脯氨酸代谢与植物抗渗透胁迫的研究进展

脯氨酸被认为是植物和细菌内的一种相容渗透剂,有助于植物和细菌抵御渗透胁迫。本文就近年来有关植物体内脯氨酸合成和代谢、脯氨酸含量受渗透胁迫的影响情况、脯氨酸合成降解有关的酶及其基因、脯氨酸在细胞中的运输和定位、ABA与脯氨酸的诱导合成以及脯氨酸和植物抗渗透胁迫关系的研究进展作了简要综述。     

脯氨酸代谢与植物抗渗透胁迫的研究进展

脯氨酸被认为是植物和细菌内的一种相容渗透剂,有助于植物和细菌抵御渗透胁迫。本文就近年来有关植物体内脯氨酸合成和代谢、脯氨酸含量受渗透胁迫的影响情况、脯氨酸合成降解有关的酶及其基因、脯氨酸在细胞中的运输和定位、ＡＢＡ与脯氨酸的诱导合成以及脯氨酸和植物抗渗透胁迫关系的研究进展作了简要综述。     

脯氨酸在植物生长和非生物胁迫耐受中的作用

脯氨酸是生物界分布最广的渗透保护物质之一,干旱、高盐、高温及重金属等非生物胁迫条件都会导致植物体内脯氨酸含量的增加,其作用是防止渗透胁迫对植物造成的伤害、清除自由基,还可以作为氮、碳以及NADPH的重要来源。近年来,在转化脯氨酸代谢相关基因提高植物胁迫抗性方面也取得了很大进展。本文概要介绍了脯氨酸在植物生长和耐受非生物胁迫中的作用、与植物脯氨酸累积有关的信号转导、胁迫条件下脯氨酸的吸收和器官间的运输途径,以及通过转基因技术过量表达脯氨酸提高植物胁迫耐性的代谢工程的进展。     

植物体内脯氨酸累积和γ-谷氨酰磷酸合成活力的关系

渗透胁迫下植物体内脯氨酸合成加强,致使脯氨酸大量累积(汤章城 1984,Stewart和Hanson 1980,Dashek等1981)。在微生物中已经证实脯氨酸合成的关键酶为γ-谷氨酰激酶,并且已得到纯化酶(Baich 1969,Smith等1984),在高等植物中尚未完全证实这种酶的存在。Ericson等(1984)认为在高等植物中测定     

外源茉莉酸甲酯对盐胁迫下小桐子幼苗渗透调节和脯氨酸代谢的影响

以小桐子幼苗为材料,采用人工气候箱内水培试验,设置不同浓度(0、20、40、60、80、100μmol·L^(-1))茉莉酸甲酯(MeJA)和150 mmol·L^(-1)NaCl胁迫处理,分析不同处理条件下小桐子幼苗叶片的组织活力、MDA含量、水势、含水量和叶片渗透调节物质脯氨酸、甜菜碱和可溶性糖的含量,以及脯氨酸代谢关键酶P5CS、OAT和甜菜碱合成关键酶BADH活性和相关基因表达水平,探讨外源茉莉酸甲酯对盐胁迫下小桐子幼苗渗透调节能力的影响及其机制。结果表明:(1)外源MeJA处理可提高盐胁迫下小桐子幼苗叶片的组织活力和叶片含水量,降低小桐子幼苗叶片的MDA含量和水势,且60μmol·L^(-1)浓度处理效果最佳。(2)外源MeJA处理可提高盐胁迫下小桐子幼苗叶片的脯氨酸、甜菜碱和可溶性糖的含量,且60μmol·L^(-1)MeJA处理显著提高了盐胁迫下小桐子幼苗内源茉莉酸、脯氨酸和甜菜碱的含量。(3)60μmol·L^(-1)MeJA也提高了盐胁迫下小桐子BADH、P5CS和OAT的活性,并上调了JcBADH、JcP5CS、JcOAT基因的表达水平,但MeJA降低了脯氨酸降解酶ProDH的活性,下调了JcProDH基因的表达。研究发现,在盐胁迫条件下,外源MeJA通过活化脯氨酸生物合成的谷氨酸和鸟氨酸途径,尤其是鸟氨酸途径,以及抑制脯氨酸降解途径来促进小桐子幼苗脯氨酸的积累,同时MeJA也激活了幼苗体内甜菜碱的生物合成过程,从而强化了盐胁迫下幼苗的渗透调节作用和耐盐性,表明MeJA诱导的渗透调节在小桐子耐盐性形成过程中发挥着重要作用。     

