以‘岩溪晚芦’椪柑(Citrus reticulata Blanco)成熟果实果皮和果肉为材料,采用抑制性差减杂交技术成功构建了果皮(tester)与果肉(driver)的差减cDNA文库。结果显示:cDNA克隆外源插入片段大小介于100～500bp之间。成功对411个克隆进行了测序,测序结果经与NCBI基因库中序列进行比较,377个EST找到了同源序列,它们分属于126个基因,涉及到色素合成、能量代谢、初级代谢、次生代谢、抗逆防御、蛋白代谢、信号转导、香味形成、果皮软化(衰老)、精油合成、抗氧化等代谢途径和生理生化过程。还有一些基因涉及到三价铁螯合物、反转录转座子gag蛋白、查尔酮合成、硼离子转运、儿茶酚氧位甲基转移酶等功能。功能已知且没有重复的EST共有63个,占总数的49.2%。有5个基因共62个克隆虽在Blast搜索时找到了较高的同源性序列,但功能目前尚未明确。另有34个基因未搜索到同源序列,可能为新发现的基因。 A subtracted cDNA library of tester and driver was successfully constructed using suppression subtractive hybridization (SSH) from the peel and pulp of ripe fruit of Citrus reticulata Blanco. The results showed that the size of exogenous insert of cDNA clone ranged from 100 to 500 bp. 411 clones were successfully sequenced. Compared with the NCBI gene sequences, 377 ESTs found homologous sequences, belonging to 126 genes, which involved in pigment synthesis, energy metabolism, primary metabolism, secondary Metabolism, anti-retrograde defense, protein metabolism, signal transduction, flavor formation, pericarp softening (aging), essential oil synthesis, antioxidant and other metabolic pathways and physiological and biochemical processes. There are also some genes related to ferric chelates, anti-transposon gag protein, chalcone synthesis, boron ion transport, catechol oxygen methyltransferase and other functions. There are 63 ESTs with known functions and no duplication, accounting for 49.2% of the total. A total of 62 clones with 5 genes found a higher homology sequence during Blast search, but the function is not yet clear. Another 34 genes were not found homologous sequences, may be newly discovered genes.