Ultrastructural Study in Leaf Cell of Brassica oleracea var. capitata Under Heat Stress

Ultrastructural changes of the leaf cells in response to heat stress in thermo-resistant cultivar and thermo-sensitive cultivar of Brassica oleracea var. capitata L. were investigated. No ultrastructural change was shown in mesophyll cell of the thermo-resistant cultivar. All membrane-bound structure remained intact. However, the cell ultrastructure of thermosensitive cultivar showed significant changes. One of the major effects of heat stress was the disruption of membrane structure. Chloroplasts and nucleus were extremely sensitive to heat stress. Chloroplasts rounded off in different extents and their outer membranes disappeared. Structural alteration of the thylakoid membrane were visualized. Large amount of plastoglobulis appeared within the chloroplast. Mitochondria were far more resistant to heat stress than chloroplasts. There was no distinct changes of mitochondria structure. Nucleus suffered from serious damage as heat caused disruption of nuclear envelope and condensation of nucleoplasm, showing, eventually, numerous fibrillar granulous material and irregularly shaped hollow spaces presented within the nucleus.     

甘蓝热胁迫叶片细胞的超微结构研究

高温胁迫引起植物体损伤是很常见的,人们已从多方面探讨高温胁迫对植物的影响,主要工作集中在生理生化、膜结构、热激蛋白及作物产量等方面［１—４］,超微结构的变化也有少量报道［５—７］。但目前要建立一个高温对植物体影响的模式还比较困难。本文探讨了甘蓝不同耐热品种叶片细胞超微结构在热胁迫后的差异,目的是建立植物耐热程度的细胞学指标,为育种工作者选育耐热性品种提供细胞学依据。材料和方法试验材料为甘蓝（Ｂｒａｓｓｉｃａｏｌｅｒａｃｅａｖａｒ．ｃａｐｉｔａｔａＬ．）耐热品种“夏光甘蓝”和感热品种“京丰甘蓝”。…     

The chloroplast-targeting domain of plastocyanin transit peptide can form a helical structure but does not have a high affinity for lipid bilayers.

Conformational properties and interactions with lipid membranes were studied for the chemically synthesized peptides PC(1-37) and PC(1-43), corresponding to the N-terminal 37 and 43 residues, respectively, of the transit peptide of the precursor to plastocyanin of Silene pratensis. PC(1-43) covers the entire chloroplast targeting domain of the transit peptide. CD spectra of PC(1-37) and PC(1-43) showed that both peptides have little ordered structure in aqueous solutions but form partially helical conformations in the presence of detergent micelles or in methanol. Vesicle disruption and direct-binding experiments revealed, however, that neither PC(1-37) nor PC(1-43) had a high affinity for lipid membranes. Since in the intact plastocyanin transit peptide the chloroplast-targeting domain is followed by a hydrophobic thylakoid-transfer domain, the plastocyanin precursor may well be transported to the chloroplast surface first with the aid of the thylakoid-transfer domain. The chloroplast-targeting domain may then form a helical structure in the lipid environments, and a chloroplast-specific motif displayed on the helical structure may be recognized by a receptor protein located at the chloroplast envelope membranes.     

Chlorophyll-Protein Complexes in Salix sp. 'Aquatica Gigantea' under Strong and Weak Light II. Effect of Water Stress on the Chlorophyll-Protein Complexes and Chloroplast Ultrastructure

The effect of slowly-induced water stress on the distributionof chlorophyll among the chlorophyll-protein complexes and onthe chloroplast ultrastructure was studied in Salix sp. ‘aquaticagigantea’ grown under two different light regimes. Underhigh irradiance the proportion of chlorophyll was largest inthe P700-chlorophyll-a protein complex (CPI). With increasingwater stress the proportion of the light-harvesting chlorophylla/b-protein complex (LHCP) and the chlorophyll a-protein complex(CPa) decreased, and these changes corresponded with inversechanges in the amount of chlorophyll in the protein free pigment(FP) band. The CPI complex remained stable throughout the dryingperiod. Under low irradiance the LHCP complex was the largeststructural component and its proportion of the total chlorophyllremained constant with increasing water stress. Under theselight conditions CPI and the CPa complex appeared to be thelabile components of the chloroplast lamellae. The integrityof the chloroplast membranes, as judged by electron microscopicobservations, was preserved well under both light regimes andwith increasing water stress. This and the relative constancyof the chlorophyll content during the experiments suggestedthat no great changes had occurred in the lamellar structureduring water stress. In the chloroplast stroma area, however,large crystal formations were observed under the strong lightregime with increasing water stress. In weak light these crystalstructures could be seen in the well-watered controls, but seemedto disappear totally as water stress became severe. (Received February 1, 1982; Accepted May 10, 1982)     

