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Disruption of a gene for rice sucrose transporter OsSUT1 impairs pollen function but pollen maturation is unaffected

机译:水稻蔗糖转运蛋白OsSUT1基因的破坏削弱了花粉的功能但花粉的成熟不受影响

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摘要

Sucrose transporters (SUTs) are known to play critical roles in the uptake of sucrose from the apoplast in various steps of sugar translocation. Because developing pollen is symplastically isolated from anther tissues, it is hypothesized that SUTs are active in the uptake of apoplastic sucrose into pollen. To investigate this possibility, a comprehensive expression analysis was performed for members of the SUT gene family in the developing pollen of rice (Oryza sativa L.) using real-time RT-PCR combined with a laser microdissection technique. Among the five SUT genes, OsSUT1 and OsSUT3 were found to be preferentially expressed and had temporal expression patterns that were distinct from each other. Expression of OsSUT1 in pollen was confirmed by a promoter–GUS fusion assay. The physiological function of OsSUT1 in pollen was further investigated using retrotransposon insertion mutant lines. While the homozygote of disrupted OsSUT1 (SUT1–/–) could not be obtained, heterozygote plants (SUT1+/–) showed normal grain filling. Their progeny segregated into SUT1+/– and SUT1+/+ with the ratio of 1:1, suggesting that the pollen disrupted for OsSUT1 is dysfunctional. This hypothesis was reinforced in vivo by a backcross of SUT1+/– plants with wild-type plants and also by in vitro pollen germination on the artificial media. However, starch accumulation during pollen development was not affected by disruption of OsSUT1, suggesting that the sugar(s) required for starch biosynthesis is supplied by other sugar transporters.
机译:已知蔗糖转运蛋白(SUT)在糖转运的各个步骤中从质外体摄取蔗糖中起关键作用。因为发育中的花粉是与花药组织共生分离的,所以可以假设SUTs在将非可塑性蔗糖摄取到花粉中具有活性。为了研究这种可能性,使用实时RT-PCR结合激光显微切割技术对水稻(Oryza sativa L.)的花粉中SUT基因家族的成员进行了全面的表达分析。在这五个SUT基因中,OsSUT1和OsSUT3被优先表达并具有彼此不同的时间表达模式。 OsSUT1在花粉中的表达已通过启动子-GUS融合测定得到证实。 OsSUT1在花粉中的生理功能使用反转录转座子插入突变株进一步研究。虽然无法获得被破坏的OsSUT1的纯合子(SUT1-/-),但杂合子植物(SUT1-/-)却显示出正常的籽粒填充。它们的后代以1:1的比例分离为SUT1 +/-和SUT1 + / +,这表明被OsSUT1破坏的花粉是功能障碍的。 SUT1 +/–植物与野生型植物的回交以及在人工培养基上的体外花粉萌发在体内增强了这一假说。但是,花粉发育过程中的淀粉积累不受OsSUT1破坏的影响,表明淀粉生物合成所需的糖是由其他糖转运蛋白提供的。

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