摘要
【目的】以解磷菌Penicillium brocae为对象,研究其在土壤中的定殖动态和解磷效果,为菌剂的开发和应用提供参考。【方法】利用携带多种筛选标记的P.brocae转化子Z3开展土壤培育试验,结合筛选标记,通过平板计数法统计该菌在土壤中存活数量随时间的变化,同时分析土壤有效磷随时间的变化。【结果】绿色荧光(gfp)、潮霉素抗性(hph)及β-葡萄糖苷酸酶(gus)基因在转化子Z3中均可正常表达,且转化子Z3和野生型菌株的生长曲线和解磷能力无显著差异。转化子Z3以1×10^(6) CFU/g接种土壤后,在定殖数量上随时间呈下降、上升和下降的总体趋势。接种后3周,转化子Z3的数量达到峰值4.3×10^(6) CFU/g,后期逐步下降并在接种后10周达到1.4×10^(3) CFU/g。土壤有效磷含量呈先升后降趋势,接种后6周达到峰值14.7 mg/L,较初始增加41.3%。接种7周后有效磷含量逐步下降,接种后10周降至12.7 mg/L,较初始值仍高出22.1%。对转化子Z3定殖数量和土壤有效磷数据的分析发现,有效磷含量上升期转化子Z3的数量维持在1×10~5 CFU/g左右或以上,当转化子Z3数量下降并低于1×10~5 CFU/g时,土壤有效磷也随之降低。【结论】转化子Z3接种到土壤后可存续10周以上,表现出良好的定殖能力,并且能够显著提高有效磷含量。另外,转化子Z3的定殖数量对维持解磷效果具有积极作用。
【Objective】The colonization dynamics and phosphate solubilization effects of Penicillium brocae in soil were studied in order to provide references for the development and application of soil phosphate solubilization bioinoculant,.【Method】A P.brocae transformant Z3 with multiple molecular screening makers was used for soil cultivation test.Combining with the screening makers,the colony forming units (CFU) of Z3 in soil were quantified with plate counting method at different time nodes and the soil available phosphate (AP) was measured at the same time.【Result】The green fluorescent protein (gfp),hygromycin B phosphotransferase (hph) and β-glucuronidase (gus) genes could expressed normally in Z3,and there was no significant difference in the growth curve and phosphate solubilization ability between Z3 and wild type (Wt) strain.The colonization number of Z3 in soil decreased from 1×10^(6) CFU/g to 1×10~5 CFU/g at the first stage,then it increased to the maximum value of 4.3×10^(6) CFU/g at the third week and followed by a gradually decreasing to 1.4×10^(3) CFU/g in soil at the tenth week.The soil AP content showed a trend of increasing first and then decreasing.At the sixth week,the soil AP reached the maximum value of 14.7 mg/L which increased by 41.3%compared to the AP of original soil.After inoculation for seven weeks,the soli AP gradually decreased to 12.7 mg/L at the tenth week which still increased by 22.1%compared to the AP of original soil.Additionally,it was also found that the colonization number of Z3 in soil were close to or higher than 1×10~5 CFU/g during the increasing of soil AP and the AP gradually decreased when the number of Z3 was significantly lower than 1×10~5 CFU/g.【Conclusion】The transformant Z3 can well colonize for ten weeks after being inoculated into soil,which shows good colonization ability and increases soil AP concentration significantly.Additionally,the colonization number of Z3 has a positive role in the maintenance of phosphate solubilization effects.
作者
许昌超
张俊涛
苏杨
冼卓慧
XU Changchao;ZHANG Juntao;SU Yang;XIAN Zhuohui(Guangzhou Institute of Forestry and Landscape Architecture,Guangzhou 510405,China)
出处
《广东农业科学》
CAS
2022年第10期79-86,共8页
Guangdong Agricultural Sciences
基金
广州市科技计划项目(202102020517,202206010058)。
关键词
解磷菌
筛选标记
定殖
平板计数
有效磷
phosphate solubilization strain
screening maker
colonization
plate counting method
available phosphate
