CAJ | 학술논문

为了定量评估生防真菌绿僵菌在土壤中的存活能力和种群数量，需要确定高效的选择性培养基及其分离效率。本文研究了不同浓度多果定对绿僵菌的选择作用和分离检出率，结果表明，在以氯霉素抑制土壤细菌的基础上，多果定浓度在50～200 mg/L时能够有效抑制土壤中的非目标真菌而选择出可清晰鉴别和计数的绿僵菌；当浓度为25 mg/L时，对杂菌的抑制不完全，残留杂菌数占总数50%左右。随着多果定浓度增高，绿僵菌生长减缓、产孢颜色变黄、分离检出率降低。在多果定50 mg/L浓度下，接种培养绿僵菌4 d和6 d的菌落检出率分别69.20%和71.43%，显著低于对照培养基的检出率95.56%和94.72%；在200 mg/L的高浓度下，4 d和6 d检出率分别只有45.45%和61.24%，又显著低于前者低浓度下的检出率。试验确定了在蔗糖基础培养基中添加50 mg/L多果定为选择剂，能够有效抑制土壤中非目标真菌99%以上，对绿僵菌的分离效率稳定在70%左右。选择剂的抑制作用和选择效率是土壤微生物定量检测中不可忽略的因子。正确地评估田间绿僵菌数量是研究其种群动态、生态效应和持续生防作用的必要基础。
위료정량평고생방진균록강균재토양중적존활능력화충군수량，수요학정고효적선택성배양기급기분리효솔。본문연구료불동농도다과정대록강균적선택작용화분리검출솔，결과표명，재이록매소억제토양세균적기출상，다과정농도재50～200 mg/L시능구유효억제토양중적비목표진균이선택출가청석감별화계수적록강균；당농도위25 mg/L시，대잡균적억제불완전，잔류잡균수점총수50%좌우。수착다과정농도증고，록강균생장감완、산포안색변황、분리검출솔강저。재다과정50 mg/L농도하，접충배양록강균4 d화6 d적균락검출솔분별69.20%화71.43%，현저저우대조배양기적검출솔95.56%화94.72%；재200 mg/L적고농도하，4 d화6 d검출솔분별지유45.45%화61.24%，우현저저우전자저농도하적검출솔。시험학정료재자당기출배양기중첨가50 mg/L다과정위선택제，능구유효억제토양중비목표진균99%이상，대록강균적분리효솔은정재70%좌우。선택제적억제작용화선택효솔시토양미생물정량검측중불가홀략적인자。정학지평고전간록강균수량시연구기충군동태、생태효응화지속생방작용적필요기출。
This paper is to determine an efficiently selective medium and its efficiency for quantitative evaluation of the fungusMetarhizium anisopliae as a biocontrol agent applied in soil. The selectivity and detection rate of dodine was investigated at different concentrations. The results showed that 50-200 mg/L of dodine in medium could effectively inhibit non-target fungi in soil and highlight M. anisopliae, while preventing bacterial growth by chloromycetin. But lower dodine concentration, 25 mg/L, could not completely preclude non-target fungi with residues accounted for about 50% of the total colonies. When increasing dodine concentration, M. anisopliae would slow growth, yellow conidiation and decrease colonies. At 50 mg/L of dodine, detection rates get to 69.20% and 71.43% after inoculation ofM. anisopliae conidia for 4 d and 6 d. The values were significantly higher than 45.45% and 61.24% detected on 200 mg/L concentration, though significantly lower than 95.56% and 94.72% detected on control medium without dodine in the same inoculation and incubation. Finally, a favorable medium including basic nutrition supplemented with 50 mg/L dodine as selective agent was determined, which can effectively inhibit non-target fungi and extract stablyM. anisopliaein 70% efficiency. Inhibition and selection efficiency of selective agent should not be ignored in quantitative detection of microorganism in soil. Properly quantitative assessment of M. anisopliae in field is a necessary foundation for analysis of population dynamics, ecological effects and sustained biocontrol.