基于罗氏沼虾染色体核型、同工酶及遗传多样性分析

doi:10.7668/hbnxb.20193764

摘要/Abstract

摘要：

为保护、评估及合理利用罗氏沼虾种质资源,为今后指导育种及养殖提供理论依据,以采自广西及江苏数丰良种场的罗氏沼虾为材料,采用PHA和秋水仙素肌肉注射、精巢或卵巢细胞直接制片法,对罗氏沼虾染色体核型进行了研究。采用聚丙烯酰胺凝胶(PAGE)电泳技术,分析了罗氏沼虾肌肉或鳃组织乳酸脱氢酶(LDH)、苹果酸脱氢酶(MDH)、酯酶(EST)、醇脱氢酶(ADH)4种同工酶的组织表达模式,并初步分析了其生化遗传学特性及遗传结构。结果表明:罗氏沼虾为二倍体,2N=118=56M+24SM+10ST+28T,臂数(NF)为198。未发现带有随体或次缢痕的染色体及性染色体。4 种酶共记录出22个等位基因 15个基因位点,位点平均有效等位基因数(Ne)为1.47;4 种酶有7个基因频率为 0.1500~0.8500的多态位点;多态位点比例 P 为46.7%;平均期望杂合度(He)值为0.191 7,平均观测杂合度(Ho)值为0.261 1,Hardy-Weinberg 遗传偏离指数(D)值为0.379 7,表明其杂合子过剩及缺失现象并存,但杂合子过剩现象更突出,说明该群体遗传变异范围较宽,变异程度较大,群体遗传多样性高,可为今后的人工育种提供良好的亲本。

关键词: 罗氏沼虾, 染色体, 核型, 同工酶, 遗传结构

Abstract:

In order to evaluate the germplasm resources of M.rosenbergii and provide reference for breeding guidance,it studied the chromosome karyotypes of M.rosenbergii using PHA,narcissin intramuscular injection and the sperms or ovaries cell direct production.The tissue expression patterns of lactate dehydrogenase(LDH),malate dehydrogenase(MDH),esterase(EST)and alcohol dehydrogenase(ADH)of muscle and gill of M.rosenbergii were analyzed by polyacrylamide gel electrophoresis(PAGE).The results showed that M.rosenbergii was diploid,2N=118=56M+24SM+10ST+28T,and arm number(NF)was 198.No sex chromosome or chromosome with satellite or secondary constriction was found.Four isozymes revealed 22 alleles and 15 genetic loci, and the effective number of alleles (Ne) were 1.47.Four enzymes had 7 polymorphic loci with gene frequencies ranging from 0.150 0 to 0.850 0.The proportion of polymorphic sites(P)was 46.7%.The average expected heterozygosity(He)value was 0.191 7,the average observed heterozygosity(Ho)value was 0.261 1,and the Hardy-Weinberg genetic deviation index(D)value was 0.379 7,indicating that there were both heterozygote excess and loss,but the heterozygote excess was more prominent,indicating that this population had a wide range of genetic variation.The degree of variation was large and the population genetic diversity was high,which could provide good parents for future artificial breeding.

Key words: Macrobrachium rosenbergii, Chromosome, Karyotype, Isoenzyme, Genetic structure

张林, 甘金华, 张涛, 何力, 周剑光. 基于罗氏沼虾染色体核型、同工酶及遗传多样性分析[J]. 华北农学报, 2023, 38(S1): 429-436. doi: 10.7668/hbnxb.20193764.

