The effects of chilling in the dark and in the light on photosynthesis of tomato: electron transfer reactions

Exposure of tomato plants (Lycopersicon esculentum Mill. cv. Floramerica) to chilling temperatures in the dark for as little as 12 h resulted in a sizable inhibition in the rate of light- and CO₂-saturated photosynthesis. However, when photosynthesis was measured at low light intensity, the inhibition disappeared and the quantum yield of CO₂ reduction was diminished only slightly. Chilling the tomato plants under strong illumination caused an even more rapid and severe decline in the rate of light- and CO₂-saturated photosynthesis, accompanied by a large decline in the quantum efficiency. Sizeable inhibition of photosystem II activity was observed only after dark exposures to low temperature of grater than 16 h. No inhibition of photosystem I electron transfer capacity was observed even after 40 h of dark chilling. Chilling under high light resulted in a rapid decline in both photosystem I and photosystem II electron transfer capacity as well as in significant reaction center inactivation.Regardless of whether the chilling exposure was in the presence or absence of illumination and regardless of its duration, the electron transfer capacity of thylakoid membranes isolated from the treated plants was always in excess of that necessary to support light- and CO₂-saturated photosynthesis. Thus, in neither case of chilling inhibition of photosynthesis does it appear that impaired electron transfer capacity represents a significant rate limitation to whole plant photosynthesis.Abbreviations BSA bovine serum albumin - DBMIB 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone - DCMU 3-(3,4-Dichlorophenyl)-1,1-dimethylurea - DHQ duroquinol - EDTA ethylene-diamine-tetraacetic acid - HEPES N-2-hydroxylpiperazine-N-2-ethanesulfonic acid - MES 2-(N-Morpholino)ethanesulfonic acid - MV methylviologen - PS I & II photosystems I and II - PD_OX p-phenylenediimine (oxidized) - TMPD N,N,N,N-tetramethyl-p-phenylenediamine     

Long-term chilling of young tomato plants under low light and subsequent recovery

The influence of unfavourable climatic conditions at the onset of the growth period on chilling-sensitive tomato (Lycopersicon esculentum Mill., cv. Abunda) was studied by exposing young plants to combinations of low temperature and low light (60–100 mol quanta · m^–2 · s^–1) for several weeks. When the temperature did not decrease below a critical point (8 ° C) no loss of developmental capacity of the plants was detected. However, while new leaves were readily formed upon return to normal growth conditions (22/18 °C, day/night, in a greenhouse), net accumulation of biomass showed a lag phase of approximately one week. This delay was accompanied by a strong, irreversible inhibition of photosynthesis in the fully expanded leaves which had been exposed to the chilling treatment. When plants were subjected to temperatures below 8 ° C, survival rates decreased after three weeks at 6 ° C and irreversible damage of apical meristematic tissue occurred. Drought-hardening prior to chilling ensured survival at 6 ° C and protected the plants against meristem loss.Abreviation Chl chlorophyll Thanks are due to G.P. Telkamp for technical assistance. This research is financially supported by the Netherlands Technology Foundation (STW, Utrecht, The Netherlands), and is coordinated by the Foundation for Biological Research (BION, 's-Gravenhage, The Netherlands).     

The ultrastructure of chilling stress

Chilling injury to crop plants was first described 70 years ago and has been systematically investigated with electron microscopy since the late 1960s. Chloroplasts are the first and most severely impacted organelle. Thylakoids swell and distort, starch granules disappear, and a peripheral reticulum (vesicles arising from inner membrane of chloroplast envelope) appears. Chloroplast disintegration follows prolonged chilling. Mitochondria, nuclei and other organelles are less susceptible to chilling injury. Organellar development and ontogeny may also be disrupted. The inherent chilling sensitivity of a plant, as well as the ability of some species to acclimate to chilling, influence the timing and appearance of ultrastructural injury with the resulting outcome being mild, moderate, or severe. Other environmental factors that exacerbate injury are irradiance, chilling duration, and water status. The physiological basis for chloroplast swelling may be linked to chilling‐stable starch‐degrading enzymes that produce soluble sugars thus lowering stromal water potential at a time when chloroplast photosynthate export is reduced. Thylakoid dilation appears to be related to photo‐oxidative conditions produced during chilling in the light. The peripheral reticulum is proposed to increase surface area of the transport‐limiting membrane (chloroplast inner membrane) in response to the chilling‐induced reduction in metabolite transport. Many of the ultrastructural symptoms appearing during moderate stress resemble those seen in programmed cell death. Future research directions are discussed.     

