水稻农垦58与农垦58s的RAPD和ISSR分析  

RAPD and ISSR Analysis between Photoperiod Sensitive Genie Male Sterile Rice Nongken 58s and its Original Variety Nongken 58

作　　者：江树业[1] 陈启锋[2] 方宣钧[1] 

机构地区：[1]中国农科院生物技术研究所,北京100081 [2]福建农业大学作物遗传育种研究所,福州350002

出　　处：《农业生物技术学报》2000年第1期63-66,共4页Journal of Agricultural Biotechnology

基　　金：国家863计划!Z16-02-03;福建省自然科学基金!C97027

摘　　要：以光敏核不育水稻农垦５８ｓ及其原始株农垦５８核ＤＮＡ为模板，进行了ＲＡＰＤ和ＩＳＳＲ多态性分析，结果表明：在可扩增的 １５４个ＲＡＰＤ引物和７８个 ＩＳＳＲ引物中， ８个ＲＡＰＤ引物和７个 ＩＳＳＲ引物的扩增产物表现出 ＤＮＡ多态性，ＲＡＰＤ－ＰＣＲ产物的多态性标记分别出现在农垦５８ｓ，农垦５８或二者同时出现，而ＩＳＳＲ－ＰＣＲ扩增产物的多态性标记只出现在农垦５８或农垦５８ｓ中。获得多态性ＲＡＰＤ标记及ＩＳＳＲ标记分别为２３个和１３个，其中１０个ＲＡＰＤ扩增片段和７个ＩＳＳＲ扩增片段为农垦５８ｓ所特有。本研究对ＲＡＰＤ－ＰＣＲ和ＩＳＳＲ－ＰＣＲ扩增产物采用非变性的聚丙烯酰胺凝胶电泳，结合快速银染法染色，与琼脂糖胶电泳，ＥＢ染色相比，大大提高了分辨率，值得推广应用。本文还对获得的ＭＰＤ及ＩＳＳＲ多态性片段进一步通过ＢＳＡ分析，或用转育前后的ｍＲＮＡ作Ｎｏｒｔｈｅｒｎ杂交分析加以鉴定等问题作了探讨。RAPD and ISSR polymorphisms between photoperiod sensitive genie male sterile rice Nongken 58s and its original variety Nongken 58 were analyzed with their genomic DNAs as PCR template. The results are as following: 154 RAPD and 78 ISSR primers generated amplified products in RAPD and ISSR analyses. Among them, 8 RAPD and 7 ISSR primers produced polymorphic bands. Among the polymorphic bands by RAPD-PCR. some of them are particular with Nongken 58s, some of them are connected with Nongken 58, and some of them appeared both in Nongken 58s and in Nongken 58. However. ISSR polymorphic bands appeared only in either Nongken58 or Nongken58s. There are 23 specific RAPD bands and 13 ISSR specific bands in total, among which 10 specific RAPD bands and 7 specific ISSR bands only appeared in Nongken58s. Undenatured polyacrylamide gel electrophoresis and rapid silver staining were employed to isolate and show RAPD and ISSR products. This method is considered suitable for being popu larized for that it is convent ent, quick and it dramatically promotes the resolution compared with agarose gel electrophoresis and EB staining. Some other relative problems are also discussed in this paper. and we think that polymorphic DNA fragments can be identified by BSA analysis or by Northern blotting with sterile and fertile mRNA produced during the transformation of pollen fertility.

关 键 词：光敏核不育 水稻 RAPD ISSR 种质资源 

分 类 号：S511.03[农业科学—作物学] S511.024