ZHANG Lin, GAN Jinhua, ZHANG Tao, HE Li, ZHOU Jianguang. Karyotype,Isozyme and Population Genetic Structure Analysis of Macrobrachium rosenbergii[J]. Acta Agriculturae Boreali-Sinica, 2023, 38(S1): 429-436. doi: 10.7668/hbnxb.20193764.

http://www.hbnxb.net/CN/Y2023/V38/IS1/429

图/表 8

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染色体编号 Chromosome No.	长臂/μm Long arm	短臂/μm Short arm	总长度/μm Total length	臂比 Arm ratio	相对长度/% Relative length	着丝粒指数 Centromeric index	类型 Classification
1	1.056	0.914	1.970	1.155	2.607	0.464	M
2	0.600	0.459	1.932	1.307	2.557	0.238	M
3	0.921	0.718	1.679	1.283	2.222	0.428	M
4	0.904	0.719	1.657	1.257	2.193	0.434	M
5	0.912	0.550	1.639	1.658	2.169	0.336	M
6	1.016	0.663	1.621	1.532	2.145	0.409	M
7	0.969	0.689	1.566	1.406	2.072	0.440	M
8	0.761	0.690	1.505	1.103	1.992	0.458	M
9	0.928	0.638	1.498	1.455	1.982	0.426	M
10	0.775	0.620	1.471	1.250	1.947	0.421	M
11	1.109	0.663	1.462	1.673	1.935	0.453	M
12	0.782	0.645	1.451	1.212	1.920	0.445	M
13	0.721	0.616	1.428	1.170	1.890	0.431	M
14	0.669	0.667	1.417	1.003	1.875	0.471	M
15	0.787	0.711	1.396	1.107	1.847	0.509	M
16	0.800	0.616	1.376	1.298	1.821	0.448	M
17	0.730	0.645	1.338	1.132	1.771	0.482	M
18	0.757	0.714	1.336	1.060	1.768	0.534	M
19	0.753	0.510	1.301	1.476	1.722	0.392	M
20	0.765	0.535	1.278	1.430	1.691	0.419	M
21	0.621	0.591	1.263	1.051	1.671	0.478	M
22	0.918	0.587	1.212	1.564	1.604	0.484	M
23	0.662	0.434	1.159	1.525	1.534	0.374	M
24	0.616	0.440	1.096	1.400	1.450	0.401	M
25	0.791	0.487	1.096	1.624	1.450	0.401	M
26	0.700	0.459	1.071	1.525	1.417	0.429	M
27	0.663	0.433	1.059	1.531	1.401	0.409	M
28	0.587	0.484	1.056	1.213	1.398	0.458	M
29	1.224	0.708	2.066	1.729	2.734	0.343	SM
30	1.065	0.555	1.620	1.919	2.144	0.343	SM
31	1.428	0.638	1.595	2.238	2.110	0.400	SM
32	1.031	0.564	1.595	1.828	2.111	0.354	SM
33	0.975	0.480	1.455	2.031	1.926	0.330	SM
34	0.975	0.420	1.395	2.321	1.846	0.301	SM
35	0.840	0.465	1.305	1.806	1.727	0.356	SM
36	0.812	0.485	1.297	1.672	1.713	0.324	SM
37	0.855	0.435	1.290	1.966	1.707	0.337	SM
38	0.840	0.405	1.245	2.074	1.648	0.325	SM
39	0.780	0.345	1.125	2.261	1.489	0.307	SM
40	0.750	0.375	1.125	2.000	1.489	0.333	SM
41	0.885	0.285	1.170	3.105	1.548	0.244	ST
42	0.840	0.255	1.095	3.294	1.449	0.233	ST
43	0.855	0.150	1.005	5.700	1.330	0.149	ST
44	0.765	0.225	0.990	3.400	1.310	0.227	ST
45	0.720	0.180	0.900	4.000	1.191	0.200	ST
46	1.245	0.000	1.245	∞	1.648	0.000	T
47	1.155	0.000	1.155	∞	1.529	0.000	T
48	1.125	0.000	1.125	∞	1.489	0.000	T
49	1.095	0.000	1.095	∞	1.449	0.000	T
50	1.080	0.000	1.080	∞	1.429	0.000	T
51	1.050	0.000	1.050	∞	1.390	0.000	T
52	0.990	0.000	0.990	∞	1.310	0.000	T
53	0.975	0.000	0.975	∞	1.290	0.000	T
54	0.975	0.000	0.975	∞	1.290	0.000	T
55	0.960	0.000	0.960	∞	1.270	0.000	T
56	0.900	0.000	0.900	∞	1.191	0.000	T
57	0.855	0.000	0.855	∞	1.131	0.000	T
58	0.795	0.000	0.795	∞	1.052	0.000	T
59	0.794	0.000	0.794	∞	1.051	0.000	T

