Proline accumulation induced by excess nickel in detached rice leaves

The regulation of proline accumulation in detached rice leaves exposed to excess NiSO₄ was investigated. NiSO₄ treatment increased proline and Ni contents but had no effect on relative water content, indicating that proline accumulation in Ni-exposed detached rice leaves was due to Ni uptake per se, rather than to water stress. Proline accumulation caused by NiSO₄ was related to protein hydrolysis, a decrease in proline dehydrogenase activity, and a decrease in proline utilization. It seems that an increase in the content of ammonia and an increase in the activities of Δ¹-pyrroline-5-carboxylate reductase and ornithine-δ-aminotransferase play minor if any role in Ni-induced proline accumulation in detached rice leaves.     

Differential responses of the enzymes involved in proline biosynthesis and degradation in drought tolerant and sensitive cotton genotypes during drought stress and recovery

The relative water content (RWC), free proline levels and the activities of enzymes involved in proline metabolism were studied in drought tolerant (Ca/H 680) and drought sensitive (Ca/H 148) genotypes of cotton (Gossypium hirsutum L.) during induction of water stress and posterior recovery. Water stress caused a significant increase in proline levels and P5CS activity in leaves of both tolerant and sensitive genotypes, whereas the activity of P5CR increased minimally and the activity of OAT remains unchanged. The activity of PDH decreased under drought stress in both the genotypes. The leaf of tolerant genotype maintained higher RWC, photosynthetic activity and proline levels, as well as higher P5CS and P5CR activities under water stress than that of drought sensitive genotype. The drought induced proline levels and activities of P5CS and P5CR declined and tend to be equal to their respective controls, during recovery, whereas the PDH activity tends to increase. These results indicate that induction of proline levels by up regulation of P5CS and down regulation of PDH may be involved in the development of drought tolerance in cotton.     

Role of nitric oxide under saline stress: implications on proline metabolism

The present work is focused on the possible relationship between nitric oxide and the induction of proline in response to salt stress. The plants were subjected to 100 mM NaCl and sodium nitroprusside (SNP; the donor of NO) at different concentrations. The plants showed lower NaCl-induced oxidative stress and proline accumulation after application of low concentrations of SNP together with the NaCl treatment. The reduction in the proline content was related to increased activity of proline dehydrogenase. These results suggest that the NO could be capable of mitigating damage associated with salt stress.     

The effect of NaCl on proline accumulation in potato seedlings and calli

The effects of salt stress were studied on the accumulation and metabolism of proline and its correlation with Na⁺ and K⁺ content in shoots and callus tissue of four potato cultivars, viz., Agria, Kennebec (relatively salt tolerant), Diamant and Ajax (relatively salt sensitive). Na⁺ and proline contents increased in all cultivars under salt stress. However, K⁺ and protein contents decreased in response to NaCl treatments. The activities of enzymes involved in proline metabolism, Δ¹-pyrroline-5-carboxylate synthetase (P5CS) and proline dehydrogenase (ProDH) increased and decreased, respectively, in response to elevated NaCl concentrations. The changes of P5CS and ProDH activities in more salt sensitive cultivars (Diamant, Ajax) were more than those in the tolerant ones. Then the stimulation of synthesis in combination with a partially increase of protein proteolysis, a decrease in proline utilization and inhibition of oxidation resulted in high proline contents in seedlings and calli under salt stress. In callus tissue, reduced growth and cell size may be partially responsible for high proline accumulation in response to high NaCl levels. However, although the basic proline contents in the seedlings of more salt tolerant cultivars were higher than the sensitive ones, a clear relationship was not generally observed between accumulation of proline and salt tolerance in potato.     

