Sekiyama Yasuyo, Okazaki Kazuyuki, Kikuchi Jun, Ikeda Seishi
Food Research Institute, National Agriculture and Food Research Organization (NARO), Tsukuba 305-8642, Japan.
RIKEN Center for Sustainable Resource Science, 1-7-22 Suehiro-cho, Yokohama 235-0045, Japan.
Metabolites. 2017 Jan 26;7(1):4. doi: 10.3390/metabo7010004.
leaf spot (CLS) is one of the most serious leaf diseases for sugar beet ( L.) worldwide. The breeding of sugar beet cultivars with both high CLS resistance and high yield is a major challenge for breeders. In this study, we report the nuclear magnetic resonance (NMR)-based metabolic profiling of field-grown leaves for a subset of sugar beet genotypes harbouring different levels of CLS resistance. Leaves were collected from 12 sugar beet genotypes at four time points: seedling, early growth, root enlargement, and disease development stages. ¹H-NMR spectra of foliar metabolites soluble in a deuterium-oxide (D₂O)-based buffer were acquired and subjected to multivariate analyses. A principal component analysis (PCA) of the NMR data from the sugar beet leaves shows clear differences among the growth stages. At the later time points, the sugar and glycine betaine contents were increased, whereas the choline content was decreased. The relationship between the foliar metabolite profiles and resistance level to CLS was examined by combining partial least squares projection to latent structure (PLS) or orthogonal PLS (OPLS) analysis and univariate analyses. It was difficult to build a robust model for predicting precisely the disease severity indices (DSIs) of each genotype; however, GABA and Gln differentiated susceptible genotypes (genotypes with weak resistance) from resistant genotypes (genotypes with resistance greater than a moderate level) before inoculation tests. The results suggested that breeders might exclude susceptible genotypes from breeding programs based on foliar metabolites profiled without inoculation tests, which require an enormous amount of time and effort.
叶斑病(CLS)是全球范围内甜菜最严重的叶部病害之一。培育兼具高抗CLS能力和高产的甜菜品种对育种者来说是一项重大挑战。在本研究中,我们报告了对田间种植的、具有不同CLS抗性水平的一部分甜菜基因型叶片进行的基于核磁共振(NMR)的代谢谱分析。在四个时间点从12个甜菜基因型中采集叶片:幼苗期、早期生长阶段、根部膨大期和病害发展阶段。获取了可溶于基于重水(D₂O)的缓冲液中的叶部代谢物的¹H-NMR谱,并进行多变量分析。对甜菜叶片的NMR数据进行主成分分析(PCA)显示,不同生长阶段之间存在明显差异。在较晚的时间点,糖和甘氨酸甜菜碱含量增加,而胆碱含量降低。通过结合偏最小二乘投影到潜在结构(PLS)或正交PLS(OPLS)分析以及单变量分析,研究了叶部代谢物谱与对CLS的抗性水平之间的关系。很难建立一个精确预测每个基因型病害严重指数(DSI)的稳健模型;然而,在接种测试之前,γ-氨基丁酸(GABA)和谷氨酰胺(Gln)能够区分易感基因型(抗性较弱的基因型)和抗性基因型(抗性大于中等水平的基因型)。结果表明,育种者可能无需接种测试(接种测试需要大量时间和精力),而是基于叶部代谢物谱从育种计划中排除易感基因型。