^(60)Coγ辐射诱导马铃薯(Solanum tuberosum)基因组变异的SRAP标记分析(英文)

SRAP Marker Analysis of Genomic Mutation Induced by ^(60)Coγ-Ray Irradiation in Potato (Solanum tuberosum)

[目的]探讨60Coγ射线对马铃薯(Solanum tuberosum)染色体的诱变效应。[方法]采用不同辐照剂量的γ射线对费乌瑞它微型薯进行辐射处理,利用SRAP分子标记,对不同处理的VM1代及VM2代基因组变异进行分析。[结果]VM1代分析结果表明,88对引物有30对表现多态性,引物多态性比率为34.1%,共获得225条扩增条带,其中64条为多态性条带,条带多态性比率为28.4%。多态性表现形式有条带增加和缺失。选择25对有多态性的引物对VM2代进行扫描,5对显带不良,20对中有9对表现多态性,获得多态性条带9条,其中仅4条在VM1中检测到,其余5条是新增的缺失条带。VM1代的多态性条带在VM2代中的检出率仅为9.8%。选择表现稳定、清晰的9条多态性条带进行回收、克隆、测序,获得6个片段的DNA序列。经比对分析,5个片段序列与马铃薯染色体组部分片段的相似性为77%~89%,其中3个序列位于抗性基因簇中;另1个片段序列与番茄第5染色体的部分序列相似性为93%。[结论]γ辐照可以导致马铃薯基因组DNA发生变异,多态性条带数与辐照剂量之间没有明显的相关性,变异的多态性条带缺失数高于条带增加数。

[Objective] This study was to explore the mutagenic effect of 60Co γ-ray on Solanum tuberosum chromosomes.[Method] The 60Co γ-ray at different irradiation doses was applied to treat the minitubers of potato variety ＇Favorita＇,and the genomic mutations in VM1 and VM2 gene rations in treatments with different doses were analyzed by SRAP markers.[Resalt] Thirty pairs of SRAP primers out of 88 pairs exhibited polymorphism with a rate of 34.1％ in the bulked selection of VM1 generation.A total of 225 bands were obtained,of which 64 were polymorphic with a rate of 28.4％.The polymorphism was reflected in the forms of deleted bands and added bands.Based on the result of the bulked selection of VM1 generation,25 primers with polymorphism were selected to scan VM2 generation.Five primers performed poorly,and nine of the rest 20 pairs revealed polymorphism and obtained nine polymorphic bands,of which only four bands were detected in VM1 generation and the other five ones were newly deleted bands.Only 9.8％ of the bands detected in VM1 generation were obtained in VM2 generation.Eventually,nine stable and dear polymorphic bands were recovered and cloned,and DNA sequences of six bands of them were acquired by sequencing.According to the comparative analysis,five fragments sequences were similar to potato chromosome with a similarity rate of 77％-89％,three of them located at the resistance gene cluster; another one fragment had a similarity of 93％ with some regions of the No.5 chromosome in tomato.[Conclusion] 60Co γ-ray irradiation can cause mutation of genomic DNA in potato;there is no significant correlation between the number of polymorphic bands and the irradiation dose; potymorphic bands are characterized by a larger number of deleted bands than that of the added ones.