雌雄红螯螯虾染色体核型比较分析

doi:10.3969/j.issn.1004-1524.20221534

浙江农业学报 ›› 2023, Vol. 35 ›› Issue (9): 2079-2089.DOI: 10.3969/j.issn.1004-1524.20221534

雌雄红螯螯虾染色体核型比较分析

刘芳芳¹(), 陈红林²^,^*(), 刘峰², 许晓军², 黄福勇², 楼宝², 钱豪杰³

1.浙江海洋大学水产学院,浙江舟山 316022
2.浙江省农业科学院水生生物研究所,浙江杭州 310021
3.浙江省海宁市鸿海养殖有限公司,浙江海宁 314400

收稿日期:2022-10-31 出版日期:2023-09-25 发布日期:2023-10-09
作者简介:刘芳芳(1997—),女,河南濮阳人,硕士研究生,主要从事水产动物遗传育种研究。E-mail: liufangfang_1022@163.com
通讯作者: 陈红林,E-mail: chenhonglin@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(2021C02069-4)

Comparative analysis of chromosome karyotype between male and female red claw crayfish (Cherax quadricarinatus)

LIU Fangfang¹(), CHEN Honglin²^,^*(), LIU Feng², XU Xiaojun², HUANG Fuyong², LOU Bao², QIAN Haojie³

1. School of Fishery, Zhejiang Ocean University, Zhoushan 316022, Zhejiang, China
2. Institute of Hydrobiology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China
3. Haining Honghai Breeding Co., Ltd., Haining 314400, Zhejiang, China

Received:2022-10-31 Online:2023-09-25 Published:2023-10-09

摘要/Abstract

摘要：

为获得精确的染色体数目、臂长信息等数据来辅助红螯螯虾(Cherax quadricarinatus)染色体水平的基因组组装,同时也为深入了解其细胞遗传学特征,特对雌、雄红螯螯虾的染色体核型差异进行分析。以红螯螯虾肝胰腺和精巢组织为材料,筛选秋水仙素最佳作用时长,采用低渗和空气干燥的方法制备染色体标本,分别从雌雄红螯螯虾的肝胰腺组织中获得有丝分裂相,从精巢组织中获得减数分裂相。共统计36个雌、雄个体有丝分裂相的染色体数目和36个雄虾减数分裂相的二价体数目。结果表明:雌、雄红螯螯虾染色体数目出现频率最高的均为200,分别占39.3%、50.0%,二价体数目出现频率最高的为100,占50%。从中选取染色体形态清晰、着丝粒可识别的有丝分裂中期相进行核型分析,发现雌、雄红螯螯虾染色体在核型分组上的结果一致,核型公式为n=100=36(m)+33(sm)+14(st)+17(t),未发现异型染色体。独立样本t检验结果显示,雌、雄红螯螯虾的各组型染色体长度和染色体臂比无显著差异(P>0.05)。本研究首次比较了雌、雄红螯螯虾染色体核型数据,丰富了红螯螯虾细胞遗传学基础研究和十足目动物染色体核型信息,为其遗传育种工作提供了基础数据,为红螯螯虾染色体水平的基因组组装提供了可靠的参考数据。

关键词: 红螯螯虾, 肝胰腺, 精巢, 染色体, 核型分析

Abstract:

In order to obtain accurate chromosome number, arm length and other data to assist chromosome level genome assembly of red claw crayfish (Cherax quadricarinatus), as well as to understand its cytogenetic characteristics, the karyotype differences between male and female red claw crayfish chromosomes were analyzed. The hepatopancreas and testis of red claw crayfish were used as materials to screen the best duration of colchicine. The chromosome specimens were prepared by low permeability and air drying methods. The mitotic phases were obtained from male and female hepatopancreas, and the meiosis phases were obtained from testis. The number of chromosomes of 36 female and male mitotic phases and the number of bivalents of 36 male meiotic phases were counted. The results showed that the highest frequency of the number of chromosomes was 200, which accounted for 39.3% and 50.0% in female and male red claw crayfish, respectively. And the highest frequency of the number of bivalents was 100, which accounted for 50%. The mitotic metaphase with clear chromosome morphology and identifiable centromere was selected for karyotype analysis. It was found that the results of karyotype grouping of female and male red claw crayfish chromosomes were consistent, and the karyotype formula was n=100=36(m)+33(sm)+14(st)+17(t). No heterotypic chromosome was found. The results of independent sample t-test showed that there was no significant difference in chromosome length and chromosome arm ratio of each type between male and female red claw cravfish (P>0.05). The present study compared the chromosome karyotype of female and male red claw crayfish for the first time, which enriched the basic research of red claw crayfish cytogenetics and the chromosome karyotype information of decapods, provided basic data for the genetic breeding of red claw crayfish and reliable reference data for its genome assembly at the chromosome level.

