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。     

水分胁迫期间及胁迫解除后桃树叶片中的碳水化合物代谢

水分胁迫期间、盆栽桃幼树叶片中淀粉和蔗糖含量降低,山梨醇、葡萄糖和果糖则升高;淀粉酶、6－磷酸醛糖还原酶（A6PR）、酸性转化酶（AI）和蔗糖合酶（SS）涤性升高,ADPG焦磷酸化酶（ADPGPPase）、中性转化酶（NI）、山梨醇脱氢酶（SDH）和磷酸蔗糖合酶（SPS）的活性降低。胁迫解除后,淀粉和葡萄糖含量恢复,山梨醇和果糖仍高于对照;A6PR、SDH和ADPGPase活性维持在原水平上;淀粉酶活性恢复;SPS活性增加而AI活性降低。     

外源钙对汞胁迫下菹草(Potamogeton crispus L.)叶片抗氧化系统及脯氨酸代谢的调节效应

以组织培养的菹草无菌苗为实验材料,研究了外源Ca(NO3)2对汞(Hg)胁迫下菹草无菌苗抗氧化酶系统、可溶性蛋白、总抗氧化能力(T-AOC)、非蛋白巯基(NP-SH)、植物络合素(PCs)以及脯氨酸代谢的调节效应。结果表明:(1)Hg胁迫使菹草体内过氧化氢酶(CAT)、抗坏血酸过氧化物酶(APX)及谷胱甘肽还原酶(GR)活性明显降低,但超氧化物歧化酶(SOD)及过氧化物酶(POD)的活性并没有明显变化。外源添加25 mmol·L-1Ca(NO3)2后,明显提升了SOD、POD、APX及GR的活性;(2)Hg胁迫使菹草体内可溶性蛋白含量及T-AOC大幅降低,并引起NP-SH和PCs大量积累。外添Ca(NO3)2则有效减缓可溶性蛋白含量及T-AOC的下降,显著降低NP-SH和PCs水平;(3)脯氨酸是植物体内重要的渗透调节物质,外源Ca添加后,脯氨酸代谢关键酶吡咯啉-5-羧酸合成酶(P5CS)及δ-鸟氨酸转氨酶(OAT)活性显著增加,脯氨酸含量显著高于对照组。以上结果表明,Hg胁迫打破了菹草无菌苗抗氧化酶系统的内在平衡,而外添Ca(NO3)2有效促进了可溶性蛋白合成,提高了抗氧化酶活性和抗氧化物质含量,使菹草无菌苗维持较高的总抗氧化能力,并通过调节脯氨酸代谢来减轻Hg胁迫对菹草的伤害,增强植物对重金属Hg的耐受能力。     

外源脯氨酸对盐胁迫下甜瓜脯氨酸代谢的影响

为探明外源脯氨酸对盐胁迫下甜瓜脯氨酸代谢的影响,以甜瓜品种‘雪美’为材料采用营养液栽培,对盐胁迫(100mmol·L-1 NaCl)、盐胁迫下添加外源脯氨酸(100mmol·L-1 NaCl+0.2mmol·L-1 Proline)以及对照3种处理后甜瓜幼苗叶片脯氨酸(Pro)含量、吡咯啉-5-羧酸合成酶(P5CS)、鸟氨酸转氨酶(OAT)和脯氨酸脱氢酶(ProDH)活性进行测定,并对OAT和ProDH基因进行克隆及半定量表达分析。结果显示:与对照相比较,盐胁迫条件下甜瓜幼苗叶片内Pro含量显著增加,P5CS活性增幅大于OAT活性,OAT基因表达量大部分时段内没有增加,ProDH活性下降,ProDH基因表达量减少;盐胁迫下添加外源脯氨酸进一步使幼苗叶片内Pro含量增加、OAT、ProDH活性提高、P5CS活性降低,并且使OAT基因表达量迅速增加、ProDH基因表达量先增加后回落。研究表明,盐胁迫条件下,甜瓜幼苗体内脯氨酸积累主要是通过增强脯氨酸的谷氨酸合成途径和抑制脯氨酸降解来实现;适量外源脯氨酸可以增强盐胁迫幼苗脯氨酸的鸟氨酸合成途径,但对谷氨酸合成途径有一定的抑制作用;通过调节合成和降解2种代谢途径进一步提高了脯氨酸含量,从而增强甜瓜幼苗耐盐胁迫能力。     

