渗透胁迫对大花萱草EMS离体诱变系统的影响  

Osmotic Stress: Effects on in vitro Mutagenesis System of EMS in Hemerocallis hybrida

作　　者：杨丽莉[1] 刘化涛[1] 杨睿[1] 常建忠[1] Yang Lili;Liu Huatao;Yang Rui;Chang Jianzhong(Institute of Dryland Farming,Shanxi Academy of Agricultural Sciences,Taiyuan 030032)

机构地区：[1]山西省农业科学院旱地农业研究中心

出　　处：《中国农学通报》2019年第30期71-77,共7页Chinese Agricultural Science Bulletin

基　　金：山西省重点研发计划“甲基磺酸乙酯(EMS)离体诱导大花萱草抗病突变体的研究”(201603D221017-1);山西省农业科学院重点项目“EMS诱变大花萱草抗病突变体的研究”(YZD1513);山西省农业科学特色农业攻关项目“EMS诱变大花萱草突变体的产生及新种质资源创制”(YGG17037)

摘　　要：旨在为萱草属植物的EMS诱变提供一种预处理技术方法。以‘红运’萱草带芽愈伤组织为材料,采用高渗透固体培养基(MS+甘露醇36 g/L+山梨醇36 g/L+蔗糖50 g/L)在1.0%EMS半致死剂量诱变前分别进行0、30、60、90 min的渗透胁迫预处理。测定24 h的相对电导率和SOD、POD、CAT酶等生理生化指标;观察统计不同生长时期愈伤组织的生长量、成活率、分化率和再生苗率等;通过40%叶枯病菌粗毒素液选择压筛选获得抗病阳性植株,计算突变体率。实验结果表明:随着胁迫时间的增长,愈伤组织的生长量、成活率、分化率和再生苗率显著下降;24 h时的细胞质膜透性和SOD、CAT酶活性升高、POD酶活性降低。胁迫时长30~60 min时,愈伤组织生长量普遍降低4.5~5.6 g,对愈伤组织成活率、分化率和再生苗率未造成显著影响;突变体率比ck提高了6.34%~9.27%。胁迫时长达90 min时,愈伤组织生长量、成活率、分化率和再生苗率极度降低,通过选择压筛选获得的2株再生苗均为阳性植株。结论:渗透胁迫对EMS诱变愈伤组织的伤害呈叠加效应,随着胁迫时间的延长伤害作用加剧。高渗透固体培养基预处理30~60 min可以作为大花萱草EMS离体诱变提高突变体率的方法。渗透胁迫时间的长短与突变率提高的幅度没有规律性。This study aims to provide a pretreatment technique for EMS mutagenesis of Hemerocallis.It adopts‘Hongyun’with bud callus as material with high-permeability solid medium(MS+mannitol 36 g/L+sorbitol 36 g/L+sucrose 50 g/L)to do osmotic stress pretreatment at 0 min,30 min,60 min and 90 min respectively before 1.0% EMS semi-lethal dose mutagenesis.The physiological and biochemical indexes of 24 h such as the relative conductivity,SOD,POD and CAT enzyme were measured.The growth,survival rate,differentiation rate and regeneration rate of callus in different growth periods were observed.Mutant rates were calculated for disease resistance plants selected through selective pressure of 40% leaf blight pathogens.The results showed that the growth,survival rate,differentiation rate and regeneration rate of callus decreased significantly with the increase of stress time.The cytoplasmic membrane permeability,SOD and CAT enzyme activity increased and POD enzyme activity decreased after 24 h.When pretreated for 30-60 min,the growth decreased by 4.5-5.6 g,with no significant effect on the survival rate,differentiation rate and regeneration rate.The mutant rate increased by 6.34%-9.27%.After 90 min pretreatment,the growth,survival rate,differentiation rate and regeneration rate were extremely reduced;the two regenerated plants obtained by selective pressure screening were positive.The osmotic stress has a superimposed effect on the damage of EMS mutagenic callus,and the damage is aggravated as the prolongation of stress time.Pretreatment of high-permeability solid medium for 30-60 min can be used as a new method to increase the mutant rate of EMS in vitro for Hemerocailis hybridai.There is no regularity in the length of stress time and the increase of mutation rate.

关 键 词：渗透胁迫 甲基磺酸乙酯(EMS) 离体诱变 大花萱草 突变体率 

分 类 号：S337[农业科学—作物遗传育种]