GFP标记的植物促生菌B96-Ⅱ-gfp的定殖能力研究

Colonization ability of plant growth promoting Bacillus B96-Ⅱ-gfp labeled with GFP

  • 摘要: 采用绿色荧光蛋白基因标记技术研究了植物促生菌B96-Ⅱ的标记菌B96-Ⅱ-gfp在盆栽黄瓜土壤中的时间、空间定殖动态以及在黄瓜植株上的分布。研究表明: B96-Ⅱ-gfp在土壤中具有持久的定殖能力, 接种180 d时在自然土和黄瓜枯萎菌病土中的定殖数量分别为2.7×104 cfu·g-1和6.6×104 cfu·g-1, 360 d时在土壤中仍可检测到B96-Ⅱ-gfp的存在。B96-Ⅱ-gfp可在盆栽植株生长土壤的表层(0~4 cm)、中层(4~8 cm)和底层(8~12 cm)定殖, 定殖数量随土壤深度的增加而增加。此外, B96-Ⅱ-gfp还可在黄瓜的根、茎和叶上定殖, 根部定殖的数量(7.2×104 cfu·g-1)显著高于茎部和叶部定殖的数量; 黄瓜植株体内定殖的数量多于体表定殖的数量。对土壤中可培养的3大微生物类群影响的研究表明: B96-Ⅱ-gfp对土壤中真菌数量具有显著抑制作用, 而对细菌和放线菌数量则没有明显影响。防病促生试验表明: B96-Ⅱ-gfp可增加黄瓜株高、鲜重和干重, 对黄瓜枯萎病有一定的防治效果, 且与未标记菌株B96-Ⅱ的防病促生作用无显著差异。

     

    Abstract: Colonization dynamics of plant growth promoting Bacillus B96-Ⅱlabeled with GFP (B96-Ⅱ-gfp) in soils and cucumber plants were investigated via Green Fluorescent Protein Gene Label technique. The results show that B96-Ⅱ-gfp successfully colonizes in soils and cucumber plants. Total B96-Ⅱ-gfp populations are respectively 2.7×104 cfu·g-1 and 6.6×104 cfu·g-1 at 180 days after incubation in natural soils and usarium wilt pathogenic soils. B96-Ⅱ-gfp is detectable in the soil even at 360 days after incubation. Spatial colonization test indicates that B96-Ⅱ-gfp can colonize in the surface (0~4 cm), middle layer (4~8 cm) and lower layer (8~12 cm) of soil, and its population density increases with soil depth. Furthermore, B96-Ⅱ-gfp colonizes in different organs of cucumber plants, with a higher population in the interior than at the exterior surface. B96-Ⅱ-gfp population in roots is highest (7.2×104 cfu·g-1). Whereas B96-Ⅱ-gfp significantly reduces the population of cultured native soil microorganisms (e.g., fungi), it does not significantly influence the population of bacteria and antinomyces. The results further show that B96-Ⅱ-gfp increases the height, fresh weight and dry weight of cucumber plants. B96-Ⅱ-gfp exhibits a certain degree of control on Fusarium wilt. Labeled B96-Ⅱ-gfp is not significantly different from un-labeled B96-Ⅱ in terms of disease control and growth promotion.

     

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