突脐蠕孢与弯孢原生质的双亲灭活融合

Protoplasts Fusion Between Exserohilum monoceras and Curvularia lunata with Biparental Inactivate Technique

采用灭活原生质体方法,对致病力强、产孢量低的突脐蠕孢EM菌株和致病力较弱、但产孢量高的弯孢CL菌株两株稗草致病菌进行原生质体融合,通过观察形态、生物学及抑草活性等特性筛选融合子。结果表明,2%溶壁酶+2%蜗牛酶处理获得两亲株的原生质体释放数量最高,CL和EM分别达1.25×106个/ml和1.21×106个/ml;再生培养基○c处理的CL和EM原生质体再生率分别达14.7%和13.2%。EM利用热灭活55℃5min;CL利用紫外灭活30s,距离20cm,照射时间4min;聚乙二醇促融,获得3个融合子。融合子在孢子形态上与亲株存在明显差异,其菌丝生长速度、孢子产量、对稗草根生长的抑制作用等也与双亲菌株有差异。

Protoplast fusion between Exserohilum monoceras （EM） ducted with biparental inactivation technique. EM is highly toxic ductivity, while CL is reversed to EM. The fusants were selected and Curvularia lunata （ CL ） was conto barnyard grass but lower spore prothrough observing the characteristics of morphology and biology, as well as the activity against weed. The results showed that co-function protoplast of the 2% Lywallzyme plus 2% Snailase was the most effective way to its yield, 1.25 ×10^6 for CL and 1.21×10^6 for EM, respectively. The regeneration ratio of CL and EM treated by R3 medium was 14.7% and 13.2% respectively. The protoplasts of CL and EM were inactivated by UV for 30 s with the radiated distance of 20 cm and by hot water at 55℃ for 5 min, respectively. 40% PEG was utilized to enhance the protoplast fusion. Three potent fusants were obtained, which were obviously different from the parental strains in spore morphology, mycelia growth rate, conidia yield and activity of inhibiting the barnyard grass root.