Cheng Jintao, Wang Zhenyu, Yao Fengzhen, Gao Lihong, Ma Si, Sui Xiaolei, Zhang Zhenxian
Beijing Key Laboratory of Growth and Developmental Regulation for Protected Vegetable Crops, College of Agronomy and Biotechnology, China Agricultural University, Beijing 100193, China.
Beijing Key Laboratory of Growth and Developmental Regulation for Protected Vegetable Crops, College of Agronomy and Biotechnology, China Agricultural University, Beijing 100193, China
Plant Physiol. 2015 Jun;168(2):635-47. doi: 10.1104/pp.15.00290. Epub 2015 Apr 17.
Efficient sugar transport is needed to support the high metabolic activity of pollen tubes as they grow through the pistil. Failure of transport results in male sterility. Although sucrose transporters have been shown to play a role in pollen tube development, the role of hexoses and hexose transporters is not as well established. The pollen of some species can grow in vitro on hexose as well as on sucrose, but knockouts of individual hexose transporters have not been shown to impair fertilization, possibly due to transporter redundancy. Here, the functions of CsHT1, a hexose transporter from cucumber (Cucumis sativus), are studied using a combination of heterologous expression in yeast (Saccharomyces cerevisiae), histochemical and immunohistochemical localization, and reverse genetics. The results indicate that CsHT1 is a plasma membrane-localized hexose transporter with high affinity for glucose, exclusively transcribed in pollen development and expressed both at the levels of transcription and translation during pollen grain germination and pollen tube growth. Overexpression of CsHT1 in cucumber pollen results in a higher pollen germination ratio and longer pollen tube growth than wild-type pollen in glucose- or galactose-containing medium. By contrast, antisense suppression of CsHT1 leads to inhibition of pollen germination and pollen tube elongation in the same medium and results in a decrease of seed number per fruit and seed size when antisense transgenic pollen is used to fertilize wild-type or transgenic cucumber plants. The important role of CsHT1 in pollen germination, pollen tube growth, and seed development is discussed.
花粉管在雌蕊中生长时,需要高效的糖分运输来支持其高代谢活性。运输失败会导致雄性不育。虽然蔗糖转运蛋白已被证明在花粉管发育中起作用,但己糖和己糖转运蛋白的作用尚未完全明确。一些物种的花粉在体外既可以在己糖上生长,也可以在蔗糖上生长,但单个己糖转运蛋白的敲除并未显示会损害受精,这可能是由于转运蛋白的冗余性。在这里,利用酵母(酿酒酵母)中的异源表达、组织化学和免疫组织化学定位以及反向遗传学相结合的方法,研究了黄瓜(Cucumis sativus)的己糖转运蛋白CsHT1的功能。结果表明,CsHT1是一种定位于质膜的己糖转运蛋白,对葡萄糖具有高亲和力,仅在花粉发育过程中转录,在花粉粒萌发和花粉管生长过程中在转录和翻译水平均有表达。在含葡萄糖或半乳糖的培养基中,CsHT1在黄瓜花粉中的过表达导致花粉萌发率高于野生型花粉,花粉管生长更长。相比之下,CsHT1的反义抑制导致在相同培养基中花粉萌发和花粉管伸长受到抑制,并且当使用反义转基因花粉对野生型或转基因黄瓜植株进行授粉时,每个果实的种子数和种子大小会减少。本文讨论了CsHT1在花粉萌发、花粉管生长和种子发育中的重要作用。