西葫芦果实CpNCED1基因3′端的克隆及其表达分析  

Clonging and analysis of CpNCED1 gene in zucchini

作　　者：吴洁芳[1] 纪凯[1] 冷平[1] 任杰[1] 赵胜利[1] 孙亮[1] 

机构地区：[1]中国农业大学农学与生物技术学院,北京100193

出　　处：《中国农业大学学报》2010年第5期25-30,共6页Journal of China Agricultural University

基　　金：北京市科委重大项目(D0706002000091)

摘　　要：采用RT-PCR和RACE-PCR的方法从西葫芦果实中克隆了ABA生物合成关键酶NCED基因保守片段及其3'端,命名为CpNCED1,并通过Real time RT-PCR方法分析了CpNCED1基因在果实发育和成熟过程中的表达。结果表明:CpNCED1基因的DNA序列及其推导的蛋白质序列与番茄、鳄梨及柑橘果实相同基因的同源性分别为61.90%(LeNCED1)、62.15%(PaNCED1)、62.55%(PaNCED3)、54.67%(CsNCED1)、56.23%(CsNCED2)和69.75%、65.74%、66.67%、68.75%、71.76%。果肉中CpNCED1基因表达共出现了3个高峰,分别在花后5 d,花后20 d及花后35 d,并分别与ABA积累相对应;果皮中CpNCED1表达在花后25 d达到最大值,而瓤和种子中CpNCED1的表达没有显著变化。果皮和种子中ABA含量与其CpNCED1的表达基本一致。西葫芦果实在生长发育过程中乙烯产生量很低,为非跃变型果实;呼吸高峰出现在花后20 d。上述结果说明,西葫芦果实的成熟主要和ABA调控有关,而乙烯对果实成熟的贡献很小。One cDNAs segments encoding NCEDs which was the critical step in regulation of abscisic acid（ABA） synthesis in higher plant（zucchini） using RT-PCR and RACE-PCR and named it as CpNCED1.Using Real time RT-PCR,the expression pattern of CpNCED1 was determined.The results suggest that the DNA sequence of CpNCED1 homology with tomato、avocado、citrus was 61.90%（LeNCED1）、62.15%（PaNCED1）、62.55%（PaNCED3）、54.67%（CsNCED1）、56.23%（CsNCED2）,and the homology of protein was 69.75%、65.74%、66.67%、68.75%、71.76%.The expression of CpNCED1 has three peak after bloom in pulp,and this consistent with the accumulation of ABA.The expression of CpNCED1 has a peak at 25 days after bloom（DAB） in peep,and the expression of CpNCED1 has not obvious change in seeds.ABA content of peep and seeds was resond consistent with the expression of CpNCED1.Ethylene production capacity was very lower during growth and development of zucchini fruit,and the respiration rates was higher at 20 DAB.The result show the ripening of zucchini fruit was mainly related with the regulation of ABA,and the role of ethylene was not important.

关 键 词：西葫芦 CpNCED1 ABA 实时定量RT-PCR 

分 类 号：S642.6[农业科学—蔬菜学]