Key words: red claw crayfish (Cherax quadricarinatus), hepatopancreas, testis, chromosome, karyotype analysis

中图分类号:

S917

刘芳芳, 陈红林, 刘峰, 许晓军, 黄福勇, 楼宝, 钱豪杰. 雌雄红螯螯虾染色体核型比较分析[J]. 浙江农业学报, 2023, 35(9): 2079-2089.

LIU Fangfang, CHEN Honglin, LIU Feng, XU Xiaojun, HUANG Fuyong, LOU Bao, QIAN Haojie. Comparative analysis of chromosome karyotype between male and female red claw crayfish (Cherax quadricarinatus)[J]. Acta Agriculturae Zhejiangensis, 2023, 35(9): 2079-2089.

图/表 9

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染色体序号 Chromosome No.	雄性Male				雌性Female
染色体序号 Chromosome No.	长度 Length/μm	相对长度 Relative length/%	臂比 Arm ratio	类型 Type	长度 Length/μm	相对长度 Relative length/%	臂比 Arm ratio	类型 Type
1	3.220	2.5	1.405	m	2.667	2.4	1.323	m
2	2.943	2.3	1.571	m	2.285	2.1	1.246	m
3	2.825	2.2	1.244	m	2.122	1.9	1.089	m
4	2.422	1.9	1.242	m	1.829	1.7	1.436	m
5	2.369	1.9	1.181	m	1.697	1.6	1.482	m
6	2.110	1.7	1.275	m	1.552	1.4	1.287	m
7	1.963	1.5	1.515	m	1.409	1.3	1.170	m
8	1.777	1.4	1.426	m	1.384	1.3	1.551	m
9	1.742	1.4	1.360	m	1.246	1.1	1.417	m
10	1.697	1.3	1.458	m	1.224	1.1	1.150	m
11	1.605	1.3	1.117	m	1.187	1.1	1.194	m
12	1.423	1.1	1.220	m	1.135	1.0	1.368	m
13	1.357	1.1	1.188	m	1.121	1.0	1.448	m
14	1.279	1.0	1.417	m	1.096	1.0	1.260	m
15	1.252	1.0	1.273	m	1.017	0.9	1.351	m
16	1.251	1.0	1.133	m	0.997	0.9	1.382	m
17	1.155	0.9	1.465	m	0.997	0.9	1.304	m
18	1.133	0.9	1.343	m	0.985	0.9	1.227	m
19	1.126	0.9	1.362	m	0.982	0.9	1.496	m
20	1.090	0.9	1.497	m	0.940	0.9	1.406	m
21	1.046	0.8	1.557	m	0.920	0.8	1.172	m
22	1.043	0.8	1.211	m	0.908	0.8	1.168	m
23	1.008	0.8	1.167	m	0.899	0.8	1.064	m
24	0.963	0.8	1.534	m	0.890	0.8	1.262	m
25	0.928	0.7	1.442	m	0.871	0.8	1.639	m
26	0.897	0.7	1.492	m	0.864	0.8	1.280	m
27	0.893	0.7	1.453	m	0.836	0.8	1.389	m
28	0.892	0.7	1.382	m	0.829	0.8	1.052	m
29	0.789	0.6	1.415	m	0.825	0.8	1.156	m
30	0.788	0.6	1.516	m	0.812	0.7	1.122	m
31	0.768	0.6	1.489	m	0.710	0.6	1.320	m
32	0.731	0.6	1.270	m	0.656	0.6	1.209	m
33	0.676	0.5	1.404	m	0.647	0.6	1.247	m
34	0.634	0.5	1.158	m	0.643	0.6	1.197	m
35	0.537	0.4	1.498	m	0.599	0.5	1.192	m
36	0.418	0.3	1.596	m	0.573	0.5	1.234	m
37	3.652	2.9	2.963	sm	2.125	1.9	1.999	sm
38	3.223	2.5	2.535	sm	1.649	1.5	1.946	sm
39	3.189	2.5	2.407	sm	1.645	1.5	1.828	sm
40	2.148	1.7	2.284	sm	1.609	1.5	1.890	sm
