马铃薯茎尖脱毒方法优化及病毒检测  

Optimization of Virus-Free of Shoot Tip and Virus Detection of Test-Tube Plantlet of Potato

作　　者：卢艳丽[1,2] 周洪友[2] 张笑宇[2] 

机构地区：[1]内蒙古自治区排污权交易管理中心,内蒙古呼和浩特010011 [2]内蒙古农业大学农学院,内蒙古呼和浩特010019

出　　处：《作物杂志》2017年第1期161-167,共7页Crops

基　　金：国家自然科学基金(31460468);公益性行业(农业)科研专项(201503109)

摘　　要：为优化马铃薯茎尖脱毒方法,得到无毒壮苗,研究生长素及L-半胱胺酸盐酸盐、培养基和马铃薯芽灭菌时间对28个马铃薯品种(系)组培苗形成的影响,比较不同p H的MS培养基上苗的生长情况,并用RT-PCR方法进行病毒检测。结果表明,28个马铃薯品种(系)中,有的很快长出愈伤组织并形成幼苗,有的不能成苗。含有萘乙酸(NAA)的MS培养基愈伤组织出愈速度快且大、成苗慢、成苗率低,但是苗较壮。含有吲哚乙酸(IAA)的MS培养基多数不形成或形成很小的愈伤组织,成苗快、成苗率较高,但苗较弱。L-半胱胺酸盐酸盐能减轻愈伤组织褐变,提高成苗率。试管分装培养基115℃条件下最佳灭菌时间是20min。采用升汞对较小的马铃薯芽灭菌5~7min效果最好,对较大的芽灭菌9~10min效果最好。苗生长的MS培养基最佳p H为5.8。采用RT-PCR检测到试管苗马铃薯S病毒(PVS)、马铃薯Y病毒(PVY)和马铃薯卷叶病毒(PLRV)的带毒率分别为18%、12%和5%。In order to optimize virus-free technology of potato shoot tip, and to obtain strong virus-free seedlings, the different potato varieties, auxin, L-cysteamine hydrochloride, sterilization time of medium and potato shoots were studied in the experiment. The pH of MS medium was compared for seedling growth and the virus was detected with RT-PCR. Results showed that among 28 potato varieties, some rapidly grew callus and developed into seedlings, others did not. In MS medium containing NAA, the callus formation was faster and bigger, seedling formation was slower and less, but seedlings were stronger. In MS medium containing IAA, no or small calluses were produced, the seedlings formed faster and more, but seedlings were weaker. L-cysteamine hydrochloride could reduce callus brown, increased seedling emergence rate. Sterilization time 20min at 115 ℃ was the best for medium in tubes. 5-7rain and 9-10min with 0.1% mercuric chloride were the best sterilization time for small shoots and big shoots, respectively. The optimum pH of MS medium for seedling growth was 5.8. PVS of 18%, PVY of 12% and PLRV of 5% were detected in different variety seedlings.

关 键 词：马铃薯 茎尖脱毒 L-半胱氨酸盐酸盐 生长素 培养基 RT-PCR 病毒检测 

分 类 号：S435.32[农业科学—农业昆虫与害虫防治]