高等植物尿素代谢及转运的分子机理

尿素广泛存在于自然界中, 是易于被许多生物(如植物)利用的生长氮源。该文通过概述尿素在不同生命系统中存在的基础生理意义及各类型尿素转运蛋白, 讨论了植物细胞中尿素合成与分解的各种途径及尿素在植物氮营养、代谢和运输中的生理作用。迄今为止, 在植物中已发现了2类转运尿素的膜蛋白, 即MIPs和DUR3, 它们分别在低亲和力、高亲和力尿素运输中发挥潜在作用。异源表达结果表明, MIPs介导了尿素的被动迁移; 而AtDUR3则参与拟南芥根系对尿素的吸收。对MIPs和DUR3转运尿素的酶学特征、亚细胞作用位点和表达调控状况等的研究表明: 它们的分子生物学功能与植物的氮营养及氮素再分配和利用相关。     

棉花DUR3基因的鉴定及进化分析

植物DUR3同源蛋白属于钠离子/溶质共运蛋白家族的尿素高亲和力运输蛋白，在植物体对外源尿素的主动吸收及内源尿素的再分配过程中具有重要作用。为明确棉花DUR3基因的结构和进化情况，基于生物信息学的方法，从全基因组水平鉴定陆地棉和雷蒙德氏棉的DUR3基因，并对基因结构、跨膜结构域、基序分布、进化关系等进行分析。结果表明：(1)从陆地棉A亚组和D亚组染色体各鉴定出1个DUR3基因，从雷蒙德氏棉基因组鉴定出1个DUR3基因。这3个棉花DUR3同源蛋白同其他植物DUR3同源蛋白一样，具有15个跨膜结构域，具有3个位置一致、高度保守的基序。(2)基因结构分析表明，双子叶植物DUR3基因的外显子个数明显多于单子叶植物，这3个棉花DUR3基因的外显子个数亦是如此。(3)根据物种间种属亲缘关系，对不同物种DUR3氨基酸序列构建的进化树显示，棉花的同双子叶植物的聚在一起。(4)DUR3直系同源基因和旁系同源基因的K_a/K_s比值普遍均大于1,说明这些基因在进化过程中主要受到正向选择的作用。该研究结果为深入研究棉花DUR3同源蛋白提供了理论基础。     

高通量筛选诱变菌株降低黄酒发酵氨基甲酸乙酯前体积累

【目的】氨基甲酸乙酯是发酵食品中普遍存在的一种潜在危害物。黄酒中氨基甲酸乙酯的前体主要是尿素和乙醇。本研究通过高通量筛选策略降低黄酒发酵过程中尿素的积累,从而降低氨基甲酸乙酯积累。【方法】以一株黄酒生产工业菌株酿酒酵母XZ-11为研究对象,采用ARTP诱变和高通量筛选策略,获得尿素积累量较低菌株。使用实时定量PCR技术检测氮代谢中尿素代谢和转运相关基因(DUR1,2和DUR3)的变化。【结果】筛选得到一株尿素高效稳定性利用菌株5-11C。其尿素积累量比酿酒酵母XZ-11降低了50.6%。实时定量荧光PCR结果表明,与尿素代谢和转运相关的基因(DUR1,2和DUR3)表达量分别提高了3.3和2.2倍。【结论】高通量筛选策略可以用于降低黄酒生产过程中氨基甲酸乙酯前体尿素的含量。由于未采用基因工程手段,避免了可能的法规问题,消费者易于接受,在发酵食品行业具有较好的应用前景。     

拟南芥养分离子转运蛋白研究进展

养分离子的跨膜转运是细胞获取养分的重要环节,亦是植物在组织和器官水平上进行 养分吸收运移的基础。文中综述了拟南芥中养分离子转运蛋白在基因克隆、序列与结构分析、功能鉴定、表达与调控方面的研究进展,其中着重讨论了这些转运蛋白在氮、磷和钾等营养元素吸收、运输、分配中的作用。     

