• ISSN 1008-505X
  • CN 11-3996/S

糖转运蛋白OsSTP1过表达调控水稻产量对氮梯度的响应

Yield response of rice materials overexpressing sugar transporter protein OsSTP1 to nitrogen gradient

  • 摘要:
    目的 糖转运蛋白OsSTP1过表达通过参与碳水化合物在韧皮部的装载和卸载过程,调控碳水化合物的源库分配,从而影响水稻产量。我们研究了不同施氮水平对OsSTP1过表达材料碳水化合物分配及产量的影响,加深了解氮素调控水稻产量的分子生物学机理。
    方法 以台北309为遗传背景的野生型(WT)和两个OsSTP1过表达株系OsSTP1-OE1OsSTP1-OE2为试验材料,在湖南农业大学进行了田间试验。试验采用裂区设计,每个遗传材料下设置5个施氮量,分别为:0 (N0)、60 (N60)、90 (N90)、120 (N120)和180 (N180) kg/hm2。在授粉后12天,取植株样测定茎鞘和籽粒中的淀粉酶(AMY)、腺苷二磷酸葡萄糖焦磷酸化酶(AGP)、蔗糖合酶(SUS)和蔗糖磷酸合成酶(SPS)活性。在抽穗期和授粉后24天,取样测定非结构性碳水化合物(NSC)含量,并计算NSC的运转率。成熟期调查水稻产量及其构成因子。
    结果 与野生型相比,各施氮处理下两个OsSTP1过表达株系茎鞘中AMY和SPS的活性显著提高23.3%—34.3%和11.9%—18.3%,增幅以N120最高,穗部AGP和SUS活性增加了8.20%—14.4%和24.1%—33.3%,以N90和N60处理增幅最大。各施氮处理两个株系茎鞘中的蔗糖和可溶性糖含量与野生型没有显著差异,但籽粒中显著高于野生型;茎鞘中的淀粉含量低于野生型而籽粒中高于野生型。同氮处理下,OsSTP1过表达株系茎鞘中NSC转运率显著高于野生型24.2%—40.2%,以N120时的增幅最大。过表达株系的产量较野生型在不同施氮处理下产量均显著增加,增幅为5.50%~14.5%,且在N90处理下增幅最高。OsSTP1过表达株系氮肥利用率与收获指数分别较野生型提高了11.9%-~17.9%和6.20%-~13.2%。拟合分析表明,OsSTP1过表达株系在70 kg/hm2施氮量与野生型在120 kg/hm2施氮量处理的产量相同,表明种植OsSTP1过表达株系具有一定的减施氮肥潜力。
    结论 OsSTP1过表达株系提高了水稻茎鞘中淀粉酶和蔗糖磷酸合成酶活性以及穗中蔗糖合酶和腺苷二磷酸葡萄糖焦磷酸化酶活性,促进茎鞘中碳水化合物的再活化,提高茎鞘非结构性碳水化合物转运率,促进了蔗糖、淀粉和可溶性糖在籽粒中的积累。糖转运蛋白OsSTP1过表达通过高效调控水稻碳水化合物的源库分配,提高了氮肥利用率,在氮素供给较低的条件下可获得野生型水稻高氮水平下的产量,因此,可作为氮高效育种重要的指示基因。

     

    Abstract:
    Objectives Overexpression of the sugar transporter protein OsSTP1 affects rice yield by participating in the loading and unloading processes of carbohydrates in the phloem and regulating the source-pool allocation of carbohydrates. We investigated the carbohydrate allocation and yield in OsSTP1 overexpression materials under different nitrogen application levels, to deepen our understanding of the molecular biological mechanisms of nitrogen regulation of rice yield.
    Methods A field trial was conducted at Hunan Agricultural University using the wild type (WT) from the genetic background of Taipei 309 and two OsSTP1 overexpression lines, OsSTP1-OE1 and OsSTP1-OE2, as test materials. The experiment was conducted in a split-zone design, and five nitrogen application rates were set under each genetic material, namely: 0 (N0), 60 (N60), 90 (N90), 120 (N120), and 180 (N180) kg/hm2. Plant samples were taken to determine amylase (AMY), adenosine diphosphate glucose pyrophosphorylase (AGP), sucrose synthase (SUS), and sucrose phosphate phosphorylase (SPS) in stem sheaths and kernels at 12 days post-pollination. SUS) and sucrose phosphate synthase (SPS) activities. Samples were taken at tasseling stage and 24 days after pollination to determine the non-structural carbohydrate (NSC) content and to calculate the NSC run rate. Rice yield and its components were investigated at maturity.
    Results Compared with the WT, the activities of AMY and SPS in the stem sheath of the two OsSTP1 overexpression lines were significantly increased by 23.3%−34.3% and 11.9%−18.3% under the five N application treatments, with the increase being the highest in the case of N120, and the activities of AGP and SUS in the spike section were increased by 8.20%−14.4% and 24.1%−33.3%, and the highest increase was observed in N90 and N60 treatments. Sucrose and soluble sugar contents in the stem sheaths of the two lines were not significantly different from those of the WT, but were significantly higher than those of WT in the kernels; starch contents in the stem sheaths were lower than those of the WT but higher than those of WT in the kernels in parallel. The NSC translocation rate in the stem sheaths of OsSTP1 overexpression lines was significantly higher than that of the WT by 24.2%−40.2% under each nitrogen homozygote treatment, with the greatest increase at N120. The yields of the overexpression lines were significantly higher than those of WT under nitrogen application treatments, with increases ranging from 5.50% to 14.5%, and the highest increase was observed at N90. The nitrogen fertilizer utilization and harvest index of the OsSTP1 overexpression lines were increased by 11.9% to 17.9% and 6.20% to 13.2%, respectively, compared with those of the WT. The fitting analysis showed that the yield of OsSTP1 overexpression lines at N 70 kg/hm2 was the same as that of the WT at 120 kg/hm2, which indicated that planting OsSTP1 overexpression strain had some potential for reducing nitrogen fertilizer.
    Conclusions OsSTP1 overexpression increased amylase and sucrose phosphate synthase activities in rice stem sheaths and sucrose synthase and adenosine diphosphate glucose pyrophosphorylase activities in spikes, promoting the reactivation of carbohydrates in stem sheaths, increasing the rate of non-structural carbohydrate transport in stem sheaths, and facilitating the accumulation of sucrose, starch, and soluble sugars in the seeds. The high allocation of carbohydrates in grains of the rice mateirals overexpressing the sugar transporter protein OsSTP1 acquired high nitrogen use efficiency, and produced the same yield at low nitrogen input rate as the wild type did at high nitrogen inpout rate, indicating the the sugar transporter protein OsSTP1 being an important gene for high-N-efficient rice breeding.

     

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