浙江农业学报
浙江農業學報
절강농업학보
ACTA AGRICULTURAE ZHEJIANGENSIS
2014年
1期
14-19
,共6页
孙凤梅%沈晓霞%沈宇峰%张启坤%王志安
孫鳳梅%瀋曉霞%瀋宇峰%張啟坤%王誌安
손봉매%침효하%침우봉%장계곤%왕지안
薏苡%分子标记%AFLP体系
薏苡%分子標記%AFLP體繫
의이%분자표기%AFLP체계
Coix%molecular markers%AFLP system
以30个不同薏苡种质为试验材料,利用AFLP分子标记技术进行遗传多样性及亲缘关系的研究。通过对AFLP常规流程中(基因组提取、酶切、连接、预扩增、选择性扩增、电泳及银染检测)关键因素进行优化,建立适合薏苡的AFLP扩增检测体系。结果表明,采用酶切与连接分开的两步法,酶切完全,每步反应时间4 h并附加15 min的酶失活时间;连接产物稀释10倍;预扩增产物稀释50倍,试验效果最佳。应用优化后体系,从64对引物中筛选出14对多态性较好的薏苡AFLP选择性扩增引物。
以30箇不同薏苡種質為試驗材料,利用AFLP分子標記技術進行遺傳多樣性及親緣關繫的研究。通過對AFLP常規流程中(基因組提取、酶切、連接、預擴增、選擇性擴增、電泳及銀染檢測)關鍵因素進行優化,建立適閤薏苡的AFLP擴增檢測體繫。結果錶明,採用酶切與連接分開的兩步法,酶切完全,每步反應時間4 h併附加15 min的酶失活時間;連接產物稀釋10倍;預擴增產物稀釋50倍,試驗效果最佳。應用優化後體繫,從64對引物中篩選齣14對多態性較好的薏苡AFLP選擇性擴增引物。
이30개불동의이충질위시험재료,이용AFLP분자표기기술진행유전다양성급친연관계적연구。통과대AFLP상규류정중(기인조제취、매절、련접、예확증、선택성확증、전영급은염검측)관건인소진행우화,건립괄합의이적AFLP확증검측체계。결과표명,채용매절여련접분개적량보법,매절완전,매보반응시간4 h병부가15 min적매실활시간;련접산물희석10배;예확증산물희석50배,시험효과최가。응용우화후체계,종64대인물중사선출14대다태성교호적의이AFLP선택성확증인물。
By taking 30 samples of Coix species as testing materials , the genetic diversity and genetic relationship was analyzed using AFLP molecular markers technology .The optimal analysis system of AFLP has been established by studying the key factors of conventional process such as gene extraction , restriction digestion and link up , pre-am-plication, selective amplification, electrophoresis and siliver staining detection .The result showed that the genome DNA was digested more completely by using two-step method compared with one-step method.In each step, the en-zyme reaction time was 4 h added with 15 min for enzyme inactivation .The connection product should be diluted for 10 times and the pre-amplification product should be diluted for 50 times to get a better result .With application of the es-tablished system, 14 selective amplification primers with better polymorphism were selected out from 64 primers.