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采用RT-PCR和RACE相结合的方法,从荔枝果实中分离得到两个内切-1,4-β-葡聚糖酶基因(EG)全长序列,分别命名为LcEG1和LcEG2。推导的LcEG1和LcEG2蛋白均含有4个半胱氨酸Cys残基和两个保守的糖基水解酶启动位点。Northernblotting分析表明,LcEG1mRNA在果皮发育过程中逐渐减弱,而在果肉发育过程中逐渐增强;LcEG2仅在果肉发育的前期表达,在果皮中检测不到其表达。LcEG1在荔枝易裂果品种‘糯米糍’果皮和果肉中的表达均明显强于在不易裂果品种‘淮枝’果皮和果肉中的表达。上述结果表明,LcEG1的表达可能参与荔枝果实发育,并且与裂果密切相关,而LcEG2可能只参与果肉的早期生长。
The full-length sequences of two endo-1,4-β-glucanase genes (EG) were isolated from the litchi fruits by RT-PCR and RACE, and named as LcEG1 and LcEG2 respectively. The deduced LcEG1 and LcEG2 proteins all contain 4 cysteine Cys residues and two conserved glycosyl hydrolase initiation sites. Northern blotting analysis showed that LcEG1mRNA decreased gradually in the process of pericarp development and gradually increased in the process of pulp development. LcEG2mRNA was only expressed in the early stage of pulp development, but not in the pericarp. The expression of LcEG1 was significantly higher in pericarp and flesh than that of the ’Huai Zhi’ peel and flesh, which were the most susceptible varieties of ’Litchi’. The above results showed that LcEG1 expression may be involved in the development of litchi fruit, and is closely related to the split fruit, and LcEG2 may only be involved in the early growth of the pulp.