高等植物蔗糖转运的分子调控

在高等植物中,蔗糖的合成、运输与分配是一个复杂的过程。蔗糖由源到库的运输不仅与植物的生长发育相关,还受到植物体内的激素水平以及外界环境条件变化等因素的影响。蔗糖转运蛋白介导了蔗糖在植物韧皮部的装载、运输和卸载,在某些库中的蔗糖转运和库组织分配的分子调控中起有重要的生理作用。此外,简要介绍了笔者实验室在橡胶树蔗糖转运蛋白基因研究方面的最新进展。     

水稻蔗糖转运蛋白研究进展

蔗糖转运蛋白是光合产物运输与分配调控网络中的重要节点,主要参与蔗糖从"源"到"库"的质外体运输,在蔗糖的感应、"源"器官装载、韧皮部长距离运输和"库"器官卸载中起重要作用。总结和分析了水稻蔗糖转运蛋白基因家族的组成、蛋白结构特点、表达与调控特性、生物学功能等方面的研究进展,在此基础上,提出了蔗糖转运蛋白基础理论和应用研究方面存在的不足及应予重视和加强的主要方向。     

植物体内糖分子的长距离运输及其分子机制

植物器官(如叶、叶鞘、绿色的茎等)可以通过光合作用将CO₂合成为碳水化合物, 并经过长距离运输到达库组织(如新生组织、花粉、果实等)中进行贮存或利用。蔗糖是高等植物长距离运输碳水化合物的主要形式。蔗糖分子从源到库的运输包括源组织韧皮部的装载、维管束的运输和库组织韧皮部的卸载3个步骤。遗传学和分子生物学研究证明, 蔗糖转运蛋白、转化酶和单糖转运蛋白在糖分子的装载和卸载过程中发挥重要作用。该文综述了目前对光合产物运输过程及其调控分子机制的最新研究进展。     

水稻茎鞘非结构性碳水化合物转运机理及栽培调控研究进展

非结构性碳水化合物(non-structural carbohydrate, NSC)是参与水稻能量代谢的主要物质和维持水稻生长发育及响应环境调控的重要因子。蔗糖作为水稻茎鞘NSC代谢中心物质,是水稻灌浆期叶片光合产物和花前茎鞘中储存的NSC向穗部转运的主要形式,是籽粒灌浆的主要同化物来源。理解水稻蔗糖分配、转运机理及栽培环境调控途径对充分利用茎鞘NSC提高水稻产量具有重要意义。该文重点综述了水稻茎鞘NSC再分配及关键酶、蔗糖转运机理和调控,以及温度、水分和氮素等栽培环境对茎鞘NSC调控的研究进展,并对未来研究方向进行了展望。     

蔗糖膜运输系统与植物整体碳分配相关（综述）

蔗糖是光合作用的主要产物,作为碳同化的产物在植物体内进行分配。蔗糖的转运机制和效率通过减弱产物抑制来影响光合产率,通过控制源／库关系和生物量分配来调控植物活性。蔗糖在细胞质合成,或通过胞间连丝进行细胞问转运,或跨膜区域化,或外输入质外体被相邻细胞吸收。作为相对大极性的化合物,蔗糖的有效膜转运需要转运蛋白协助。跨液泡膜运输机制可能通过异化扩散、质子对向运输和同向运输;而跨质膜的运输则可能通过质子同向运输和异化扩散类似机制。近几十年仅在分子水平对质子同向运输进行了较为详尽的研究。这篇综述旨在综合介绍最近和过去关于蔗糖跨膜转运与植物整体碳分布机制。     

植物蔗糖转运蛋白表达的调控因素与分子机制

植物光合作用的产物主要以蔗糖的形式在植物体内进行从源到库的运输。蔗糖转运蛋白是此过程的重要参与者,其表达和调控与植物中光合作用产物的分配紧密关联,从而调控着植物的生长发育、结果结实、抗逆抗病等性状。蔗糖转运蛋白的表达受到植物发育时期、外界环境条件及激素的影响。蔗糖转运蛋白的调控机制有转录因子的调节、基因内部序列调控、蛋白质的磷酸化、蛋白之间的相互作用及质子转运体的活性调节等。综述了国内外对蔗糖转运蛋白表达与活性的调控因素及机制等最新的研究内容,以期为从多角度上探索植物蔗糖转运蛋白的功能和调控机制提供相关研究信息和思路。     