41	1.986	1.6	1.867	sm	1.437	1.3	1.944	sm
42	1.711	1.3	2.169	sm	1.409	1.3	2.004	sm
43	1.589	1.2	1.972	sm	1.389	1.3	1.954	sm
44	1.373	1.1	2.272	sm	1.388	1.3	1.995	sm
45	1.338	1.0	2.521	sm	1.361	1.2	2.044	sm
46	1.214	1.0	1.949	sm	1.267	1.2	2.343	sm
47	1.211	0.9	2.068	sm	1.262	1.2	2.328	sm
48	1.201	0.9	1.935	sm	1.119	1.0	1.925	sm
49	1.189	0.9	2.175	sm	1.111	1.0	2.216	sm
50	1.183	0.9	1.874	sm	1.097	1.0	2.010	sm
51	1.167	0.9	1.745	sm	1.036	0.9	2.144	sm
52	1.127	0.9	1.934	sm	1.015	0.9	2.676	sm
53	1.090	0.9	2.481	sm	1.009	0.9	1.782	sm
54	1.089	0.9	1.835	sm	1.006	0.9	2.386	sm
55	1.085	0.8	1.750	sm	1.002	0.9	2.968	sm
56	1.069	0.8	1.854	sm	0.978	0.9	2.912	sm
57	1.039	0.8	2.485	sm	0.957	0.9	2.467	sm
58	0.987	0.8	1.848	sm	0.924	0.8	2.955	sm
59	0.940	0.7	1.918	sm	0.847	0.8	2.101	sm
60	0.920	0.7	1.997	sm	0.839	0.8	2.803	sm
61	0.888	0.7	2.024	sm	0.837	0.8	1.860	sm
62	0.885	0.7	1.968	sm	0.816	0.7	2.593	sm
63	0.860	0.7	1.976	sm	0.799	0.7	2.975	sm
64	0.860	0.7	1.709	sm	0.791	0.7	2.485	sm
65	0.859	0.7	1.784	sm	0.785	0.7	2.058	sm
66	0.756	0.6	1.940	sm	0.742	0.7	2.047	sm
67	0.732	0.6	1.846	sm	0.681	0.6	2.000	sm
68	0.671	0.5	1.947	sm	0.670	0.6	1.735	sm
69	0.573	0.4	1.974	sm	0.572	0.5	1.886	sm
70	2.531	2.0	3.496	st	2.471	2.3	3.284	st
71	2.169	1.7	3.508	st	2.066	1.9	3.480	st
72	1.863	1.5	4.628	st	1.372	1.3	3.042	st
73	1.775	1.4	3.498	st	1.174	1.1	3.103	st
74	1.676	1.3	4.710	st	1.123	1.0	3.806	st
75	1.492	1.2	4.007	st	1.065	1.0	5.507	st
76	1.396	1.1	3.274	st	1.019	0.9	3.687	st
77	1.252	1.0	3.052	st	1.016	0.9	3.231	st
78	1.024	0.8	3.621	st	1.001	0.9	3.488	st
79	1.017	0.8	3.468	st	0.991	0.9	4.730	st
80	0.979	0.8	3.418	st	0.946	0.9	3.645	st
81	0.916	0.7	3.024	st	0.858	0.8	3.108	st
82	0.915	0.7	3.463	st	0.815	0.7	4.461	st
83	0.814	0.6	3.229	st	0.628	0.6	5.009	st
84	2.025	1.6	∞	t	1.709	1.6	∞	t
85	1.488	1.2	8.727	t	1.473	1.3	∞	t
86	1.181	0.9	∞	t	1.238	1.1	∞	t
87	1.169	0.9	∞	t	1.151	1.1	∞	t
88	1.091	0.9	9.192	t	1.105	1.0	∞	t
89	1.008	0.8	∞	t	1.078	1.0	∞	t
90	0.921	0.7	∞	t	1.030	0.9	∞	t
91	0.915	0.7	∞	t	1.024	0.9	∞	t
92	0.891	0.7	∞	t	0.989	0.9	∞	t
93	0.863	0.7	∞	t	0.873	0.8	∞	t
94	0.863	0.7	∞	t	0.842	0.8	∞	t
95	0.701	0.5	∞	t	0.838	0.8	∞	t
96	0.613	0.5	∞	t	0.805	0.7	∞	t
97	0.529	0.4	∞	t	0.731	0.7	∞	t
98	0.459	0.4	∞	t	0.675	0.6	∞	t
99	0.436	0.3	∞	t	0.473	0.4	∞	t
100	0.161	0.1	∞	t	0.250	0.2	∞	t

