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中国农学通报

中国农学通报 ›› 2013, Vol. 29 ›› Issue (18): 168-174.doi: 10.11924/j.issn.1000-6850.2012-3001

所属专题： 生物技术

• 植物保护 农药 • 上一篇    下一篇

两株抗香蕉枯萎病菌链霉菌固态发酵条件的优化及混合菌载体的选择

莫坤联 曾涛 陈汉清 郭刚   

  • 收稿日期:2012-09-03 修回日期:2012-10-15 出版日期:2013-06-25 发布日期:2013-06-25

Optimization of Solid-state Fermentation Conditions for Two Streptomyces stains Against Banana Fusarium wilt and Selection of Additive.

  • Received:2012-09-03 Revised:2012-10-15 Online:2013-06-25 Published:2013-06-25

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摘要： 将具有抗香蕉枯萎病菌活性链霉菌通过固体发酵的优化，提高其菌丝生长以及孢子产量，并选择合适的吸附载体，为后期的香蕉枯萎病大田生物防治研究提供技术基础。采用单因子和4因子3水平正交试验法优化白刺链霉菌BWL15-4菌株及不吸水链霉菌BWL58菌株的固态法发酵培养基配方，通过单因子试验探讨了初始pH、接种量以及培养温度对2株菌生长情况的影响，并以孢子存活量多少确定了混合菌株的最适载体。BWL58优化后的固态发酵条件为：pH 7.0左右，接种量2%，发酵温度28℃，培养基配方为：无机盐水占大米重量的150%，黄豆粉占大米重量的5%，麸皮占大米重量的20%，沙土占大米重量的30%。BWL15-4优化后的固体发酵条件为：初始pH 6.3左右，接种量3%，发酵温度29.5℃。培养基配方为无机盐水占大米重量的130%，黄豆粉占大米重量的10%，麸皮占大米重量的20%，沙土占大米重量的20%。非耕作层土与混合菌以2:1的比例混合时，在常温下保存3个月后，孢子数只降低一个数量级。按照此优化条件进行固态发酵，白刺链霉菌BWL58菌株及不吸水链霉菌BWL15-4菌株孢子含量可分别达到1013 cfu/g和1012 cfu/g数量级以上。通过筛选得到了混合菌的最佳填料为非耕作层土。

关键词: 离体培养, 离体培养

Abstract: In this study, the solid-state fermentation conditions of two Streptomyces spp. strains that against Fusarium oxysporum f. sp. Cubense causing banana panama wilt were optimized. The purpose of this project was to increase the yield of mycelium and spores; the additive was optimized as well, for the biological control of banana panama wilt. The optimization of medium to Streptomyces albospinus Strain BWL15-4 and Streptomyces ahygroscopicus Strain BWL58 was designed by single factor test and the 4 factors 3 levels orthogonal experiments. First, the single factors, including the effect of initial pH, inoculum volume and fermentation temperature, on the mycelium growth of the two Streptomyces spp. strains were investigated. Secondly, the carrier of the bio-control agents was concluded based on the yield of the spores. The optimized conditions for the solid-state fermentation of Streptomyces spp. Strain BWL58 are as follows: pH 7.0 as initial, amount of inoculum 2%, fermentation temperature 28℃ and the optimized medium including water, soy flour, wheat bran, sandy soil and rice, and their ratio is 150:5:20:30:100, respectively by wight. The optimized conditions for the solid-state fermentation of Actinomycetes spp. Strain BWL15-4 are as follows: pH 6.3 as initial, amount of inoculum 3%, fermentation temperature 29.5℃ and the optimized medium also including water, soy flour, wheat bran, sandy soil and rice, and their ratio is 130:10:20:20:100 respectively by wight. After 3 months, the amount of the spores was reduced for 10% when the non-cultivated loess was selected as the carrior of the bio-control agent and the loess/microorganism mixture ratio is 2/1. Under the above optimized fermentation condition, the two test strains could produce large amount of living spores, which were up to 1013 cfu/g and 1012 cfu/g ,respectively. Additionally, the results show that the non-cultivated loess was found to be the best additive among the three candidate additives, including the Kaolin soil, flour and non-cultivated loess.