Mouille Grégory, Witucka-Wall Hanna, Bruyant Marie-Pierre, Loudet Olivier, Pelletier Sandra, Rihouey Christophe, Lerouxel Olivier, Lerouge Patrice, Höfte Herman, Pauly Markus
Laboratoire de Biologie Cellulaire, Institut Jean-Pierre Bourgin, Institute National de la Recherche Agronomique, 78026 Versailles, France.
Plant Physiol. 2006 Jul;141(3):1035-44. doi: 10.1104/pp.106.079384. Epub 2006 May 19.
Quantitative trait loci (QTL) analysis was used to identify genes underlying natural variation in primary cell wall composition in Arabidopsis (Arabidopsis thaliana). The cell walls of dark-grown seedlings of a Bay-0 x Shahdara recombinant inbred line population were analyzed using three miniaturized global cell wall fingerprinting techniques: monosaccharide composition analysis by gas chromatography, xyloglucan oligosaccharide mass profiling, and whole-wall Fourier-transform infrared microspectroscopy. Heritable variation and transgression were observed for the arabinose-rhamnose ratio, xyloglucan side-chain composition (including O-acetylation levels), and absorbance for a subset of Fourier-transform infrared wavenumbers. In total, 33 QTL, corresponding to at least 11 different loci controlling dark-grown hypocotyl length, pectin composition, and levels of xyloglucan fucosylation and O-acetylation, were identified. One major QTL, accounting for 51% of the variation in the arabinose-rhamnose ratio, affected the number of arabinan side chains presumably attached to the pectic polysaccharide rhamnogalacturonan I, paving the way to positional cloning of the first gene underlying natural variation in pectin structure. Several QTL were found to be colocalized, which may have implications for the regulation of xyloglucan metabolism. These results demonstrate the feasibility of combining fingerprinting techniques, natural variation, and quantitative genetics to gain original insight into the molecular mechanisms underlying the structure and metabolism of cell wall polysaccharides.
数量性状基因座(QTL)分析被用于鉴定拟南芥(Arabidopsis thaliana)初生细胞壁组成自然变异背后的基因。利用三种小型化的全细胞壁指纹图谱技术,对Bay-0与Shahdara重组自交系群体的黑暗生长幼苗的细胞壁进行了分析:气相色谱法分析单糖组成、木葡聚糖寡糖质量谱分析以及全壁傅里叶变换红外显微光谱分析。观察到阿拉伯糖与鼠李糖的比例、木葡聚糖侧链组成(包括O-乙酰化水平)以及傅里叶变换红外波数子集的吸光度存在遗传变异和超亲分离现象。总共鉴定出33个QTL,它们对应于至少11个不同的位点,这些位点控制着黑暗生长下胚轴长度、果胶组成以及木葡聚糖岩藻糖基化和O-乙酰化水平。一个主要的QTL,占阿拉伯糖与鼠李糖比例变异的51%,影响了可能连接到果胶多糖鼠李半乳糖醛酸聚糖I上的阿拉伯聚糖侧链的数量,为果胶结构自然变异背后的首个基因的定位克隆铺平了道路。发现几个QTL共定位,这可能对木葡聚糖代谢的调控有影响。这些结果证明了结合指纹图谱技术、自然变异和数量遗传学来深入了解细胞壁多糖结构和代谢潜在分子机制的可行性。