Institute of Horticultural Plants, College of Horticulture, China Agricultural University, Beijing, 100193, People's Republic of China.
Plant Mol Biol. 2019 Apr;99(6):561-573. doi: 10.1007/s11103-019-00836-8. Epub 2019 Feb 8.
SlPIN8 is expressed specifically within tomato pollen, and that it is involved in tomato pollen development and intracellular auxin homeostasis. The auxin (IAA) transport protein PIN-FORMED (PIN) plays key roles in various aspects of plant development. The biological role of the auxin transporter SlPIN8 in tomato development remains unclear. Here, we examined the expression pattern of the SlPIN8 gene in vegetative and reproductive organs of tomato. RNA interference (RNAi) transgenic lines specifically silenced for the SlPIN8 gene were generated to identify the role of SlPIN8 in pollen development. We found that SlPIN8 mRNA is expressed specifically within tomato pollen. In the anthers, the highest mRNA expression and β-glucuronidase (GUS) activity of promoter-SlPIN8-GUS was detected during late stages of anther development, when pollen maturation occurred. The downregulation of SlPIN8 did not drastically affect the vegetative growth of tomato. However, in SlPIN8-RNAi transgenic plants, approximately 80% of the pollen grains were identified to be abnormal and lack viability; they were shriveled and flattened. Furthermore, the downregulation of SlPIN8 affected the gene expression of some anther development-specific proteins. SlPIN8-RNAi transgenic plants induced seedless fruits because of defective pollen function rather than defective female gametophyte function. In addition, SlPIN8 was found to localize to the endoplasmic reticulum, consistent with the changes in the auxin levels of SlPIN8-RNAi lines, whereas the level of free IAA was increased in SlPIN8-overexpressing protoplasts, indicating that SlPIN8 is involved in intracellular auxin homeostasis.
SlPIN8 特异性表达于番茄花粉中,参与番茄花粉发育和细胞内生长素稳态。生长素(IAA)转运蛋白 PIN 形成(PIN)在植物发育的各个方面发挥关键作用。生长素转运体 SlPIN8 在番茄发育中的生物学作用尚不清楚。在这里,我们研究了 SlPIN8 基因在番茄营养和生殖器官中的表达模式。为了鉴定 SlPIN8 在花粉发育中的作用,我们生成了特异性沉默 SlPIN8 基因的 RNA 干扰(RNAi)转基因系。我们发现 SlPIN8 mRNA 特异性表达于番茄花粉中。在花药中,当花粉成熟时,SlPIN8 启动子-SlPIN8-GUS 的 mRNA 表达和 β-葡萄糖醛酸酶(GUS)活性在花药发育的后期达到最高水平。SlPIN8 的下调并没有严重影响番茄的营养生长。然而,在 SlPIN8-RNAi 转基因植物中,大约 80%的花粉粒被鉴定为异常且缺乏活力;它们皱缩和平坦。此外,SlPIN8 的下调影响了一些花药发育特异性蛋白的基因表达。由于花粉功能缺陷而不是雌性配子体功能缺陷,SlPIN8-RNAi 转基因植物诱导无籽果实。此外,发现 SlPIN8 定位于内质网,与 SlPIN8-RNAi 系中生长素水平的变化一致,而 SlPIN8 过表达原生质体中的游离 IAA 水平增加,表明 SlPIN8 参与细胞内生长素稳态。