糜子抗旱及复水相关基因的cDNA-AFLP差异显示

以糜子抗旱种质为实验材料,采用cDNA-AFLP技术分析四叶期糜子幼苗叶片在干旱、复水与对照等条件下的基因表达差异。选取了36个引物组合(6个荧光标记TaqⅠ引物×6个非荧光标记MseⅠ引物)进行选择性扩增,结果表明,这些引物组合都有很好的扩增效果,在三种处理之间均能扩增出表达模式不同的差异片段。随机选取了8个干旱特有和4个复水特有的差异片段进行了克隆测序。利用NCBI的Blastn进行序列的dbEST比对结果表明,12个EST序列中有2个分别与小麦、玉米的EST序列有较高的相似性,其余10个与已知EST序列的同源性都很低,可能为新的糜子抗旱相关基因。用Blastx与Genebank的非冗余蛋白数据库进行比较结果表明,其中的一个序列(DR007245)与N-乙酰葡萄糖胺-N-乙酰胞壁酸五肽(UDP-N-acetylglucosamine--N-acetylmuramyl-(Pentapeptide))的同源性较高,两个序列(DR007249,DR007250)与水稻的反转录转座子蛋白具有较高的同源性,反转录转座蛋白在植物抗旱、耐盐等抗逆境方面有重要作用。另外还有两个序列(DR007251,DR007252)与两种假定蛋白同源性较高。

Analysis on the responsive genes during drought and rehydration in Broomcorn millet(Panicum miliaceum L.) by means of cDNA-AFLP

cDNA-AFLP(cDNA Amplified Fragment Length Polymorphism) was used to analyze genes differential expressed in broomcorn millet(Panicum miliaceum L.) seedling under the condition of normal watering,serious drought and rehydration.All of the thirty-six primer pairs used in the selective amplification could amplify differential expression profile among the samples.Twelve fragments only presented in serious drought or rehydration were selected,cloned and sequenced.Blastn search to NCBI dbEST showed that only two of the fragments had significant homologous nucleotide sequence with wheat and rice ESTs,others could not found significant homologous nucleotide sequence.Based on the Blastx search to NCBI non-redundant protein database,one fragment was similar to UDP-N-acetylglucosamine-N-acetylmuramyl-(Pentapeptide),two fragments had significant homologous protein sequence with rice retrotransposon,another two had significant homologous protein sequence with two hypothetical proteins.