百喜草体细胞耐盐突变体筛选技术研究  

Screening technique of salt-tolerant somatic mutant in Paspalum notatum Flugge.

作　　者：叶晓青[1] 佘建明[1] 贾新平[1] 邓衍明[1] 梁丽建[1] 童红玉[1] 沈生元 沈耀明 

机构地区：[1]江苏省农业科学院农业生物技术研究所,江苏省农业生物学重点实验室,江苏南京210014 [2]江苏吴江市农业委员会,江苏吴江215200

出　　处：《江苏农业学报》2012年第6期1247-1252,共6页Jiangsu Journal of Agricultural Sciences

基　　金：江苏省科技厅科技支撑项目(BE2010339)

摘　　要：为了提高体细胞耐盐突变体筛选的效率,本试验研究不同浓度氯化钠(NaCl)对百喜草愈伤组织分化和植株再生的影响。在琼脂浓度为8 g/L和16 g/L的继代培养基上,以百喜草品种太阳花的成熟种子分别诱导产生湿润、粘连和干燥、松散2种类型的颗粒状愈伤组织。在NaCl浓度为0(对照)、1.2%、1.6%、2.0%的培养基上分别筛选30 d,干燥、松散颗粒状愈伤组织的绿芽分化率分别为36.1%、20.4%、19.0%和9.3%,绿芽的植株再生率分别为52.6%、46.3%、30.0%和20.6%;湿润、粘连颗粒状愈伤组织的绿芽分化率分别为15.3%、3.6%、0.7%和0,绿芽的植株再生率分别为25.8%(对照)、16.7%(1.2%NaCl)和0(1.6%NaCl)。表明干燥、松散的颗粒状胚性愈伤组织适宜被选做体细胞耐盐突变体筛选材料。SSR分子标记检测结果显示,479对引物组合中有8对引物在百喜草田间实生苗、组织培养再生植株和不同NaCl浓度(1.2%、1.6%、2.0%)筛选得到的再生植株混合株之间PCR扩增存在多态,其中2对引物WXE-30和WXE-246适用于耐盐突变体分子鉴定。In order to improve the screening efficiency of somatic mutant with salt tolerance,the effect of NaCl concentrations on callus differentiation and plant regeneration were studied.The mature seed of Paspalum notatum Flugge.cv.Taiyanghua was used as the explant material.The dry-and-loose pellet callus were induced in the subculture medium with 16 g/L agar,and the wet-and-sticky pellet callus in 8 g/L agar.The two types of pellet callus were salt-stressed in the callus subculture medium with 0,1.2%,1.6% and 2.0% NaCl for 30 d.The results showed that the rates of bud regeneration of dry-and-loose pellet callus were 36.1%,20.4%,19.0% and 9.3%,and the frequencies of shoot regeneration from bud were 52.6%,46.3%,30.0% and 20.6%.The rates of bud regeneration of the wet-and-sticky pellet callus were 15.3%,3.6%,0.7% and 0,and the frequencies of shoot regeneration from bud were 25.8%（control）,16.7%（1.2% NaCl）,and 0（1.6% NaCl）.The dry-and-loose pellet callus was more suitable for screening of somatic mutant with salt tolerance.Among 479 pairs of specific simple sequence repeat（SSR） primer,eight pairs were associated with the salt-tolerance of somatic mutants,and two pairs,WXE-30 and WXE-246,were suitable for identification of the mutant with salt tolerant in Paspalum notatum.

关 键 词：百喜草 颗粒状愈伤组织 体细胞突变体 耐盐 SSR标记 

分 类 号：S567.23[农业科学—中草药栽培]