Exchange of a Conjugative Plasmid At Different Soil Moisture Levels Between Streptomyces Species Colonizing Artificial Soil Aggregates

B. Bleakley, D. Crawford
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Abstract

Spores of a Streptomyces lividans strain (donor) bearing the recombinant conjugative plasmid pIJ303, which codes for thiostrepton resistance, and spores of a plasmid-free Streptomyces parvulus strain (recipient) were added together to finelysieved, sterile silt loam soil. This bulk soil was adjusted to either 20%, 35% or 45% of the soil’s water holding capacity. To this inoculated bulk soil were added sterile, nutrientamended, artificial soil aggregates. After incubation to allow streptomycete growth and aggregate colonization, both the sieved bulk soil and the aggregates were assayed for numbers of transconjugants by spreadplanting on a thiostrepton – agar selective medium. This allowed estimation of parental ability to colonize nutrient – rich soil sites at different soil moisture levels, and comparison of plasmid exchange frequency in the nonamended bulk soil versus the nutrient – rich soil aggregates. Plasmid exchange was detected only on aggregates of about 20% water holding capacity or less (4.2% wt/wt H20). The soil moisture threshold for heterotrophic streptomycete activity appeared to lie between 2.87% and 4.2% wt/wt H20 for the nutrientamended soil aggregates.
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不同土壤湿度下链霉菌种间定殖人工土壤团聚体的共轭质粒交换
将带有硫链菌抗性重组质粒pIJ303的lividans链霉菌菌株(供体)和无质粒的parvulus链霉菌菌株(受体)的孢子一起添加到细筛的无菌粉壤土中。这种散装土壤被调整为土壤持水量的20%、35%或45%。在接种后的块状土壤中加入无菌、营养改良的人工土壤团聚体。经过孵育,使链霉菌生长和聚集体定植后,通过在硫链菌-琼脂选择性培养基上铺开,检测筛选过的大块土壤和聚集体的转偶联菌数量。这样就可以估计在不同土壤水分水平下亲本在富营养化土壤中的定殖能力,并比较未改良的散装土壤与富营养化土壤团聚体的质粒交换频率。质粒交换仅在持水量约为20%或更少(4.2% wt/wt H20)的聚集体上检测到。异养链霉菌活性的土壤水分阈值在2.87% ~ 4.2% wt/wt H20之间。
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