外源ABA对楸树幼苗NaCl胁迫的缓解效应及其生长生理响应特征

以2年生楸树(苏楸1号和008-1)扦插苗为材料,采用盆栽试验法,分析盐胁迫(0.5%NaCl)处理下楸树幼苗生长、生理的变化,并分析不同浓度外源ABA(15、25、35 mg/L)对盐胁迫(30 d)楸树幼苗的缓解效应及其生理生化特性,以探索重度盐胁迫下适合楸树幼苗生长的适宜外源ABA浓度,为增强盐碱地楸树的耐盐性、提高盐碱地的利用提供理论依据。结果显示:(1)0.5%NaCl胁迫下,两品种楸树幼苗叶片表现出不同程度的盐害症状,且‘苏楸1号’叶片盐害症状较‘008-1’严重;随胁迫时间延长,两品种楸树幼苗的相对电导率(REC)均呈先上升后下降的变化趋势,叶绿素(Chl)、相对含水量(RWC)均呈降低趋势,可溶性糖(SS)、可溶性蛋白(SP)、脯氨酸(Pro)以及超氧化物歧化酶(SOD)活性均呈先上升后下降趋势,但‘008-1’的REC显著低于‘苏楸1号’,Chl、RWC、SS、SP、Pro、SOD均显著高于苏楸1号,表明‘008-1’的耐盐性较‘苏楸1号’更强。(2)喷施外源ABA使得盐胁迫下‘008-1’楸树的苗高显著增加、新叶提前萌发,表明外源ABA在一定程度上能够缓解盐胁迫对楸树生长的影响;喷施外源ABA降低了盐胁迫下‘008-1’楸树幼苗叶片的REC,提高了Chl、RWC、SS、SP、Pro、SOD、过氧化物酶(POD)以及过氧化氢酶(CAT)活性,促进了内源激素生长素(IAA)、脱落酸(ABA)、赤霉素(GA3)以及玉米素核苷(ZR)的积累。研究表明,楸树品种‘008-1’的耐盐性更强;外源喷施适宜浓度ABA能够缓解盐胁迫对楸树幼苗生长的影响,降低幼苗叶片细胞膜透性,促进幼苗渗透调节物质的积累,增强渗透调节能力,并提高盐胁迫下幼苗的抗氧化酶活性,促进植物对内源激素含量的调节,从而提高楸树的耐盐性,且以25 mg/L ABA处理的效果最好。     

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.     

Genetic engineering of chickpea (Cicer arietinum L.) with the P5CSF129A gene for osmoregulation with implications on drought tolerance

Abiotic stresses including water deficit severely limits crop yields in the semi-arid tropics. In chickpea, annual losses of over 3.7 million tones have been estimated to be due to water deficit conditions alone. Therefore, major efforts are needed to improve its tolerance to water deficit, and genetic engineering approaches provide an increasing hope for this possibility. We have used transgenic technology for the introduction of an osmoregulatory gene P5CSF129A encoding the mutagenized Δ¹-pyrroline-5-carboxylate synthetase (P5CS) for the overproduction of proline. A total of 49 transgenic events of chickpea were produced with the 35S:P5CSF129A gene through Agrobacterium tumefaciens-mediated gene transfer through the use of axillary meristem explants. Eleven transgenic events that accumulated high proline (2–6 folds) were further evaluated in greenhouse experiments based on their transpiration efficiency (TE), photosynthetic activity, stomatal conductance, and root length under water stress. Almost all the transgenic events showed a decline in transpiration at lower values of the fraction of transpirable soil water (dryer soil), and extracted more water than their untransformed parents. The accumulation of proline in the selected events was more pronounced that increased significantly in the leaves when exposed to water stress. However, the overexpression of P5CSF129A gene resulted only in a modest increase in TE, thereby indicating that the enhanced proline had little bearing on the components of yield architecture that are significant in overcoming the negative effects of drought stress in chickpea.     

The Effect of Polyethylene Glycol on Proline Accumulation in Rice Leaves

The regulation of proline accumulation in polyethylene glycol (PEG, –1.5 MPa) treated rice leaves was investigated. PEG treatment resulted in a decrease in relative water content, indicating that PEG treatment caused water stress in rice leaves. Proline accumulation caused by PEG was related to protein hydrolysis, an increase in ornithine--amino- transferase activity, an increase in the content of ammonia, and an increase in the contents of the precursors of proline biosynthesis, glutamic acid, ornithine, and arginine. Results also show that abscisic acid accumulation is not required for proline accumulation in PEG-treated rice leaves.     

Cadmium and nickel accumulation in rice plants. Effects on mineral nutrition and possible interactions of abscisic and gibberellic acids

Rice plants accumulate high quantities of Cd and Ni when grown for 10 days in a medium containing these heavy metals. Accompanying Cd and Ni uptake, a decrease in shoot and root length was observed, though dry matter accumulation was not affected accordingly. Metal treatments also induced a decrease in K, Ca and Mg contents in the plants, particularly in the shoots, indicating that Cd and Ni interfered not only with nutrient uptake but also with nutrient distribution into the different plant parts. Addition of abscisic acid (ABA) or gibberellic acid (GA₃) to the external solution could not overcome the depressing effects of the metals on nutrient acquisition, and even induced a further decrease of Ca content in Ni-treated plants. Both hormones also reduced, significantly, heavy metal incorporation into the plants. Additionally, hormonal applications affected the transport of Cd and Ni to the shoots, resulting in a higher percentage of the metals taken up remaining in the roots.     

