龙眼成花逆转相关基因表达的cDNA-AFLP分析

摘要：以龙眼正常成花花芽和成花逆转花芽为试材，应用cDNA-AFLP技术，研究龙眼正常成花与成花逆转花芽cDNA的差异表达，并利用RT-PCR技术验证，探讨了龙眼成花逆转的分子机制。通过64对引物组合的cDNA-AFLP分析获得了2 560条扩增片段。选择其中有明显差异的片段进行回收、测序，获得了14条核苷酸序列。其中有13个片段在GenBank数据库中发现同源序列，1个功能未知。通过核酸和氨基酸比对分析，与龙眼成花逆转相关的112基因片段同源于银灰杨（Populus tremula x Populus alba）中编码Nek激酶家族(NIMA related protein kinases)基因（83 ％）， 331的核苷酸序列（80％）同源于与拟南芥（Arabidopsis thaliana）中编码内切-1,4-β-葡聚糖酶(EGases)的基因，313的核苷酸序列（78%）同源于与柑橘（Citrus sinensis）中的几丁质酶（chitinase CHI1）基因。

cDNA-AFLP Analysis of Gene Expression between Normal Flowering and Flowering Reversion Buds in Longan

In order to investigate the molecular mechanism of flowering reversion in Longan(Dimocarpus longan Lour.cv.Longyou),differential expression profile of cDNA between the longan normal flowering and flowering reversion was analyzed by cDNA-AFLP.Sixty-four pairs ofprimer were used for the eDNA-AFLP amplification and totally 2 560 differentially expressed fragments were detected.Fragments with obvious difference were selected to re-amplification and purification,sequenced,and 14 sequences were obtained.According to the alignment of Blastn and Biastx,differentially expressed 112 gene fragments were 83%identical in nucleotides to Populus tremula×Populus alba NIMA-related protein kinase mRNA and 331 were 80% identical in nucleotides to Arabidopsis thaliana xyloglucan endo-1,4-beta-D-glucanase precursor mRNA and 313 were 78%identical in nucleotides to Citrus sinensis chitinase CHI1(chil)mRNA,indicating that these gene fragments relate to organ fall off,procreaction growth and nutrition growth during the flowering reversion in longan.