染色体编号 Chromosome No.	长臂/μm Long arm	短臂/μm Short arm	总长度/μm Total length	臂比 Arm ratio	相对长度/% Relative length	着丝粒指数 Centromeric index	类型 Classification
1	1.056	0.914	1.970	1.155	2.607	0.464	M
2	0.600	0.459	1.932	1.307	2.557	0.238	M
3	0.921	0.718	1.679	1.283	2.222	0.428	M
4	0.904	0.719	1.657	1.257	2.193	0.434	M
5	0.912	0.550	1.639	1.658	2.169	0.336	M
6	1.016	0.663	1.621	1.532	2.145	0.409	M
7	0.969	0.689	1.566	1.406	2.072	0.440	M
8	0.761	0.690	1.505	1.103	1.992	0.458	M
9	0.928	0.638	1.498	1.455	1.982	0.426	M
10	0.775	0.620	1.471	1.250	1.947	0.421	M
11	1.109	0.663	1.462	1.673	1.935	0.453	M
12	0.782	0.645	1.451	1.212	1.920	0.445	M
13	0.721	0.616	1.428	1.170	1.890	0.431	M
14	0.669	0.667	1.417	1.003	1.875	0.471	M
15	0.787	0.711	1.396	1.107	1.847	0.509	M
16	0.800	0.616	1.376	1.298	1.821	0.448	M
17	0.730	0.645	1.338	1.132	1.771	0.482	M
18	0.757	0.714	1.336	1.060	1.768	0.534	M
19	0.753	0.510	1.301	1.476	1.722	0.392	M
20	0.765	0.535	1.278	1.430	1.691	0.419	M
21	0.621	0.591	1.263	1.051	1.671	0.478	M
22	0.918	0.587	1.212	1.564	1.604	0.484	M
23	0.662	0.434	1.159	1.525	1.534	0.374	M
24	0.616	0.440	1.096	1.400	1.450	0.401	M
25	0.791	0.487	1.096	1.624	1.450	0.401	M
26	0.700	0.459	1.071	1.525	1.417	0.429	M
27	0.663	0.433	1.059	1.531	1.401	0.409	M
28	0.587	0.484	1.056	1.213	1.398	0.458	M
29	1.224	0.708	2.066	1.729	2.734	0.343	SM
30	1.065	0.555	1.620	1.919	2.144	0.343	SM
31	1.428	0.638	1.595	2.238	2.110	0.400	SM
32	1.031	0.564	1.595	1.828	2.111	0.354	SM
33	0.975	0.480	1.455	2.031	1.926	0.330	SM
34	0.975	0.420	1.395	2.321	1.846	0.301	SM
35	0.840	0.465	1.305	1.806	1.727	0.356	SM
36	0.812	0.485	1.297	1.672	1.713	0.324	SM
37	0.855	0.435	1.290	1.966	1.707	0.337	SM
38	0.840	0.405	1.245	2.074	1.648	0.325	SM
39	0.780	0.345	1.125	2.261	1.489	0.307	SM
40	0.750	0.375	1.125	2.000	1.489	0.333	SM
41	0.885	0.285	1.170	3.105	1.548	0.244	ST
42	0.840	0.255	1.095	3.294	1.449	0.233	ST
43	0.855	0.150	1.005	5.700	1.330	0.149	ST
44	0.765	0.225	0.990	3.400	1.310	0.227	ST
45	0.720	0.180	0.900	4.000	1.191	0.200	ST
46	1.245	0.000	1.245	∞	1.648	0.000	T
47	1.155	0.000	1.155	∞	1.529	0.000	T
48	1.125	0.000	1.125	∞	1.489	0.000	T
49	1.095	0.000	1.095	∞	1.449	0.000	T
50	1.080	0.000	1.080	∞	1.429	0.000	T
51	1.050	0.000	1.050	∞	1.390	0.000	T
52	0.990	0.000	0.990	∞	1.310	0.000	T
53	0.975	0.000	0.975	∞	1.290	0.000	T
54	0.975	0.000	0.975	∞	1.290	0.000	T
55	0.960	0.000	0.960	∞	1.270	0.000	T
56	0.900	0.000	0.900	∞	1.191	0.000	T
57	0.855	0.000	0.855	∞	1.131	0.000	T
58	0.795	0.000	0.795	∞	1.052	0.000	T
59	0.794	0.000	0.794	∞	1.051	0.000	T