Importance of phosphatidylcholine on the chloroplast surface

In plant cells, phosphatidylcholine (PC) is a major glycerolipid of most membranes but practically lacking from the plastid internal membranes. In chloroplasts, PC is absent from the thylakoids and the inner envelope membrane. It is however the main component of the outer envelope membrane, where it exclusively distributes in the outer monolayer. This unique distribution is likely related with operational compartmentalization of plant lipid metabolism. In this review, we summarize the different mechanisms involved in homeostasis of PC in plant cells. The specific origin of chloroplast PC is examined and the involvement of the P4-ATPase family of phospholipid flippases (ALA) is considered with a special attention to the recently reported effect of the endoplasmic reticulum-localized ALA10 on modification of chloroplast PC desaturation. The different possible roles of chloroplast PC are then discussed and analyzed in consideration of plant physiology.     

外源GSH对盐胁迫下水稻叶绿体活性氧清除系统的影响

研究了外源GSH对盐胁迫下耐盐性不同的水稻品种Pokkali(耐盐)和Peta(盐敏感)叶绿体中抗氧化酶活性和抗氧化剂含量的影响.结果表明:盐胁迫下,外源GSH可以提高水稻叶绿体中活性氧清除系统中SOD、APX、GR的活性以及AsA、GSH的含量,降低叶绿体中H2O2和MDA的含量,从而降低了叶绿体膜脂过氧化的水平,缓解盐胁迫对叶绿体膜的伤害.外源GSH对盐胁迫下盐敏感品种Peta叶绿体中上述指标增加或减少的幅度大于耐盐品种Pokkali.     

Suppression of Oxygen Evolving System in Spinach Chloroplast Membranes due to Release of Manganese on Exposure to Osmotic Stress in vitro in Presence of Magnesium

When spinach chloroplast membranes were exposed to osmotic stress in vitro, by incubation in 1.0 M sorbitol + 10 mM MgCl₂ their oxygen evolving system was suppressed. The possible reasons for such inactivation of PS II mediated oxygen evolution were examined. There were conformational changes in the chloroplast membranes, as indicated by their absorption spectra. The pattern of sensitivity to DCMU was not altered. The sensitivity of PS II to water stress remained, even after a pre-wash treatment with NaCI (which removed 18 and 24 kD proteins) but not when the thylakoids were pretreated with NH₂0H or CaCl₂ (removed manganese and 33 kD). The manganese content of thylakoid membranes was markedly reduced under osmotic stress in presence of magnesium. We suggest that exposure of chloroplasts to 1.0 M sorbitol in presence of Mg₂₊ released manganese from thylakoid membranes, thereby leading to a suppression in oxygen evolution.     

Differences in physiological characteristics between two wheat cultivars exposed to field water deficit conditions

We investigated various physiological characteristics of two wheat (Triticum aestivum L.) cultivars differing in drought tolerance, i.e., Shannong16 (a drought-tolerant cultivar) and Weimai8 (a high-yield wheat cultivar under well-watered conditions), under field drought conditions. The experiments were conducted over a two-year period. Drought stress (DS) was imposed by controlling irrigation and sheltering the plants from rain. Compared with Weimai8, Shannong16 exhibited the better water balance, the higher osmotic adjustment, the slower degradation of chlorophyll, and the higher net photosynthetic rate under drought-stress conditions. At the same time, we observed that Shannong16 maintained more integrated chloroplast and thylakoid ultrastructure in flag leaves than Weimai8 under field drought stress. The different levels of antioxidant competence, indicated by MDA content, antioxidant enzyme activities, and the level of superoxide radicals observed in the two wheat cultivars may be involved in the different levels of drought resistance of these cultivars.     

