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从热胁迫和非胁迫小麦根际土壤中分离植物促生根际细菌的特性及其作为生物接种剂的应用。

Characterisation of plant growth-promoting rhizobacteria from rhizosphere soil of heat-stressed and unstressed wheat and their use as bio-inoculant.

机构信息

Department of Plant Sciences, Quaid-i-Azam University, Islamabad, Pakistan.

Department of Biosciences, University of Wah, Wah Cantt, Pakistan.

出版信息

Plant Biol (Stuttg). 2019 Jul;21(4):762-769. doi: 10.1111/plb.12972. Epub 2019 Apr 1.

Abstract

High temperature induces several proteins in plants that enhance tolerance to high temperature shock. The fate of proteins synthesised in microbial cells or secreted into culture media by interacting microbes has not been fully elucidated. The present investigation aimed to characterise plant growth-promoting rhizobacteria (PGPR) isolated from the rhizosphere of wheat genotypes (differing in tolerance to high temperature stress) and evaluate their performance as bioinoculant for use in wheat. Four bacterial strains, viz. Pseudomonas brassicacearum, Bacillus thuringiensis, Bacillus cereus strain W6 and Bacillus subtilis, were isolated from the rhizosphere of heat-stressed and unstressed wheat genotypes. The wheat genotypes were exposed to high temperature stress at 45 °C for 10 days (3 h daily) at pre-anthesis phase. Isolates were identified on the basis of morphology and biochemical characteristics, 16S rRNA gene sequencing and whole cell protein profiles. Results were further complemented by size exclusion chromatography (SEC) with fast protein liquid chromatography (FPLC) and SDS PAGE of 80% ammonium sulphate precipitates of the cell-free supernatants. Isolates were positive for catalase, oxidases and antimicrobial activity. P. brassicacearum from the rhizosphere of the heat-tolerant genotype was more efficient in phosphate solubilisation, bacteriocin production, antifungal and antibacterial activity against Helminthosporium sativum, Fusarium moniliforme and Klebsiella pneumonia, respectively. The inoculated seedlings had significantly higher root and shoot fresh weight, enhanced activity of antioxidant enzymes, proline and protein content. Total profiling of the culture with SDS-PAGE indicated expression of new protein bands in 95 kDa in P. brassicacearum. Temperature-induced changes in PGPR isolates are similar to those in the host plant. P. brassicacearum may be a good candidate for use in biofertiliser production for plants exposed to high temperature stress.

摘要

高温会诱导植物产生几种蛋白质,从而增强其对高温冲击的耐受性。微生物细胞内合成的蛋白质或与微生物相互作用后分泌到培养基中的蛋白质的命运尚未完全阐明。本研究旨在从小麦基因型(对高温胁迫的耐受性不同)的根际中分离出具有促植物生长的根际细菌(PGPR),并评估它们作为生物接种剂在小麦中的应用效果。从热胁迫和非胁迫小麦基因型的根际中分离到了 4 种细菌菌株,分别为:野油菜黄单胞菌、苏云金芽孢杆菌、蜡样芽孢杆菌 W6 和枯草芽孢杆菌。在开花前阶段,将小麦基因型暴露在 45°C 的高温胁迫下 10 天(每天 3 小时)。根据形态和生化特征、16S rRNA 基因测序和全细胞蛋白图谱对分离物进行鉴定。结果还通过排阻色谱(SEC)与快速蛋白液相色谱(FPLC)以及无细胞上清液的 80%硫酸铵沉淀的 SDS-PAGE 进一步补充。分离物对过氧化氢酶、氧化酶和抗菌活性呈阳性。耐热基因型根际的野油菜黄单胞菌在溶解磷酸盐、产生细菌素、抑制真菌和抑制对禾谷镰刀菌、串珠镰刀菌和肺炎克雷伯氏菌的抗菌活性方面更为有效。接种的幼苗的根和茎鲜重显著增加,抗氧化酶、脯氨酸和蛋白质含量的活性增强。SDS-PAGE 对培养物的总分析表明,野油菜黄单胞菌在 95kDa 处表达了新的蛋白质条带。PGPR 分离物的温度诱导变化与宿主植物的变化相似。野油菜黄单胞菌可能是用于生产在高温胁迫下暴露的植物的生物肥料的良好候选物。

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