诱变结合小孢子培养创制大麦黄花叶病不同抗性突变体

Mutants Resistant to Barley Yellow Mosaic Virus(BaYMV) Created by Mutagenesis Combined With Microspore Culture

为了创制具有不同黄花叶病抗性的大麦突变体材料,对大麦品种花30的干种子和游离小孢子分别采用物理和化学诱变处理,结合小孢子培养技术获得了一批纯合稳定的加倍单倍体(double haploid,DH)株系,并利用黄花叶病种植鉴定圃进行突变体DH株系的大田抗性筛选与鉴定,比较2种诱变方式结合小孢子培养的诱变效果。结果表明:以400 Gy剂量60Co-γ射线处理干种子诱变方式对小孢子培养的愈伤组织诱导能力和绿苗成活率有较大影响,获得的大部分株系(84.2%)对黄花叶病表现感病(感病或高感),未筛选到比亲本抗性增强(高抗)的材料;以3 mg·L-1剂量EMS处理游离小孢子诱变方式对小孢子培养相对影响较小,筛选到少量(5.5%)比亲本抗性增强(高抗)的株系,只有近1/3的材料(31.1%)表现为感病(感病或高感),进一步鉴定筛选到2份高感和1份高抗黄花叶病的大麦突变体材料。综上所述,EMS处理游离小孢子结合小孢子培养诱变效果较好。本研究为利用大麦黄花叶病抗性突变体材料进行其抗病机理研究奠定了基础。

In order to create mutants resistant to Barley Yellow Mosaic Virus（ Ba YMV）,the dry seeds and isolated microspores from barley cultivar Hua- 30 were mutagenized with60Co-γ ray and ethyl methane sulfonate（ EMS）respectively,and the homogeneous double haploid（ DH） lines were produced from microspore culture,which were planted in Ba YMV-infected field for identification of Ba YMV resistance. The mutagenic effect of two mutagenesis methods combined with microspore culture was also evaluated. The results showed that^60 Co irradiation on dry seeds at400 Gy greatly affected callus induction and the survival rate of green plants in microspore culture. Comparing to the wild type,most of produced DH lines（ 84. 2%） were more susceptible to Ba YMV（ susceptible or high susceptible）,and none was found with enhanced resistance（ high resistant）. EMS treatment on isolated microspores at 3 mg·L^- 1had little impact on microspore culture,minor of produced DH lines（ 5. 5%） showed enhanced resistance（ high resistant） to Ba YMV,and only about one-third（ 31. 1%） of them were more susceptible（ susceptible or high susceptible）. Two high susceptible（ HS） mutation lines and one high resistant（ HR） mutation lines were further identified. Overview,EMS treatment on isolated microspores combined with microspore culture achieved more positive effect on mutation induction. It will facilitate further studies on the mechanism of Ba YMV resistance by using the mutants obtained in this study.