等渗NaCl、干旱胁迫对番茄幼苗光合特性及叶绿体超微结构的影响

为分析等渗盐分和水分胁迫对番茄叶片光合功能的影响,选用耐盐性不同的4种基因型番茄(小果型的辽园红玛瑙、野生醋栗番茄、大果型的金田粉冠、超402)进行等渗的140 mmol·L^-1NaCl和15%PEG6000模拟盐分和水分胁迫.结果表明: 在光合特性方面,处理12 d后,两种胁迫导致4种不同基因型番茄的叶绿素含量(叶绿素a、叶绿素b、叶绿素总量)、光合速率、气孔导度、胞间CO₂浓度、蒸腾效率降低,气孔限制值升高.两种胁迫导致的光合下降由气孔性因素所致.等渗的盐分胁迫对番茄的光合系统损伤大于水分胁迫,这是因为盐分胁迫除了渗透胁迫还会导致离子伤害.4种不同基因型番茄中,耐盐型的辽园红玛瑙具有高光合特性,金田粉冠光合效率最差.在叶绿体超微结构方面,两种胁迫会造成番茄叶片的气孔密度增加,气孔张开率降低,叶绿体长度增加,宽度变小,长宽比增大,叶绿体内基粒数减少,嗜锇颗粒数增多.盐分胁迫下两种番茄叶绿体结构的影响大于水分胁迫,耐盐种醋栗番茄气孔变化小于盐敏感品种金田粉冠.     

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.     

Changes in lipid composition, water relations and gas exchange in leaves of two young ‘Chemlali’ and ‘Chetoui’ olive trees in response to water stress

The comparative responses of two young olive trees (Olea europaea L. ‘Chemlali’ and ‘Chetoui’) to drought stress were investigated during 1 month. Three-month-old own-rooted plants were subjected to two irrigation treatments: WW (well watered plants that were irrigated with fresh water to maintain a soil water content close to field capacity), and WS (water stressed plants by withholding water). Leaf water potential, gas exchange and leaf lipid composition were studied. ‘Chemlali’ was able to maintain higher leaf CO₂ assimilation rate and leaf stomatal conductance throughout the drought cycle compared to ‘Chetoui’. Water stress induced a larger decrease in the total lipid content in ‘Chetoui’ than in ‘Chemlali’. Interestingly, the constitution of different lipid classes was highly altered in ‘Chetoui’. Lipid changes in Chemlali, a drought tolerant cultivar, revealed more stability of its cellular membranes to drought stress as compared to the drought susceptible olive cultivar, Chétoui. Furthermore, in comparison to the controls, drought stressed plants showed an increase in the degree of unsaturation of leaf lipids in the two olive cultivars. Moreover, the results observed in Chemlali showed that besides changes in lipids composition this cultivar may have an efficient defence strategy which can be related on antioxidative production against oxidative stress.     

Chloroplast downsizing under nitrate nutrition restrained mesophyll conductance and photosynthesis in rice (Oryza sativa L.) under drought conditions

The phenomenon whereby ammonium enhances the tolerance of rice seedlings (Oryza sativa L., cv. 'Shanyou 63' hybrid indica China) to water stress has been reported in previous studies. To study the intrinsic mechanism of biomass synthesis related to photosynthesis, hydroponic experiments supplying different nitrogen (N) forms were conducted; water stress was simulated by the addition of polyethylene glycol. Water stress decreased leaf water potential (Ψ(leaf)) under nitrate nutrition, while it had no negative effect under ammonium nutrition. The decreased Ψ(leaf) under nitrate nutrition resulted in chloroplast downsizing and subsequently decreased mesophyll conductance to CO(2) (g(m)). The decreased g(m) and stomatal conductance (g(s)) under nitrate nutrition with water stress restrained the CO(2) supply to the chloroplast and Rubisco. The relatively higher distribution of leaf N to Rubisco under ammonium nutrition might also be of benefit for photosynthesis under water stress. In conclusion, chloroplast downsizing induced a decline in g(m), a relatively higher decrease in g(s) under nitrate nutrition with water stress, restrained the CO(2) supply to Rubisco and finally decreased the photosynthetic rate.     

