【作者】 鲁云风；

【导师】 原新明； 原国辉； 文祯中；

【作者基本信息】 河南农业大学 ， 农业昆虫与害虫防治， 2005， 硕士

【摘要】 本实验利用毛色、尾色形态标记和RAPD分子标记两种手段对南阳市境内南阳黄牛种质资源现状随机抽样调查,进行遗传分析,并依据调查结果和实地走访资料制定科学的保护策略。 选取2头纯种南阳牛为标准对照,随机抽取新野境内牛35头,方城境内牛45头,邓州境内牛32头为调查样本,对每个调查样本均做毛色、尾色记录,并数码拍照备案,静脉抽血加抗凝剂,-70℃保存。 利用毛色、尾色形态标记,以纯种南阳牛为标准,分别与3市（县）内调查样本聚类,凡毛色、尾色均与纯种南阳牛一致的个体牛,可以认为是南阳本地牛,否则为杂交牛,结果显示,新野境内南阳本地牛仅占14.29%,方城境内南阳本地牛只占33.33%,邓州境内南阳本地牛只占37.50%。 提取所调查牛基因组DNA,紫外分光光度法检测A260/A280值为1.75,表明DNA纯度较好,用Marker λDNA/HindⅢ作为分子量标准,琼脂糖凝胶电泳检测DNA片段大小为23Kbp左右,符合RAPD扩增的要求。选20条随机引物对南阳种牛、新野境内牛、方城境内牛及邓州境内牛共114个个体基因组DNA进行了随机扩增多态DNA（RAPD）分析,每次RAPD扩增,均设空白对照,避免了实验的假阳性,重复2-3次,其中16个引物得到的扩增结果比较稳定,重复性较好,且均具有多态性,分别是:OPG_01-04,OPG_06-10,OPG_12-18,单个引物获得的RAPD标记在1-6之间,在南阳种牛、新野境内牛、方城境内牛和邓州境内牛个体间可检测到不同扩增带型。16条具多态性随机引物RAPD扩增纯种南阳牛和调查样本牛基因组DNA,根据RAPD电泳条带,凡条带清晰或条带较模糊但重复性好的记为“1”,否则记为“0”,运用SPSS软件,分别输入每个调查市（县）个体牛和南阳种牛对应扩增条带“1”和“0”进行聚类,生成系统聚类图,3市（县）境内牛分别与纯种南阳牛进行聚类分析,和纯种南阳牛聚在一类的,可认为是南阳本地牛,否则为杂交牛,聚类图显示,新野境内南阳本地牛仅占14.29%,方城境内调查样本牛遗传背景较为一致,邓州境内南阳本地牛只占18.79%. 两种调查手段得出的结果虽然有些差异,但均揭示南阳境内本地牛数量偏少,受外来种影响严重,保种形势十分严峻,为保护南阳黄牛这一优良品种资源,依据实地走访资料及调查结果,针对南阳黄牛种质资源的现状,参考相关资料,提出建立南阳黄牛纯种繁育体系、建立南阳黄牛保种体系、建立南阳牛保种群、建立社会保种区、建立南阳牛冻精冻胚保种基因库、开展本品种选育、加强品系繁育推广工作、建立开放式育种体系、大力发展生态养牛业等9条保种策略。更多还原

【Abstract】 The status quo of NanYang yellow cattle breed was investigated in the way of random sample using the morphological markers such as fur color, tail color and RAPD and the scientific strategies for NanYang yellow cattle was established according to the investigation results and the correlative data. 35 cattle in XinYe,45 cattle in FangCheng, 32 cattle in DengZhou was sampled randomly and 2 head of NanYang breed cattle was selected as control .Every cattle was taken photo and it’s fur color and tail color was recorded; The cattle’s blood was exsanguinated from the vain and anticoagulant was added into it .At last it was stored at -70℃ .The cattle in 3 cities (county) was clustered respectively with 2 NanYang breed cattle using the morphological markers. The cattle can be seen as NanYang cattle if the color of fur and tail is all accord with NanYang breed cattle ,or else, it is crossbreed cattle. The result shows that NanYang cattle’s percentage in XinYe is onlyl4.29 percent, NanYang cattle’s percentage in FangCheng is 33.33 percent and 37.50 percent in DengZhou.The cattle’s DNA was extracted and it’s A260/280 is 1.75 examined through ultraviolet spectrophotometer. The number of DNA length is 23Kbp by agar gels electrophoresis, it shows that the DNA is according with the request of RAPD. The genetic DNA of random samples and NanYang breed cattle was amplified using 20 order of random primers. If the strip is clear or repeatedly well "1"was recorded; If there is no strip"0" was recorded. The random sample cattle in 3 cities (county) was clustered respectively with 2 NanYang breed cattle using SPSS.The cattle can be seen NanYang cattle if it is clustered with NanYang breed cattle in the dengrogram,or else, it is crossbreed cattle. The dengrogram shows that更多还原