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。     

外源ABA对低温胁迫下小桐子幼苗脯氨酸积累及其代谢途径的影响

本文以小桐子为材料,研究了外源脱落酸（ABA）对低温胁迫下小桐子幼苗脯氨酸积累的影响。结果表明,低温胁迫（5℃）可促进小桐子幼苗体内脯氨酸的积累,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）的活性,上调P5CS基因（JcP5CS）的表达,及抑制脯氨酸降解酶脯氨酸脱氢酶（Pro DH）的活性。用外源ABA处理低温胁迫下的小桐子幼苗,发现150μmol·L^-1的ABA可显著提高其低温胁迫下的脯氨酸含量,上调P5CS的活性和JcP5CS的表达水平,及抑制ProDH的活性。表明外源ABA可通过活化脯氨酸合成的谷氨酸途径和抑制脯氨酸的降解途径来促进低温胁迫下小桐子幼苗脯氨酸的积累。     

水分胁迫对植物线粒体结构和脯氨酸氧化酶活性的影响

水分胁迫期间,小麦幼苗芽鞘和棉花幼苗胚轴细胞内游离脯氨酸浓度增加;但复水后又恢复正常。电镜观察发现线粒体肿大,嵴消失。胞质中出现脂肪滴。气相层析技术分析,发现水分胁迫使线粒体脂肪酸组成及含量有明显变化;不饱和脂肪酸含量增加。随着水分胁迫时间的延长,脯氨酸氧化酶活性也明显下降。设想水分胁迫使线粒体结构和组分发生了不利于脯氨酸氧化酶活性表达的变化,因而抑制了酶活性。     

水分胁迫对荔枝叶片氮和核酸代谢的影响及其与抗旱性的关系

水分胁迫下,荔枝叶片蛋白酶活性和Ｐｒｏ含量增加,ＰＤＨ活性与可溶性蛋白质含量下降,抗旱性强的品种蛋白酶活性增加的幅度蛋白质含量下降的幅度小于抗旱性弱的品种,而ＰＤＨ活性下降的幅度和Ｐｒｏ含量上升的幅度均大于抗旱性弱的品种。水分胁迫引起荔枝叶片核酸,ＤＮＡ和ＲＮＡ含量下降,ＤＮＡ含量下降的幅度小于ＲＮＡ含量下降的幅度;ＤＮａｓｅ和ＲＮａｓｅ活性上升,ＤＮａｓｅ活性上升的幅度小于ＲＮａｓｅ活性上升的幅     

Ammonium in relation to proline accumulation in detached rice leaves

Ammonium accumulation in relation to prolineaccumulation in detached rice leaves under stressconditions was investigated. Ammonium accumulation indark-treated detached rice leaves preceded prolineaccumulation. Ammonium accumulation caused by waterstress coincided closely with proline accumulation indetached rice leaves. Exogenous NH₄Cl andmethionine sulfoximine (MSO), which caused anaccumulation of ammonium in detached rice leaves,increased proline content. It was found that prolinein NH₄Cl- or MSO-treated rice leaves is lessutilized than in water-treated rice leaves (controls). These results are in agreement with the observationthat a decrease in proline utilization contributes tothe accumulation of proline in dark-treated and waterstressed rice leaves. Although ammonium contentincreased in Cd- and Cu-treated rice leaves, theincrease in ammonium content was only observed afterthe increase in proline content.     