In vivo regulation of thylakoid proton motive force in immature leaves

In chloroplast, proton motive force (pmf) is critical for ATP synthesis and photoprotection. To prevent photoinhibition of photosynthetic apparatus, proton gradient (ΔpH) across the thylakoid membranes needs to be built up to minimize the production of reactive oxygen species (ROS) in thylakoid membranes. However, the regulation of thylakoid pmf in immature leaves is little known. In this study, we compared photosynthetic electron sinks, P700 redox state, non-photochemical quenching (NPQ), and electrochromic shift (ECS) signal in immature and mature leaves of a cultivar of Camellia. The immature leaves displayed lower linear electron flow and cyclic electron flow, but higher levels of NPQ and P700 oxidation ratio under high light. Meanwhile, we found that pmf and ΔpH were higher in the immature leaves. Furthermore, the immature leaves showed significantly lower thylakoid proton conductivity than mature leaves. These results strongly indicated that immature leaves can build up enough ΔpH by modulating proton efflux from the lumenal side to the stromal side of thylakoid membranes, which is essential to prevent photoinhibition via thermal energy dissipation and photosynthetic control of electron transfer. This study highlights that the activity of chloroplast ATP synthase is a key safety valve for photoprotection in immature leaves.     

Effect of drought stress on proteolytic activities in Phaseolus and Vigna leaves from sensitive and resistant plants

Although much information is available concerning the effect of senescence on cell proteolytic activities, few reports are devoted to the impact of drought stress. Our aim was to study the influence of water deficit on the cell proteolytic potential, and to determine whether or not it could be used as a physiological parameter for screening varieties for tolerance to drought. We have used Phaseolus and Vigna species differing in their senstivity to water deficit: V. unguiculata L. Walp. cv. EPACE (resistant), V. unguiculata L. Walp. cv. IT83D (moderately senstive) and P. vulgaris L. cv. Carioquinha (sensitive). The plants were subjected to controlled water conditions. Proteolytic activities were assayed using azocasein in the case of leaf extracts and [¹⁴C]-methylated casein in the case of cell compartments: soluble fraction, membrane fraction and isolated, purified chloroplasts. The results indicate that the leaf extracts contained 3 groups of proteinases with optimum pH at 4.5, 5-6 and 8.5 for the Vigna cultivars and 5.0, 6.5 and 9.0 for the Phaseolus cultivar. The sensitive P. vulgaris Carioquinha showed higher caseinolytic activities than the other tow cultivars in response to water deficit. As regards cell fractions, proteolytic activities were determined for pH values of 4.5, 6.0 and 9.0. In soluble fractions of stressed plants, proteolytic activities increased at all the pH values tested; this clearly correlated with the drought sensitivity level of the plants, especially at pH 4.5. The same phenomenon was observed in the case of membranes and purified chloroplasts of the sensitive cultivar. Under drought stress, the proteolytic potential of the cell increased especially in the vaculoar sap (soluble fraction). The higher activities observed for all the cell compartments in the sensitive cultivar could be responsible, at least partly, for the rapid degradation of leaf and chloroplast proteins under drought. The use of [¹⁴C]-methylated casein and soluble cell fractions seem to allow a clear differentiation between cultivars with respect to the drought tolerance at the cellular level.     

根施甜菜碱对水分胁迫下烟草幼苗光合机构的保护

以烟草品种大黄金5210（抗旱性强）和中烟100（抗旱性弱）为材料,研究了水分胁迫对烟草叶片光合机构的影响,并通过根部施用甜菜碱的方法,探讨了甜菜碱对烟草光合机构的保护作用。结果表明：水分胁迫导致烟草幼苗光合机构损伤,表现在叶绿素含量、PSII光化学效率、希尔反应活力以及类囊体膜ATPase活性下降,且对抗旱性弱的中烟100损伤更加严重。外源甜菜碱处理减轻了水分胁迫对以上指标的降低程度,特别是对干旱敏感型烟草品种中烟100的效果更加明显。甜菜碱的这种保护作用可能与它能够维持叶片中各种抗氧化酶活性、减轻活性氧的积累、保护类囊体膜上各种色素蛋白的功能以及缓解水分胁迫对膜的破坏作用有关。     

Effects of water stress on photochemical function and protein metabolism of photosystem II in wheat leaves