The chilling sensitivity of root ammonium influx in a cultivated and wild tomato

A chilling episode of a few hours damaged root ammonium absorption in a cultivated tomato ( Lycopersicon esculentum cv. T-5), but not in a wild congener from high altitudes ( Lycopersicon hirsutum LA1778). In the cultivar, ammonium influx was strongly temperature dependent and showed the residual effects of chilling, whereas ammonium efflux was nearly temperature invariant and showed no persistent effects. A 2 h exposure to 5 °C significantly depressed subsequent ammonium absorption at 20 °C, and about 12 h at 20 °C was required for recovery. For both the cultivated and wild species, rerooted cuttings were slightly less sensitive to chilling than seedlings. The relative inhibition (mean ± SE) of ammonium absorption before and after chilling was 58·4 ± 2·5% for the cultivated species and 29·0 ± 9·1% for the wild species. The F₁ hybrid between the species showed a relative inhibition of 52·4 ± 3·6%, suggesting that chilling sensitivity may be dominant. In a backcross of the hybrid to L. esculentum , the phenotypic distribution of the relative inhibition of ammonium absorption indicated that this trait is segregating.     

Response of tomato plants to chilling stress in association with nutrient or phosphorus starvation

The experiments were conducted on two tomato cultivars: Garbo and Robin. Mineral starvation due to plant growth in 20-fold diluted nutrient solution (DNS) combined with chilling reduced the rate of photosynthesis (P _N) and stomatal conductance (g) to a greater extent than in plants grown in full nutrient solution (FNS). In phosphate-starved tomato plants the P _N rate and stomatal conductance decreased more after chilling than in plants grown on FNS. In low-P plants even 2 days after chilling the recovery of CO₂ assimilation rate and stomatal conductance was low. A resupply of phosphorus to low-P plants (low P + P) did not improve the rate of photosynthesis in non-chilled plants (NCh) but prevented P_N inhibition in chilled (Ch) plants. The greatest effect of P resupply was expressed as a better recovery of photosynthesis and stomatal conductance, especially in non-chilled low P + P plants. The F _v/F _m (ratio of variable to maximal chlorophyll fluorescence) decreased more during P starvation than as an effect of chilling. Supplying phosphorus to low-P plants caused the slight increase in the F _v/F _mratio. In conclusion, after a short-term chilling in darkness a much more drastic inhibition of photosynthesis was observed in nutrient-starved or P-insufficient tomato plants than in plants from FNS. This inhibition was caused by the decrease in both photochemical efficiency of photosystems and the reduction of stomatal conductance. The presented results support the hypothesis that tomato plants with limited supply of mineral nutrients or phosphorus are more susceptible to chilling.     

低温胁迫下外源褪黑素对番茄幼苗光抑制的缓解效应

为了探究外源褪黑素对低温逆境条件下番茄光合系统破坏的缓解机制,设置常温+水(NW)、常温+褪黑素(NM)、低温+水(CW)、低温+褪黑素(CM)4个处理,对番茄幼苗光合与叶绿素荧光参数、叶绿体抗坏血酸-谷胱甘肽(AsA-GSH)循环效率进行分析。结果表明: 与NW相比,CW的光合速率下降了50.3%～72.6%,叶绿体中的丙二醛含量升高了17.5%～132.7%,超氧阴离子产生速率、H₂O₂含量分别升高了86.5%～235.9%、96.6%～208.4%;而低温下施用褪黑素显著缓解了这一趋势,CM处理光合速率比CW提升了22.7%～24.7%,丙二醛含量、超氧阴离子产生速率、H₂O₂含量分别降低了16.6%～29.0%、14.9%～22.7%、10.7%～27.1%。与CW相比,CM处理光系统Ⅱ光化学能转化系数和调节性能量耗损系数分别升高了15.8%和7.2%,非调节性能量耗损系数下降了24.7%,AsA-GSH循环中关键代谢酶活性均有不同程度的增强。综上,低温下施用外源褪黑素可以平衡光系统Ⅱ的能量分布,增强叶绿体中AsA-GSH循环的活性氧清除效率,进而缓解低温引起的光抑制。     

The inheritance of chilling tolerance in tomato (Lycopersicon spp.)

During the past 25 years, chilling tolerance of the cultivated (chilling-sensitive) tomato Lycopersicon esculentum and its wild, chilling-tolerant relatives L. peruvianum and L. hirsutum (and, less intensively studied, L. chilense) has been the object of several investigations. The final aim of these studies can be seen in the increase in chilling tolerance of the cultivated genotypes. In this review, we will focus on low-temperature effects on photosynthesis and the inheritance of these traits to the offspring of various breeding attempts. While crossing L. peruvianum (male symbol) to L. esculentum (female symbol) so far has brought the most detailed insight with respect to physiological questions, for practical purposes, e.g., the readily cross ability, crossing programmes with L. hirsutum as pollen donor at present seem to be a promising way to achieve higher chilling-tolerant genotypes of the cultivated tomato. This perspective is due to the progress that has been made with respect to the genetic basis of chilling tolerance of Lycopersicon spp. over the past five years.     

