【作者】 龚韡斌；

【导师】 刘君昂； 何雪香；

【作者基本信息】 中南林业科技大学 ， 森林保护学， 2009， 硕士

【摘要】 植物根际有益微生物菌剂研究及应用技术是倍受关注的国际性研究课题,其在根际定居(或定殖)是十分重要的,新剂型的开发有助于解决这一问题。随着根际微生物菌剂研究的不断深入,根际微生物对林木成活和生长的重要性愈来愈被世人所认识,应用研究普遍受到世界各国的重视。本论文以广东沿海地区本地红树林根际促生菌为研究对象,主要研究本地红树林根际促生菌,开展其分离筛选、培养基条件优化及胶囊剂型研制,使之成为一种存活率高、易于工业化生产和运输的新型菌剂。本地红树林根际促生菌分离筛选。以广东南沙湿地公园采回的野生红树白骨壤小苗为材料,分离筛选出了具有溶P、固N能力的菌株,并以广东省林科院实验室从本地红树林根际分离筛选的溶P菌3株,编号为：P7#、07PCK、07pqwp1-5#；固N菌4种,编号为：NGQQ②D、NGWB③+④B、MLN2#、07Nhs-7#为供试菌株,在室温盆栽条件下研究了7种菌株对红树木榄幼苗(广东南沙湿地公园提供)的接种效应,结果表明：固N菌接种的木榄幼苗苗高比对照增加20.4%～31.8%,生物量比对照增加19.4%～21.6%,对木榄幼苗苗高及生物量进行显著性分析表明,苗高、生物量与对照相比均达极显著值；溶P菌接种的木榄幼苗苗高比对照增加22.8%～28.5%,生物量比对照增加19.4%～22.6%,对木榄幼苗苗高及生物量进行显著性分析表明,苗高、生物量与对照相比均达极显著值。在盆栽试验过程中,对促生菌的根部定殖情况进行了检测,溶P菌和固N菌根部定殖菌落数数量级均达109以上,结果表明,促生菌根部定殖良好。根际促生菌培养基条件优化。鉴于不同环境下筛选出来的促生菌对培养基的适应性不一样,本研究以实验室自行筛选的促生菌为供试菌株,进行了单因素对促生菌生长影响研究,结果表明,促生菌最佳培养基条件为：盐度15‰,pH 8.0,营养源C源为葡萄糖,N源为蛋白胨。本地红树林根际促生菌胶囊剂型研制。试验研制了本地红树林促生菌新剂型-微胶囊剂,选用壁材为海藻酸钠,采用锐孔-凝固浴法制做胶囊剂型。试验对微胶囊胶液配方进行了优化,通过对15种不同的胶液配方制作的微胶囊进行形态、硬度的比较,筛选出了最佳胶液配方：海藻酸钠(2.5%)：菌液混=3：1。试验将微胶囊剂型(干品、湿品)与液体剂型(4℃保存、-20℃保存)菌株的存活率进行比较,结果表明,常温下液体剂型对菌种的保存能力最差,2个月后存活率不足10万,供试溶P菌和固N菌活菌数仅为5.59x104、7.37×105,而胶囊剂干品常温保存2个月后,存活率仍能达到1000万,供试溶P菌和固N菌活菌数分别为11.37×106、10.13×107。在室温盆栽条件下研究了以本试验筛选出的促生菌制作的大胶囊、微胶囊和常规液体剂型(混合菌,单菌)对红树秋茄幼苗(广东南沙湿地公园提供)的接种效应,试验结果表明,接种促生菌菌剂的秋茄幼苗苗高比对照增加12.6%～23.0%,生物量比对照增加16.2%～17.5%。对秋茄幼苗苗高及生物量进行差异显著性分析表明,接种促生菌菌剂的秋茄苗高与对照相比均达极显著差异,生物量与对照相比,除NGQQ②D单菌液接种的秋茄幼苗外,均达极显著差异。各剂型促生菌根部定殖情况良好,尤以微胶囊效果最佳,而且胶囊剂干品携带、运输均比液体剂型方便,其优越性显而易见。更多还原

