菜心小孢子胚再生体系优化及植株倍性鉴定  

Optimization of Microspore Embryo Regeneration System and Ploidy Identification of Chinese Flowering Cabbage

作　　者：庞强强[1] 周曼[1] 蔡兴来 张文[1] 孙晓东 陈贻诵 Pang Qiangqiang;Zhou Man;Cai Xinglai;Zhang Wen;Sun Xiaodong;Chen Yisong(Hainan Provincial Engineering Research Center for Melon and Vegetable Breeding,Hainan Provincial Key Laboratory for Vegetable and Biology,Hainan Vegetable Academician Team Innovation Center,Vegetable Research Institute for Hainan Provincial Academy of Agricultural Sciences,Haikou,571100)

机构地区：[1]海南省农业科学院蔬菜研究所,海南省蔬菜学院士团队创新中心,海南省蔬菜生物学重点实验室,海南省瓜菜育种工程技术研究中心,海口571100

出　　处：《分子植物育种》2021年第14期4745-4751,共7页Molecular Plant Breeding

基　　金：海南省自然科学基金项目(318QN293);国家现代农业产业技术体系建设专项资金项目(CARS-23-G52);海南省重大科技计划项目(ZDKJ2017001)共同资助。

摘　　要：为进一步优化菜心游离小孢子胚再生技术体系,本研究以3份不同基因型的菜心小孢子胚状体为试验材料,研究不同培养基类型、培养基中不同琼脂浓度和不同胚龄对胚再生的影响,并对再生植株进行倍性鉴定。结果表明,使用B5培养基时各基因型的成苗率均高于MS培养基,褐化率低于MS培养基,B5培养基是菜心小孢子胚状体再生的适宜培养基;培养基中添加适宜浓度的琼脂可提高再生植株的成苗率,随着琼脂浓度的增加,出苗率呈先升后降的趋势,琼脂浓度为1.0%时,各基因型的出苗率达到最高;胚状体再生的最佳胚龄是25~29 d,胚龄时间过长或过短均不利于胚状体出苗,其中40 d以上胚龄的胚状体几乎不出苗;倍性鉴定发现菜心小孢子再生植株中同时存在单倍体、二倍体和多倍体,3个基因型的二倍体自然加倍率在62.22%~71.11%之间;75 mg/L秋水仙素处理可提高菜心再生株系的二倍体率。本研究为菜心游离小孢子胚再生体系优化及再生植株倍性鉴定提供了技术基础。In order to optimize the microspore embryo regeneration technology system,the effects of different medium,agar concentration and embryonic age on the regeneration of embryonic were studied with 3 genotypes of Chinese flowering cabbage,and to identify the ploidy of the regenerated plants.The results showed that the seedling formation rate of each genotype was higher than MS medium,and the browning rate was lower when using B5 medium.B5 medium was an appropriate medium for the regeneration of microspore embryoids in Chinese flowering cabbage.The seedling rate of regenerated plants could be increased by adding appropriate concentration of agar in the medium.The emergence rate showed a trend of first increasing and then decreasing with the increase of agar concentration.The emergence rate of each genotype reached the highest when the agar concentration was 1.0%.The optimal embryonic age for embryoid regeneration was 25~29 d.Too long or too short the embryonic age was not conducive to the emergence of embryoid seedlings,and the embryoids with more than 40 days of embryo age hardly appeared.Ploidy identification showed that haploid,diploid and polyploid in the microspore regenerated plants of Chinese flowering cabbage,and the rates of natural diploid doubling were 62.22%to 71.11%.75 mg/L colchicine treatment could improve the diploid rate of regenerated lines.This study provided a technical basis for the optimization of the regeneration system and the ploidy identification of regenerated plants in Chinese flowering cabbage.

关 键 词：菜心(Brassica campestris L.) 小孢子培养 再生体系 倍性鉴定 

分 类 号：S634.5[农业科学—蔬菜学]