一种利用微创采血快速制备鳖科动物染色体核型的方法

doi:10.3969/j.issn.1004-1524.20240311

浙江农业学报 ›› 2025, Vol. 37 ›› Issue (1): 32-38.DOI: 10.3969/j.issn.1004-1524.20240311

一种利用微创采血快速制备鳖科动物染色体核型的方法

葛健辉¹^,²(), 管辛帅¹, 刘金殿¹, 任晋东¹, 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

收稿日期:2024-04-01 出版日期:2025-01-25 发布日期:2025-02-14
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: *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)

A method for rapidly preparing the karyotype of Trionychidae species by minimally invasive blood sampling

GE Jianhui¹^,²(), GUAN Xinshuai¹, LIU Jindian¹, REN Jindong¹, 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

Received:2024-04-01 Online:2025-01-25 Published:2025-02-14

摘要/Abstract

摘要：

为建立一种适用于鳖科动物的微创染色体核型制备方法,以中华鳖(Pelodiscus sinensis)为例,采用抽血刺激与植物血凝素(phytohemagglutinin, PHA)体内注射相结合的方法促进造血细胞增殖。以椎管内微创采集的血液为组织材料,通过秋水仙素处理和分离白细胞开展染色体制备,并对其中期分裂相进行核型分析。结果表明,该方法可获得足量中期分裂相;中华鳖染色体数量为33对,其中6对为大型染色体,27对为小型染色体;其核型公式为2n=66=4M+6SM+2T+54m。本研究提供了一种微创、简便快捷的鳖科动物染色体核型制备方法,可以为鳖科动物遗传育种及种质鉴定工作提供有力工具。

关键词: 鳖科动物, 微创, 染色体核型

Abstract:

In order to establish a minimally invasive chromosome karyotype preparation method for Trionychidae species, a combination of blood drawing stimulation and in vivo injection of phytohemagglutinin (PHA) on Pelodiscus sinensis was used to promote hematopoietic cell proliferation. The blood collected from the spinal canal was used as initial tissue material. Chromosome preparation was carried out by colchicine treatment and leukocyte isolation, and karyotype analysis was performed for the mitotic metaphase cells. The results showed that adequate chromosomes of mitotic metaphase could be obtained by this method. In 33 pairs of chromosomes, six were large chromosomes, and the rest 27 were small chromosomes. The karyotype formula was 2n=66=4M+6SM+2T+54m. This study would provide a minimally invasive, simple and fast karyotype method of Trionychidae species, which could provide a powerful tool for the genetic breeding and germplasm identification of Trionychidae species.

Key words: Trionychidae species, minimally invasive, karyotype

中图分类号:

S917

葛健辉, 管辛帅, 刘金殿, 任晋东, 许晓军. 一种利用微创采血快速制备鳖科动物染色体核型的方法[J]. 浙江农业学报, 2025, 37(1): 32-38.

GE Jianhui, GUAN Xinshuai, LIU Jindian, REN Jindong, XU Xiaojun. A method for rapidly preparing the karyotype of Trionychidae species by minimally invasive blood sampling[J]. Acta Agriculturae Zhejiangensis, 2025, 37(1): 32-38.

图/表 5

表1 中华鳖染色体计数

Table 1 Counts of the chromosome in the cells of P. sinensis

染色体数量 Number of chromosome	≤64	65	66	67	≥68	总和 Sum
分裂相数量 Number of metaphase	8	14	35	2	1	60
所占百分比 Percentage of metaphase/%	13.33	23.33	58.33	3.33	1.68	100.00

图1 中华鳖染色体中期分裂相

Fig.1 Metaphase chromosomes of P. sinensis

图2 中华鳖染色体核型模式图

Fig.2 Karyotype pattern of P. sinensis

表2 中华鳖大型染色体长度与臂比

Table 2 Large chromosomes length and arm ratio of P. sinensis

染色体序号 Chromosome No.	长度 Length/μm	臂比指数 Arm ratio	染色体类型 Chromosome type
1	11.995	1.499	M
2	8.267	1.033	M
3	7.120	1.883	SM
4	5.035	1.874	SM
5	5.720	2.305	SM
6	5.055	9.215	T

图3 中华鳖各染色体相对长度

Fig.3 Chromosome relative length of P. sinensis

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一种利用微创采血快速制备鳖科动物染色体核型的方法

A method for rapidly preparing the karyotype of Trionychidae species by minimally invasive blood sampling

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