中国野生葡萄抗黑痘病基因RAPD标记的克隆及测序

Cloning and sequence of RAPD marker linked to anthracnose resistant gene in wild grapes native to China

用 Wizard DNA Clean- up System试剂盒将与中国野生葡萄抗黑痘病基因相连锁的 RAPD遗传标记 OPV0 2 - 6 0 0回收纯化 ,连接于 T- Easy Vector并克隆测序 ,得到了 OPV0 2 - 6 0 0的全序列 ,其长度为 6 16 bp。将两端序列设计特异 PCR扩增引物作为合成中国野生葡萄抗黑痘病检测探针的基础 ,用于检测葡萄抗黑痘病种质和标记辅助育种。

Randomly amplified polymorphic DNA (RAPD) was employed to detect molecular markers linked to anthracnose ( Spheceloma ampelinum de Bary) resistant gene in the wild grapes ( vitis L.) native to China.RAPD marker OPV02 600 was linked to anthracnose resistant gene using 90-3 cross F 1 V.quinquangularis Rehd (shang 24)× V.vinifera (Longyan).OPV02 600 fragment was reclaimed by Wizard DNA Clean up System from electrophoresis gel and cloned in T easy Vector,then sequenced from two sides.The DNA fragment OPV02 600 was actually 616 bp.It is suggested that the sequence of RAPD marker OPV02 600 might be used as a basis for synthesizing the specific PCR primers and the probe for detecting grape anthracnose resistant in disease resistant breeding and molecular marker assisted selection(MAS).