皖西白鹅和雁鹅的遗传多样性研究
摘要
养鹅业是我国农民的传统副业，我国鹅种具有产蛋多、羽绒品质好和肉质佳等优势。实验在分子水平上对皖西白鹅和雁鹅进行了RAPD分析，为皖西白鹅和雁鹅品种资源特性的研究，选育以及推广提供科学依据。
实验提取了46只皖西白鹅和48只雁鹅的血液基因组DNA，DNA的OD260/OD280值介于1.4~1.8之间，用作RAPD扩增基因组DNA的模板。对RAPD反应条件和程序逐个进行筛选，建立了最适宜的RAPD反应体系和反应程序。用RAPD技术分析皖西白鹅和雁鹅的基因组DNA的遗传多样性。研究结果表明:
1．用实验提取的基因组DNA对皖西白鹅和雁鹅的每个个体进行扩增，可以产生清晰，稳定的RAPD图谱。
2．筛选并建立了适合于皖西白鹅和雁鹅的扩增反应体系和程序：反应体积25μl，内含100ng的模板DNA，MgCl2（20mM）2.5μl，dNTP（5mM）1μl，随机引物（20μM）1μl，1U TaqDNA聚合酶。扩增程序为：94℃预变性4min，94℃变性1min，36℃退火1min，72℃延伸1min，42个循环后72℃延伸10min，于4℃结束反应。
3．用随机扩增多态性DNA技术对皖西白鹅和雁鹅进行遗传多样性分析，从50个随机引物中筛选出了12个引物对两个品种的所有个体的基因组DNA进行PCR扩增，扩增结果表明：50个随机引物中有12个引物在皖西白鹅及雁鹅基因组中分别扩增共获得83和85条条带，其中多态性条带分别为51条和57条，多态性条带率分别为61.44%和67.06%；碱基对数分别为0.21~1.52kb 和0.196~1.55kb之间，每个引物产生的条带数在2~9和3~9之间。
4．皖西白鹅和雁鹅扩增出来的条带的频率最低只有11.11%，所有引物的带频率的平均值分别为0.8236、0.7442，可以看出两群体的选育程度较高，皖西白鹅要略高于雁鹅；皖西白鹅及雁鹅群体内的遗传相似度分别平均为0.8645和0.8205，群体内的遗传多样性指数分别平均为0.7250和 1.1649，可见皖西鹅群体内个体间遗传差异较雁鹅小；两群体的遗传分化指数平均为0.7957，两群体遗传分化指数变化范围在0.6785~0.8770之间，平均值为0.7957，表明，在总遗传变异中，存在于群体间的变异达79.57%，群体内的变异仅为20.43%，显示，两群体的遗传变异主要发生在群体间，而不是群体内。
Goose breeding is a traditional homework in our country. The breeds of goose in our country have many merits such as producing many eggs, good feather and meat quality. This experiment analyzed on Wanxi White goose and Yan goose of RAPD markers in molecule level, to further provide scientific evidence for the breed resources research, selection and application on Wanxi White goose and Yan goose.
An experiment was conducted with 46 Wanxi White goose and 48 Yan goose.The OD value of Genomic DNA is between 1.4~1.8. Genomic DNA extracted from the blood of goose, which were directly used as template of PCR. RAPD reaction was carried out and programs were filtrated one by one, Optimal PCR conditions and programs were developed. RAPD is used to amplify genomic DNA of Wanxi White goose and Yan goose, their genetic diversity were also analyzed. The research results indicated:
1. Using conventional method to extract genomic DNA. The OD value of all the individual genomic DNA between 1.4~1.8. Amplified results indicated that it can produced explicit and steady RAPD bands, and testified that RAPD reaction did not request for template strictly.
2．The influencing factors of RAPD of Wanxi White goose and Yan goose genetic diversity research were studied and the experimental parameters were optimized. The optimal RAPD conditions are: 25μl reaction volume, containing 100ng template DNA, MgCl2(20mM)2.5μl，dNTP(5mM)1μl，primer (20μM)1μl，1UTaqDNA polymerase. Amplification program is : After initial denaturation at 94℃ for 4min, the amplification was performed by using 42 cycles of denaturation at 94℃ for 1min, annealing at 36℃ 1min, and extension at 72℃ for 1min. A final period of extention was carried out for 10min and final holding at 4℃.
3．Genetic diversity of Wanxi White goose and Yan goose was assessed through
Random Amplified Polymorphic DNA (RAPD) analysis. The total genomic DNA of all the individuals of both breeds were amplified through PCR method with 12 selected primers out of 50 arbitrary primers. The results suggested: Total 83，85 bands and 51, 57 polymorphic bands were acquired, which polymorphic rate is 61.44%, 67.06% respectively, The amplified base pair size were 0.21~1.52kb, 0.196~1.55kb and the number in each track were 2~9, 3~9 respectively, it also indicated that RAPD is a kind of higher polymorphic DNA marker method .
4．The lowest amplified band frequency of Wanxi White goose and Yan goose is only 11.11% of all, the average of band frequency of all primes is 0.8236, 0.7442 separately. It is obvious that higher selecting (choosing) in both populations, which Wanxi White goose is a little higher than Yan goose. The average genetic comparability is 0.8645, 0.8205 and genetic diversity is 0.7250, 1.1649 respectively, it is seen that Wanxi White goose has a smaller genetic difference in population than Yan goose’s. The average genetic differentiation index is 0.7957 in both population, whose range is between 0.6785~0.8770, its average is 0.7957, the data indicated, in total genetic variation, over 79.57% variation consist in both population, 20.43% variation consist in individual population, therefore, genetic variation does not occur in individual population mostly, but in both population.
