秋水仙素离体双层诱导同源四倍体青花菜的研究  

Study on in vitro double-deck induction of autotetraploids in broccoli by colchicines

作　　者：郑于莉 张蜀宁[1] 王雅美[1] 徐丽娟[1] 侯喜林[1] 

机构地区：[1]南京农业大学园艺学院/作物遗传与种质创新国家重点实验室,江苏南京210095

出　　处：《南京农业大学学报》2014年第6期13-18,共6页Journal of Nanjing Agricultural University

基　　金：江苏省科技支撑计划项目(BE2011313)

摘　　要：研究秋水仙素离体双层培养诱导对3种青花菜染色体加倍的诱导效应,建立青花菜四倍体的离体双层培养诱导体系,为植物倍性育种提供新途径。以子叶期保留下胚轴0.4-0.6 cm的子叶为外植体,接入含不同浓度的秋水仙素离体双层培养基（上层为含秋水仙素的液体培养基,下层为不含秋水仙素的固体培养基）中诱导24、48和96 h。结果表明：以300mg·L^-1秋水仙素处理48 h诱导获得四倍体的效果最佳,LQ-2、S139-2和LQ-23三个材料外植体存活率达100.00%,增殖系数分别为2.00、2.33和2.67,四倍体诱导率分别为66.67%、71.42%和62.50%。与二倍体相比,四倍体植株在株高、叶片、花球等性状上增大,气孔大小和密度呈现显著差异,LQ-2、S139-2和LQ-23三个材料的花粉粒增大,增幅分别为18.85%×63.71%、25.00%×34.68%和3.36%×61.86%。四倍体根尖染色体为2n=4x=36。流式细胞仪鉴定发现二倍体DNA相对含量为300,四倍体为600。结论：通过大量试验,找到了秋水仙素双层诱导青花菜的最佳秋水仙素浓度和诱导时间,提高了诱导率,获得了3个同源四倍体青花菜新材料。We studied the effects of the double-deck induction with different colchicine concentration and treatment time duration in three species of broccoli,established the in vitro double-deck induction system of tetraploid broccoli,provided a new way for the ploidy breeding,meanwhile it was also significant to increase the diversity of germplasm resources. In this study,diploid cotyledons with hypocoty of 0. 4-0. 6 cm length of the broccoli were used as explants. After the explants were pre-cultured for two days,they were transferred to the double-deck media which combined liquid medium including different colchicine concentrations with solid media without colchicine to induce autotetraploids. During the process,the colchicine concentration was 100,300 and 450 mg·L^-1,at the same time,treatment time duration was 24,48 and 96 h. After treatment for the specified period,explants were washedthree times by distilled water and transferred to solidified media to proliferation. Morphological characters,chromosome number and flow cytometry techniques were used to identified ploidy. The results showed that 239 of 243 treated explants were alive and the survival rate was 98. 35%. The propagation coefficients of LQ-2,S139-2 and LQ-23 were 1. 11-3. 38,1. 00-3. 22,1. 00-3. 33. 178 of the 511 plants derived from the double-deck induction were tetraploid. Treating with 300 mg·L^-1colchicines for 48 h could get the best results,explants from LQ-2,S139-2and LQ-23 had high survival rate,which reached to 100. 00%,and coefficient propagation were 2. 00,2. 33 and 2. 67 respectively,meanwhile the rate of induced tetraploids reached to 66. 67%,71. 42% and 62. 50%. Compared with the diploid,the tetraploid exhibited larger,rounder and thicker leaves. The size of tetraploid leaves for LQ-2,LQ-23 and S139-2 was 43. 33 cm×23. 83 cm,40. 40 cm×25. 17 cm and 39. 13 cm×22. 33 cm. Plant height,ball-flower diameter and single head weight of tetraploid increased in different degrees,and plant height,ball-flower diameter and single head weight of te

关 键 词：青花菜 秋水仙素 双层培养 离体诱导 四倍体 

分 类 号：S635.3[农业科学—蔬菜学]