植物体内蔗糖转运蛋白的功能和调控

蔗糖转运蛋白（sucrose transporter,SUT）负责蔗糖的跨膜运输,在韧皮部介导的源-库蔗糖运输和为库组织供应蔗糖的生理活动中起关键作用。本文介绍植物体内蔗糖转运蛋白基因家族、细胞定位与功能调节以及高等植物的蔗糖感受机制的研究进展。     

甘薯蔗糖转运蛋白的功能分析

甘薯(Ipomoea batatas)是重要的粮食和工业加工原料作物。蔗糖是植物体内碳水化合物长距离转运的主要形式,蔗糖转运蛋白(sucrose transporter,SUT)在植物的生长代谢中调控蔗糖的跨膜运输和分配,在韧皮部介导的源-库蔗糖运输和为库组织供应蔗糖的生理活动中起关键作用。本研究根据不同淀粉性状甘薯块根中差异表达的2个SUT基因转录本,进行cDNA末端快速扩增(rapid amplification of cDNA ends,RACE)克隆,获得IbSUT62788和IbSUT81616的全长cDNA序列;通过系统发育分析明确其分类;通过在本氏烟草(Nicotiana benthamiana)中瞬时表达明确其亚细胞定位;通过酵母功能互补系统鉴定IbSUT62788和IbSUT81616是否具有吸收、转运蔗糖和己糖的能力。通过实时荧光定量PCR(real-time fluorescence quantitative polymerase chain reaction,RT-qPCR)分析IbSU62788和IbSUT81616在甘薯各器官中的表达特征;通过蘸花法得到外源表达IbSUT62788和IbSUT81616基因的拟南芥(Arabidopsis thaliana)植株,比较与野生型拟南芥的淀粉和糖含量的差异。结果表明,IbSUT62788和IbSUT81616分别编码505个和521个氨基酸的SUT蛋白,均属于SUT1亚家族。IbSUT62788和IbSUT81616均定位于细胞膜,在酵母系统中具有转运蔗糖、葡萄糖和果糖的能力。此外,IbSUT62788还具有转运甘露糖的能力。IbSUT62788在甘薯叶片、侧枝和茎中的表达量更高,IbSUT81616在侧枝、茎和块根中表达量更高。IbSUT62788和IbSUT81616在拟南芥中异源表达后,植株可以正常生长,但生物量增加。IbSUT62788的异源表达增加了拟南芥植株叶片可溶性糖含量、叶片大小和种子千粒重;IbSUT81616的异源表达增加了拟南芥植株叶片、根尖的淀粉积累量和种子千粒重,但减少了可溶性糖含量。本研究结果表明,IbSUT62788和IbSUT81616可能是调控甘薯蔗糖和糖含量性状的重要基因,在细胞膜上进行着蔗糖的跨膜运输、蔗糖进出库组织、韧皮部蔗糖的运输与卸载等生理功能,在拟南芥中异源表达造成的性状改变说明其在提高其他植物或作物产量中的应用潜力。本研究为揭示甘薯淀粉和糖代谢及重要品质性状形成机制提供了重要信息。     

植物氨基酸转运子研究进展

氨基酸是高等植物氮素同化产物长距离运输及在组织间分配的主要形式,通过跨膜转运的方式在植物体内进行运输。氨基酸转运子是位于生物膜上吸收及转运氨基酸的蛋白家族,对植物氮素营养具有重要贡献。本文对植物氨基酸转运子的表达、调控及其与氮素利用效率、植物产量与品质形成、抗逆性及适应性等方面的研究进展进行了综述。     

水稻水孔蛋白RWC3的分布及其受GA和蔗糖的调控(英文)

水孔蛋白能介导水分的跨膜运输,在植物的生长发育过程中起重要作用。对RWC3启动子-GUS转基因水稻的组织化学染色表明,水稻(Oryza sativa L.)水孔蛋白RWC3可能在包括营养和生殖器官在内的各部位中广泛表达。同时发现,赤霉素(GA)能提高转基因植物的愈伤组织、悬浮细胞和叶片中的GUS活性,而GA合成的抑制剂ancymidol降低了GUS活性。进一步研究发现,蔗糖能抑制GA对GUS活性的提高,说明GA和蔗糖在对RWC3的表达调控的信号传递过程中可能存在着相互作用。     