同工酶 Isozyme	基因位点 Genetic locus	等位基因 Allele	基因频率 Gene frequcy	期望杂合度(He) Expected heterozygosity	观测杂合度(Ho) Observed heterozygosity
LDH	LDH-1	a	1.000 0	0.000 0	0.000 0
	LDH-2	a	1.000 0	0.000 0	0.000 0
	LDH-3	a	0.500 0	0.500 0	1.000 0
		b	0.500 0
	LDH-4	a	1.000 0	0.000 0	0.000 0
	LDH-5	a	1.000 0
EST	EST-1	a	0.368 4	0.465 4	0.315 8
		b	0.631 6
	EST-2	a	0.842 1	0.265 9	0.315 8
		b	0.157 9
	EST-3	a	1.000 0	0.000 0	0.000 0
MDH	MDH-1	a	1.000 0	0.000 0	0.000 0
	MDH-2	a	0.357 1	0.459 2	0.526 3
		b	0.642 9
	MDH-3	a	0.578 9	0.487 6	0.842 1
		b	0.421 1
	MDH-4	a	0.666 7	0.444 4	0.666 7
		b	0.333 3
	ADH-1	a	1.000 0	0.000 0	0.000 0
ADH	ADH-2	a	0.150 0	0.255 0	0.300 0
		b	0.850 0
	ADH-3	a	1.000 0	0.000 0	0.000 0
合计Total	15	22		2.875 1	3.966 7
平均值Average				0.191 7	0.261 1

同工酶 Isozyme	基因位点 Genetic locus	等位基因 Allele	基因频率 Gene frequcy	期望杂合度(He) Expected heterozygosity	观测杂合度(Ho) Observed heterozygosity
LDH	LDH-1	a	1.000 0	0.000 0	0.000 0
	LDH-2	a	1.000 0	0.000 0	0.000 0
	LDH-3	a	0.500 0	0.500 0	1.000 0
		b	0.500 0
	LDH-4	a	1.000 0	0.000 0	0.000 0
	LDH-5	a	1.000 0
EST	EST-1	a	0.368 4	0.465 4	0.315 8
		b	0.631 6
	EST-2	a	0.842 1	0.265 9	0.315 8
		b	0.157 9
	EST-3	a	1.000 0	0.000 0	0.000 0
MDH	MDH-1	a	1.000 0	0.000 0	0.000 0
	MDH-2	a	0.357 1	0.459 2	0.526 3
		b	0.642 9
	MDH-3	a	0.578 9	0.487 6	0.842 1
		b	0.421 1
	MDH-4	a	0.666 7	0.444 4	0.666 7
		b	0.333 3
	ADH-1	a	1.000 0	0.000 0	0.000 0
ADH	ADH-2	a	0.150 0	0.255 0	0.300 0
		b	0.850 0
	ADH-3	a	1.000 0	0.000 0	0.000 0
合计Total	15	22		2.875 1	3.966 7
平均值Average				0.191 7	0.261 1

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