Increased Rubisco content in maize mitigates chilling stress and speeds recovery

Many C₄ plants, including maize, perform poorly under chilling conditions. This phenomenon has been linked in part to decreased Rubisco abundance at lower temperatures. An exception to this is chilling‐tolerant Miscanthus, which is able to maintain Rubisco protein content under such conditions. The goal of this study was to investigate whether increasing Rubisco content in maize could improve performance during or following chilling stress. Here, we demonstrate that transgenic lines overexpressing Rubisco large and small subunits and the Rubisco assembly factor RAF1 (RAF1‐LSSS), which have increased Rubisco content and growth under control conditions, maintain increased Rubisco content and growth during chilling stress. RAF1‐LSSS plants exhibited 12% higher CO₂ assimilation relative to nontransgenic controls under control growth conditions, and a 17% differential after 2 weeks of chilling stress, although assimilation rates of all genotypes were ~50% lower in chilling conditions. Chlorophyll fluorescence measurements showed RAF1‐LSSS and WT plants had similar rates of photochemical quenching during chilling, suggesting Rubisco may not be the primary limiting factor that leads to poor performance in maize under chilling conditions. In contrast, RAF1‐LSSS had improved photochemical quenching before and after chilling stress, suggesting that increased Rubisco may help plants recover faster from chilling conditions. Relatively increased leaf area, dry weight and plant height observed before chilling in RAF1‐LSSS were also maintained during chilling. Together, these results demonstrate that an increase in Rubisco content allows maize plants to better cope with chilling stress and also improves their subsequent recovery, yet additional modifications are required to engineer chilling tolerance in maize.     

Light-dependent damage to photosynthesis in olive leaves during chilling and high temperature stress

Abstract The leaves of olive are long lived and likely to experience both chilling and high temperature stress during their life. Changes in photosynthetic CO₂ assimilation resulting from chilling and high temperature stress, in both dim and high light, are investigated. The quantum yield (φ) of photosynthesis at limiting light levels was reduced following chilling (at 5°C for 12 h), in dim light by approximately 10%, and in high light by 75%; the difference being attributed to photoinhibition. Similar reductions were observed in the light-saturated rate of CO₂ uptake (A_max). Decrease in A_max correlated with a halving of the leaf internal CO₂ concentration (c_i), suggesting an increased limitation by stomata following photoinhibition. Leaves were apparently more susceptible to photoinhibitory damage if the whole plant, rather than the leaf alone, was chilled. On return to 26 °C, I he photosynthetic capacity recovered to pre-stress levels within a few hours if leaves had been chilled in high light for 8 h or less, but did not fully recover from longer periods of chilling when loss of chlorophyll occurred. Leaves which were recovering from chilling in high light showed far more damage on being chilled a second time in high light. Three hours in high light at 38 °C reduced φ by 80%, but φ recovered within 4h of return to 26 °C. Although leaves of Olive are apparently less susceptible to photoinhibitory damage during chilling stress than the short-lived leaves of chilling-sensitive annual? crops, the results nevertheless show that photoinhibition during temperature stress is potentially a major factor influencing the photosynthetic productivity of Olive in the field.     

Low growth temperature-induced changes to pigment composition and photosynthesis in Zea mays genotypes differing in chilling sensitivity

The effects on pigment composition and photosynthesis of low temperature during growth were examined in the third leaf of three chilling-tolerant and three chilling-sensitive genotypes of Zea mays L. The plants were grown under a controlled environment at 24 or 14 °C at a photon flux density (PFD) of 200 or 600 μ mol m^–2 s^–1. At 24 °C, the two classes of genotypes showed little differences in their photosynthetic activity and their composition of pigments. At 14 °C, photosynthetic activity was considerably reduced but the chilling-tolerant genotypes displayed higher photosynthetic rates than the chilling-sensitive ones. Plants grown at 14 °C showed a reduced chlorophyll (Chl) a + b content and a reduced Chl a / b ratio but an increased ratio of total carotenoids to Chl a + b . These changes in pigment composition in plants grown at low temperature were generally more pronounced in the chilling-sensitive genotypes than in the tolerant ones, particularly at high PFD. Furthermore, at 14 °C, all the genotypes showed increased ratios of lutein, neoxanthin and xanthophyll-cycle carotenoids to Chl a + b but a reduced ratio of β -carotene to Chl a + b , especially at high PFD. At 14 °C, the chilling-tolerant genotypes, when compared with the sensitive ones, were characterized by higher contents of β -carotene and neoxanthin, a lower content of xanthophyll-cycle carotenoids, a lower ratio of xanthophylls to β -carotene, and less of their xanthophyll-cycle carotenoid pool in the form of zeaxanthin. These differences between the two classes of genotypes were more pronounced at high PFD than at low PFD. The results are discussed in terms of the relationship that may exist in maize between pigment composition and the capacity to form an efficient photosynthetic apparatus at low growth temperature.     

