Zhao Yan, Zhao Qian, Ao Guangming, Yu Jingjuan
State Key Laboratory for Agricultural Biotechnology, College of Biological Sciences, China Agricultural University, No. 2 Yuanmingyuan West Road, 100094, Beijing, People's Republic of China.
Planta. 2006 Jul;224(2):405-12. doi: 10.1007/s00425-006-0226-z. Epub 2006 Feb 1.
A pollen-specific gene, sb401, which was isolated from a cDNA library of in vitro geminated pollen of the diploid potato species Solanum berthaultii, belongs to the class of genes expressed late during pollen development. Using sb401 as a probe, a pollen-specific gene st901 was isolated from the genomic library of a potato species Solanum tuberosum cv. Desiree. Sequencing and RT-PCR analysis showed that the st901 genomic gene is 2,889 bp long, contains three exons and two introns, and encodes a putative polypeptide of 217 residues. The predicted protein sequence contains four imperfect repeated motifs of V-V-E-K-K-N/E-E; the core sequence of the repeats (K-K-N/E-E) resembles a microtubule-binding domain of the microtubule-associated protein MAP1B from mouse. The examination of a promoter-reporter construct in transgenic potato plants revealed that the st901 is expressed exclusively in mature pollen grains, which is consistent with the results of Northern blot and RT-PCR. For analysis of the function of st901, transgenic plants harboring antisense copies of st901 cDNA driven by a native st901 promoter were generated. Suppression of st901 gene in potato resulted in aberrant pollen at maturation and pollen viability of transgenic plants ranged from 4.4 to 14.8%, while that of control plants were more than 90%. These results strongly suggest that st901 has an essential role in pollen development.
一个花粉特异性基因sb401,是从二倍体马铃薯物种伯氏马铃薯(Solanum berthaultii)体外萌发花粉的cDNA文库中分离得到的,属于在花粉发育后期表达的基因类别。以sb401为探针,从马铃薯品种Desiree的基因组文库中分离出一个花粉特异性基因st901。测序和逆转录聚合酶链反应(RT-PCR)分析表明,st901基因组基因长2889 bp,包含三个外显子和两个内含子,编码一个由217个残基组成的假定多肽。预测的蛋白质序列包含四个不完全重复基序V-V-E-K-K-N/E-E-E;重复序列的核心序列(K-K-N/E-E)类似于来自小鼠的微管相关蛋白MAP1B的微管结合结构域。对转基因马铃薯植株中启动子-报告基因构建体的检测表明,st901仅在成熟花粉粒中表达,这与Northern印迹和RT-PCR的结果一致。为了分析st901的功能,构建了由天然st901启动子驱动的携带st901 cDNA反义拷贝的转基因植株。马铃薯中st901基因的抑制导致花粉成熟异常,转基因植株的花粉活力在4.4%至14.8%之间,而对照植株的花粉活力超过90%。这些结果强烈表明st901在花粉发育中起重要作用。