橡胶树组培苗常见污染源的鉴定及防除方法

Predominant Contamination Agents in Rubber Tree Tissue Culture and Its Elimination

摘要: 植物组织培养技术是一门实用性非常强的现代生物技术。自根幼态无性系是橡胶树新一代种植材料，与传统芽接苗相比具有高产速生的特点，其推广应用正是建立在规模化组培繁殖体系上的。然而，组培生产过程中杂菌污染是造成生产成本高的主要原因之一。本研究通过分子鉴定的方法确认橡胶树组织培养中最主要的污染源是黑曲霉、杂色曲霉等。我们发现苯甲酸钠和山梨酸钾以及益生菌哈茨木霉菌株13-3-2可以用于降低污染率。添加3%的苯甲酸钠或山梨酸钾的培养基可有效防止细菌及部分真菌(包括黑曲霉)污染；在后期驯化练苗过程中，接种的哈茨木霉菌13-3-2可有效杀死多种污染真菌，使污染苗的成活率提高43%以上。该研究结果将用于降低生产橡胶树组培苗的生产成本，加快橡胶树优良品种或材料在橡胶树生产中的应用。

Abstract: Tissue culture is a practical technique of modern biotechnology. Self-rooting juvenile clones of rubber tree is a new type material for field planting, which has characters of higher yielding and faster growth, compared with previous budded rubber trees, and its quicker propagations are based on tissue culture. However, contamination during tissue culture progress has been one key reason for higher cost. In this study, the main causal agents were identified by molecular method to be Aspergillus niger and Aspergillus versicolor. Chemicals such as potassium sorbate or sodium benzoate and biological agents as Trichoderma harzianum strain 13-3-2 were suggested to reduce the rate of contamination. 3% sodium benzoate or potassium sorbate were found effective in killing or inhibiting contaminated bacteria and some of the contaminated fungi, including the pre- dominant A. niger. During the later periods when seedlings acclimated, Tri. harzianum 13-3-2 was found so useful of killing miscellaneous agents that the live rate of contaminated seedlings could be enhanced by 43%. These results will be used for reducing cost in mass tissue culture production and fastening the propagation or utilization of elite rubber tree clones or materials.