CAJ | 학술논문

目的:对准确分离植物内生真菌的方法和适宜的培养条件进行研究.方法:采用75%乙醇、次氯酸钠溶液(活性氯含量1.3%～5.2%)浸泡和无菌水洗涤工艺对植物材料进行表面火菌,以火菌强度逐步递减的方式,确定最佳分离植物内生真菌的表而灭菌强度;设置超净工作台无菌状态检测对照、漂洗液无菌检测对照和组织印迹无菌检测对照3种对照处理,确保适度的表面灭菌强度;采用不同的培养基分离培养内生真菌.结果:高浓度杀菌剂短时间处理比低浓度杀菌剂长时间处理更适合植物内生真菌的分离;使用10%马铃薯葡萄糖双抗培养基、22℃低温培养,能抑制快生型菌株的疯长,从而使慢生型菌株拥有一定的生长空间,最大限度地分离所有植物内生真菌;同时设置3种对照处理可以确保植物内生真菌分离的准确性.结论:设计合适的表面灭菌方法、表面灭菌效果检测方法和分离培养方法对植物内生真菌的分离至关重要.
목적:대준학분리식물내생진균적방법화괄의적배양조건진행연구.방법:채용75%을순、차록산납용액(활성록함량1.3%～5.2%)침포화무균수세조공예대식물재료진행표면화균,이화균강도축보체감적방식,학정최가분리식물내생진균적표이멸균강도;설치초정공작태무균상태검측대조、표세액무균검측대조화조직인적무균검측대조3충대조처리,학보괄도적표면멸균강도;채용불동적배양기분리배양내생진균.결과:고농도살균제단시간처리비저농도살균제장시간처리경괄합식물내생진균적분리;사용10%마령서포도당쌍항배양기、22℃저온배양,능억제쾌생형균주적풍장,종이사만생형균주옹유일정적생장공간,최대한도지분리소유식물내생진균;동시설치3충대조처리가이학보식물내생진균분리적준학성.결론:설계합괄적표면멸균방법、표면멸균효과검측방법화분리배양방법대식물내생진균적분리지관중요.
The isolation and culture conditions of endophytes from branch and leaves of Pseudolarix kaempferi Gord. were investigated. Plant materials were sterilized by surface treatment with 75% ethanol, sodium hypochlorite solution and sterilized water. The optimal surface sterilization intensity for isolating endophytes was determined in a progressive decrease way with biological safety cabinet sterilization detection control, washing liquid sterilization detection control and tissue blotting steriliza- tion detection control. The effects of three various media on isolation of endophytes were compared. A short-time sterilization with high concentration sodium hypoehlorite solution was more suitable for the isolation of endophytes than a long-time sterilization with low concentration. The growth of fast-growing endophytes was inhibited in a 10% PDA medium supple- merited with potassium benzylpenicillin and streptomycin sulfate at 22 ℃ and thus slow-growing ones could obtain some growing space, which was beneficial to maximize the separation of endophytes. The accuracy of isolation of endophytes was guaranteed by the setting of three detection controls.