染色体序号 Chromosome No.	雄性Male				雌性Female
染色体序号 Chromosome No.	长度 Length/μm	相对长度 Relative length/%	臂比 Arm ratio	类型 Type	长度 Length/μm	相对长度 Relative length/%	臂比 Arm ratio	类型 Type
1	3.220	2.5	1.405	m	2.667	2.4	1.323	m
2	2.943	2.3	1.571	m	2.285	2.1	1.246	m
3	2.825	2.2	1.244	m	2.122	1.9	1.089	m
4	2.422	1.9	1.242	m	1.829	1.7	1.436	m
5	2.369	1.9	1.181	m	1.697	1.6	1.482	m
6	2.110	1.7	1.275	m	1.552	1.4	1.287	m
7	1.963	1.5	1.515	m	1.409	1.3	1.170	m
8	1.777	1.4	1.426	m	1.384	1.3	1.551	m
9	1.742	1.4	1.360	m	1.246	1.1	1.417	m
10	1.697	1.3	1.458	m	1.224	1.1	1.150	m
11	1.605	1.3	1.117	m	1.187	1.1	1.194	m
12	1.423	1.1	1.220	m	1.135	1.0	1.368	m
13	1.357	1.1	1.188	m	1.121	1.0	1.448	m
14	1.279	1.0	1.417	m	1.096	1.0	1.260	m
15	1.252	1.0	1.273	m	1.017	0.9	1.351	m
16	1.251	1.0	1.133	m	0.997	0.9	1.382	m
17	1.155	0.9	1.465	m	0.997	0.9	1.304	m
18	1.133	0.9	1.343	m	0.985	0.9	1.227	m
19	1.126	0.9	1.362	m	0.982	0.9	1.496	m
20	1.090	0.9	1.497	m	0.940	0.9	1.406	m
21	1.046	0.8	1.557	m	0.920	0.8	1.172	m
22	1.043	0.8	1.211	m	0.908	0.8	1.168	m
23	1.008	0.8	1.167	m	0.899	0.8	1.064	m
24	0.963	0.8	1.534	m	0.890	0.8	1.262	m
25	0.928	0.7	1.442	m	0.871	0.8	1.639	m
26	0.897	0.7	1.492	m	0.864	0.8	1.280	m
27	0.893	0.7	1.453	m	0.836	0.8	1.389	m
28	0.892	0.7	1.382	m	0.829	0.8	1.052	m
29	0.789	0.6	1.415	m	0.825	0.8	1.156	m
30	0.788	0.6	1.516	m	0.812	0.7	1.122	m
31	0.768	0.6	1.489	m	0.710	0.6	1.320	m
32	0.731	0.6	1.270	m	0.656	0.6	1.209	m
33	0.676	0.5	1.404	m	0.647	0.6	1.247	m
34	0.634	0.5	1.158	m	0.643	0.6	1.197	m
35	0.537	0.4	1.498	m	0.599	0.5	1.192	m
36	0.418	0.3	1.596	m	0.573	0.5	1.234	m
37	3.652	2.9	2.963	sm	2.125	1.9	1.999	sm
38	3.223	2.5	2.535	sm	1.649	1.5	1.946	sm
39	3.189	2.5	2.407	sm	1.645	1.5	1.828	sm
40	2.148	1.7	2.284	sm	1.609	1.5	1.890	sm
41	1.986	1.6	1.867	sm	1.437	1.3	1.944	sm
42	1.711	1.3	2.169	sm	1.409	1.3	2.004	sm
43	1.589	1.2	1.972	sm	1.389	1.3	1.954	sm
44	1.373	1.1	2.272	sm	1.388	1.3	1.995	sm
45	1.338	1.0	2.521	sm	1.361	1.2	2.044	sm
46	1.214	1.0	1.949	sm	1.267	1.2	2.343	sm
47	1.211	0.9	2.068	sm	1.262	1.2	2.328	sm
48	1.201	0.9	1.935	sm	1.119	1.0	1.925	sm
49	1.189	0.9	2.175	sm	1.111	1.0	2.216	sm