The effects of osmotic dehydration in wheat leaves ( Triticum aestivum L. cv. Longchun No. 10) on the photochemical function and protein metabolism of PSII were studied with isolated thylakoid and PSII membranes. The results indicated that PSII was rather resistant to water stress as mild water deficit in leaves did nut significantly affect its activity. However, extreme stress conditions such as 40% decrease in relative water content (RWC) or 1.8 MPa in water potential (Ψ) caused ca 50% reduction in O₂ evolution and ca 25% inhibition of DCIP (2.6-dichlorophenol indophenol) photoreduction of PSII. In addition, it was found that the inhibited DCIP photoreduction of PSII could not be reversed by DPC (2.2-diphenylcarbazide), a typical electron donor to PSII, suggesting that water stress did not affect electron donation to PSII. Urea-SDS-PAGE and western blot analysis showed that the steady slate levels of major PSII proteins, including the D1 and D2 proteins in the PSII reaction center, declined on a chlorophyll basis with increasing water stress, possibly as a result of increased degradation. In vitro translation experiments and quantitative analysis of chloroplast RNAs indicated that the potential synthesis of chloroplast proteins from their mRNAs was impaired by water stress. From the results it is concluded that the effects of water stress on PSII protein metabolism, especially on the reaction center proteins, may account for the damage to PSII photochemistry.     

Involvement of cytosolic ascorbate peroxidase and Cu/Zn-superoxide dismutase for improved tolerance against drought stress

In order to understand the role of cytosolic antioxidant enzymes in drought stress protection, transgenic tobacco (Nicotiana tabacum cv. Xanthi) plants overexpressing cytosolic Cu/Zn-superoxide dismutase (cytsod) (EC 1.15.1.1) or ascorbate peroxidase (cytapx) (EC 1.11.1.1) alone, or in combination, were produced and tested for tolerance against mild water stress. The results showed that the simultaneous overexpression of Cu/Znsod and apx or at least apx in the cytosol of transgenic tobacco plants alleviates, to some extent, the damage produced by water stress conditions. This was correlated with higher water use efficiency and better photosynthetic rates. In general, oxidative stress parameters, such as lipid peroxidation, electrolyte leakage, and H(2)O(2) levels, were higher in non-transformed plants than in transgenic lines, suggesting that, at the least, overexpression of cytapx protects tobacco membranes from water stress. In these conditions, the activity of other antioxidant enzymes was induced in transgenic lines at the subcellular level. Moreover, an increase in the activity of some antioxidant enzymes was also observed in the chloroplast of transgenic plants overexpressing cytsod and/or cytapx. These results suggest the positive influence of cytosolic antioxidant metabolism on the chloroplast and underline the complexity of the regulation network of plant antioxidant defences during drought stress.     

The effects of water stress on the development of the photosynthetic apparatus in greening leaves

The effects of low and high relative humidity and of polyethylene glycol-induced root water stress on chlorophyll accumulation, on formation of the lamellar chlorophyll-protein complexes, and on the development of photosynthetic activity during chloroplast differentiation were examined. Low relative humidity or polyethylene glycol-induced root water stress (stress conditions) resulted in a 3 to 4 hour lag in chlorophyll accumulation, retarded the rate of chlorophyll b accumulation, and reduced the rate of formation of the light-harvesting chlorophyll a/b protein. All of these effects could be overcome by high relative humidity (nonstress) conditions. Concomitant measurement of leaf water potential showed that under stress conditions greening leaves were subjected to initial water deficits of −8 bars which decreased to −5 bars after 3 to 4 hours of illumination corresponding to the end of the lag phase. Leaves greening under nonstress conditions did not experience leaf water deficits greater than about −5 bars. It seems that the attainment of a minimum leaf water potential of −5 bars may be critical in the control of early chloroplast development. These results demonstrate that the lag phase is not indicative of a programmed event in chloroplast development, but rather is attributable to environmental conditions prevailing during leaf development and greening.     

水分胁迫条件下枇杷若干生理指标的变化

随着水分胁迫程度的加强,枇杷叶片脯氨酸含量增加,可溶性蛋白质含量下降。轻度及中度水分胁迫均使枇杷叶片各细胞器H⁺-ATPase活性增强;严重水分胁迫时,细胞胞质H⁺-ATPase活性急剧下降到对照以下水平,而线粒体及叶绿体的H⁺-ATPase活性仍保持高于对照的水平。同一程度的水分胁迫条件下,长红3号叶片可溶性蛋白质含量降幅小于解放钟,而线粒体和叶绿体H⁺-ATPase活性增幅则大于解放钟,说明长红3号抵御干旱的能力比解放钟强。