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ลักษณะทางสรีรวิทยาและชีวเคมี การกําหนดลักษณะทางสรีรวิทยาได้ดําเนินการเพื่อกําหนดความสามารถของเชื้อราที่แยกได้ในการเติบโตในที่ที่มีความเข้มข้นของ nacl ต่างกัน ((w/v) 0%, 5%, 10% และ 20%) และในช่วงอุณหภูมิที่แตกต่างกัน (25 ̊c และ 40 ̊c) การเจริญเติบโตขึ้นอยู่กับเส้นผ่านศูนย์กลางของอาณานิคมหลังจากฟักตัว 7 วันหรือหลังจากฟักตัว 15 วันสําหรับไอโซเลทที่เติบโตช้า
the ability of the isolates to produce extracellular enzymes was examined. amylase activity was tested on nutrient agar containing 2 g/l of soluble starch. cultures were incubated for 2 to 5 days, depending on the growth rate of each isolate, and then flooded with an iodine solution. a clear zone around the colony indicated the presence of amylase [34]. cellulase activity was tested on a solid medium containing 1% cellulose (7.0 g/l kh2po4, 2.0 g/l k2hpo4, 0.1 g/l mgso4.7h2o, 1.0 g/l (nh4)2so4, 0.6 g/l yeast extract, 10 g/l microcrystalline cellulose and 15 g/l agar) [35]. after incubation, the cultures were further incubated at 50 ̊c for 16 h to accelerate enzyme activity. the cultures were then flooded with 5 ml of 1% congo red solu- tion and rinsed with distilled water to detect a hydrolysis zone. protease activity was tested on casein agar medium containing 30% skim milk and 2% agar. degradation of casein was indi- cated by a clear zone around the colony [36]. lipase activity was tested on a solid medium con- taining tween 80 (10 g/l peptone, 5 g/l nacl, 0.1 g/l cacl2.2h2o, 17 g/l agar and 10 ml/l tween 80). tween 80 was sterilized before addition to the sterile medium. cultures were placed at 4 ̊c for 12 h after incubation to observe opaque precipitation around the colony [34]. each enzyme activity was evaluated by an enzymatic index (ei) where ei = r/r (r being the diameter of the clear zone and r the diameter of the colony) [32]. mycovirus dsrnas were extracted using licl fractionation as described by hull and covey [37]. dsrna samples were purified by phenol-chloroform extraction. dna and ssrna contamination were removed by sequential dnase i and si nuclease treatment. the viral genome was proved to be dsrna by digestion with rnase iii. the presence of the dsrna was checked by electrophoresis in a 1% (w/v) agarose gel in 1x tae buffer.