寒兰离体化学诱变效果的ISSR分析  

ISSR analysis of the mutagenic effect on Cymbidium kanranin vitro

作　　者：郑改笑 李夏 熊冬金[1,2] 杨柏云 

机构地区：[1]南昌大学生命科学学院 [2]江西省植物资源重点实验室,江西南昌330031

出　　处：《南昌大学学报（理科版）》2017年第3期255-258,共4页Journal of Nanchang University(Natural Science)

基　　金：国家自然科学基金资助项目(31260485);江西省农业科技支撑项目(20123BBF6076);江西省自然科学基金(20122BAB204010)

摘　　要：诱变育种中诱变剂的筛选与诱变效果评估是诱变育种成功的关键步骤之一。用7种化学诱变剂甲基磺酸乙酯(EMS)、硫酸二乙酯(DES)、秋水仙素(colchicines)、咖啡碱(caffeine)、5-溴尿嘧啶(5-BU)、马来酰肼(MH)、亚硫酸钠(Na_2SO_3)处理寒兰(Cymbidium kanran)的原球茎,经培养成再生植物株后,对其叶片进行ISSR分子标记分析。结果表明:诱变处理后的再生植株和正常植株之间的基因组ISSR扩增条带均有不同程度的增加和缺失,其中DES、EMS、5-BU三种诱变剂处理的再生植株扩增条带数变幅较秋水仙素、咖啡碱、马来酰肼和亚硫酸钠四种诱变剂丰富,EMS、DES和5-BU均缺失了条带1、3、4,EMS增加条带6,DES和5-BU均增加了条带6、7、10,DES还增加了条带8、9,这表明DES、EMS、5-BU 3种诱变剂诱变效果明显,比较适合作为寒兰的离体化学诱变剂,其中DES更适合,研究结果可为寒兰的诱变育种及其分子鉴定提供参考。The screening of mutagenic agents and the evaluation of mutagenic effects are key steps to the success of mutation breeding.Protocorms for Cymbidium kanran were treated with seven chemical mutagens,such as ethyl methanesulfonate（EMS）,diethyl sulfate（DES）,colchicines,caffeine,5-bromouracil（5-BU）,maleic hydrazide（MH）and sodium sulfite（Na2SO3）.The leaves of regeneration plants for the protocorms were analyzed by ISSR molecular markers.The results showed that there were increase and deletion for ISSR amplified bands between regeneration plants and CK plants in different degrees.The number of amplified bands of the regeneration plants of DES,EMS and 5-BU mutants was richer than that one of colchicine,caffeine,maleic hydrazide and sodium sulfite.More specifically,EMS,DES and 5-BU all missed the bands 1,3,4while EMS increased the band 6;DES and 5-BU increased the bands 6,7,10 while DES also increased the bands 8,9.This indicates that the effect of mutagen for DES,EMS,5-BU mutagen mutation was more suitable as an isolated chemical,and that the mutagenic effect of the three mutagen,DES,EMS,and 5-BU is obviously and are relatively more suitable for chemical mutagens for C.kanranin vitro.The results can provide reference for mutagenesis breeding and molecular identification.

关 键 词：寒兰 化学诱变 原球茎 ISSR 

分 类 号：Q949[生物学—植物学]