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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。
Using litchi fruit of ’Feizixiao’ as material, the differential expression of genes during postharvest litchi pericarp browning was studied by SSH and cDNA microarray. The total RNA of pericarp collected at 0 h and 32 h postharvest were used as driving and testing groups respectively to construct forward and reverse SSH libraries, and 282 and 76 positive clones were obtained respectively. 17 cDNA clones were found in the pericarp 32 h after harvesting, 49 clones were down-regulated, 32 of which were up-regulated and 17 down-regulated in 32 h post-harvest, respectively There are more heat shock protein genes, glucose metabolism related genes, cell wall metabolism related genes. RT-PCR detection of gene expression results and cDNA microarray hybridization results.