干旱胁迫下雷竹叶片叶绿素的高光谱响应特征及含量估算

植物叶片的反射光谱特征与叶绿素含量密切相关。以重要的笋用竹种雷竹(Phyllostachys violascens)为研究对象,采用盆栽及控水试验方法研究了2年生雷竹在干旱胁迫条件下冠层叶片反射光谱的响应特征,分析了叶片叶绿素含量与不同波段光谱反射率一阶微分值以及光谱特征参数之间的相关关系,并以雷竹叶绿素含量敏感波段及构建的植被指数与叶绿素含量进行了拟合。结果表明,重度缺水处理后雷竹叶片叶绿素含量显著降低,在可见光区叶片光谱反射率随叶绿素含量的降低而增加,以波长493、639、693、756 nm等处的光谱反射率一阶微分值与叶绿素含量的相关性较高。雷竹叶片叶绿素含量与光谱特征参数如绿峰反射率、红谷反射率、蓝边面积、绿峰面积之间的相关性较高。与已有的植被指数相比基于雷竹叶绿素含量敏感波段修正后的植被指数与叶绿素含量相关性优于原植被指数。基于反射率一阶微分值构建的多元回归方程以及修正的绿色归一化植被指数(m GNDVI)构建的回归方程拟合效果较好,为雷竹叶绿素含量的较优估算方程。研究结果可以为雷竹叶绿素含量的快速无损测定以及季节性干旱条件下雷竹林的科学经营及灾后评估提供依据。     

干旱胁迫下紫花苜蓿幼苗非结构性碳水化合物代谢对NO的响应

以紫花苜蓿（Medicago sativa）为材料,采用盆栽试验方法,用聚乙二醇（PEG-6000）作为渗透介质模拟干旱胁迫,外源喷施NO供体硝普钠,NO清除剂（carboxy-PTIO,cPTIO）,对紫花苜蓿幼苗叶片、根系中非结构性碳水化合物含量及相关酶活性的变化进行研究,探讨NO对紫花苜蓿耐旱机制的作用。结果表明：外源NO促进了紫花苜蓿叶片中淀粉的分解、根系中淀粉的积累,提高叶片及根系中可溶性糖（蔗糖、果糖和葡萄糖）含量,降低了渗透势,促进细胞吸水,缓解干旱造成的损伤。此外,外源NO能提高干旱胁迫下紫花苜蓿叶片中蔗糖合成酶（SS）、酸性转化酶（AI）和中性转化酶（NI）活性,降低了蔗糖磷酸合成酶（SPS）的活性,提高根系中SS、SPS和转化酶活性,使蔗糖的合成与分解处于高水平的动态平衡,增强了紫花苜蓿的抗旱性。而NO清除剂cPTIO则会不同程度的抑制紫花苜蓿幼苗中非结构性碳水化合物（NSC）及其相关酶活性。因此,NO可以通过调控NSC的代谢响应干旱胁迫,缓解干旱胁迫造成的不利影响,在紫花苜蓿的抗旱中扮演着重要的角色。     

Separately and simultaneously induced dark chilling and drought stress effects on photosynthesis, proline accumulation and antioxidant metabolism in soybean

The effects of separately or simultaneously induced dark chilling and drought stress were evaluated in two Glycine max (L.) Merrill cultivars. For the separately induced dark chilling treatment (C), plants were incubated at 8 °C during 9 consecutive dark periods. During the days, plants were kept at normal growth temperatures. For the separately induced drought treatment (D), plants were maintained at normal growth temperatures without irrigation. For the simultaneously induced dark chilling and drought stress treatment (CD), plants were dark chilled without irrigation. All treatments caused similar decreases in pre-dawn leaf water potential, but resulted in distinct physiological and biochemical effects on photosynthesis. In Maple Arrow, where C had the smallest effect on photosynthesis, prolonged CD caused less inhibition of photosynthesis compared to D. Compared to Fiskeby V, the photosynthetic apparatus of Maple Arrow appears to possess superior dark chilling tolerance, a property which probably also conveyed enhanced protection against CD. Proline accumulation was prevented by CD at the ψ_PD where D already resulted in considerable accumulation. The superior capacity for proline accumulation in Maple Arrow would seem to be an important factor in its stress tolerance. Antioxidant activity evoked by CD and D was higher than for C alone. In Fiskeby V, the small increase in ascorbate peroxidase (EC 1.11.1.7) activity, which was in most cases not accompanied by increased gluthatione reductase (EC 1.6.4.2) activity, could impact negatively on its stress tolerance. These results demonstrate large genotypic differences in response to chilling and drought stress, even between soybean cultivars regarded as chilling tolerant.     

