葡萄风信子黄酮醇合酶基因克隆和表达分析
Cloning and Expression Analysis of FLS Gene in Muscari armeniacum

以葡萄风信子‘亚美尼亚’的根、茎、叶、花为材料，采用RT-PCR技术克隆得到一条FLS基因，命名为MaFLS2。生物信息学分析表明，该基因cDNA全长为1 152 bp，含有完整的开放读码框1 056 bp，编码353个氨基酸，MaFLS2蛋白是一种亲水性蛋白，氨基酸序列含有典型的DIOX_N结构域(2-酮戊二酸-双加氧酶活性的N端区域)和2OG-FeII_Oxy超家族结构域，属于2-酮戊二酸-双加氧酶(2-ODDs)蛋白家族。实时定量PCR结果显示，MaFLS2基因在蓝白葡萄风信子中的表达存在组织特异性，蓝色品种‘亚美尼亚’中在花蕾未着色时表达量最高，而白色品种‘白丽人’在根中表达量最高， 在花发育的不同阶段，MaFLS2基因在‘亚美尼亚’中的表达峰值出现在花蕾未着色时期，基因表达总体呈现出先降低后升高再降低的趋势，而在‘白丽人’中的表达峰值出现在转色中期，总体呈现出先升后降的趋势。研究结果为开展单子叶植物花色育种提供了理论依据。
In this study,we cloned a flavonol synthase gene (FLS) from grape hyacinth Muscari armeniacum (MaFLS2) by using PCR technique.Bioinformatics analysis showed that the length of MaFLS2 cDNA was 1 152 bp,including a complete open reading frame with 1 056 bp.MaFLS2 codes 353 amino acids and belonging to hydrophilic protein.NCBI analysis showed that MaFLS2 protein had the typical DIOX_N and 2OG-FeII_Oxy structure domains,therefore,MaFLS2 belonged to 2-oxoglutarate-dioxygenase protein families.Real time PCR showed that MaFLS2 gene was a tissue specific gene in grape hyacinths,and it had the highest expression level in flower buds at the period of starting to pigment in M.armeniacum while in M.uscariaucheri ‘White Beauty’,its expression peak value appeared in roots.At different flowering developmental phases,MaFLS2 gene expressed the highest in uncolored flower buds of M.armeniacum,whose flower was blue in color,its expression pattern showed a overall tread of reducing firstly,increasing before decreasing again to its minimum level.However,in M. ‘White Beauty’,whose flower was white in color,MaFLS2 gene presented a different expression pattern as its expression peak value appeared in flower buds staying at the middle period of its changing color,and it showed a general expression trend of increasing before declining.This study could be used to explore the function of FLS gene in flower coloring