Lo Tiffany Chien Ting, Kang Ming Wei, Wang Bor Cheh, Chang C Allen
Department of Biological Science and Technology, National Chiao Tung University, Hsinchu, Taiwan, ROC.
Anal Chim Acta. 2007 Jun 5;592(2):146-53. doi: 10.1016/j.aca.2007.04.021. Epub 2007 Apr 20.
We report here the first combined amplified fragment length polymorphism (AFLP) analysis of genomic DNA fingerprinting data and cluster analysis of the exo-polysaccharide glycosyl linkage data of 10 regionally different strains of Lentinula edodes to compare their genetic and structural similarities and differences. In addition, the monosaccharide compositions, molecular weights, glycosyl structural linkages were investigated for the exo-polysaccharides extracted from these different phylogenetic groups of regionally different L. edodes. All exo-polysaccharides had similar molecular weight distribution between 1x10(4) and 3x10(6) Da and the monosaccharide composition analysis revealed the presence of heterogeneous materials containing glucose, mannose, xylose, galactose, fucose, rhamnose and arabinose in different ratios. Among these monosaccharides, the glucose contents are the highest for all but one strain, indicating that glucose probably is the building block of the backbones of these exo-polysaccharides. The AFLP assay data helped to classify the 10 L. edodes strains into three distinct genetic groups. Gas chromatographic and mass spectrometric (GC-MS) data revealed five different glycosyl linkage types for these exo-polysaccharides. Most of the exo-polysaccharide backbone structures contain (1-->4)-linked-D-glucopyranosyl and (1-->6)-linked-D-glucopyranosyl moieties. Arabinose 1-->4 linkages and mannose 1-->2 linkages also exist in all strains. The only differences among these linkages are their monosaccharide compositions leading to different degree of backbone and branch formations. Cluster analyses of the GC-MS data of the exo-polysaccharides of the 10 strains resulted in 10 dendrograms. However, four of the 10 dendrograms were identical and were obtained using the average, Ward and weighted linkage type method of Manhattan distance and using the Ward method of Euclidean distance. The results of cluster analyses were not very much different from that of the AFLP assay and allowed the comparison of genetic and structural similarities and differences.
我们在此报告首次对10个来自不同地区的香菇菌株进行基因组DNA指纹图谱数据的联合扩增片段长度多态性（AFLP）分析以及胞外多糖糖基连接数据的聚类分析，以比较它们的遗传和结构异同。此外，还对从这些不同系统发育组的地区性不同香菇中提取的胞外多糖的单糖组成、分子量、糖基结构连接进行了研究。所有胞外多糖的分子量分布相似，在1×10⁴至3×10⁶ Da之间，单糖组成分析表明存在含有葡萄糖、甘露糖、木糖、半乳糖、岩藻糖、鼠李糖和阿拉伯糖的异质物质，比例各不相同。在这些单糖中，除一个菌株外，所有菌株的葡萄糖含量最高，这表明葡萄糖可能是这些胞外多糖主链的组成单元。AFLP分析数据有助于将10个香菇菌株分为三个不同的遗传组。气相色谱和质谱（GC-MS）数据显示这些胞外多糖有五种不同的糖基连接类型。大多数胞外多糖主链结构包含（1→4）连接的-D-吡喃葡萄糖基和（1→6）连接的-D-吡喃葡萄糖基部分。所有菌株中也存在阿拉伯糖1→4连接和甘露糖1→2连接。这些连接之间的唯一差异在于它们的单糖组成，导致主链和分支形成程度不同。对10个菌株的胞外多糖的GC-MS数据进行聚类分析得到了10个树状图。然而，10个树状图中有4个是相同的，它们是使用曼哈顿距离的平均、沃德和加权连锁类型方法以及欧几里得距离的沃德方法获得的。聚类分析结果与AFLP分析结果差异不大，能够比较遗传和结构的异同。