拟南芥养分离子转运蛋白研究进展

养分离子的跨膜转运是细胞获取养分的重要环节,亦是植物在组织和器官水平上进行养分吸收运移的基础。文中综述了拟南芥中养分离子转运蛋白在基因克隆、序列与结构分析、功能鉴定、表达与调控方面的研究进展,其中着重讨论了这些转运蛋白在氮、磷和钾等营养元素吸收、运输、分配中的作用。     

与高等植物吸收(NO_3)~-/(NH_4)~+、(PO_4)~(3-)和K~+有关的膜转运蛋白编码基因的分子生物学研究现状

高等植物对土壤中营养元素的吸收是其一切生命活动过程的基础,尤其在营养元素缺乏的状态下,更与其抗营养饥饿等特性息息相关。兼于土壤中Ｎ、Ｐ、Ｋ元素缺乏的严重性与普遍性,以及Ｎ、Ｐ、Ｋ对高等植物生长和发育的重要性,有关高等植物吸收营养元素的膜转运蛋白编码基因的分子生物学研究已引起有关学者的高度重视。ＮＯ－３／ＮＨ＋４、ＰＯ３－４与Ｋ＋膜转运蛋白均有低亲和力和高亲和力系统（ＬｏｗＡｆｉｎｉｔｙＴｒａｎｓｐｏｒｔｅｒ＆ＨｉｇｈＡｆｉｎｉｔｙＴｒａｎｓｐｏｒｔｅｒ）。对ＰＯ４３－和Ｋ＋而言,低亲和力系统是组成性表达的系统,在正常营养状态下对根系吸收营养起重要作用。而高亲和力系统是受营养缺乏而诱导表达的系统,对于植物的抗逆性、耐营养饥饿至关重要。迄今为止,与之有关的基因的全长ｃＤＮＡ或全基因已在几种植物中被克隆。此外,对基因的表达特性亦有广泛研究。本文简要概述这三大营养元素的膜转运蛋白编码基因的分子生物学研究现状。     

黄色条纹/黄色条纹样蛋白与植物中的铁转运功能

黄色条纹/黄色条纹样蛋白（yellow stripe/yellow stripe like,YS/YSL）在植物金属转运,尤其是在微量营养元素铁的分配和利用过程中发挥重要作用。近年来,植物的铁长距离运输机制已成为研究的热点之一。YS/YSL转运蛋白涉及铁的转移、运输、装载和卸载。鉴定并精确解析YS/YSL的功能对于阐明铁的长距离运输机制具有重要的意义。本文综述了YS/YSL的蛋白质性质、金属转运活性、底物特异性及其功能表达,以期为植物YS/YSL相关铁转运机制研究奠定基础,并为提高作物籽粒铁营养提供理论依据。     

拟南芥COPT家族蛋白研究进展

铜(Cu)是植物必需的微量元素, 作为多种酶的辅因子参与许多植物生理生化反应。Cu缺乏和过量均影响植物正常生长发育, 因此植物进化出精妙复杂的调控网络来严格控制植物体内的Cu含量。植物Cu转运蛋白COPT家族成员与Cu有很高的亲和力, 能够调节植物对Cu的吸收和转运, 在维持植物体内Cu稳态平衡过程中发挥重要作用。COPT蛋白涉及不同的Cu转运功能, 如从外界环境中摄取Cu、从细胞器中输出Cu、长距离运输Cu以及在不同器官间动用和再分配Cu。此外, COPT蛋白在其它离子的稳态平衡维持、昼夜节律性生物钟调控、植物激素合成和植物对激素信号的感受过程中也发挥重要作用。该文综述了模式植物拟南芥(Arabidopsis thaliana) COPT家族各成员的表达和定位、调控机制以及生物学功能等方面的最新进展。     

