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定殖于小麦(普通小麦)根际的固氮螺菌的RNA测序转录谱分析

RNA-seq transcriptional profiling of Herbaspirillum seropedicae colonizing wheat (Triticum aestivum) roots.

作者信息

Pankievicz V C S, Camilios-Neto D, Bonato P, Balsanelli E, Tadra-Sfeir M Z, Faoro H, Chubatsu L S, Donatti L, Wajnberg G, Passetti F, Monteiro R A, Pedrosa F O, Souza E M

机构信息

Department of Biochemistry and Molecular Biology, Universidade Federal do Paraná, Curitiba, PR, Brazil.

Department of Biochemistry and Biotechnology, Universidade Estadual de Londrina, Londrina, PR, Brazil.

出版信息

Plant Mol Biol. 2016 Apr;90(6):589-603. doi: 10.1007/s11103-016-0430-6. Epub 2016 Jan 22.

Abstract

Herbaspirillum seropedicae is a diazotrophic and endophytic bacterium that associates with economically important grasses promoting plant growth and increasing productivity. To identify genes related to bacterial ability to colonize plants, wheat seedlings growing hydroponically in Hoagland's medium were inoculated with H. seropedicae and incubated for 3 days. Total mRNA from the bacteria present in the root surface and in the plant medium were purified, depleted from rRNA and used for RNA-seq profiling. RT-qPCR analyses were conducted to confirm regulation of selected genes. Comparison of RNA profile of root attached and planktonic bacteria revealed extensive metabolic adaptations to the epiphytic life style. These adaptations include expression of specific adhesins and cell wall re-modeling to attach to the root. Additionally, the metabolism was adapted to the microxic environment and nitrogen-fixation genes were expressed. Polyhydroxybutyrate (PHB) synthesis was activated, and PHB granules were stored as observed by microscopy. Genes related to plant growth promotion, such as auxin production were expressed. Many ABC transporter genes were regulated in the bacteria attached to the roots. The results provide new insights into the adaptation of H. seropedicae to the interaction with the plant.

摘要

固氮螺菌是一种能进行固氮作用的内生细菌,与具有重要经济价值的禾本科植物共生,促进植物生长并提高产量。为了鉴定与细菌定殖植物能力相关的基因,将在霍格兰氏培养基中进行水培的小麦幼苗接种固氮螺菌,并培养3天。从根表面和植物培养基中的细菌中纯化总mRNA,去除rRNA后用于RNA测序分析。进行RT-qPCR分析以确认所选基因的调控。对附着在根上的细菌和浮游细菌的RNA谱进行比较,发现它们对附生生活方式有广泛的代谢适应性。这些适应性包括特定黏附素的表达和细胞壁重塑以附着在根上。此外,新陈代谢适应了微氧环境,固氮基因得以表达。聚羟基丁酸酯(PHB)合成被激活,通过显微镜观察到PHB颗粒被储存起来。与促进植物生长相关的基因,如生长素产生相关基因被表达。许多ABC转运蛋白基因在附着于根的细菌中受到调控。这些结果为固氮螺菌适应与植物相互作用提供了新的见解。

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