丛枝菌根提高宿主植物抗旱性分子机制研究进展

丛枝菌根(arbuscular mycorrhiza, AM)对于植物适应各种逆境胁迫具有重要生态学意义。有关菌根共生体对植物抵御干旱胁迫的积极作用已有较多文献报道:无论在植物个体层面——AM调节植物水分生理,还是在生态层面——干旱条件下菌根真菌和宿主植物之间的互动关系,人们都已有一定的认识。然而,目前对于菌根植物适应干旱胁迫的生理和分子机制还缺乏系统深入的研究。综述了近年来相关研究成果,从干旱胁迫相关植物基因入手,讨论了AM对晚期胚胎富集蛋白(LEA)、脯氨酸合成限速酶△¹-吡咯啉-5-羧酸合成酶(P5CS)、水孔蛋白(MIPs),及脱落酸合成途径重要酶9-顺式-环氧类胡萝卜素双加氧酶(NCED)编码基因的可能调控机制,旨在揭示AM共生体提高植物抗旱性的分子基础和实质贡献,同时通过分析当前研究工作薄弱之处及未来研究热点,期望推动相关研究进展。     

渗透胁迫下白花柽柳cDNA文库的构建及EST分析

目的：寻找渗透胁迫相关基因EST片断,方法：构建白花柽柳cDNA文库,对文库阳性克隆随机测序,并进行网上比对分析。结果：该cDNA文库克隆的重组率大于95%,插入片段主要集中在250～1000bp之间,并获得了如脯氨酸丰富蛋白基因,钙依赖蛋白激酶基因,丝氨酸／苏氨酸激酶基因,过氧化氢酶基因,膜转运蛋白等基因。结论：渗透胁迫基因是多基因参与的系统调控过程,它们涉及了植物细胞壁合成、信号传递、基因调控、氧化物质清除、渗透调节物质的转运等方面。     

外源水杨酸对盐胁迫下小桐子幼苗脯氨酸代谢的影响

以小桐子幼苗为材料,设置盐胁迫(200mmol·L-1 NaCl)和外源水杨酸处理(0~2.0mmol·L-1 SA)水培试验,通过检测幼苗叶片脯氨酸含量、脯氨酸代谢关键酶活性及相关代谢酶基因的表达水平,研究了外源水杨酸对盐胁迫下小桐子幼苗脯氨酸代谢机理的影响。结果显示:(1)外源0.9mmol·L-1 SA处理可显著提高盐胁迫下小桐子幼苗的脯氨酸含量,上调脯氨酸合成关键酶Δ1-吡咯琳-5-羧酸合成酶(P5CS)和鸟氨酸转氨酶(OAT)活性,以及上调JcP5CS和JcOAT基因的表达水平。(2)SA也显著抑制了脯氨酸降解酶ProDH的活性及JcProDH基因的表达水平。(3)SA处理还显著提高了盐胁迫下小桐子幼苗的组织活力,降低了叶片电解质渗漏率和丙二醛(MDA)含量。研究发现,外源SA可通过活化脯氨酸合成的谷氨酸途径和鸟氨酸途径,以及抑制脯氨酸的降解途径来促进盐胁迫下小桐子幼苗脯氨酸的积累;外源SA处理也可提高小桐子幼苗的耐盐性,且这种提高可能与SA诱导脯氨酸的积累密切相关。     

植物盐胁迫的信号调控机制

盐胁迫会引起渗透胁迫、离子毒害和次级胁迫,极大危害到植物的生存。之前的研究中,借助于分子生物学和遗传学手段,已经鉴定出多个基因参与到盐胁迫信号通路的调控,其中SOS途径是主要的植物抗盐调节机制。氨基酸、甜菜碱、可溶性糖等渗透调节物质,脯氨酸、多胺等信号小分子,SOD、CAT等抗氧化酶,HKT等离子通道蛋白均参与到了植物对盐胁迫的响应。本研究总结了盐胁迫条件下植物调控离子平衡、氧化胁迫、生长发育的最新研究进展,为之后的研究提供依据。     

植物抗旱基因工程研究进展

综述了渗透保护物质生物合成关键酶基因、胚胎后期发生丰富蛋白(Lea)基因或Lea相关基因、编码转录因子的调节基因、抗氧化胁迫相关的酶等植物抗旱基因工程研究进展,并对其研究中存在的问题及今后的应用前景进行了讨论.     

Regulation of proline utilization in Salmonella typhimurium: How do cells avoid a futile cycle?