Genotype differences in the metabolism of proline and polyamines under moderate drought in tomato plants

Water stress is one of the most important factors limiting the growth and productivity of crops. The implication of compatible osmolytes such as proline and polyamines in osmotic adjustment has been widely described in numerous plants species under stress conditions. In the present study, we investigated the response of five cherry tomato cultivars (Solanum lycopersicum L.) subjected to moderate water stress in order to shed light on the involvement of proline and polyamine metabolism in the mechanisms of tolerance to moderate water stress. Our results indicate that the most water stress‐resistant cultivar (Zarina) had increased degradation of proline associated with increased polyamine synthesis, with a higher concentration of spermidine and spermine under stress conditions. In contrast, Josefina, the cultivar most sensitive to water stress, showed a proline accumulation associated with increased synthesis after being subjected to stress. In turn, in this cultivar, no rise in polyamine synthesis was detected. Therefore, all the data appear to indicate that polyamine metabolism is more involved in the tolerance response to moderate water stress.     

干旱胁迫下白刺花种子大小与萌发对策

种子大小与种子萌发及其与环境因子的关系是植物种子萌发对策研究中的重要科学问题之一。采用PEG模拟干旱法研究不同干旱胁迫强度(0,5%,10%,15%,20%)下,白刺花(Sophora davidii)种子萌发进程、种子大小与种子萌发及种子命运的关系。结果表明:不同干旱胁迫下,白刺花种子具有相似的萌发进程,但中度干旱处理(10%PEG)萌发率显著高于零干旱(0%PEG)和重度干旱处理(P0.05),重度干旱处理(20%PEG)种子萌发开始时间晚于零干旱和中度干旱处理;种子大小与种子萌发开始时间的关系表现为零干旱处理下呈极显著负线性关系,中度干旱处理(5%PEG,10%PEG)下无相关关系,重度干旱处理(15%PEG,20%PEG)下呈负二次曲线关系;种子大小对种子命运的影响表现为零干旱处理有利于大、小种子萌发和小种子休眠,中度干旱处理(10%PEG)增加中等种子萌发、大种子休眠和小种子死亡风险,重度干旱处理(15%PEG,20%PEG)增加大种子死亡风险、中等种子和小种子休眠。综合分析表明,白刺花种子大小与萌发行为及种子命运的关系具有较强的环境依赖性,即种子萌发行为表现为顺境下种子越大萌发越快,逆境下小种子和大种子较中等种子萌发更快;种子命运表现为顺境增加种子死亡的风险,中度干扰有利于种子萌发,逆境则有利于种子休眠。     

