中华鳖性别分子鉴定方法开发与验证

doi:10.3969/j.issn.1004-1524.20230385

浙江农业学报 ›› 2023, Vol. 35 ›› Issue (11): 2594-2599.DOI: 10.3969/j.issn.1004-1524.20230385

中华鳖性别分子鉴定方法开发与验证

葛健辉¹^,²(), 任晋东¹, 关文志¹, 牛宝龙¹, 王伟², 楼宝¹, 于瑾³^,^*(), 许晓军¹^,^*()

1.浙江省农业科学院水生生物研究所,浙江杭州 310021
2.浙江万里学院生物与环境学院,浙江宁波 315100
3.龙游县养殖业发展中心,浙江衢州 324000

收稿日期:2023-03-22 出版日期:2023-11-25 发布日期:2023-12-04
作者简介:葛健辉(1999—),男,浙江天台人,硕士研究生,研究方向为水产动物遗传育种。E-mail: 2233167022@qq.com
通讯作者: * 于瑾,E-mail: 5653135@qq.com;许晓军,E-mail: xuxj@zaas.ac.cn
基金资助:
浙江省农业(水产)新品种选育重大科技专项(2021C02069-8)

Development and validation of molecular sex identification method in Pelodiscus sinensis

GE Jianhui¹^,²(), REN Jindong¹, GUAN Wenzhi¹, NIU Baolong¹, WANG Wei², LOU Bao¹, YU Jin³^,^*(), XU Xiaojun¹^,^*()

1. Institute of Hydrobiology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China
2. College of Biological and Environmental Sciences, Zhejiang Wanli University, Ningbo 315100, Zhejiang, China
3. Longyou County Breeding Development Center, Quzhou 324000, Zhejiang, China

Received:2023-03-22 Online:2023-11-25 Published:2023-12-04

摘要/Abstract

摘要：

为开发中华鳖(Pelodiscus sinensis)性别分子鉴定方法,分析比对了雌性中华鳖基因组中SBNO1基因2个拷贝的序列信息,筛选出1处内含子长度差异,设计引物进行雌雄个体PCR扩增检测和测序验证;并进一步以48只雌雄中华鳖基因组为样本进行准确率检验。结果显示,雌性中华鳖扩增产物为2 031 bp和1 624 bp双带,雄性中华鳖扩增产物为2 031 bp单一带。结合解剖学性别鉴定记录,48只雌雄中华鳖性别分子鉴定的判别准确率为100%。本研究提供了一种微创、简便快捷的中华鳖性别分子鉴定方法,为中华鳖单性养殖和性控育种奠定了基础。

关键词: 中华鳖, 性别分子鉴定, 聚合酶链式反应

Abstract:

In order to develop a molecular sex identification method for Pelodiscus sinensis, an intron length difference of SBNO1 gene was screened by analyzing and comparing the sequences of two copies of SBNO1 gene in female P. sinensis genome. And primers design, PCR verification, sequencing were performed. The accuracy of this sex identification method was further tested with 48 P. sinensis individuals (24 male and 24 female respectively). The results showed that the amplification products of the female individual contained 2 bands (2 031 bp and 1 624 bp), and that of the male individual had a single band (2 031 bp). Combined with anatomical sex identification records, the accuracy of sex identification was 100%. This study provided a minimally invasive, simple and fast molecular sex identification method for P. sinensis, which laid a foundation for the monosex cultivation and sex controlled breeding.

Key words: Pelodiscus sinensis, molecular sex identification, polymerase chain reaction

中图分类号:

S917.4

葛健辉, 任晋东, 关文志, 牛宝龙, 王伟, 楼宝, 于瑾, 许晓军. 中华鳖性别分子鉴定方法开发与验证[J]. 浙江农业学报, 2023, 35(11): 2594-2599.

GE Jianhui, REN Jindong, GUAN Wenzhi, NIU Baolong, WANG Wei, LOU Bao, YU Jin, XU Xiaojun. Development and validation of molecular sex identification method in Pelodiscus sinensis[J]. Acta Agriculturae Zhejiangensis, 2023, 35(11): 2594-2599.

图/表 2

图1 中华鳖日本品系雌鳖13号与雄鳖1号引物扩增验证电泳结果 M,DL2000 DNA分子量标准;13,中华鳖日本品系雌鳖扩增产物;1,中华鳖日本品系雄鳖扩增产物。

Fig.1 Banding patterns of female and male Japanese Pelodiscus sinensis individuals M, DL2000 DNA marker; 13, Female Japanese Pelodiscus sinensis individual; 1, Male Japanese Pelodiscus sinensis individual.

图2 四十八只雌雄中华鳖PCR性别检验电泳结果 A:M,DL2000分子量标准;1~12号样本为中华鳖日本品系雄鳖扩增产物;13~24号样本为中华鳖日本品系雌鳖扩增产物。B:M,DL2000分子量标准;25~36号样本为清溪乌鳖雄鳖扩增产物;37~48号样本为清溪乌鳖雌鳖扩增产物。

Fig.2 Banding patterns of 48 female and male Pelodiscus sinensis individuals A: M, DL2000 marker; Sample 1-12, Male Japanese Pelodiscus sinensis individuals; Sample 13-24, Female Japanese Pelodiscus sinensis individuals. B: M, DL2000 marker; Sample 25-36, Male Qingxi Pelodiscus sinensis individuals; Sample 37-48, Female Qingxi Pelodiscus sinensis individuals.

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中华鳖性别分子鉴定方法开发与验证

Development and validation of molecular sex identification method in Pelodiscus sinensis

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