Apoplastic Phloem Unloading in the Stem of Bean

Sucrose has been found in the apoplast of bean stems at a concentrationof 25–60 mM with an axial concentration gradient in theappropriate direction for Munch translocation. Removal of theepidermis from a 50 mm length of stem enabled the washout oflabelled photosynthate from the apoplast. The rate of labelwashout was strongly dependent on temperature, and the rateincreased on blockage of phloem pathways to the main sink forthat assimilate. Washout did not reduce when the bathed tissuewas plasmolyzed. We propose that sucrose is unloaded from thephloem into the apoplast, and a sucrose concentration is maintainedthere by a balance of sucrose uptake into sink tissue or reloadinginto the phloem. It is proposed that the apoplastic pool ofphotosynthate can act to buffer sudden changes in phloem contentswhen there are rapid changes in source-sink configuration. Key words: Sucrose, Phaseolus vulgaris, Apoplast, Phloem unloading     

Germination Behaviour of Solanum nigrum Seeds

Sucrose has been found in the apoplast of bean stems at a concentrationof 25–60 mM with an axial concentration gradient in theappropriate direction for Munch translocation. Removal of theepidermis from a 50 mm length of stem enabled the washout oflabelled photosynthate from the apoplast. The rate of labelwashout was strongly dependent on temperature, and the rateincreased on blockage of phloem pathways to the main sink forthat assimilate. Washout did not reduce when the bathed tissuewas plasmolyzed. We propose that sucrose is unloaded from thephloem into the apoplast, and a sucrose concentration is maintainedthere by a balance of sucrose uptake into sink tissue or reloadinginto the phloem. It is proposed that the apoplastic pool ofphotosynthate can act to buffer sudden changes in phloem contentswhen there are rapid changes in source-sink configuration. Key words: Sucrose, Phaseolus vulgaris, Apoplast, Phloem unloading     

Evidence for an essential role of the sucrose transporter in phloem loading and assimilate partitioning.

Sucrose is the principal transport form of assimilates in most plants. In many species, translocation of assimilates from the mesophyll into the phloem for long distance transport is assumed to be carrier mediated. A putative sucrose proton cotransporter cDNA has been isolated from potato and shown to be expressed mainly in the phloem of mature exporting leaves. To study the in vivo role and function of the protein, potato plants were transformed with an antisense construct of the sucrose transporter cDNA under control of the CaMV 35S promoter. Upon maturation of the leaves, five transformants that expressed reduced levels of sucrose transporter mRNA developed local bleaching and curling of leaves. These leaves contained > 20-fold higher concentrations of soluble carbohydrates and showed a 5-fold increase in starch content as compared with wild type plants, as expected from a block in export. Transgenic plants with a reduced amount of sucrose carrier mRNA show a dramatic reduction in root development and tuber yield. Maximal photosynthetic activity was reduced at least in the strongly affected transformants. The effects observed in the antisense plants strongly support an apoplastic model for phloem loading, in which the sucrose transporter located at the phloem plasma membrane represents the primary route for sugar uptake into the long distance distribution network.     

The Breakdown of Sucrose During Translocation: With two Plates and one Figure in the Text

When a vine petiole is carrying labelled sucrose away from thelamina, the quantity of labelled carbon dioxide lost from thepetiole bears a constant relation to the quantity of labelledsucrose inside the petiole. Sucrose is virtually the only labelledsugar in the petiole, and the labelled sucrose is confined topart of the phloem. Calculations based on these measurementsand some assumptions suggest that the rate of breakdown of thetranslocated sucrose is about 0·5 mg. per c.c. of phloemper hour. The bearings of these findings on the problem of energysupply to translocation are discussed.     

Role of free space in translocation in sugar beet

The involvement of the free space in phloem loading of sucrose was studied in sugar beet source leaves (Beta vulgaris, L.). Sucrose, supplied exogenously to the abraded upper surface of leaves at a concentration of 20 mm, was available for translocation at rates similar to those obtained with photosynthesis. The exogenous sucrose substituted as a source of translocate for assimilate derived from photosynthesis when the latter process was disrupted by plasmolysis of the leaf with 0.8 M mannitol. The mesophyll symplast was not completely disrupted by this treatment, however. Data from the sugar uptake experiments indicate that phloem loading can occur from the free space.     