Summary Salmonella typhimurium can degrade proline for use as a carbon, nitrogen, or energy source. To determine whether a futile cycle occurs which degrades the proline accumulated by proline biosynthesis, we studied the expression and enzymatic activity of the proline utilization (put) pathway under conditions which increase the concentration of the intracellular proline pools: catabolism of the dipeptide glycyl-proline, overproduction of proline due to a mutation which prevents feedback inhibition of proline biosynthesis, and accumulation of proline due to osmotic stress. The results indicate that: (i) internal proline induces the put genes, but only when accumulated to concentrations greater than the normal proline biosynthetic pool; and (ii) degradation of proline pools accumulated under high osmotic pressure is limited because proline oxidase is directly inhibited under these conditions.     

Oscillation and regulation of proline content by P5CS and ProDH gene expressions in the light/dark cycles in Arabidopsis thaliana L

The fluctuation of proline content, and protein and mRNA levels of delta1-pyrroline-5-carboxylate synthetase (P5CS) and proline dehydrogenase (ProDH), both of which are involved in proline biosynthesis and degradation, in the shoots of Arabidopsis grown in light/dark cycles were demonstrated under salt-stressed and unstressed conditions. Proline content, as well as proteins and mRNAs of these enzymes, clearly oscillated in the light/dark cycles under the stressed and unstressed conditions. A reciprocal relationship between P5CS and ProDH was observed. Protein levels of P5CS and ProDH were well synchronized with their mRNA levels, although the fluctuation of protein levels was not as significant as that of their mRNA levels. Both mRNA and protein levels of the two enzymes as well as the proline content did not oscillate under the continuous light or the dark conditions. Thus, P5CS and ProDH gene expressions seemed to be involved in light irradiation. Moreover, relative water content (RWC) in the plants oscillated in the light/dark cycles. The fluctuations of proline content in shoot reversely responded to that of RWC. It is suggested that the expression of two genes responds sensitively to a subtle change of cellular water status, and accumulated proline keeps the osmotic balance between cells and the outer environment.     

Stress-induced synthesis of proline confers tolerance to water deficit in transgenic wheat

Water deficit is one of the main abiotic factors that affect spring wheat planted in subtropical regions. Accumulation of proline appears to be a promising approach to maintain the productivity of plants under stress condition. However, morphological alterations and growth reduction are observed in transgenic plants carrying genes coding for osmoprotectants controlled by constitutive promoters. We report here the effects of water deficit on wheat plants transformed with the Vigna aconitifolia Delta(1)-pyrroline-5-carboxylate synthetase (P5CS) cDNA that encodes the key regulatory enzyme in proline biosynthesis, under the control of a stress-induced promoter complex-AIPC. Transgenic wheat plants submitted to 15 days of water shortage presented a distinct response. We have found that drought resulted in the accumulation of proline. The tolerance to water deficit observed in transgenic plants was mainly due to protection mechanisms against oxidative stress and not caused by osmotic adjustment.     

Bacterial endophytes contribute to abiotic stress adaptation in pepper plants (Capsicum annuum L.)

Endophytes are nonpathogenic plant-associated bacteria that can play an important role in plant vitality and may confer resistance to abiotic or biotic stress. The effects of 5 endophytic bacterial strains isolated from pepper plants showing 1-aminocyclopropane-1-carboxylate deaminase activity were studied in sweet pepper under in vitro conditions. Four of the strains tested showed production of indole acetic acid. Plant growth, osmotic potential, free proline content, and gene expression were monitored in leaves and roots under control and mild osmotic stress conditions. All indole acetate producers promoted growth in Capsicum annuum L. 'Ziegenhorn Bello', from which they were isolated. Osmotic stress caused an increase in the content of free proline in the leaves of both inoculated and noninoculated plants. Inoculated control plants also revealed higher proline levels in comparison with noninoculated control plants. Differential gene expression patterns of CaACCO, CaLTPI, CaSAR82A, and putative P5CR and P5CS genes during moderate stress were observed, depending on the bacterium applied. Inoculation with 2 bacterial strains, EZB4 and EZB8 (Arthrobacter sp. and Bacillus sp., respectively), resulted in a significantly reduced upregulation or even downregulation of the stress-inducible genes CaACCO and CaLTPI, as compared with the gene expression in noninoculated plants. This indicates that both strains reduced abiotic stress in pepper under the conditions tested.     

