大龙虎泡野生日本沼虾遗传多样性分析
Genetic Diversity Analysis of Macrobrachium nipponense from Dalonghupao
为了深入了解本地日本沼虾的遗传特性,为育种选育提供理论支撑,利用微卫星分子标记技术,对大龙虎泡日本沼虾进行遗传多样性比较分析。结果显示,12个微卫星位点均属于高多态性位点(PIC>0.5);太湖2号养殖群体的等位基因数为4~11,有效等位基因为3.686 4~7.480 5,观测杂合度为0.318 2~0.833 3,期望杂合度为0.744 9~0.884 8;各位点的多态性为0.691 9~0.851 7,属于高遗传多样性的群体。大龙虎泡野生群体等位基因数为1~2,有效等位基因为1.00 0 0~2.000 0,观测杂合度为0.000 0~1.000 0,期望杂合度为0.000 0~0.517 2,各位点的多态性为0.000 0~0.375 0,属于遗传多样性较低的群体。2个群体均有7个位点显著偏离Hardy-Weinberg平衡,大龙虎泡野生群体另出现3个位点无法进行Hardy-Weinberg平衡检验。太湖2号养殖群体的7个位点表现出来的是杂合子缺失,大龙虎泡野生群体的7个表现的是杂合子过剩。分子方差分析显示,有36.15%(P<0.01)的遗传变异来源于群体间,而63.85%(P<0.01)的变异来源于群体内。与太湖2号养殖群体的遗传相似度为0.276 9,遗传距离为1.284 3,这2个群体遗传相似度低,遗传距离远,存在很大的遗传差异。2个群体的遗传分化指数FST为0.361 53(P<0.01),遗传分化程度明显。种群遗传结构分析结果显示,最佳K值为3,说明这2个种群有3个可能的祖先。大龙虎泡野生群体遗传多样性水平较低,提示对大龙虎泡野生群体种质资源的保护刻不容缓。
To learn more about the genetic characteristics of Macrobrachium nipponense and provide theoretical support for breeding selection,it used microsatellite molecular marking technology to compare the genetic diversity of Macrobrachium nipponense in Dalonghupao. The results showed that all 12 microsatellite loci were highly polymorphic loci (PIC>0.5). In the population of Taihu 2,the observed number of alleles ranged from 4 to 11,the effective number of alleles ranged from 3.686 4 to 7.480 5,the observed heterozygosity was 0.318 2—0.833 3,the expected heterozygosity was 0.744 9—0.884 8,the PIC of each locus was 0.691 9—0.851 7. The population of Taihu 2 was with high genetic diversity. In the population of Dalonghupao,the observed number of alleles ranged from 1—2,the effective number of alleles was 1.000 0—2.000 0,the observed heterozygosity was 0.000 0—1.000 0,the expected heterozygosity was 0.000 0—0.517 2,the PIC of each locus was 0.000 0—0.375 0. The population of Dalonghupao was with low genetic diversity. Seven loci in both populations deviated significantly from Hardy-Weinberg equilibrium,and three loci in the population of Dalonghupao could not be tested for Hardy-Weinberg equilibrium. Seven loci in the population of Taihu 2 showed heterozygous deletion,while seven loci in the population of Dalonghupao showed heterozygous excess. AMOVA analysis showed that 36.15% (P<0.01)of the total genetic variation was among populations and 63.85% (P<0.01)was among individuals. The genetic similarity between the population of Dalonghupao and the population of Taihu 2 was 0.276 9 and the genetic distance was 1.284 3,which showed that the genetic similarity between the two populations was low,and the genetic distance was far,and there were great genetic differences. The genetic differentiation index (FST)of the two populations was 0.361 53 (P<0.01),and the degree of genetic differentiation was obvious. The population genetic structure analysis showed an optimal K value of 3,indicating that the two populations had three possible ancestors. The level of genetic diversity of Dalonghupao wild population was low,which prompting us to protect the germplasm resources of the population of Dalonghupao.