Transport of organic solutes in Phloem and xylem of a nodulated legume

Collections of xylem exudate of root stumps or detached nodules, and of phloem bleeding sap from stems, petioles, and fruits were made from variously aged plants of Lupinus albus L. relying on nodules for their N supply. Sucrose was the major organic solute of phloem, asparagine, glutamine, serine, aspartic acid, valine, lysine, isoleucine, and leucine, the principal N solutes of both xylem and phloem. Xylem sap exhibited higher relative proportions of asparagine, glutamine and aspartic acid than phloem sap, but lower proportions of other amino acids. Phloem sap of petioles was less concentrated in asparagine and glutamine but richer in sucrose than was phloem sap of stem and fruit, suggesting that sucrose was unloaded from phloem and amides added to phloem as translocate passed through stems to sinks of the plant. Evidence was obtained of loading of histidine, lysine, threonine, serine, leucine and valine onto phloem of stems but the amounts involved were small compared with amides. Analyses of petiole phloem sap from different age groups of leaves indicated ontogenetic changes and effects of position on a shoot on relative rates of export of sucrose and N solutes. Diurnal fluctuations were demonstrated in relative rates of loading of sucrose and N solutes onto phloem of leaves. Daily variations in the ability of stem tissue to load N onto phloem streams were of lesser amplitude than, or out of phase with fluctuations in translocation of N from leaves. Data were related to recent information on C and N transport in the species.     

Role of phloem in sucrose transport by Ricinus cotyledons

Sucrose is taken up and accumulated by cotyledons of Ricinus communis L. Autoradiographic studies reveal a predominant accumulation of sucrose in the phloem of the cotyledons. The export of sucrose from the cotyledons to hypocotyl and roots proceeds in the phloem by mass flow. These results, taken together with previous data, are experimental evidence for proton-sucrose symport as the mechanism of phloem loading.     

Inorganic ions modulate the path of phloem loading of sucrose in Ricinus communis L. seedlings

The impact of inorganic ions on sucrose fluxes in the cotyledons and on the pathway of phloem loading was studied in Ricinus communis L. seedlings. The cotyledons were incubated in defined solutions which contained either potassium, sodium, magnesium or calcium as chloride salts, or the sodium salts of sulphate or phosphate. Sucrose uptake from the medium into the cotyledons was only slightly affected by the salts. Sucrose efflux to the medium was increased by phosphate and sulphate and to a lesser extent by sodium and potassium. Phloem loading from the apoplasm and the symplasm was analysed by addition of labelled sucrose to the medium, determination of the specific radioactivity of sucrose in sieve-tube exudate and quantification of export into the seedling axis. Potassium and sodium stimulated the apoplasmic route of phloem loading of sucrose, mostly at the expense of loading from cotyledon sucrose pools. In contrast, sulphate and phosphate strongly inhibited the apoplasmic route whereas the (small) symplasmic flux from the cotyledon sucrose pools was less affected. Magnesium ions inhibited phloem loading by both pathways. The potential of ions in modulating the pathways of sucrose export in day to day operation of plants is discussed.     

Sucrose carrier RcSCR1 is involved in sucrose retrieval, but not in sucrose unloading in growing hypocotyls of Ricinus communis L

The transport of assimilates from source to sink tissues is mediated by the phloem. Along the vascular system the phloem changes its physiological function from loading phloem to transport and unloading phloem. Sucrose carrier proteins have been identified in the transport phloem, but it is unclear whether the physiological role of these transporters is phloem unloading of sucrose or retrieval of apoplasmic sucrose back into the sieve element/companion cell complex. Here, we describe the dynamic expression of the Ricinus communis sucrose carrier RcSCR1 in the hypocotyl at different sink strengths. Our results indicate that phloem unloading in castor bean is not catalysed by the phloem loader RcSCR1. However, this sucrose carrier represents the molecular basis of the sucrose retrieval mechanism along the transport phloem, which is dynamically adjusted to the sink strength. As a consequence, we assume that other release carrier(s) exist in sink tissues, such as the hypocotyl, in R. communis.