拟南芥COPT家族蛋白研究进展

铜(Cu)是植物必需的微量元素, 作为多种酶的辅因子参与许多植物生理生化反应。Cu缺乏和过量均影响植物正常生长发育, 因此植物进化出精妙复杂的调控网络来严格控制植物体内的Cu含量。植物Cu转运蛋白COPT家族成员与Cu有很高的亲和力, 能够调节植物对Cu的吸收和转运, 在维持植物体内Cu稳态平衡过程中发挥重要作用。COPT蛋白涉及不同的Cu转运功能, 如从外界环境中摄取Cu、从细胞器中输出Cu、长距离运输Cu以及在不同器官间动用和再分配Cu。此外, COPT蛋白在其它离子的稳态平衡维持、昼夜节律性生物钟调控、植物激素合成和植物对激素信号的感受过程中也发挥重要作用。该文综述了模式植物拟南芥(Arabidopsis thaliana) COPT家族各成员的表达和定位、调控机制以及生物学功能等方面的最新进展。     

植物重金属超富集机理研究进展

植物超富集重金属机理主要涉及植物对金属离子高的吸收、运输能力,区域化作用及螯合作用等方面,其中跨膜运载蛋白的表达、调控对重金属超富集这一特性起了关键作用。金属阳离子运载蛋白家族主要包括CDF家族、NRAMP家族和ZIP家族等,在超富集植物中已克隆出多个家族的金属运载蛋白基因,这些基因的过量表达对重金属在细胞中的运输、分布和富集及提高植物的抗性方面发挥了重要作用。综述了近年来研究重金属超富集植物吸收、转运和贮存Zn、Ni、Cd等重金属的生理和分子机制所取得的主要进展。     

Molecular and physiological interactions of urea and nitrate uptake in plants

While nitrate acquisition has been extensively studied, less information is available on transport systems of urea. Furthermore, the reciprocal influence of the two sources has not been clarified, so far. In this review, we will discuss recent developments on plant response to urea and nitrate nutrition. Experimental evidence suggests that, when urea and nitrate are available in the external solution, the induction of the uptake systems of each nitrogen (N) source is limited, while plant growth and N utilization is promoted. This physiological behavior might reflect cooperation among acquisition processes, where the activation of different N assimilatory pathways (cytosolic and plastidic pathways), allow a better control on the nutrient uptake. Based on physiological and molecular evidence, plants might increase (N) metabolism promoting a more efficient assimilation of taken-up nitrogen. The beneficial effect of urea and nitrate nutrition might contribute to develop new agronomical approaches to increase the (N) use efficiency in crops.     

Molecular mechanisms of urea transport in plants

Urea is a soil nitrogen form available to plant roots and a secondary nitrogen metabolite liberated in plant cells. Based on growth complementation of yeast mutants and “in-silico analysis”, two plant families have been identified and partially characterized that mediate membrane transport of urea in heterologous expression systems. AtDUR3 is a single Arabidopsis gene belonging to the sodium solute symporter family that cotransports urea with protons at high affinity, while members of the tonoplast intrinsic protein (TIP) subfamily of aquaporins transport urea in a channel-like manner. The following review summarizes current knowledge on the membrane localization, energetization and regulation of these two types of urea transporters and discusses their possible physiological roles in planta.     

Strategies to identify transport systems in plants

Since the first molecular structures of plant transporters were discovered over a decade ago, considerable advances have been made in the study of plant membrane transport, but we still do not understand transport regulation. The genes encoding the transport systems in the various cell membranes are still to be identified, as are the physiological roles of most transport systems. A wide variety of complementary strategies are now available to study transport systems in plants, including forward and reverse genetics, proteomics, and in silico exploitation of the huge amount of information contained in the completely known genomic sequence of Arabidopsis.     

