拐枣七内生细菌溶磷相关基因的鉴定

发布时间：2018-03-15 06:26

本文选题：溶磷能力　切入点：pqqE　出处：《西北植物学报》2017年08期 　论文类型：期刊论文

【摘要】：为了鉴定拐枣七内生细菌Pseudomonas sp.GZJR-8的pqqE和GDH基因在溶磷方面的功能,该研究通过同源重组技术分别获得pqqE和GDH基因的缺失突变体ΔpqqE和ΔGDH,采用电转法获得它们的互补菌株ΔpqqE(pqqE)和ΔGDH(GDH);难溶性无机磷培养基(PKO)定性检测结果显示,与野生型菌株(WT)相比,ΔpqqE和ΔGDH不能产生溶磷圈,ΔpqqE(pqqE)和ΔGDH(GDH)能够产生溶磷圈;抗坏血酸-钼蓝显色定磷法定量检测结果显示,WT、ΔpqqE、ΔpqqE(pqqE)、ΔGDH、ΔGDH(GDH)产生的有效磷总量分别为1 939.000 mg/L、1 279.000mg/L、1 999.000mg/L、439.000mg/L、2 314.000mg/L,与WT产生的有效磷总量相比,ΔpqqE降低1.52倍,ΔpqqE(pqqE)增加1.03倍,ΔGDH降低4.42倍,ΔGDH(GDH)增加1.19倍;培养液离心后上清的pH测定结果显示,WT、ΔpqqE、ΔpqqE(pqqE)、ΔGDH、ΔGDH(GDH)的pH分别为4.08、4.34、4.03、4.71、4.00,与WT的pH相比,ΔpqqE上升1.06倍,ΔpqqE(pqqE)降低0.27倍,ΔGDH上升1.15倍,ΔGDH(GDH)降低1.02倍。研究表明:pqqE和GDH基因具有溶磷能力,其中ΔpqqE和ΔGDH较WT的溶磷能力下降,但并未完全丧失溶磷能力,而ΔpqqE(pqqE)和ΔGDH(GDH)可以恢复到WT的溶磷能力,拐枣七内生细菌Pseudomonas sp.GZJR-8是通过产生酸性物质来溶磷,在农业生产方面具有潜在应用价值。
[Abstract]:In order to identify the function of the pqqE and GDH genes of endophytic bacteria Pseudomonas sp.GZJR-8 in phosphorus solubilization, In this study, the deletion mutants 螖 pqqE and 螖 GDHof pqqE and GDH genes were obtained by homologous recombination technique, and their complementary strains 螖 pqqE) and 螖 GDHH GDHH were obtained by electroporation. Compared with wild-type strain, 螖 pqqE and 螖 GDH could not produce phosphorus solubilization cycle, and 螖 pqqE (pqqE) and 螖 GDH (GDH) could produce phosphorus solubilizing cycle. The total amount of available phosphorus produced by ascorbic acid-molybdenum blue colorimetric determination method was 1.52 times lower than that of total available phosphorus produced by WTT, 螖 pqqE, 螖 pqqE, 螖 GDH, 螖 GDHH, respectively. The total amount of available phosphorus produced was 1.03 times higher, 螖 GDH decreased 4.42 times, 螖 GDH decreased 4.42 times, 螖 GDH decreased 4.42 times, 螖 GDH decreased 1.19 times, compared with total available phosphorus produced by WT, 螖 pqqE decreased 1.52 times, 螖 pqqEpqqE increased 4.42 times, 螖 GDH decreased 4.42 times, 螖 GDH decreased 4.42 times, 螖 GDH decreased 4.42 times, 螖 GDH decreased 4.42 times, 螖 GDH decreased 4.42 times, 螖 GDH decreased 4.42 times, 螖 GDH decreased 4.42 times, 螖 GDH decreased 4.42 times, 螖 GDH decreased 4.42 times, 螖 GDH decreased 4.42 times, 螖 GDH decreased 4.42 times, 螖 GDH decreased 4.42 times, 螖 GDH decreased 4.42 times, 螖 GDH decreased 1.19 times. The results of pH determination of supernatant after centrifugation showed that the pH values of WTH, 螖 pqqE, 螖 pqqE, 螖 GDH and 螖 GDHN were 4.08 ~ 4.34 ~ 4.03 ~ 4.71 ~ 4.00, respectively. Compared with the pH of WT, 螖 pqqE increased 1.06 times, 螖 pqqqE) decreased 0.27 times, 螖 GDH increased 1.15 times, 螖 GDH-GDH) decreased 1.02 times. The results showed that 螖 pqqE and GDH gene had the ability to dissolve phosphorus, and 螖 pqqqE increased 0.27 times, 螖 GDH increased 1.15 times, 螖 GDH-GDH decreased 1.02 times, compared with WT pH, 螖 pqqE increased 1.06 times, 螖 GDH increased 1.15 times, 螖 GDH-GDH decreased 1.02 times. The phosphorus solubility of 螖 pqqE and 螖 GDH was lower than that of WT, but not completely lost, and 螖 pqqE (pqqE) and 螖 GDH (GDH) could recover to WT's. Pseudomonas sp.GZJR-8, an endophytic bacterium, could dissolve phosphorus by producing acidic substances. It has potential application value in agricultural production.
【作者单位】：西北农林科技大学生命科学学院;
【基金】：陕西省自然科学基金(2016JM3014)
【分类号】：Q933

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