Effects of Proline and Betaine on Heat Inactivation of Ribulose-1,5-Bisphosphate Carboxylase/Oxygenase in Crude Extracts of Rice Seedlings

Activity of ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBPCO) in heated crude extracts from seedlings of the rice cultivars Hitomebore and IR28 was investigated in the presence of proline and betaine. Both solutes retarded the inactivation of the enzyme extracted from the leaves of both cultivars at temperature-stress from 35 to 45 °C. At 50 °C, however, betaine was effective in both cultivars. Stabilization of RuBPCO activity was independent of the added solute from 1 to 2 M concentration. This revised version was published online in June 2006 with corrections to the Cover Date.     

Effect of Increasing Concentrations of Lead and Cadmium on Cucumber Seedlings

The effect of lead and cadmium on biomass accumulation of cucumber seedlings (Cucumis sativus L.) as well as the contents of abscisic acid (ABA), free proline and soluble proteins in leaves were studied. Seedlings were subjected to lead nitrate or cadmium bromide in low concentrations (1 – 5 µM) for 1, 4 or 7 d, and then to the action of the same substances in high concentrations (500 – 1000 µM). The pretreatments of the seedlings with heavy metals in low concentrations enabled them to tolerate the subsequent high concentrations of cadmium and lead without injury. The plant responses to heavy metal treatment were accompanied by the accumulation of ABA, free proline and soluble proteins in leaf tissues.     

内源抗坏血酸对水稻种子萌发及幼苗生长的影响

为了解内源抗环血酸在水稻(Oryza sativa)种子中的作用,以野生型品种‘中花11’(ZH-11)、抗坏血酸合成关键酶GLDH基因的上调(超表达)株系GO-2及下调(干涉)株系GI-2为材料,研究了抗环血酸含量对其种子萌发和幼苗生长的影响。结果表明,GO-2株系的种子萌发率比野生型ZH-11高6%,比干涉型GI-2高60%。外源添加1 mmol L–1抗坏血酸后,干涉型GI-2萌发率提高了22%,GO-2及ZH-11则没有明显增加。GO-2株系在幼苗的根长、株高、分蘖数和鲜重等指标上均高于ZH-11和GI-2。实时荧光定量测定结果表明,GO-2株系叶片的GLDH基因表达量显著上调,而GI-2株系则显著下调。这说明抗坏血酸有助于维持水稻种子活力和促进水稻种子发芽和幼苗生长。     

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     

镉胁迫对西瓜幼苗生长及其叶片解剖结构和生理特性的影响

该研究以‘北抗1号’西瓜幼苗为试验材料,采用实验室盆栽方法,考察在不同浓度镉胁迫(0、60、120、180和240 mg/L)处理下西瓜幼苗的生长及其叶片生理特性和解剖结构的变化特征,初步探讨西瓜耐受镉胁迫的生理机制。结果显示：(1)在镉胁迫条件下,西瓜幼苗的生长受到抑制,随着镉胁迫浓度的增加,西瓜幼苗的叶片形态黄化现象逐渐加重,根系形态逐渐纤弱,株高、茎粗、茎节数和叶片数均呈现下降趋势。(2)镉胁迫下,西瓜幼苗叶片主脉中细胞受损,主脉直径显著减小,叶肉组织疏密度显著降低。(3)随着镉胁迫浓度的升高,西瓜幼苗的含水量、净光合速率、SOD活性等显著降低,丙二醛和游离脯氨酸含量显著升高,POD活性和可溶性蛋白含量先升高后降低,生理特性受到显著影响。研究发现,‘北抗1号’西瓜幼苗对镉有一定的适应性,在低浓度(60 mg/L)镉胁迫处理下,西瓜幼苗的形态特征、生理特性变化不显著,但在高浓度(180 mg/L)镉胁迫下,幼苗的生长受到严重抑制,渗透调节系统以及生物膜保护系统严重受损。     

Hydrogen sulfide donor sodium hydrosulfide-improved heat tolerance in maize and involvement of proline

Hydrogen sulfide (H₂S) has long been considered as a phytotoxin, but nowadays as a cell signal molecule involved in growth, development, and the acquisition of stress tolerance in higher plants. In the present study, hydrogen sulfide donor, sodium hydrosulfide (NaHS), pretreatment markedly improved germination percentage of seeds and survival percentage of seedlings of maize under heat stress, and alleviated an increase in electrolyte leakage of roots, a decrease in tissue vitality and an accumulation of malondialdehyde (MDA) in coleoptiles of maize seedlings. In addition, pretreatment of NaHS could improve the activity of Δ¹-pyrroline-5-carboxylate synthetase (P5CS) and lower proline dehydrogenase (ProDH) activity, which in turn induced accumulation of endogenous proline in maize seedlings. Also, application of proline could enhance endogenous proline content, followed by mitigated accumulation of MDA and increased survival percentage of maize seedlings under heat stress. These results suggest that sodium hydrosulfide pretreatment could improve heat tolerance of maize and the acquisition of this heat tolerance may be involved in proline.     

