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Maleness-on-the-Y (MoY) orthologue is a key regulator of male sex determination in Zeugodacus cucurbitae (Diptera: Tephritidae)
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FAN Zi-zhen1, 2, MA Qin1, 2, MA Si-ya3, CAO Feng-qin1, 2, 4, YAN Ri-hui1, 2, 4, LIN Xian-wu1, 2
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1 Key Laboratory of Green Prevention and Control of Tropical Plant Diseases and Pests of Ministry of Education, Hainan University, Haikou 570228, P.R.China
2 School of Plant Protection, Hainan University, Haikou 570228, P.R.China
3 School of Life Sciences, Hainan University, Haikou 570228, P.R.China
4 Hainan Yazhou Bay Seed Lab, Sanya 572000, P.R.China
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摘要
昆虫性别决定初始信号是调控昆虫性别分化的关键。在地中海实蝇中Y染色体连锁的雄性决定因子Maleness-on-the-Y (MoY)是决定雄性性别发育的初始基因，该基因调控性别决定的功能在部分实蝇科昆虫中保守，然而目前仍未鉴定出瓜实蝇（Zeugodacus cucurbitae）性别决定初始信号。为了探究瓜实蝇的性别决定初始信号，我们克隆并鉴定了MoY在瓜实蝇中的同源基因，ZcMoY。ZcMoY全长438 bp，编码70个氨基酸。同源比对发现ZcMOY蛋白与桔小实蝇同源蛋白BdMOY显示出72.6%的序列保守性，MOY的两个区域（KXNSRT和EXXKL）在实蝇科各物种之间保守。通过RT-PCR技术探究ZcMoY在瓜实蝇发育过程以及不同组织中的表达模式，发现ZcMoY在胚胎发育早期1 h开始转录，3 h达到顶峰，之后逐渐减弱；幼虫期、蛹期、成虫期均没有检测到ZcMoY的表达；检测ZcMoY在羽化第一天的雄性成虫各组织中的表达情况，发现ZcMoY在精巢中的表达量最高，头部、中肠、马氏管等组织中有微弱表达。通过RNA 干扰下调早期胚胎中ZcMoY的表达水平，发现雄性发育受到抑制，性别比例发生偏移，超过70%的个体发育为雌性或间性；间性个体呈现出异常的内外生殖器，同时兼具雌性和雄性的特征。随机选取部分RNAi胚胎发育而来的雌性成虫进行生殖能力测定，发现部分雌虫不能产卵；进一步通过PCR对这部分雌虫进行基因型分析，发现其基因型为XY型。本研究通过对瓜实蝇MoY同源基因ZcMoY的鉴定，证明了性别决定初始信号MoY在实蝇科昆虫中的保守性。基因表达模式和沉默ZcMoY的结果表明，ZcMoY在瓜实蝇早期胚胎雄性性别发育中起关键作用。我们的研究鉴定了瓜实蝇的性别决定初始信号并对它进行了功能阐释，不仅丰富了瓜实蝇性别决定机制的研究，并为利用该基因开发瓜实蝇的遗传防控品系奠定了基础。
Abstract
The initiation of sex differentiation in insects is regulated by primary sex determination signals. In the Medfly Ceratitis capitata and other Tephritids, Maleness-on-the-Y (MoY) is the master gene for male sex determination. However, the primary signal in Zeugodacus cucurbitae (Coquillett), a very destructive Tephritid pest across the world, remains ambiguous. In this study, we have isolated and characterized the Medfly MoY homolog in Z. cucurbitae, ZcMoY. ZcMOY protein shows high sequence conservation to its homologs in Bactrocera species. ZcMoY transcription begins and peaks at very early embryonic stages and then becomes undetectable except the testes and heads of day 1 male adults. Silencing ZcMoY in early embryos by RNAi causes abnormal external genitalia and interior reproductive organs, giving rise to intersexes and feminization of XY individuals. The expression pattern and knockdown phenotypes of ZcMoY indicate that ZcMoY plays a key role in regulating sex determination of Z. cucurbitae males. Our findings will help the understanding of sex determination in Z. cucurbitae and facilitate the development of genetic sexing strains in its biological control.
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Received: 07 December 2021
Accepted: 10 May 2022
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This study was supported by the the National Natural Science Foundation of China (31702059, 31860523, and 31660339), and the Hainan Provincial Natural Science Foundation of China (321CXTD435).
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| About author: FAN Zi-zhen, Tel: 0898-66180419, E-mail: 294257042@qq.com; Correspondence LIN Xian-wu, Tel: 0898-66256215, E-mail: linxianw@hainanu.edu.cn; YAN Ri-hui, Tel: 0898-66275675, E-mail: ryan1@hainanu.edu.cn |
Cite this article:
FAN Zi-zhen, MA Qin, MA Si-ya, CAO Feng-qin, YAN Ri-hui, LIN Xian-wu.
2023.
Maleness-on-the-Y (MoY) orthologue is a key regulator of male sex determination in Zeugodacus cucurbitae (Diptera: Tephritidae)
. Journal of Integrative Agriculture, 22(2): 505-513.
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