）为靶点。UGT参与昆虫体内外源物质解毒和内源生物量调节。利用bmnpv敏感品种“HYB”的早期胚胎，获得了在IE1启动子下过表达BmUGT41A3的转基因系HYB- ugt41a3。qPCR结果显示，与野生型对照‘HYB’相比，HYB- ugt41a3从蚕卵到5龄幼虫的个体发育阶段，BmUGT41A3上调；3龄幼虫表达量最高，表达量上调最显著。个体组织qPCR结果显示，BmUGT41A3在HYB- ugt41a3的血细胞中表达最高，其次是中肠，而在HYB中的表达非常低。二龄第1天BmNPV对HYB- ugt41a3和对照“HYB”幼虫梯度投食。结果表明，HYB- ugt41a3的LC50为4.040 × 107粒/mL，是HYB的20倍。在病毒接种后48 h， HYB- ugt41a3的BmNPV载量比HYB的下降更为显著。上述结果表明，BmUGT41A3过表达可抑制家蚕BmNPV的增殖，提高家蚕对BmNPV的抗性。

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来源期刊

Transgenic Research 生物-生化研究方法

CiteScore

5.40

自引率

0.00%

发文量

审稿时长

4-8 weeks

期刊介绍： Transgenic Research focusses on transgenic and genome edited higher organisms. Manuscripts emphasizing biotechnological applications are strongly encouraged. Intellectual property, ethical issues, societal impact and regulatory aspects also fall within the scope of the journal. Transgenic Research aims to bridge the gap between fundamental and applied science in molecular biology and biotechnology for the plant and animal academic and associated industry communities. Transgenic Research publishes -Original Papers -Reviews: Should critically summarize the current state-of-the-art of the subject in a dispassionate way. Authors are requested to contact a Board Member before submission. Reviews should not be descriptive; rather they should present the most up-to-date information on the subject in a dispassionate and critical way. Perspective Reviews which can address new or controversial aspects are encouraged. -Brief Communications: Should report significant developments in methodology and experimental transgenic higher organisms