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以‘新台糖22号’甘蔗为材料,在其伸长初期以200mg/L的赤霉素进行叶面喷施处理,对照喷清水,分别提取对照和处理(0、6、12、24h)的总RNA进行等量混合,获得2种不同处理的样品池,采用cDNA-AFLP技术对赤霉素诱导下甘蔗节间伸长的差异表达进行分析。结果显示:(1)通过169对引物组合的选择性扩增,共得到了约11 000条大小在50～700bp的cDNA片段,获得186条差异条带。(2)经反向Northern杂交,于结果显示阳性的TDFs中选取56个片段进行克隆和序列分析,获得56条大小在80～500bp的差异条带。(3)经BLAST分析,按功能可将获得的56条TDFs分为7类,分别为能量与代谢相关基因、未知功能蛋白、未知基因、植物抗性相关基因、细胞壁生物合成与修饰相关基因、信号传导相关基因和转录因子相关基因。该研究获得了一些与甘蔗节间伸长相关的差异基因片段,为进一步研究甘蔗节间伸长的分子机理奠定了基础。
The sugar cane of ’Xintai No.22’ was used as the material and foliar sprayed with gibberellic acid 200mg / L in the initial stage of elongation. The control was sprayed with water and the control and untreated (0, 6, 12, 24h) The total RNA was mixed in the same amount to obtain two different sample pools. The differential expression of internode elongation induced by gibberellin was analyzed by cDNA-AFLP. The results showed that: (1) A total of about 11 000 cDNA fragments ranging in size from 50 to 700 bp were obtained by selective amplification of 169 pairs of primers, and 186 differential bands were obtained. (2) By reverse Northern blotting, 56 fragments were selected from TDFs that showed positive results for cloning and sequence analysis, and 56 differential bands ranging from 80 to 500 bp in size were obtained. (3) According to the BLAST analysis, 56 TDFs could be divided into 7 categories according to their functions, namely energy and metabolism related genes, unknown function proteins, unknown genes, plant resistance related genes, cell wall biosynthesis and modification related genes, Signal transduction related genes and transcription factor related genes. In the present study, some differential gene fragments related to internode elongation were obtained, which laid the foundation for further study on the molecular mechanism of internode elongation in sugarcane.