Govender Nisha T, Mahmood Maziah, Seman Idris A, Mui-Yun Wong
Institute of Tropical Agriculture, Universiti Putra Malaysia, Selangor D.E., Malaysia.
Institute of Tropical Agriculture, Universiti Putra Malaysia, Selangor D.E., Malaysia; Department of Biochemistry and Biomolecular Sciences, Faculty of Biotechnology, Universiti Putra Malaysia, Selangor D.E., Malaysia.
Pol J Microbiol. 2016 Aug 26;65(3):383-388. doi: 10.5604/17331331.1215619.
Ganoderma boninense, a phytopathogenic white rot fungus had sought minimal genetic characterizations despite huge biotechnological potentials. Thus, efficient collection of fruiting body, basidiospore and protoplast of G. boninense is described. Matured basidiocarp raised under the glasshouse conditions yielded a total of 8.3 × 104 basidiospores/ml using the low speed centrifugation technique. Mycelium aged 3-day-old treated under an incubation period of 3 h in lysing enzyme from Trichoderma harzianum (10 mg/ml) suspended in osmotic stabilizer (0.6 M potassium chloride and 20 mM dipotassium phosphate buffer) yielded the highest number of viable protoplasts (8.9 × 106 single colonies) among all possible combinations tested (regeneration media, age of mycelium, osmotic stabilizer, digestive enzyme and incubation period).
邦氏灵芝是一种植物病原性白腐真菌，尽管具有巨大的生物技术潜力，但对其进行的遗传特征研究却很少。因此，本文描述了高效收集邦氏灵芝子实体、担孢子和原生质体的方法。在温室条件下培养成熟的担子果，采用低速离心技术，每毫升可产生8.3×10⁴个担孢子。在所有测试的可能组合（再生培养基、菌丝体年龄、渗透稳定剂、消化酶和孵育时间）中，将3日龄的菌丝体在含有哈茨木霉裂解酶（10mg/ml）的渗透稳定剂（0.6M氯化钾和20mM磷酸氢二钾缓冲液）中孵育3小时处理后，获得了最高数量的活原生质体（8.9×10⁶个单菌落）。
