甘蓝游离小孢子培养及再生植株倍性鉴定研究  

Isolated Microspore Culture and Determination of Ploidy Level of Microspore-derived Plants in Cabbage( Brassica oleracea var. capitata)

作　　者：祁魏峥 颉建明[1] 郁继华[1] 康俊根[2,3] 

机构地区：[1]甘肃农业大学园艺学院,甘肃兰州730070 [2]北京市农林科学院蔬菜研究中心,农业部华北地区园艺作物生物学与种质创制重点实验室,北京100097 [3]北京京研益农科技发展中心,北京100097

出　　处：《西南农业学报》2015年第6期2381-2388,共8页Southwest China Journal of Agricultural Sciences

基　　金：‘十二五’国家科技支撑计划项目(2012BAD02B01;2012BAD50G01;2014BAD01B08);北京市科委(Z141105002314020)

摘　　要：对108个甘蓝品种进行游离小孢子培养,计算出胚率,分析表型与出胚难易的关系。经流式细胞仪对其再生植株进行倍性鉴定,计算自然加倍率。以流式细胞仪的结果为基准对再生植株叶片气孔保卫细胞周围叶绿体数目,保卫细胞长、宽以及周长规律进行研究。结果表明：具有圆球、早熟、春甘蓝3个特性的甘蓝品种易出胚,而扁球、中熟、晚熟以及越冬甘蓝不易出胚;再生植株的自然加倍率会因基因型的不同而有较大差异;不同倍性的再生植株叶片保卫细胞周围叶绿体平均个数所得规律为：单倍体6~9个,二倍体9~12个,四倍体12~23个,且随着倍性的增加,叶绿体个数分布范围更加广泛。保卫细胞长度变化范围为：单倍体18~23μm,二倍体25~35μm,四倍体35~45μm。保卫细胞周长变化范围为：单倍体60~75μm,二倍体75~95μm,四倍体100~115μm。保卫细胞宽度随气孔大小变化而差异较大,规律不明显。在保卫细胞长度和周长均能说明倍性的情况下,显然通过测量保卫细胞的长度来判定倍性的方法更为简便。The 108 cabbage varieties were used for the isolated microspore culture, the embryogenesis ratio of each variety was calculated, and the relationship between the phenotype and the embryogenesis was further analyzed. Flow cytometry were used to identify the ploidy level of microspore-derived plants, and then the natural doubling rate was calculated. The number of chloroplast surrounding the leaf stoma guard cells, the length, the width and the circumference of stoma guard cell in different ploidy level of microspore-derlved plants were studied. The results showed that the microspores of cabbage with the characteristics of spherical type, early-maturing variety, spring cabbage were easy to produce embryoids, but the cabbage with another characteristics like the oblate spheroidal style, middle-maturing and late-maturing variety, winter cabbage were difficult to produce embryoids; The natural doubling rate of the microspore-derived plant were largely different because of different 8enotypes ; The rule formed by the number of chlorep]asts surrounding the stoma guard cells in different mlcmspere-derlved plants was： the number of chloroplasts in haploid, diploid and tetmploid was 6 - 9, 9 - 12 and 12 - 23, respectively. The distribution range of the number of chloroplasts is more extensive as ploidy increased. The length range of guard cells in haploid, diploid and tetraploid was 18 - 23, 25 - 35 and 35 - 45 um, respectively. The perimeter range of guard cells in haploid, diploid and tetraploid was 60 - 75,75 - 95 and 100 -115 μm, respectively. The width of guard cells largely changes as stoma size changes, but there is no obvious rule here. The method for measurement of the length of stoma guard cells was proved to be a simple way of estimating ploidy of the microspore-derived plants in the situation which the length and the perimeter of the stoma guard cells both could estimate the ploidy.

关 键 词：甘蓝 游离小孢子培养 叶绿体 气孔保卫细胞 倍性鉴定 

分 类 号：S635.9[农业科学—蔬菜学]