荔枝采后果皮褐变过程中差异表达基因的SSH分析

 以‘妃子笑’荔枝果实为材料，采用抑制差减杂交（SSH）与cDNA 微阵列技术相结合，研究了采后荔枝果皮褐变过程中的基因差异表达。分别以采后0 h 与32 h 的果皮总RNA 为驱动组与检测组，构建了正向与反向SSH 文库，分别获得了282 个与76 个阳性克隆。通过cDNA 微阵列杂交筛选获得了在采后32 h 果皮中上调表达克隆17 个，下调表达克隆49 个，分别代表了在采后32 h 果皮中上调表达基因16 个和下调表达基因17 个，其中有较多的热击蛋白基因、糖代谢相关基因、细胞壁代谢相关基因等。RT-PCR 检测基因表达结果与cDNA 微阵列杂交结果一致。

Analysis of Genes Differentially Expressed During Pericarp Browning of Postharvest Litchi Fruit Using Suppression Subtractive Hybridization Technology

The differentially expressed genes between pericarp of‘Feizixiao’fruit at 0 h and 32 h afterharvest（HAH）were identified by the combination of suppression subtractive hybridization（SSH）andcDNA microarray technology. The reciprocal，forward and reverse，SSH libraries were constructed withRNA extracted from the pericarp at 0 HAH and 32 HAH as tester and driver，respectively. Then 282 and76 positive clones were obtained from the forward and reverse library. However，only 17 and 49 cloneswith enhanced or repressed expression at 32 HAH were identified by cDNA microarray hybridization，representing 16 and 17 genes both with enhanced and repressed expression in the pericarp at 32 HAH，respectively. Among the tested genes，genes encoding enzymes involved in carbohydrate metabolism，cellwall metabolism and genes encoding heat shock proteins had higher frequencies than other genes. Theresults of cDNA microarray hybridization were confirmed by RT-PCR.