Li Xue-Bao, Cai Lin, Cheng Ning-Hui, Liu Jian-Wei
Temasek Life Sciences Laboratory, 1 Research Link, National University of Singapore, Singapore 117604.
Plant Physiol. 2002 Oct;130(2):666-74. doi: 10.1104/pp.005538.
Each fiber of cotton (Gossypium hirsutum) is a single epidermal cell that rapidly elongates to 2.5 to 3.0 cm from the ovule surface within about 16 d after anthesis. A large number of genes are required for fiber differentiation and development, but so far, little is known about how these genes control and regulate the process of fiber development. To investigate gene expression patterns in fiber, a cDNA, GhTUB1, encoding beta-tubulin was isolated from a cotton fiber cDNA library. The analyses of RNA northern-blot hybridization and reverse transcriptase-polymerase chain reaction demonstrated that GhTUB1 transcripts preferentially accumulated at high levels in fiber, at low levels in ovules at the early stage of cotton boll development, and at very low levels in other tissues of cotton. The corresponding GhTUB1 gene including the promoter region was isolated by screening a cotton genomic DNA library. To demonstrate the specificity of the GhTUB1 promoter, the 5'-flanking region including the promoter and 5'-untranslated region was fused with the beta-glucuronidase reporter gene. The expression of the reporter chimera was examined in a large number of transgenic cotton plants. Histochemical assays demonstrated that GhTUB1::beta-glucuronidase fusion genes were expressed preferentially at high levels in fiber and primary root tip of 1- to 3-d-old seedlings and at low levels in other tissues such as ovule, pollen, seedling cotyledon, and root basal portion. The results suggested that the GhTUB1 gene may play a distinct and required role in fiber development. In addition, the GhTUB1 promoter may have great potential for cotton improvement by genetic engineering.
每个棉纤维(陆地棉)都是一个单一的表皮细胞,在开花后约16天内从胚珠表面迅速伸长至2.5到3.0厘米。纤维分化和发育需要大量基因,但到目前为止,对于这些基因如何控制和调节纤维发育过程知之甚少。为了研究纤维中的基因表达模式,从棉花纤维cDNA文库中分离出一个编码β-微管蛋白的cDNA,即GhTUB1。RNA Northern杂交和逆转录聚合酶链反应分析表明,GhTUB1转录本在纤维中优先高水平积累,在棉铃发育早期的胚珠中低水平积累,在棉花的其他组织中极低水平积累。通过筛选棉花基因组DNA文库分离出包括启动子区域在内的相应GhTUB1基因。为了证明GhTUB1启动子的特异性,将包括启动子和5'-非翻译区的5'-侧翼区域与β-葡萄糖醛酸酶报告基因融合。在大量转基因棉花植株中检测了报告嵌合体的表达。组织化学分析表明,GhTUB1::β-葡萄糖醛酸酶融合基因在1至3日龄幼苗的纤维和初生根尖中优先高水平表达,在胚珠、花粉、幼苗子叶和根基部等其他组织中低水平表达。结果表明,GhTUB1基因可能在纤维发育中发挥独特且必要的作用。此外,GhTUB1启动子在通过基因工程改良棉花方面可能具有巨大潜力。