类萌发素蛋白(germin-like protein, GLPs)是一类与小麦萌发素序列相似性较高、位于胞外基质的可溶性糖蛋白, 在植物的生长发育阶段以及对生物和非生物胁迫的应答中起着重要的作用。为了研究GLP13基因的生理功能, 我们分离并鉴定了GLP13的敲减突变体glp13, 同时构建了其超表达植株35S::GLP13。用甲基紫精(methyl viologen, MV)处理2种不同基因型和野生型(WT)植株, 结果发现, 与野生型相比, 突变体glp13子叶变绿率较低, 主根生长受抑制较明显; 而超表达植株35S::GLP13子叶变绿率较高, 主根生长的受抑制程度较WT轻。用MV处理2周的35S::GLP13植株, 其叶绿素荧光参数Fv/Fm 的下降较野生型对照缓慢。半定量RT-PCR分析结果表明, 与野生型相比, 经MV处理4小时后的35S::GLP13中抗氧化酶系基因FSD1的表达上调, 而CAT1、CSD1和UGT71C1的表达水平在35S::GLP13、glp13和野生型植株三者之间没有明显差异。以上结果表明GLP13基因在拟南芥抗氧化胁迫响应中起重要作用。
Germin-like proteins (GLPs) comprise a large family of soluble extracellular matrix glycoprotein, which is similar to wheat germin and plays an important role in growth and development and in biotic and abiotic stress responses of plants. To understand the role of GLP13 in planta, we investigated its expression patterns; isolated and characterized a knock-down mutation in the GLP13 gene (named glp13); and constructed overexpression GLP13 plants (named 35S::GLP13). Compared to the wild type, the glp13 mutant showed a lower cotyledon green ratio and more restrained root growth when 3 different types of plants were treated with methyl viologen (MV); however, the cotyledon green ratio was higher and the root growth less restrained in 35S::GLP13 plants. The chlorophyll fluorescence parameter Fv/Fm of 35S::GLP13 plants decreased slower than in the wild type after 2-week MV treatment of seedlings. The expression of oxidative stress response genes (CSD1, FSD1, UGT71C1 and CAT1) were analyzed by semi-quantitative RT-PCR; the expression of FSD1 was increased in 35S::GLP13 plants, with no differences in the expression level of CAT1, CSD1 and UGT71C1 among 3 different types of plants after 4-hr treatment with MV. Our results indicate that GLP13 plays an important role in response to oxidative stress in Arabidopsis.
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