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基于核磁共振代谢组学分析的马铃薯叶片代谢变化与田间晚疫病抗性关系的研究

A survey of metabolic changes in potato leaves by NMR-based metabolic profiling in relation to resistance to late blight disease under field conditions.

作者信息

Tomita Satoru, Ikeda Seishi, Tsuda Shogo, Someya Nobutaka, Asano Kenji, Kikuchi Jun, Chikayama Eisuke, Ono Hiroshi, Sekiyama Yasuyo

机构信息

Food Research Institute, NARO, Tsukuba, Ibaraki, Japan.

Hokkaido Agricultural Research Center, NARO, Memuro, Hokkaido, Japan.

出版信息

Magn Reson Chem. 2017 Feb;55(2):120-127. doi: 10.1002/mrc.4506. Epub 2016 Sep 6.

DOI:10.1002/mrc.4506
PMID:27549366
Abstract

Non-targeted nuclear magnetic resonance (NMR)-based metabolic profiling was applied to potato leaves to survey metabolic changes associated with late blight resistance under field conditions. Potato plants were grown in an experimental field, and the compound leaves with no visible symptoms were collected from 20 cultivars/lines at two sampling time points: (i) the time of initial presentation of symptoms in susceptible cultivars and (ii) 12 days before this initiation. H NMR spectra of the foliar metabolites soluble in deuterium oxide- or methanol-d -based buffers were measured and used for multivariate analysis. Principal component analysis for six cultivars at symptom initiation showed a class separation corresponding to their levels of late blight resistance. This separation was primarily explained by higher levels of malic acid, methanol, and rutin and a lower level of sucrose in the resistant cultivars than in the susceptible ones. Partial least squares regression revealed that the levels of these metabolites were strongly associated with the disease severity measured in this study under field conditions. These associations were observed only for the leaves harvested at the symptom initiation stage, but not for those collected 12 days beforehand. Subsequently, a simple, alternative enzymatic assay for l-malic acid was used to estimate late blight resistance, as a model for applying the potential metabolic marker obtained. This study demonstrated the potential of metabolomics for field-grown plants in combination with targeted methods for quantifying marker levels, moving towards marker-assisted screening of new cultivars with durable late blight resistance. Copyright © 2016 John Wiley & Sons, Ltd.

摘要

基于非靶向核磁共振(NMR)的代谢谱分析应用于马铃薯叶片,以调查田间条件下与晚疫病抗性相关的代谢变化。马铃薯植株种植在试验田中,在两个采样时间点从20个品种/品系中采集无可见症状的复叶:(i)感病品种出现症状的初始时间;(ii)症状出现前12天。测量了可溶于氧化氘或基于氘代甲醇缓冲液的叶片代谢物的1H NMR光谱,并用于多变量分析。对六个品种在症状出现时进行的主成分分析显示出与它们的晚疫病抗性水平相对应的类别分离。这种分离主要是由于抗病品种中苹果酸、甲醇和芦丁的含量高于感病品种,而蔗糖含量低于感病品种。偏最小二乘回归显示,这些代谢物的水平与本研究在田间条件下测量的病情严重程度密切相关。仅在症状出现阶段收获的叶片中观察到这些关联,而在提前12天采集的叶片中未观察到。随后,使用一种简单的替代酶法测定L-苹果酸,以估计晚疫病抗性,作为应用获得的潜在代谢标记的模型。本研究证明了代谢组学在田间种植植物中的潜力,结合靶向方法定量标记水平,朝着标记辅助筛选具有持久晚疫病抗性的新品种迈进。版权所有© 2016约翰威立父子有限公司。

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