日本沼虾 / 遗传多样性 / 微卫星 / 太湖2号 / 分子标记 {{custom_keyword}} /
Macrobrachium nipponense / Genetic diversity / Microsatellite / Taihu 2 / Molecular marker {{custom_keyword}} /
表1 12对引物信息Tab.1 Information of twelve pairs of microsatellite primers |
| 位点 Locus | 基因序列号 GenBank accession number | 引物来源文献 Source literature of primers |
|---|---|---|
| WXM25 | GU189621 | Qiao等[10] |
| WXM33 | GU189629 | |
| Mni001 | EU130924 | Feng等[11] |
| Mni004 | EU130926 | |
| Mni006 | EU130928 | |
| Mni013 | EU130932 | |
| Mni034 | GQ257531 | Ma等[12] |
| Mni040 | GQ257537 | |
| Mni051 | GQ257548 | |
| Mni058 | GQ257555 | |
| Mni076 | GQ257573 | |
| X781 | KF601809 | Zhao等[13] |
| XZ339 | KF601824 |
表2 12个微卫星位点在2个日本沼虾群体中的遗传多样性Tab.2 Genetic diversity of twelve microsatellite loci in two Macrobrachium nipponense populations |
| 位点 Locus | 等位基因数 Allele number | 有效等位基因数 Number of effective allele | 观测杂合度 Observed heterozygosity | 期望杂合度 Expected heterozygosity | 多态信息含量 Polymorphism information content |
|---|---|---|---|---|---|
| WXM25 | 7 | 5.643 8 | 0.769 2 | 0.833 5 | 0.799 0 |
| WXM33 | 6 | 3.846 4 | 0.708 3 | 0.747 8 | 0.702 9 |
| Mni001 | 9 | 5.358 1 | 0.750 0 | 0.821 9 | 0.791 6 |
| Mni004 | 5 | 4.161 3 | 0.739 1 | 0.768 0 | 0.720 8 |
| Mni006 | 8 | 4.169 5 | 0.847 8 | 0.768 5 | 0.725 9 |
| Mni013 | 9 | 5.992 5 | 0.675 0 | 0.843 7 | 0.813 1 |
| Mni034 | 6 | 2.733 9 | 0.191 5 | 0.641 0 | 0.602 4 |
| Mni051 | 9 | 2.878 2 | 0.191 5 | 0.659 6 | 0.614 9 |
| Mni058 | 11 | 3.400 7 | 0.416 7 | 0.713 4 | 0.687 6 |
| Mni076 | 10 | 4.170 1 | 0.666 7 | 0.768 2 | 0.726 5 |
| X781 | 4 | 3.013 7 | 0.541 7 | 0.675 2 | 0.619 1 |
| XZ339 | 7 | 4.951 1 | 0.666 7 | 0.807 0 | 0.769 8 |
| 平均Mean | 7.583 3 | 4.193 3 | 0.597 0 | 0.754 0 | 0.714 5 |
表3 2个群体遗传多样性参数Tab.3 Genetic diversity parameters of the two populations |
| 位点 Loci | 等位基因数 Allele number | 有效等位基因数 Number of effective allele | 观测杂合度 Observed heterozygosity | 期望杂合度 Expected heterozygosity | P值 P-value | 多态信息含量 Polymorphism information content | ||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| 太湖 2号 TH | 大龙 虎泡 DM | 太湖 2号 TH | 大龙 虎泡 DM | 太湖 2号 TH | 大龙 虎泡 DM | 太湖 2号 TH | 大龙 虎泡 DM | 太湖 2号 TH | 大龙 虎泡 DM | 太湖 2号 TH | 大龙 虎泡 DM | |
| WXM25 | 7 | 2 | 5.142 9 | 2.000 0 | 0.625 0 | 1.000 0 | 0.822 7 | 0.517 2 | 0.028 3 | 0.000 3** | 0.778 0 | 0.375 0 |
