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利用酶处理桑黄(Phellinus igniarius)菌丝体,分离到原生质体并对其进行紫外线诱变,确定诱变后的致死率,以利于桑黄原生质体诱变及融合的合理实施。菌龄为10 d的桑黄菌丝体在1.5%溶壁酶、100 r·min~(-1)、30℃条件下酶解3 h,获得原生质体;再分别经紫外线照射10 s、20 s、30 s、40 s、50 s、60 s、70 s、80 s、90s后;利用血球计数板对美兰染色后的活性原生质体计数,计算桑黄原生质体紫外诱变后的致死率。结果显示,原生质体致死率随着照射时间的延长而增大,在40 s~50 s诱变时间内,曲线的变化最为剧烈;诱变时间为45 s时,致死率达到73%;诱变时间达到90 s时,致死率为100%。该结果能够为桑黄原生质体诱变育种及融合育种的实施提供切实的理论依据。
The mycelium of Phellinus igniarius was treated with enzyme, the protoplasts were isolated and mutagenized by UV light to determine the lethality after mutagenesis, in order to facilitate the mutagenesis and fusion of Phellinus igniarius. The mycelium of P. mori with the age of 10 d was protolyzed with 1.5% lywallzyme, 100 r · min -1, 30 ℃ for 3 h, then the protoplasts were obtained by UV irradiation for 10 s, 20 s, 30 s, 40 s, 50 s, 60 s, 70 s, 80 s and 90 s, respectively. The count of active protoplasts stained with Melanie was calculated by using a hemocytometer, and the lethality after ultraviolet mutagenesis of P. moris protoplast was calculated . The results showed that the protoplast lethality increased with the extension of irradiation time, and the curve changed most rapidly during the 40 s-50 s mutagenesis time; the lethality reached 73% when the mutagenesis time was 45 s; When the time reaches 90 s, the lethality rate is 100%. The results can provide a practical theoretical basis for the mutagenesis breeding of Phellinus igniarius and the implementation of fusion breeding.