杨伟克, 唐芬芬, 刘增虎, 董占鹏. 2021: 家蚕β-呋喃果糖苷酶基因BmSuc1表达特征及其对激素的响应. 南方农业学报, 52(7): 1991-1997. DOI: 10.3969/j.issn.2095-1191.2021.07.029
引用本文: 杨伟克, 唐芬芬, 刘增虎, 董占鹏. 2021: 家蚕β-呋喃果糖苷酶基因BmSuc1表达特征及其对激素的响应. 南方农业学报, 52(7): 1991-1997. DOI: 10.3969/j.issn.2095-1191.2021.07.029
YANG Wei-ke, TANG Fen-fen, LIU Zeng-hu, DONG Zhan-peng. 2021: Expression profiles and response to exogenous hormones of β-fructofuranosidase gene BmSuc1 in silkworm(Bombyx mori). Journal of Southern Agriculture, 52(7): 1991-1997. DOI: 10.3969/j.issn.2095-1191.2021.07.029
Citation: YANG Wei-ke, TANG Fen-fen, LIU Zeng-hu, DONG Zhan-peng. 2021: Expression profiles and response to exogenous hormones of β-fructofuranosidase gene BmSuc1 in silkworm(Bombyx mori). Journal of Southern Agriculture, 52(7): 1991-1997. DOI: 10.3969/j.issn.2095-1191.2021.07.029

家蚕β-呋喃果糖苷酶基因BmSuc1表达特征及其对激素的响应

Expression profiles and response to exogenous hormones of β-fructofuranosidase gene BmSuc1 in silkworm(Bombyx mori)

  • 摘要: 【目的】探究BmSuc1基因在家蚕不同组织及不同时期的表达特征及经昆虫激素处理后的表达变化规律,明确昆虫激素对BmSuc1基因的调控作用,为深入解析BmSuc1基因的功能及其表达调控机制提供参考依据。【方法】利用实时荧光定量PCR检测BmSuc1基因在家蚕发育过程中不同时期和不同组织及外源激素处理后的表达特征,并通过双链RNA (dsRNA)干扰试验检测家蚕20-羟基蜕皮素(20E)受体基因(USP)干扰效果及BmSuc1基因的表达变化。【结果】BmSuc1基因在家蚕中肠中的相对表达量最高,其次是丝腺、表皮和血淋巴,在其他组织中的相对表达量很低或几乎不表达;BmSuc1基因呈脉冲型的表达模式,在家蚕每个龄期的将眠时、五龄后期(上簇前)及预蛹时的相对表达量较高。利用外源激素20E和保幼激素(JH)分别处理五龄第2 d发育良好的家蚕,发现经20E处理后12和18 h,BmSuc1基因相对表达量极显著高于注射0.1%二甲基亚砜(DMSO)的对照组(P<0.01,下同),但在测定时间范围内JH处理组的BmSuc1基因相对表达量与对照组间无显著差异(P>0.05)。以体外转录合成USP基因的dsRNA注射五龄第3 d家蚕,注射后24和36 h,USP基因相对表达量极显著下调,即dsRNA干扰成功。利用RNAi技术干扰USP基因能阻断20E信号转导,致使BmSuc1基因表达受抑制,其相对表达量在干扰USP基因后24和36 h显著下调(P<0.05)。【结论】BmSuc1基因主要在家蚕蜕皮和变态时高表达,暗示其可能参与家蚕的变态发育过程。添加外源20E可诱导BmSuc1基因转录,而阻断20E信号转导途径会抑制BmSuc1基因表达,即BmSuc1基因作为20E信号转导通路中的下游靶基因,直接或间接受到20E信号调控。

     

    Abstract: 【Objective】 In order to clarify the regulatory effects of insect hormones on the expression of BmSuc1 and provide a reference for further analysis of the function and expression regulation mechanism of BmSuc1, this study explored the expression profiles of BmSuc1 in different tissues and periods of silkworm and the expression changes of BmSuc1 after treatment with exogenous hormones.【Method】 By using real-time fluorescence quantitative PCR technology, the expression characteristics of BmSuc1 were detected in different periods, different tissues and after treatment with exogenous hormones during the development of silkworm larvae. The expression of BmSuc1 and 20 E receptor gene USP were detected after RNA interference with double-stranded RNA(dsRNA) of USP.【Result】 The relative expression of BmSuc1 gene in midgut was the highest, followed by silk glands, epidermis and hemolymph. However, there was much lower or almost no expression in other tissues. In addition, the BmSuc1 expression profile exhibited a pulse-like pattern in silkworm larvae, the expression level of BmSuc1 was higher at each instar stage before molting, late fifth instar before cocooning and prepupal stage. The day 2 of fifth-instar silkworm larvae were treated with 20-hydroxyecdysone(20 E) and juvenile hormone(JH), it was found that the expression of BmSuc1 was extremely significantly higher at 12 and 18 h after20 E treatment than the control group injected with 0.1% dimethyl sulfoxide(DMSO) (P<0.01, the same below). But there were no significant difference of the BmSuc1 expression between the JH treatment and the control group during the measurement time range(P>0.05). The dsRNA of USP was synthesized in vitro and injected into silkworm larvae at day 3 of the fifth instar. It was showed that the USP relative expression were extremely significantly down-regulated at 24 and 36 h after injection, which indicated that dsRNA interference was successful. RNAi of USP would block 20 E signal transduction, and the expression of BmSuc1 was inhibited and significantly down-regulated at 24 and 36 h after injection of dsRNA of USP(P<0.05). 【Conclusion】 The BmSuc1 expression peaks appear in the molting of silkworm larvae and the metamorphosis of larvae to pupae, which suggests that BmSuc1 may be involved in the metamorphic development process of the silkworm. Treatment with exogenous ecdysone 20 E can activate the expression of BmSuc1, but blocking the 20 E signal transduction pathway may suppress expression of BmSuc1. It indicates that BmSuc1 as a downstream target gene in 20 E signal transduction pathway is directly or indirectly regulated by 20 E signal.

     

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