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本研究以油茶品种‘长林4号’无性系扦插苗为材料,采用RT-PCR技术分离出一个油茶铝激活苹果酸转运体基因。该基因的c DNA全长1 761 bp,编码586个氨基酸,分子量为65.59 k D,理论等电点(PI)为6.27。与其他植物的ALMT蛋白高度同源,将该基因命名为CoALMT(Gen Bank登录号:KT932706)。CoALMT蛋白含有6个跨膜区,可能定位在细胞质膜上。CoALMT蛋白二级结构中α-螺旋(Alpha helix)、β折叠(Extended strand)、β转角(Beta turn)、无规则卷曲(Random coil)分别占52.05%、16.89%、8.70%、22.35%。以‘长林4号’和‘长林166号’无性系扦插苗为材料,利用qRT-PCR技术分析了沙培条件下两个不同无性系在不同磷水平下不同器官组织中CoALMT基因的表达。结果表明,低磷处理下,两个无性系油茶根系CoALMT的表达量均上升,说明油茶根系中CoALMT基因的表达受到低磷诱导。茎和叶中的表达模式与根中存在差异。低磷处理下不同组织中,‘长林166号’中CoALMT基因的表达量均比‘长林4号’高。综上结果可知CoALMT基因参与油茶对低磷的响应,其可能会影响油茶的磷吸收与利用效率。
In this study, a Camellia oleifera aluminum-activated malate transporter gene was isolated from Camellia oleifera ’Chang lin 4’ clones using RT-PCR technique. The cDNA of this gene was 1 761 bp in length and encoded 586 amino acids with a molecular mass of 65.59 kD and a theoretical isoelectric point (PI) of 6.27. It is highly homologous to the ALMT protein of other plants and is named CoALMT (Gen Bank Accession Number: KT932706). The CoALMT protein contains six transmembrane domains, possibly located on the plasma membrane. Alpha helix, Extended strand, Beta turn and Random coil in CoALMT secondary structure accounted for 52.05%, 16.89%, 8.70% and 22.35%, respectively. Using clonal cuttings of ’Changlin 4’ and ’Changlin 166’ as materials, the expression of CoALMT gene in different organs and tissues of two different clones under different culture conditions was analyzed by qRT-PCR . The results showed that under low phosphorus treatments, the expression of CoALMT in roots of two clones was increased, which indicated that the expression of CoALMT gene in Camellia oleifera roots was induced by low-level phosphorus. The expression patterns in stem and leaf differed from those in roots. In different tissues under low phosphorus treatment, the expression of CoALMT gene in ’Chang Lin 166’ was higher than that in ’Changlin 4’. In conclusion, CoALMT gene is involved in the response of Camellia oleifera to low-P, which may affect the phosphorus absorption and utilization efficiency of Camellia oleifera.