硝态和铵态氮配比对水培油麦菜苗期生长及生理特性的影响

采用水培技术,以油麦菜幼苗为材料,研究不同硝铵态氮配比（NO₃^-∶NH₄⁺）对油麦菜苗期地上部和根系生长及生理特性的影响。结果表明：（1）油麦菜地上部和根系硝酸盐含量皆与营养液中NO₃^--N比例呈正相关关系,且各处理均达到无公害蔬菜的标准。（2）随着营养液中NH₄⁺-N比例的增加,油麦菜地上部有机酸含量先降低后升高,且在硝铵态氮配比为5∶5时最低,可溶性糖含量呈上升趋势,而可溶性蛋白质含量先升高后降低,在硝铵态氮配比为5∶5时最高;油麦菜根系有机酸和可溶性糖含量先升高后降低,两者分别在硝铵态氮配比为5∶5和7.5∶2.5时最高,而可溶性蛋白质含量呈下降趋势,在全NO₃^--N时最高。（3）随着营养液中NH₄⁺-N比例的增加,油麦菜地上部和根系中SOD活性先升后降,并分别在硝铵态氮配比为5∶5和7.5∶2.5时最高,而地上部和根系中MDA、脯氨酸含量和POD、CAT活性的变化趋势则与其相反。（4）随着营养液中NH₄⁺-N比例的增加,油麦菜地上部和根系干重皆先升后降,根冠比则逐渐减小;在硝铵态氮配比为7.5∶2.5时干重最大,根冠比适宜且稳定。研究表明,水培油麦菜苗期地上部和根系生长及生理特性受到氮素形态配比的显著影响,且根系的生理响应更敏感;营养液中硝铵态氮配比为7.5∶2.5时,油麦菜受胁迫程度最低,地上部和根系生长较协调,油麦菜生长和生理状况最佳。     

硝酸盐水平对芹菜幼苗生理及生长特性的影响

以西芹‘文图拉’幼苗为试材,通过水培方式研究了不同浓度NO3-(0、1、5、10、15、50、100、150mmol·L-1)对芹菜植株生物量、养分含量、叶绿素、丙二醛(MDA)及两种渗透调节剂含量的影响。结果显示:(1)随着NO3-浓度的增加(1～50mmol·L-1),芹菜株高、生物产量、根冠比以及叶面积显著增加,植株对N、P、K养分的吸收和叶绿素明显提高,同时植株的MDA含量上升,POD和CAT的活性增强,可溶性糖和可溶性蛋白含量显著增加。(2)当NO3-浓度等于或大于100mmol·L-1时,芹菜的生物产量和叶绿素含量下降,植株对K、P养分的吸收降低,膜脂过氧化物MDA、可溶性糖和可溶性蛋白含量及SOD活性达到最高峰值,POD和CAT活性有所下降。研究表明,NO3-浓度为15mmol·L-1时最有利于芹菜植株的生长;NO3-浓度为100mmol·L-1时对植株生长产生了硝酸盐胁迫,导致膜脂过氧化伤害,但芹菜植株能通过调节抗氧化物酶活性及渗透调节剂的合成代谢抵御环境胁迫,从而表现出一定的硝酸盐耐受性。     

Agrobacterium-mediated Transformation of Brassica juncea with a Cyanobacterial (Synechocystis PCC6803) Delta-6 Desaturase Gene Leads to Production of Gamma-linolenic Acid

Genetic manipulation of the oil-yielding crop plants for better oil quality through biotechnological methods is an important aspect of crop improvement. Due to the inherent absence of the Δ⁶-desaturase (d6D) function, Brassica juncea, an oil-yielding crop plant, is unable to synthesize γ-linolenic acid (GLA), a nutritionally important fatty acid although the crop plant synthesizes the precursor fatty acids required for GLA production. Cyanobacterial d6D introduces carbon–carbon double bond onto linoleic acid (C18:2) and α-linolenic acid (C18:3) by desaturation processes for production of GLA and octadecatetraenoic acid (OTA) respectively. In the present investigation, d6D coding sequence from Synechocystis sp. PCC6803 was cloned by polymerase chain reaction and introduced into B. juncea through Agrobacterium-mediated transformation technique. Both cytosolic as well as seed-specific expression of d6D were attempted. The transformed plants show production of GLA and OTA in contrast to their absence in the untransformed control plants adducing evidence for introgression and functional expression of the cyanobacterial d6D gene in B. juncea.