茶树菇原生质体的分离和再生以及单核菌株的筛选

Protoplast Isolation and Colony Regeneration in Agrocye chaxingo Huang and the Screening of a Protoplast-Derived Monokaryon Strain

研究了利用珍稀食用菌茶树菇双核菌丝进行原生质体分离和再生，以及原生质体单核菌株的筛选过程．利用培养4～6d的茶树菇双核菌丝，以0．6mol／mLMgSO4·7H2O作稳定剂，在溶壁酶浓度为40mg／mL，温度30～32℃的摇床振荡（70r／min）条件下进行酶解4h，制备原生质体，然后，将原生质体在下层高渗固体培养基和上层高渗半固体培养基上再生，从该再生菌落中筛选到茶树菇单核菌株1株．

The preparation of monokaryon protoplasts and the screening of protoplast-derived monokaryon strains are important parts in mushroom research and breeding. rectly from mycelia without the process of chromosome meiosis As monok instead of aryon protoplasts are derived di from spores, the genetic charac teristics of the donor parent can be conserved. Agrocye chaxingo Huang is a rare and precious edible mushroom species. This paper reports the protocol of isolation and regeneration of A. chaxingo protoplasts, and the selection of protoplast-derived monokaryon strains. One monokaryon strain in A. chaxingo was obtained by the following steps. Fi lywallzyme at a concentration of 40 rst, mL the mycelia pre-cultured for 4-6 days were hydrolyzed with and with 0.6 mol/mL MgSO4 ·7H2O as the osmotic stabilizer on the shaker at 30-32 ℃ and 70 r/min for 4 hours. Then, the protoplasts were inoculated on a solid medium with high osmotic pressure and covered by a semi-solid medium with high osmotic pressure to regenerate, and colonies which grew slower and were smaller were picked one by one and checked under a microscope to observe the mycelia morphology and clamp-like structures. Consequently, one monokaryon strain was screened from the regenerated colonies.