中华卵索线虫的体外培养  

IN VITRO CULTIVATION OF THE ENTOMOGENOUS NEMATODE OVOMERMIS SINENSIS

作　　者：王国秀[1] 陈曲侯[1] 陈果[2] 

机构地区：[1]华中师范大学生命科学院,武汉430079 [2]中国农业科学院生物防治研究所,北京100081

出　　处：《动物学报》2001年第2期235-239,T001,共6页ACTA ZOOLOGICA SINICA

基　　金：国家自然科学基金资助项目!(NO .3 95 70 10 1)&&

摘　　要：在研究中华卵索线虫的体外培养方法的同时 ,对其在不同培养基中的生长发育情况进行了观察。结果表明 :以培养基TC 199加 2 0 %热灭活胎牛血清的培养效果较为理想 ,大多数线虫可存活 3个月 ,最大虫体长5 5 1mm、宽 2 0 4 13μm ,其发育程度大致与该种索线虫在宿主粘虫体内寄生 8～ 9天的情况相近。培养期间观察到 2次蜕皮 :第一次蜕皮在卵内 ,第二次在培养 6～ 8天之后。口针消失 ,虫体内滋养物体发育明显 ,尾部附器已经形成 ,没有观察到生殖原基的发育。The insect parasitic nematode, Ovomermis sinensis Chen et al ., 1991 has the potential for the biological control of a range of insect pests due to many insect sepecies are susceptible to this nematode. For example Leucania separata, Heliothis armigera, Prodenia litura, Spodoptera exigua, Plutella xylostella, Pieris rapae, Agrotis ypsilon, Euxoa segetum etc. The insect host invariably dies when the juvenile nematode completes its parasitic development and exits from the host's hemocoel. There is an important significance for studying the mass cultivation of O. sinensis for field trials. This paper describes the techniques of in vitro cultivation of O. sinensis. Gravid females of O. sinensis collected from sand were washed three times with an antibiotic solution of 200 U penicillin/ml and 200 μg/ml streptomycin/ml per ml solution. Surface washed gravid female were placed in 25 ml culture tubes containing 1～10 ml deionized water, 100 U penicillin/ml, and 100 μg streptomycin /ml for oviposition. Eggs were collected and their surface were sterilized by immersing them for 5～10 min in 0 13%～0 5% sodium hypochlorite solution and rinsing four times with sterile distilled water. Surface sterilized eggs were incubated at 26±1℃ in the culture media. The preparasites which emerged from those eggs were utilized for culture work. The four media were tested and they were TC 199, TC 199+MK, TC 100 or Grace supplemented with 20% heat inactivated fetal bovine serum respectively. The pH of the media were adjusted to pH6 8～7 0. The culture medium was changed or replenished every 3～6 days by withdrawing the supernatant and replacing it with a portion of fresh medium. In vitro growth of O. sinensis parasites was monitored in TC 199 supplemented with 20% heat inactivated fetal bovine serum under differing condition of crowding. Crowded cultures (150～200 parasites per tube) were set up in early days of in vitro (length of nematodes<10 mm), whereas crowed culture (50～7

关 键 词：中华卵索线虫 索科 体外培养 

分 类 号：Q969[生物学—昆虫学]