实验提取了46只皖西白鹅和48只雁鹅的血液基因组DNA，DNA的OD260/OD280值介于1.4~1.8之间，用作RAPD扩增基因组DNA的模板。对RAPD反应条件和程序逐个进行筛选，建立了最适宜的RAPD反应体系和反应程序。用RAPD技术分析皖西白鹅和雁鹅的基因组DNA的遗传多样性。研究结果表明:
1．用实验提取的基因组DNA对皖西白鹅和雁鹅的每个个体进行扩增，可以产生清晰，稳定的RAPD图谱。
2．筛选并建立了适合于皖西白鹅和雁鹅的扩增反应体系和程序：反应体积25μl，内含100ng的模板DNA，MgCl2（20mM）2.5μl，dNTP（5mM）1μl，随机引物（20μM）1μl，1U TaqDNA聚合酶。扩增程序为：94℃预变性4min，94℃变性1min，36℃退火1min，72℃延伸1min，42个循环后72℃延伸10min，于4℃结束反应。
3．用随机扩增多态性DNA技术对皖西白鹅和雁鹅进行遗传多样性分析，从50个随机引物中筛选出了12个引物对两个品种的所有个体的基因组DNA进行PCR扩增，扩增结果表明：50个随机引物中有12个引物在皖西白鹅及雁鹅基因组中分别扩增共获得83和85条条带，其中多态性条带分别为51条和57条，多态性条带率分别为61.44%和67.06%；碱基对数分别为0.21~1.52kb 和0.196~1.55kb之间，每个引物产生的条带数在2~9和3~9之间。
4．皖西白鹅和雁鹅扩增出来的条带的频率最低只有11.11%，所有引物的带频率的平均值分别为0.8236、0.7442，可以看出两群体的选育程度较高，皖西白鹅要略高于雁鹅；皖西白鹅及雁鹅群体内的遗传相似度分别平均为0.8645和0.8205，群体内的遗传多样性指数分别平均为0.7250和 1.1649，可见皖西鹅群体内个体间遗传差异较雁鹅小；两群体的遗传分化指数平均为0.7957，两群体遗传分化指数变化范围在0.6785~0.8770之间，平均值为0.7957，表明，在总遗传变异中，存在于群体间的变异达79.57%，群体内的变异仅为20.43%，显示，两群体的遗传变异主要发生在群体间，而不是群体内。
Goose breeding is a traditional homework in our country. The breeds of goose in our country have many merits such as producing many eggs, good feather and meat quality. This experiment analyzed on Wanxi White goose and Yan goose of RAPD markers in molecule level, to further provide scientific evidence for the breed resources research, selection and application on Wanxi White goose and Yan goose.
An experiment was conducted with 46 Wanxi White goose and 48 Yan goose.The OD value of Genomic DNA is between 1.4~1.8. Genomic DNA extracted from the blood of goose, which were directly used as template of PCR. RAPD reaction was carried out and programs were filtrated one by one, Optimal PCR conditions and programs were developed. RAPD is used to amplify genomic DNA of Wanxi White goose and Yan goose, their genetic diversity were also analyzed. The research results indicated:
1. Using conventional method to extract genomic DNA. The OD value of all the individual genomic DNA between 1.4~1.8. Amplified results indicated that it can produced explicit and steady RAPD bands, and testified that RAPD reaction did not request for template strictly.
2．The influencing factors of RAPD of Wanxi White goose and Yan goose genetic diversity research were studied and the experimental parameters were optimized. The optimal RAPD conditions are: 25μl reaction volume, containing 100ng template DNA, MgCl2(20mM)2.5μl，dNTP(5mM)1μl，primer (20μM)1μl，1UTaqDNA polymerase. Amplification program is : After initial denaturation at 94℃ for 4min, the amplification was performed by using 42 cycles of denaturation at 94℃ for 1min, annealing at 36℃ 1min, and extension at 72℃ for 1min. A final period of extention was carried out for 10min and final holding at 4℃.
3．Genetic diversity of Wanxi White goose and Yan goose was assessed through
Random Amplified Polymorphic DNA (RAPD) analysis. The total genomic DNA of all the individuals of both breeds were amplified through PCR method with 12 selected primers out of 50 arbitrary primers. The results suggested: Total 83，85 bands and 51, 57 polymorphic bands were acquired, which polymorphic rate is 61.44%, 67.06% respectively, The amplified base pair size were 0.21~1.52kb, 0.196~1.55kb and the number in each track were 2~9, 3~9 respectively, it also indicated that RAPD is a kind of higher polymorphic DNA marker method .
4．The lowest amplified band frequency of Wanxi White goose and Yan goose is only 11.11% of all, the average of band frequency of all primes is 0.8236, 0.7442 separately. It is obvious that higher selecting (choosing) in both populations, which Wanxi White goose is a little higher than Yan goose. The average genetic comparability is 0.8645, 0.8205 and genetic diversity is 0.7250, 1.1649 respectively, it is seen that Wanxi White goose has a smaller genetic difference in population than Yan goose’s. The average genetic differentiation index is 0.7957 in both population, whose range is between 0.6785~0.8770, its average is 0.7957, the data indicated, in total genetic variation, over 79.57% variation consist in both population, 20.43% variation consist in individual population, therefore, genetic variation does not occur in individual population mostly, but in both population.
引文
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