干旱胁迫对蒙古黄芪生长及根部次生代谢物含量的影响

以山西道地药材黄芪一年生幼苗为试验材料,设置常规水分条件(CK)、轻度干旱胁迫(A1)、中度干旱胁迫(A2)、重度干旱胁迫(A3)4个不同处理,研究土壤干旱胁迫对黄芪生长及生理的影响。结果表明:黄芪茎叶快速生长集中在出苗后80—120d,以后生长减缓;当茎叶枯萎时,根中生物量短期快速积累。与常规水分条件相比,干旱胁迫处理显著降低了黄芪苗高及茎叶生物量,但对抗氧化能力、根系生长及次生代谢物积累产生了不同的影响。轻度干旱胁迫下SOD、POD、CAT 3种抗氧化酶活性升高,丙二醛(MDA)含量和细胞膜透性降低,同时根长与根生物量增加、多糖与皂苷两种次生代谢物积累增多,黄芪药材的质量得到显著提高(P0.05);胁迫上升到中度、重度时,SOD酶活性逐渐降低,重度胁迫下低于对照,而POD及CAT酶活性、MDA含量、细胞膜透性均随胁迫增强而升高,相反,根长、根生物量、多糖与皂苷含量降低,导致黄芪药材的质量显著降低(P0.01)。综上表明,干旱胁迫下,SOD酶表现较为敏感,可能在清除活性氧中起主要作用;轻度水分胁迫能有效启动黄芪体内抗氧化酶系统和次生代谢,它们相互协作共同对抗胁迫对细胞产生的伤害,通过降低地上部分的生长,将营养物质优先运往根部,促进根产量及药材质量的提高。这一结论,可在黄芪多糖和皂苷次生代谢物定向培育的水分管理中加以利用。     

Osmotic stress and water stress have opposite effects on putrescine and proline production in excised rice leaves

The effects of water stress and osmotic stress (sorbitol treatment) on the production of putrescine and proline in excised rice leaves were compared. Osmotic stress and water stress were found to affect differentially the levels of putrescine and proline in excised rice leaves. Putrescine accumulation is induced by osmotic stress, whereas proline accumulation is induced by water stress. The effects of ABA on the levels of proline and putrescine are similar to those of water stress, whereas the effects of jasmonic acid methyl ester (JA-Me) are similar to those of osmotic stress. Water stress results in an increase of endogenous ABA is excised rice leaves. However, neither osmotic stress nor JA-Me has effect on endogenous ABA levels in excised rice leaves. Of particular interest is the finding that proline levels increase when putrescine levels induced by osmotic stress or JA-Me are reduced by D-arginine and -methylornithine. L-arginine and L-ornithine applied exogenously also cause an increase in proline levels. It seems that L-arginine and L-ornithine are preferentially utilized as precursors for putrescine accumulation in excised rice leaves treated with osmotic stress and JA-Me, and for proline accumulation in excised rice leaves exposed to water stress and ABA.Abbreviations ABA abscisic acid - BSA bovine serum albumin - ELISA enzyme-linked immunosorbent assay - HPLC high performance chromatography - JA-Me jasmonic acid methyl ester - PVP poly-vinylpyrrolidone     

Alterations in the activities of the enzymes of proline metabolism in Ragi (Eleusine coracana) leaves during water stress

Free proline content in Ragi (Eleusine coracana) leaves increased markedly (6 to 85 fold) as the degree of water stress, created by polyethylene gylcol treatment, was prolonged There was also a marginal increase in soluble proteins in the stressed leaves as compared to that in the controls. Water stress stimulated the activities of ornithine aminotransferase and pyrroline-5-carboxylate reductase, the enzymes of proline biosynthesis and markedly inhibited the enzymes involved in proline degradation viz., proline oxidase and pyrroline-5-carboxylate dehydrogenase. These results suggest that increase in free proline content of Ragi leaves could be due to enhanced activities of the enzymes synthesizing proline but more importantly due to severe inhibition of the enzymes degrading proline. These observations establish for the first time, the pathway of proline metabolism in plants by way of detection of the activities of all the enzymes involved and also highlight the role of these enzymes in proline accumulation during water stress.     

Effect of water stress on proline catabolism in tobacco leaves

Proline-[¹⁴C] infiltrated into leaf disks of tobacco (Nicotiana tabacum cv BY-4) in the dark was converted to glutamic acid and then metabolized through the TCA cycle. A smaller amount of proline-[¹⁴C] was metabolized when the leaf disks were wilted than when turgid. During a 6 hr period following rehydration, disks converted a larger amount of proline-[¹⁴C] to oxidized products than when wilted, although the proline content of rehydrated disks had not declined. These results indicate that proline oxidation is inhibited by water stress.