Proline as a biochemical marker in relation to the ecology of two halophytic Juncus species

Aims Osmolytes, used for maintaining osmotic balance and as 'osmoprotectants', are synthesized in plants as a general, conserved response to abiotic stress, although their contribution to stress-tolerance mechanisms remains unclear. Proline, the most common osmolyte, accumulates in many plant species in parallel with increased external salinity and is considered a reliable biochemical marker of salt stress. We have measured proline levels in two halophytic, closely related Juncus species under laboratory and field conditions to assess the possible relevance of proline biosynthesis for salt tolerance and therefore for the ecology of these two taxa.Methods Proline was quantified in plants treated with increasing NaCl concentrations and in plants sampled in two salt marshes located in the provinces of Valencia and Alicante, respectively, in southeast Spain. Electrical conductivity, pH, Na + and Cl ? concentrations were measured in soil samples collected in parallel with the plant material.Important findings Treatment with NaCl inhibited growth of J. acutus plants in a concentration-dependent manner, but only under high salt conditions for J. maritimus. Salt treatments led to proline accumulation in both species, especially in the more salt-tolerant J. maritimus. The results, obtained under laboratory conditions, were confirmed in plants sampled in the field. In all the samplings, proline contents were significantly lower in J. acutus than in the more tolerant J. maritimus growing in the same area. No direct correlation between soil salinity and proline levels could be established, but seasonal variations were detected, with increased proline contents under accentuated water deficit conditions. Our results suggest that proline biosynthesis is not only an induced, general response to salt stress but also an important contributing factor in the physiological mechanisms of salt tolerance in Juncus, and that it therefore correlates with the ecology of both species.     

Transgenic Medicago truncatula plants that accumulate proline display nitrogen-fixing activity with enhanced tolerance to osmotic stress

Legume root nodule nitrogen-fixing activity is severely affected by osmotic stress. Proline accumulation has been shown to induce tolerance to salt stress, and transgenic plants over-expressing Delta(1)-pyrroline-5-carboxylate synthetase (P5CS), which accumulates high levels of proline, display enhanced osmotolerance. Here, we transformed the model legume Medicago truncatula with the P5CS gene from Vigna aconitifolia, and nodule activity was evaluated under osmotic stress in transgenic plants that showed high proline accumulation levels. Nitrogen fixation was significantly less affected by salt treatment compared to wild-type (WT) plants. To our knowledge, this is the first time that transgenic legumes have been produced that display nitrogen-fixing activity with enhanced tolerance to osmotic stress. We studied the expression of M. truncatula proline-related endogenous genes M. truncatulaDelta(1)-pyrroline-5-carboxylate synthetase 1 (MtP5CS1), M. truncatulaDelta(1)-pyrroline-5-carboxylate synthetase 2 (MtP5CS2), M. truncatula ornithine delta-aminotransferase (MtOAT), M. truncatula proline dehydrogenase (MtProDH) and a proline transporter gene in both WT and transgenic plants. Our results indicate that proline metabolism is finely regulated in response to osmotic stress in an organ-specific manner. The transgenic model allowed us to analyse some of the biochemical and molecular mechanisms that are activated in the nodule in response to high salt conditions, and to ascertain the essential role of proline in the maintenance of nitrogen-fixing activity under osmotic stress.     

Assessment of proline function in higher plants under extreme temperatures

Climate change and abiotic stress factors are key players in crop losses worldwide. Among which, extreme temperatures (heat and cold) disturb plant growth and development, reduce productivity and, in severe cases, lead to plant death. Plants have developed numerous strategies to mitigate the detrimental impact of temperature stress. Exposure to stress leads to the accumulation of various metabolites, e.g. sugars, sugar alcohols, organic acids and amino acids. Plants accumulate the amino acid ‘proline’ in response to several abiotic stresses, including temperature stress. Proline abundance may result from de novo synthesis, hydrolysis of proteins, reduced utilization or degradation. Proline also leads to stress tolerance by maintaining the osmotic balance (still controversial), cell turgidity and indirectly modulating metabolism of reactive oxygen species. Furthermore, the crosstalk of proline with other osmoprotectants and signalling molecules, e.g. glycine betaine, abscisic acid, nitric oxide, hydrogen sulfide, soluble sugars, helps to strengthen protective mechanisms in stressful environments. Development of less temperature-responsive cultivars can be achieved by manipulating the biosynthesis of proline through genetic engineering. This review presents an overview of plant responses to extreme temperatures and an outline of proline metabolism under such temperatures. The exogenous application of proline as a protective molecule under extreme temperatures is also presented. Proline crosstalk and interaction with other molecules is also discussed. Finally, the potential of genetic engineering of proline-related genes is explained to develop ‘temperature-smart’ plants. In short, exogenous application of proline and genetic engineering of proline genes promise ways forward for developing ‘temperature-smart’ future crop plants.