Hua Li, Feng Ming-Gung
Institute of Microbiology, College of Life Sciences, Zhejiang University, Hangzhou, Zhejiang, 310029, Peoples' Republic of China.
Mycol Res. 2005 Mar;109(Pt 3):319-25. doi: 10.1017/s095375620500239x.
The shelled grains of glutinous broomcorn millet, Panicum miliaceum, were used as solid substrate to prepare granular cultures of Zoophthora radicans, an entomophthoralean biocontrol agent against numerous insect pests. Steamed millet grains were inoculated by mixing 15 g millet grains (D.W.) with mashed pieces of half a 60-mm-dish colony in 3 ml modified Sabouraud dextrose broth and incubated at 15 degrees C and L:D 12:12 for up to 24 d. 20 grains were sampled at 3 d intervals from day six onwards and individually assessed for their sporulation capacity using a self-designed device for spore collection. The millet cultures after > or = 12 d incubation produced 12.0 - 14.9 x 10(4) spores grain(-1) during a 7 d period. The maximal sporulation capacity associated with the 21 d-old culture was about half of that of Z. radicans-killed Plutella xylostella larvae (28.7 x 10(4) spores cadaver(-1)), which individually were at least three times larger than the millet grains. Based on the time-concentration-mortality responses of second-instar P. xylostella larvae to Z. radicans in three independent bioassays, the spores ejected from the cultured millet grains, from the mycelial mats from liquid culture, and from larval cadavers displayed insignificant variations in infectivity to the host species, and yielded similar LC50 and LT50 estimates. Conclusively, the millet-based technology for production of granular cultures of Z. radicans was easy, inexpensive and highly efficient, and it could be superior to previous methods used in mass production of mycelium-based preparations of Entomophthorales since this new approach requires no special additives, drying, freezing and milling. This technology may suit to mass production of culturable but nutritionally fastidious entomopathogens from the Entomophthorales.
黍稷(Panicum miliaceum)去壳的谷粒被用作固体基质,来制备根虫瘟霉(Zoophthora radicans)的颗粒培养物,根虫瘟霉是一种防治多种害虫的虫霉目生防制剂。将15克黍稷谷粒(干重)与半个60毫米培养皿菌落的捣碎块在3毫升改良沙氏葡萄糖肉汤中混合,对蒸煮过的谷粒进行接种,并在15℃、光周期12:12条件下培养24天。从第6天起,每隔3天取样20粒谷粒,并使用自行设计的孢子收集装置单独评估其产孢能力。培养12天及以上的黍稷培养物在7天内产生了12.0 - 14.9×10⁴个孢子/粒。与21天龄培养物相关的最大产孢能力约为被根虫瘟霉杀死的小菜蛾幼虫(28.7×10⁴个孢子/虫尸)的一半,小菜蛾幼虫个体至少是黍稷谷粒的三倍大。基于在三项独立生物测定中,二龄小菜蛾幼虫对根虫瘟霉的时间 - 浓度 - 死亡率反应,从培养的黍稷谷粒、液体培养的菌丝垫以及幼虫虫尸中弹射出来的孢子,对宿主物种的感染力差异不显著,并且产生了相似的半数致死浓度(LC50)和半数致死时间(LT50)估计值。总之,基于黍稷的根虫瘟霉颗粒培养物生产技术简便、廉价且高效,并且可能优于以前用于大规模生产虫霉目基于菌丝体制剂的方法,因为这种新方法不需要特殊添加剂、干燥、冷冻和研磨。该技术可能适合大规模生产来自虫霉目的可培养但营养要求苛刻的昆虫病原真菌。