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接种定植于泛菌属和假单胞菌属菌株中的 pqqE 基因可减少化肥用量,同时促进小麦生长和提高产量。

Inoculation of pqqE gene inhabiting Pantoea and Pseudomonas strains improves the growth and grain yield of wheat with a reduced amount of chemical fertilizer.

机构信息

Department of Environmental Sciences, COMSATS University Islamabad, Vehari Campus, Pakistan.

Department of Bioinformatics and Biotechnology, Government College University, Faisalabad, Pakistan.

出版信息

J Appl Microbiol. 2020 Sep;129(3):575-589. doi: 10.1111/jam.14630. Epub 2020 Mar 30.

Abstract

OBJECTIVE

The present study was performed to examine the role of pqqE inhabiting rhizobacteria in organic acid production and relationship of the organic acids with phosphate solubilization by the bacteria in vitro as well as in vivo.

METHODS AND RESULTS

The pqqE gene was PCR amplified and sequenced in genomic DNA of Pantoea sp. WP-5 and Pseudomonas sp. NN-4. Nucleotide sequence obtained from WP-5 and NN-4 showed maximum sequence similarity (88 and 89%, respectively) with the pqqE gene of Pseudomonas fluorescens strain CMR12a (KM251420). Deduced amino acid sequence from pqqE gene of Pseudomonas sp. NN-4 and Pantoea sp. WP-5 showed 75 and 93% similarity, respectively, with protein pyrroloquinoline quinone. Phosphate solubilization and acid production assay were quantified on spectrophotometer and high-profile liquid chromatograph, respectively, by each bacterial strain. Both strains produced organic acids such as acetic, citric, gluconic, succinic and malic acid and lowered the pH of Pikovskaya broth medium under laboratory conditions. Phosphate solubilization by Pantoea sp. WP-5 was 311 ± 4 and 204 ± 3 µg ml in the culture medium supplemented with glucose and sucrose as carbon source, respectively. Pseudomonas sp. NN-4 solubilized 176 ± 3 and 298 ± 5 µg ml phosphate in Pikovskaya broth medium under similar conditions. In field experiments conducted during two consecutive years, the concentration of acetic acid and gluconic acid was higher in root exudates of plants treated with Pantoea sp. WP-5 at 30% reduced doses of nitrogen (N)- and phosphorus (P)-based chemical fertilizers as compared to non-inoculated plants. Values of chlorophyll contents, crop growth rate, leaf area index, straw yield and P contents were recorded higher in plants inoculated with Pantoea sp. WP-5 and Pseudomonas sp. NN-4 as compared to non-inoculated control. Grain yield was increased by 10-12% due to inoculation with Pantoea sp. WP-5 and Pseudomonas sp. NN-4 over non-inoculated control in the field experiments.

CONCLUSIONS

These results lead to the conclusions that the rhizobacteria inhabiting pqqE gene produced organic acids and solubilized the phosphate in vitro. On inoculation to wheat plants in field experiments, these strains produced the organic acids, solubilized the phosphate, and improved the P uptake and productivity of wheat.

SIGNIFICANCE AND IMPACT OF THE STUDY

The Pantoea sp. WP-5 and Pseudomonas sp. NN-4 are the potential candidates for inoculation to wheat as phosphate solubilizer even with reduced chemical fertilizer dose. The inoculation of the strains may enhance grain yield and net income of the farmer even with less chemical fertilizer application. This practice will be helpfull inminimizing environmental pollution.

摘要

目的

本研究旨在探讨栖息在根际的 pqqE 菌在体外和体内产生有机酸和溶解磷酸盐的作用,以及细菌产生的有机酸与磷酸盐溶解之间的关系。

方法和结果

从 Pantoea sp. WP-5 和 Pseudomonas sp. NN-4 的基因组 DNA 中扩增并测序了 pqqE 基因。从 WP-5 和 NN-4 获得的核苷酸序列与荧光假单胞菌 CMR12a (KM251420)的 pqqE 基因具有最大的序列相似性(分别为 88%和 89%)。从 Pseudomonas sp. NN-4 和 Pantoea sp. WP-5 的 pqqE 基因推导的氨基酸序列分别与吡咯喹啉醌蛋白具有 75%和 93%的相似性。通过每种细菌菌株在分光光度计和高效液相色谱仪上定量测定了溶磷和产酸试验。这两种菌株在实验室条件下产生了乙酸、柠檬酸、葡萄糖酸、琥珀酸和苹果酸等有机酸,并降低了 Pikovskaya 肉汤培养基的 pH 值。在添加葡萄糖和蔗糖作为碳源的培养基中,Pantoea sp. WP-5 的溶磷量分别为 311±4 和 204±3µgml。在类似条件下,Pseudomonas sp. NN-4 在 Pikovskaya 肉汤培养基中溶解了 176±3 和 298±5µgml 的磷酸盐。在连续两年的田间试验中,与未接种植物相比,用 30%减少的氮(N)和磷(P)基化肥处理的植物的根分泌物中乙酸和葡萄糖酸的浓度更高。接种 Pantoea sp. WP-5 和 Pseudomonas sp. NN-4 的植物的叶绿素含量、作物生长速度、叶面积指数、秸秆产量和磷含量记录较高。与未接种对照相比,田间试验中接种 Pantoea sp. WP-5 和 Pseudomonas sp. NN-4 的谷物产量增加了 10-12%。

结论

这些结果表明,栖息在 pqqE 基因上的根际细菌在体外产生有机酸并溶解磷酸盐。在田间试验中接种到小麦植物上,这些菌株产生有机酸,溶解磷酸盐,提高了小麦对磷的吸收和生产力。

研究的意义和影响

Pantoea sp. WP-5 和 Pseudomonas sp. NN-4 是潜在的候选菌株,可作为接种小麦的解磷剂,即使减少了化肥用量。菌株的接种可以提高农民的粮食产量和净收入,即使减少了化肥的使用。这种做法有助于减少环境污染。

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