大杯蕈原生质体菌株筛选的研究  

A Study on the Construction of Protoplast Regenerating Strains in a Wild Mushroom Clitocybe maxima

作　　者：彭智华[1] 曾广文[1] 寿诚学 

机构地区：[1]浙江大学园艺系,杭州310029

出　　处：《园艺学报》2000年第3期193-197,共5页Acta Horticulturae Sinica

摘　　要：对野生大杯蕈 (Clitocybemaxima)原生质体的分离再生及再生菌株的构建进行研究 ,以纤维素酶、溶壁酶、崩溃酶、β -葡萄糖苷酸酶的去壁效果最好 ,而新生酶 ,几丁质酶 ,蜗牛酶的去壁效果较差。 2种以上的酶液混合使用能提高去壁效果。酶解液在 0 .1%～2 .0 %浓度范围内原生质体释放量随酶解液的浓度升高几乎呈直线上升。酶解 3～ 5h释放的原生质体最多。酶解液pH值在 5.0～ 6.0范围内原生质体的产量较高。渗透压稳定剂以MgSO40 .6mol/L及甘露醇 0 .6mol/L处理原生质体释放量最高。培养 2～ 5d的菌丝酶解后原生质体释放量较多。培养基对原生质体释放量的影响不明显。大杯蕈的原生质体再生成活率仅 5%左右 ,所得 9个原生质体再生克隆菌株中多数菌株生长慢于对照 ,且菌丝体颜色较淡白 ,只有P0 4菌株无论母种、原种或栽培种的菌丝体生长速度均快于对照 ,且子实体的单菇重、菌盖重、菌盖直径、菌盖厚及盖柄比等均优于对照。Protoplast isolation and regeneration from the dikariotic mycelia of the wild mushroom Clitocybe maxima(Garten et mey.ex Fr.)Quel were carried out in present study.A standard set of conditions was determined for the reproducible high yields of stable protoplasts.Protoplasts were released in large amount by using cellulase,driselase,β-glucuronidase,lysase as lytic enzymes,and in less amount by using Novozym 234(Novo Biolabs,Denmark),chitinase,helicase as lytic enzymes. Stable protoplast release was obtained after enzyme digestion carried out in pH 5.5 at 25℃ for 2-6 hours,lytic enzyme solution.Among five osmotic stabilizers tested,MgSO 40.6mol/L and mannitol 0.6mol/L gave higher protoplasts yield.Culture media had no significant effect on the protoplast release.The protoplast regenerating ratio was comparatively low,about 5%.9 regenerating strains obtained were compared with the normal strain from mycelial growth to fruiting.2 strains showed more rapid mycelial growth rate,and P04 was the only one with some improvement of fruiting characteristics.

关 键 词：大杯蕈 食用菌 原生质体融合 再生植株 

分 类 号：S646.190.3[农业科学—蔬菜学] S336[农业科学—园艺学]