关键词: 龙眼, 体胚发生, Remorin基因家族, 克隆, qRT-PCR

Abstract: The RT-PCR combined with RACE method was used to clone the complete cDNA sequences of 5 members of Remorin family from embryogenic callus in Dimocarpus longan. The complete cDNA sequences of DlRemorin1–5 were 2 127，1 829，1 937，1 743 and 1 043 bp，DlRemorin1–5 encoding 572，444，541，466 and 181 amino acids，respectively. The DNA sequences of DlRemorin1–5 were 4 054，3 097，3 505，4 690 and 1 935 bp，and all the splice sites of the introns contained were obeyed to the“GT-AG”rule. DlRemorin1–5 belong to the instability and hydrophilous proteins，they all had the Remorinconserved domain，and had no signal peptide，only DlRemorin1 had transmembrane structure. The sequences of both nucleotides and amino acids of the five members were high homologous with those of the known Remorin genes in other species. Anglicizing phylogenetic tree of Remorin in plants indicated that DlRemorin5 belonged to the classical Remorin，DlRemorin1 and DlRemorin2 belonged to the long Remorin. The results of qRT-PCR indicated that DlRemorin1 showed approximately an“N”curve，it expressed at the highest levels in the heart embryos and cotyledonary embryos cultures. DlRemorin2 expressed at the lowest levels in heart embryos and cotyledonary embryos cultures. The expression of DlRemorin3 increased rapidly at the middle and late developmental stages and peaked at the cotyledonary embryos cultures. DlRemorin4 expressed at the lowest levels in globular embryos and topedo embryos cultures. DlRemorin5 expressed at the lowest levels in incomplete compact pro-embryogenic cultures. Suggesting that DlRemorin1–5 expressed with tissue specific and sequential characteristics during longan somatic embryogenesis. psRNA Target projections show DlRemorin may be regulated by miRNAs.

Key words: longan, somatic embryogenesis, Remorin family, cloning, qRT-PCR