Yang Chin-Ying, Wu Chih-Hsien, Jauh Guang Yuh, Huang Jong-Chin, Lin Chin-Chung, Wang Co-Shine
Graduate Institute of Biotechnology, National Chung Hsing University, Taichung, Taiwan.
Protoplasma. 2008 Nov;233(3-4):241-54. doi: 10.1007/s00709-008-0016-5. Epub 2008 Sep 5.
We have isolated the LLA23 gene in the pollen of Lilium longiflorum. The LLA23 gene encodes an ASR (named after abscisic acid, stress and ripening) protein that has a nuclear localization sequence at the C terminus. The gene is interrupted by one single intron and possesses a long 5'-untranslated region. Southern blots of lily genomic DNA indicated that LLA23 is a member of a small gene family. We examined the link between LLA23 location and the desiccation that naturally occurs in developing anthers using immunogold labeling. When pollen reached maturity, a significant increase in LLA23 labeling was observed in the nuclei of both vegetative and generative cells from 10- to 12-cm buds and thereafter. This clearly demonstrates that a marked increase in LLA23 translocation from the cytoplasm to both nuclei of pollen grains occurs in 12-cm buds, a stage shortly before the commencement of desiccation during anther development. In addition, microarray analysis showed that 410 (206 up-regulated and 204 down-regulated) genes have altered expression in LLA23-overexpressing plants. Quantitative PCR analysis confirmed the changes in mRNA levels observed in our microarray analysis. This genome-wide overview of gene expression supports the theory that LLA23 acts as a regulator.
我们已经从麝香百合的花粉中分离出LLA23基因。LLA23基因编码一种ASR(以脱落酸、胁迫和成熟命名)蛋白,该蛋白在C末端具有一个核定位序列。该基因被一个单一内含子打断,并拥有一个长的5'非翻译区。百合基因组DNA的Southern杂交表明LLA23是一个小基因家族的成员。我们使用免疫金标记检查了LLA23定位与发育中的花药中自然发生的干燥之间的联系。当花粉成熟时,在10至12厘米长的花蕾及之后的营养细胞和生殖细胞的细胞核中观察到LLA23标记显著增加。这清楚地表明,在12厘米长的花蕾中,即花药发育过程中干燥开始前不久的阶段,LLA23从细胞质向花粉粒的两个细胞核的易位显著增加。此外,微阵列分析表明,在LLA23过表达植物中,有410个基因(206个上调和204个下调)的表达发生了改变。定量PCR分析证实了我们在微阵列分析中观察到的mRNA水平的变化。这种全基因组范围的基因表达概述支持了LLA23作为一种调节因子的理论。