Plant Metabolic Engineering Laboratory, Biotechnology Division, CSIR-Institute of Himalayan Bioresource Technology, Council of Scientific and Industrial Research, Palampur, India.
PLoS One. 2011;6(12):e28315. doi: 10.1371/journal.pone.0028315. Epub 2011 Dec 1.
Flavonoids are synthesized by phenylpropanoid pathway. They are known to participate in large number of physiological and biochemical processes in plants. Parthenocarpy and male sterility has earlier been reported by silencing chalcone synthase (CHS) encoding gene. Silencing of CHS has blocked the synthesis of most of useful flavonoids including flavan-3-ols and flavonols. Also, these studies could not identify whether parthenocarpy/male sterility were due to lack of flavan-3-ols or flavonols or both. Flavonol synthase (FLS) is an important enzyme of flavonoid pathway that catalyzes the formation of flavonols. In this article, we propose a novel strategy towards the generation of seedless or less-seeded fruits by downregulation of flavonol biosynthesis in tobacco (Nicotiana tabacum cv Xanthi) through post-transcriptional gene silencing (PTGS) of FLS encoding mRNA. The FLS silenced lines were observed for 20-80% reduction in FLS encoding gene expression and 25-93% reduction in flavonol (quercetin) content. Interestingly, these FLS silenced tobacco lines also showed reduction in their anthocyanidins content. While the content of flavan-3-ols (catechin, epi-catechin and epi-gallocatechin) was found to be increased in FLS silenced lines. The delayed flowering in FLS silenced lines could be due to decrease in level of indole acetic acid (IAA) at apical region of their shoots. Furthermore, the pollen germination was hampered and pollens were unable to produce functional pollen tube in FLS silenced tobacco lines. Pods of FLS silenced lines contained significantly less number of seeds. The in vitro and in vivo studies where 1 µM quercetin was supplied to germination media, documented the restoration of normal pollen germination and pollen tube growth. This finding identified the role of flavonols particularly quercetin in pollen germination as well as in the regulation of plant fertility. Results also suggest a novel approach towards generation of seedless/less-seeded fruits via PTGS of FLS encoding gene in plants.
类黄酮通过苯丙烷途径合成。它们被认为参与植物中大量的生理和生化过程。以前曾有报道称,沉默查尔酮合酶(CHS)编码基因会导致单性结实和雄性不育。沉默 CHS 会阻止大多数有用的类黄酮的合成,包括黄烷-3-醇和黄酮醇。此外,这些研究还无法确定单性结实/雄性不育是由于缺乏黄烷-3-醇还是黄酮醇,或者两者都缺乏。黄酮醇合酶(FLS)是类黄酮途径中的一种重要酶,它催化黄酮醇的形成。在本文中,我们提出了一种通过烟草(Nicotiana tabacum cv Xanthi)中转录后基因沉默(PTGS)下调黄酮醇生物合成来产生无籽或少籽果实的新策略。FLS 沉默系观察到 FLS 编码基因表达减少 20-80%,黄酮醇(槲皮素)含量减少 25-93%。有趣的是,这些 FLS 沉默的烟草系也显示出其花青素含量减少。而 FLS 沉默系中的黄烷-3-醇(儿茶素、表儿茶素和表没食子儿茶素)含量增加。FLS 沉默系开花延迟可能是由于其茎顶端吲哚乙酸(IAA)水平下降所致。此外,花粉萌发受阻,花粉在 FLS 沉默的烟草系中无法产生功能花粉管。FLS 沉默系的豆荚中种子数量明显减少。在体外和体内研究中,将 1µM 槲皮素添加到萌发培养基中,记录到正常花粉萌发和花粉管生长的恢复。这一发现确定了类黄酮,特别是槲皮素在花粉萌发以及植物生育力调节中的作用。研究结果还表明,通过植物中 FLS 编码基因的 PTGS,可以为无籽/少籽果实的产生提供一种新方法。