【Abstract】 The research and applied technology of rhizosphere beneficial microbial inoculums is a repeatedly interested international research subject, which is very important to the field planting, new dosage forms can help to solve this problem. With study in a deep going way, the essentiality of rhizosphere microorganism to forest survival and growth was well-known by more and more people, applied research is generally well-focused by all countries in the world. In this paper, we considered rhizosphere growth-promoting bacteria of local mangrove in the coastal areas of Guangdong as object of study, focused on separating and selecting growth-promoting bacteria from local mangrove and developed a new bacterial agent of high survival rate and convenient in industrial production and transportation.Isolation of growth-promoting bacteria from local mangrove. Used wild mangrove Avicennia marina seedlings that picked in Guangdong Nansha Wetland Park as experimental materials, we separated and selected bacterias which have the capacity of phosphate-solubilizing and nitrogen fixation, these seven kinds of growth-promoting bacteria,there are three phosphate-solubilizing bacterias selected by GuangDong Academy of Forestry, we number them P7#、07PCK、07pqwp1-5#; and also four kinds of Nitrogen-fixing bacteria, number them NGQQ②D、NGWB③+④B、MLN2#、07Nhs-7#. In pot experiments, we studied inoculation effect of seven kinds of strains on mangrove seedlings (offered by Guangdong NanSha wetland park) and the results showed comparing with contrast seedlings, the height of inoculated seedlings increased 20.4%～31.8% and the biomass increased 19.4%-21.6%, carried out significance analysis of the mass growth and biomass of mangrove seedlings, the results showed both of them reached extreme marked value; the height of phosphate-solubilizing bacteria inoculated seedlings increased 22.8%-28.5% and the biomass increased 19.4%-22.6%, carried out significance analysis of the mass growth and biomass of mangrove seedlings, the results showed both of them reached extreme marked value. In pot experiments, we tested the situation of field planting of rhizosphere growth-promoting bacteria, phosphate-Solubilizing Bacteria and Azotobacteria colonizing the Department of magnitude the number of each colony for more than 109, experimental results showed the situation of it was well. Optimization of culture medium of rhizosphere growth-promoting bacteria. Because wetland forest ecosystem is more complex than the dry land, and the growth conditions of growth-promoting were also relative complex, so optimize the standard medium of Phosphate-Solubilizing Bacteria. Via a series of single factor experiment, the best culture condition of Phosphate-Solubilizing Bacteria was:salinity 15%o, pH 8.0, carbon source dextrose, nitrogen source peptone.Develop the capsule formulation of rhizosphere growth-promoting bacteria of local mangrove. In this paper, we optimized the recipes of microcapsule, compared the configuration and rigidity of microcapsule in different recipes and selected the best recipe:2.5% sodium alginate mixed with the bacteria liquid as 3:1. Compared the capacity of bacteria with conventional liquid dosage forms in different conditions, the preserving ability of liquid form in normal temperature was the worst, the survival rate was less than 10 million after two months, the viable counts of test Phosphate-Solubilizing Bacteria and Azotobacteria were only 5.59×104 and 7.37×105, but the survival rate of dry capsule can reach 10 million in normal temperature after preserving two months later; the viable counts of test Phosphate-Solubilizing Bacteria and Azotobacteria were 11.37×106 and 10.13×107 respectively. In pot experiments, we studied inoculation effect of large capsules, micro-capsules and liquid formulations of conventional (mixed bacteria, single bacteria) that are made by selected growth-promoting bacteria on mangrove kandelia candel seedlings (offered by Guangdong NanSha wetland park) and the results showed comparing with contrast seedlings, the height of inoculated seedlings increased 12.6%～23.0% and the biomass increased 16.2%～17.5%, carried out significance analysis of the mass growth and biomass of mangrove kandelia candel seedlings, the results showed the height of seedlings reached extreme marked value; the biomass of seedlings reached extreme marked value, excluded the seedlings that inoculated by single bacteria. We tested the situation of field planting of all kinds of dosages of rhizosphere growth-promoting bacteria, experimental results showed the situation of all dosages were well, especially the microcapsule. Also the dry capsules were convenient goods than conventional liquid formulations in carrying and storage, so the advantages of capsules are obvious.更多还原