Plant aquaporins with non-aqua functions: deciphering the signature sequences

Research in recent years on plant Major Intrinsic Proteins (MIPs), commonly referred to as 'aquaporins', has seen a vast expansion in the substrates found to be transported via these membrane channels. The diversity in sizes, chemical nature and physiological significance of these substrates has meant a need to critically analyse the possible structural and biochemical properties of MIPs that transport these, in order to understand their roles. In this work we have undertaken a comprehensive analysis of all plant MIPs, coming from different families, that have been proven to transport ammonia, boron, carbon dioxide, hydrogen peroxide, silicon and urea. The sequences were analysed for all primary selectivity-related motifs (NPA motifs, ar/R filter, P1-P5 residues). In addition, the putative regulatory phosphorylation and glycosylation sites and mechanistic regulators such as loop lengths have been analysed. Further, nine specificity-determining positions (SDPs) were predicted for each group. The results show the ar/R filter residues, P2-P4 positions and some of the SDPs are characteristic for certain groups, and O-glycosylation sites are unique to a subgroup while N-glycosylation was characteristic of the other MIPs. Certain residues, especially in loop C, and structural parameters such as loop lengths also contribute to the uniqueness of groups. The comprehensive analysis makes significant inroads into appraising the intriguing diversity of plant MIPs and their roles in fundamental life processes, and provides tools for plant selections, protein engineering and transgenics.     

高等植物钾转运蛋白

钾在植物生长发育过程中具有许多重要的作用。以模式植物拟南芥中克隆和鉴定的钾通道和转运体为基础,全面介绍了高等植物中钾转运体系家族,包括Shaker通道、KCO通道、KUP/HAK/KT转运体、HKT转运体和其它转运体。同时,分析了在高等植物中存在多种钾吸收和转运机制的可能原因。     

Enigma variations for peptides and their transporters in higher plants

BACKGROUND: Two families of proteins that transport small peptides, the oligopeptide transporters (OPTs) and the peptide transporters (PTRs), have been recognized in eukaryotes. Higher plants contain a far greater number of genes for these transporters than do other eukaryotes. This may be indicative of the relative importance of (oligo)peptides and their transport to plant growth and metabolism. RECENT PROGRESS: Recent studies are now allowing us to assign functions to these transporters and are starting to identify their in-planta substrates, revealing unexpected and important contributions of the transporters to plant growth and developmental processes. This Botanical Briefing appraises recent findings that PTRs and OPTs have key roles to play in the control of plant cell growth and development. Evidence is presented that some of these transporters have functions outside that of nitrogen nutrition and that these carriers can also surprise us with their totally unexpected choice of substrates.     

Sugar transporters in higher plants--a diversity of roles and complex regulation

Sugar-transport proteins play a crucial role in the cell-to-cell and long-distance distribution of sugars throughout the plant. In the past decade, genes encoding sugar transporters (or carriers) have been identified, functionally expressed in heterologous systems, and studied with respect to their spatial and temporal expression. Higher plants possess two distinct families of sugar carriers: the disaccharide transporters that primarily catalyse sucrose transport and the monosaccharide transporters that mediate the transport of a variable range of monosaccharides. The tissue and cellular expression pattern of the respective genes indicates their specific and sometimes unique physiological tasks. Some play a purely nutritional role and supply sugars to cells for growth and development, whereas others are involved in generating osmotic gradients required to drive mass flow or movement. Intriguingly, some carriers might be involved in signalling. Various levels of control regulate these sugar transporters during plant development and when the normal environment is perturbed. This article focuses on members of the monosaccharide transporter and disaccharide transporter families, providing details about their structure, function and regulation. The tissue and cellular distribution of these sugar transporters suggests that they have interesting physiological roles.     

Plant cation/H+ exchangers (CAXs): biological functions and genetic manipulations

Inorganic cations play decisive roles in many cellular and physiological processes and are essential components of plant nutrition. Therefore, the uptake of cations and their redistribution must be precisely controlled. Vacuolar antiporters are important elements in mediating the intracellular sequestration of these cations. These antiporters are energized by the proton gradient across the vacuolar membrane and allow the rapid transport of cations into the vacuole. CAXs (for CAtion eXchanger) are members of a multigene family and appear to predominately reside on vacuoles. Defining CAX regulation and substrate specificity have been aided by utilising yeast as an experimental tool. Studies in plants suggest CAXs regulate apoplastic Ca(2+) levels in order to optimise cell wall expansion, photosynthesis, transpiration and plant productivity. CAX studies provide the basis for making designer transporters that have been used to develop nutrient enhanced crops and plants for remediating toxic soils.