| WXM33 | 6 | 2 | 4.282 5 | 1.916 8 | 0.625 0 | 0.791 7 | 0.782 8 | 0.488 5 | 0.008 8 | 0.002 2* | 0.729 0 | 0.363 9 |
| Mni001 | 9 | 2 | 7.480 5 | 1.986 2 | 0.583 3 | 0.916 7 | 0.884 8 | 0.507 1 | 0.000 0** | 0.000 0** | 0.851 7 | 0.373 3 |
| Mni004 | 5 | 2 | 4.100 8 | 2.000 0 | 0.478 3 | 1.000 0 | 0.772 9 | 0.511 1 | 0.004 2 | 0.000 0** | 0.715 8 | 0.375 0 |
| Mni006 | 8 | 2 | 5.866 7 | 1.986 2 | 0.772 7 | 0.916 7 | 0.848 8 | 0.507 1 | 0.274 8 | 0.000 0** | 0.807 5 | 0.373 3 |
| Mni013 | 9 | 2 | 6.471 9 | 1.969 2 | 0.541 7 | 0.875 0 | 0.863 5 | 0.508 1 | 0.000 4** | 0.004 2 | 0.829 7 | 0.371 1 |
| Mni034 | 6 | 1 | 4.723 2 | 1.000 0 | 0.391 3 | 0.000 0 | 0.805 8 | 0.000 0 | 0.000 0** | - | 0.758 7 | 0.000 0 |
| Mni051 | 9 | 1 | 3.686 4 | 1.000 0 | 0.391 3 | 0.000 0 | 0.744 9 | 0.000 0 | 0.000 0** | - | 0.707 9 | 0.000 0 |
| Mni058 | 11 | 1 | 7.432 3 | 1.000 0 | 0.833 3 | 0.000 0 | 0.883 9 | 0.000 0 | 0.023 9 | - | 0.851 0 | 0.000 0 |
| Mni076 | 10 | 2 | 6.816 6 | 2.000 0 | 0.333 3 | 1.000 0 | 0.871 5 | 0.510 6 | 0.000 0** | 0.000 0** | 0.837 4 | 0.375 0 |
| X781 | 4 | 2 | 3.852 8 | 1.704 1 | 0.500 0 | 0.583 3 | 0.756 2 | 0.422 0 | 0.002 2* | 0.066 2 | 0.691 9 | 0.327 8 |
| XZ339 | 7 | 2 | 4.913 7 | 2.000 0 | 0.318 2 | 1.000 0 | 0.815 0 | 0.511 1 | 0.000 0** | 0.000 0** | 0.766 6 | 0.375 0 |
| 平均Mean | 7.583 3 | 1.713 5 | 5.397 5 | 0.509 3 | 0.532 8 | 0.626 4 | 0.821 1 | 0.364 7 | - | - | 0.777 1 | 0.275 8 |
| 注:P.偏离Hardy-Weinberg平衡的显著性;*.经Bonferroni多重校正后统计学显著水平(P < 0.05);**.经Bonferroni多重校正后统计学极显著水平(P<0.01)。 | |
| Note:P.The significance of deviation from Hardy-weinberg equilibrium;*.The statistically significant level after Bonferroni multiple correction(P<0.05);**.The extremely significant level after Bonferroni multiple correction (P<0.01). |
表4 2个日本沼虾群体的AMOVA分析Tab.4 AMOVA of two Macrobrachium nipponense populations |
| 变异来源 Source of variation | 自由度 df | 平方和 Sum of square | 方差分量 Variance component | 变异百分比 Percentage of variation |
|---|---|---|---|---|
| 组内群体间Among populations | 1 | 69.938 | 1.405 33 | 36.15** |
| 群体内Within populations | 94 | 233.292 | 2.481 83 | 63.85** |
| 总计Total variation | 95 | 303.229 | 3.887 15 |
| 注:**.1023次模拟检验后显示为极显著(P<0.01)。 | |
| Note:**.1023 simulation tests showed extremely significant (P<0.01). |
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