利用PPT或Kan叶面喷施法建立番茄转基因阳性苗筛选技术研究  

Studies on Establishing Technology for Screening Transgenic Positive Tomato Seedlings Using Foliar Spraying Method PPT or Kan

作　　者：邓瑞 潘玉朋[1] 杨羽清 崔海兵 郑玉洁 程智慧[1] DENG Rui;PAN Yupeng;YANG Yuqing;CUI Haibing;ZHENG Yujie;CHENG Zhihui(College of Horticulture,Northwest A&F University,Yangling 712100,Shaanxi,China)

机构地区：[1]西北农林科技大学园艺学院,陕西杨凌712100

出　　处：《中国蔬菜》2023年第5期56-64,共9页China Vegetables

基　　金：国家自然科学基金项目(31471865)。

摘　　要：以三叶一心期Micro-Tom幼苗为试材,叶面喷施不同浓度PPT或Kan后采用RLA法评估叶片受伤害程度,划分分级标准,确定PPT或Kan筛选的适宜浓度,建立转基因抗性苗筛选体系,并用带有bar或nptⅡ的转基因T_(1)番茄幼苗进行抗性筛选验证。结果表明,PPT和Kan对Micro-Tom番茄幼苗的伤害均可分为7级;PPT浓度为1.50~1.75mg·L^(-1)、Kan浓度为120~150mg·L^(-1)时,番茄植株反应明显,且不会产生不可逆伤害,可作为转基因植株抗性苗筛选的适宜浓度;T_(1)转基因番茄幼苗的叶面喷施PPT或Kan抗性筛选结果与利用标记基因bar或nptⅡ的PCR克隆验证结果高度一致。Taking 3-leaf stage Micro-Tom tomato seedlings as test material,this experiment assessed the damage extent of leaf blade by using RLA method after foliar spraying diff erent concentrations PPT or Kan solutions;classifi ed grading standard;determined the appropriate concentration for PPT or Kan screening;established a screening system for transgenic resistant seedlings;and conducted resistant screening verifi cation using T_(1) transgenic tomato seedlings with bar or nptⅡ.The results showed that the damages on Micro-Tom tomato seedlings by PPT and Kan all could be divided into 7 grades.When PPT concentration was 1.50-1.75 mg·L^(-1),Kan concentration was 120-150 mg·L^(-1),the response of tomato plants was obvious.Moreover,it would not cause irreversible damage and could be the suitable concentration for screening resistant seedlings of transgenic plants.The screening results of foliar PPT or Kan on T_(1) transgenic tomato seedlings were highly consistent with PCR clone verifi cation results using marker gene bar or npt Ⅱ.

关 键 词：Micro-Tom番茄 转基因 PPT Kan 阳性株筛选 

分 类 号：S641.2[农业科学—蔬菜学]