四倍体大白菜游离小孢子培养技术  

The Culture Technique of Isolated Microspore in Tetraploid Chinese Cabbage

作　　者：刘晓东[1] 吴芳 孟川 牟金贵[1] 王玉海[1] 王明秋[1] LIU Xiaodong;WU Fang;MENG Chuan;MU Jingui;WANG Yuhai;WANG Mingqiu(Institute of Cash Crops of Hebei Academy of Agricultural and Forestry Sciences,Hebei Vegetable Engineering Technology Centre,Shijiazhuang 050051,China)

机构地区：[1]河北省农林科学院经济作物研究所,河北省蔬菜工程技术中心,石家庄050051

出　　处：《园艺学报》2023年第12期2641-2652,共12页Acta Horticulturae Sinica

基　　金：河北省重点研发计划项目(21326311D-1);河北省农林科学院创新工程项目(2022KJCXZX-JZS-3);现代农业产业技术体系建设专项(CARS-23-G17)。

摘　　要：四倍体大白菜减数分裂过程中产生的配子具有多样性,开展四倍体大白菜游离小孢子培养,旨在获得更多变异程度较大的纯合二倍体新资源。从14份早熟四倍体大白菜中筛选出4份易生胚的材料,通过花蕾表型、热激温度、生根激素、倍性鉴定等方面开展研究,建立了四倍体大白菜游离小孢子培养技术体系。研究结果表明:14份不同基因型四倍体大白菜,盛花期花蕾的花瓣长度与花药长度的比值介于0~0.5时,其单核靠边期的小孢子占比高,此时的出胚率最高;热激温度为33.5℃时暗培养24 h有利于小孢子胚胎发生;在生根培养基MS中添加吲哚丁酸(IBA)0.3 mg·L^(-1)和萘乙酸(NAA)0.1 mg·L^(-1),生根效果好。利用流式细胞仪、染色体镜检及杂交授粉3种方法检测倍性,39株再生植株只有二倍体和四倍体2种倍性,二倍体为37株,占94.87%,四倍体为2株,占5.13%。The gametes produced during the meiosis process of tetraploid Chinese cabbage are diverse.The purpose of carrying out free microspore culture of tetraploid Chinese cabbage is to obtain more new homozygous diploid resources with a large degree of variation.In this study,four easy embryogenic materials were selected from 14 early-maturing tetraploid Chinese cabbages.The technical system of isolated microspore culture of tetraploid Chinese cabbage was established in the aspects of flower bud phenotype,heat shock temperature,rooting hormone and ploidy identification.The results were as follows:For different genotypes of tetraploid Chinese cabbage,when petal/anther of flower buds was 0-0.5,the microspore was in the stage of mononuclear sideline,and the embryo rate was the highest at this time.When the heat shock temperature was 33.5℃and dark culture for 24 h,they were beneficial to microspore embryogenesis.For rooting medium,the combination of indole butyric acid(IBA)and naphthylacetic acid(NAA)was better than single hormone.The optimum concentration was IBA 0.3 mg·L^(-1) and NAA 0.1 mg·L^(-1).The ploidy was detected by flow cytometry,chromosome microscopy and cross pollination.The results showed that the probability of diploid was 94.87%and the probability of tetraploid was 5.13%in 39 regenerated plants.

关 键 词：大白菜 小孢子培养 四倍体 资源创制 

分 类 号：S634.1[农业科学—蔬菜学]