50	1.183	0.9	1.874	sm	1.097	1.0	2.010	sm
51	1.167	0.9	1.745	sm	1.036	0.9	2.144	sm
52	1.127	0.9	1.934	sm	1.015	0.9	2.676	sm
53	1.090	0.9	2.481	sm	1.009	0.9	1.782	sm
54	1.089	0.9	1.835	sm	1.006	0.9	2.386	sm
55	1.085	0.8	1.750	sm	1.002	0.9	2.968	sm
56	1.069	0.8	1.854	sm	0.978	0.9	2.912	sm
57	1.039	0.8	2.485	sm	0.957	0.9	2.467	sm
58	0.987	0.8	1.848	sm	0.924	0.8	2.955	sm
59	0.940	0.7	1.918	sm	0.847	0.8	2.101	sm
60	0.920	0.7	1.997	sm	0.839	0.8	2.803	sm
61	0.888	0.7	2.024	sm	0.837	0.8	1.860	sm
62	0.885	0.7	1.968	sm	0.816	0.7	2.593	sm
63	0.860	0.7	1.976	sm	0.799	0.7	2.975	sm
64	0.860	0.7	1.709	sm	0.791	0.7	2.485	sm
65	0.859	0.7	1.784	sm	0.785	0.7	2.058	sm
66	0.756	0.6	1.940	sm	0.742	0.7	2.047	sm
67	0.732	0.6	1.846	sm	0.681	0.6	2.000	sm
68	0.671	0.5	1.947	sm	0.670	0.6	1.735	sm
69	0.573	0.4	1.974	sm	0.572	0.5	1.886	sm
70	2.531	2.0	3.496	st	2.471	2.3	3.284	st
71	2.169	1.7	3.508	st	2.066	1.9	3.480	st
72	1.863	1.5	4.628	st	1.372	1.3	3.042	st
73	1.775	1.4	3.498	st	1.174	1.1	3.103	st
74	1.676	1.3	4.710	st	1.123	1.0	3.806	st
75	1.492	1.2	4.007	st	1.065	1.0	5.507	st
76	1.396	1.1	3.274	st	1.019	0.9	3.687	st
77	1.252	1.0	3.052	st	1.016	0.9	3.231	st
78	1.024	0.8	3.621	st	1.001	0.9	3.488	st
79	1.017	0.8	3.468	st	0.991	0.9	4.730	st
80	0.979	0.8	3.418	st	0.946	0.9	3.645	st
81	0.916	0.7	3.024	st	0.858	0.8	3.108	st
82	0.915	0.7	3.463	st	0.815	0.7	4.461	st
83	0.814	0.6	3.229	st	0.628	0.6	5.009	st
84	2.025	1.6	∞	t	1.709	1.6	∞	t
85	1.488	1.2	8.727	t	1.473	1.3	∞	t
86	1.181	0.9	∞	t	1.238	1.1	∞	t
87	1.169	0.9	∞	t	1.151	1.1	∞	t
88	1.091	0.9	9.192	t	1.105	1.0	∞	t
89	1.008	0.8	∞	t	1.078	1.0	∞	t
90	0.921	0.7	∞	t	1.030	0.9	∞	t
91	0.915	0.7	∞	t	1.024	0.9	∞	t
92	0.891	0.7	∞	t	0.989	0.9	∞	t
93	0.863	0.7	∞	t	0.873	0.8	∞	t
94	0.863	0.7	∞	t	0.842	0.8	∞	t
95	0.701	0.5	∞	t	0.838	0.8	∞	t
96	0.613	0.5	∞	t	0.805	0.7	∞	t
97	0.529	0.4	∞	t	0.731	0.7	∞	t
98	0.459	0.4	∞	t	0.675	0.6	∞	t
99	0.436	0.3	∞	t	0.473	0.4	∞	t
100	0.161	0.1	∞	t	0.250	0.2	∞	t

雌雄红螯螯虾染色体核型比较分析

Comparative analysis of chromosome karyotype between male and female red claw crayfish (Cherax quadricarinatus)

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