Genetic engineering of glycinebetaine synthesis in tomato protects seeds, plants, and flowers from chilling damage

Tomato (Lycopersicon esculentum Mill.) plants, which normally do not accumulate glycinebetaine (GB), are susceptible to chilling stress. Exposure to temperatures below 10 degrees C causes various injuries and greatly decreases fruit set in most cultivars. We have transformed tomato (cv. Moneymaker) with a chloroplast-targeted codA gene of Arthrobacter globiformis, which encodes choline oxidase to catalyze the conversion of choline to GB. These transgenic plants express codA and synthesize choline oxidase, while accumulating GB in their leaves and reproductive organs up to 0.3 and 1.2 micromol g(-1) fresh weight (FW), respectively. Their chloroplasts contain up to 86% of total leaf GB. Over various developmental phases, from seed germination to fruit production, these GB-accumulating plants are more tolerant of chilling stress than their wild-type counterparts. During reproduction, they yield, on average, 10-30% more fruit following chilling stress. Endogenous GB contents as low as 0.1 micromol g(-1) FW are apparently sufficient to confer high levels of tolerance in tomato plants, as achieved via transformation with the codA gene. Exogenous application of either GB or H2O2 improves both chilling and oxidative tolerance concomitant with enhanced catalase activity. These moderately increased levels of H2O2 in codA transgenic plants, as a byproduct of choline oxidase-catalyzed GB synthesis, might activate the H2O2-inducible protective mechanism, resulting in improved chilling and oxidative tolerances in GB-accumulating codA transgenic plants. Thus, introducing the biosynthetic pathway of GB into tomato through metabolic engineering is an effective strategy for improving chilling tolerance.     

Effect of sulphate on photosynthesis in greenhouse–grown tomato plants

Sulphate accumulates in the rhizosphere of plants grown in hydroponic systems. To avoid such sulphate accumulation and promote the use of environmentally sound hydroponic systems, we examined the effects of four sulphate concentrations (0.1, 5,2, 10.4 and 20.8 m M ) on photosynthesis, ribulose-l,5-bisphosphate carboxylase/oxygenase (Rubisco, EC 4.1.1.39) activities and related physiological processes in greenhouse–grown tomato plants ( Lycopersicon esculentum Mill. cv. Trust). The lowest sulphate concentration (0.1 m M ) significantly decreased photosynthetic capacity (P_c) and Rubisco activities on a leaf area basis. This result was supported by our data for dry matter per plant, which was low for plants in the 0.1 m M treatment. The photosynthesis-related variables such as leaf conductance, chlorophyll and soluble protein were lowest for the 0.1 m M treatment. Both total Rubisco activity and the activated ratio were reduced with this treatment. However, Rubisco activities expressed per g of protein or per g of chlorophyll were not significantly affected. These results suggest that sulphur deficiency depressed P_c– by reducing the amount of both Rubisco and chlorophyll and by causing an inactivation of Rubisco. The ratio of organic sulphur vs organic nitrogen (S/N) in plants of the 0.1 m M treatment was far below the normal values. This low S/N ratio might be accountable for the negative effect of low sulphate on P_c and plant growth. P_c and dry matter were not affected until sulphate concentration in the nutrient solution reached a high level of 20.8 m M .     

干旱对番茄幼苗光合和某些生理指标的影响

干旱缺水已成为植物光合作用和生长发育主要的限制因素,在干旱胁迫下,作物的生长发育受到影响,依据作物的形态变化进行浇灌属于延后性灌溉,未必能完全补偿对作物生长造成的影响。确定灌溉时间点,既确保植物正常生长不受影响,也可以提高水分利用效率,减少水资源浪费,从而达到节水灌溉的目的。该研究以温室土槽栽培番茄幼苗为材料,设定土壤含水量为30.00%(对照)、21.00%、18.00%、15.00%、12.00%、9.00%,研究了干旱胁迫对番茄叶片光合特性、抗氧化酶(超氧化物歧化酶、过氧化物酶、过氧化氢酶)、碳酸酐酶活性变化的影响,并以此表征番茄幼苗需水信息。结果表明:随着干旱胁迫程度的增加,叶片水势逐渐降低。超氧化物歧化酶、过氧化物酶及过氧化氢酶等抗氧化酶在番茄幼苗耐受水分胁迫中起到重要的作用;超氧化物歧化酶、过氧化物酶在干旱胁迫条件下反应更迅速,但过氧化氢酶相对于超氧化物歧化酶、过氧化物酶对干旱胁迫的耐受能力更强;干旱胁迫条件下抗氧化酶活性的转折点在15.00%土壤含水量左右;水分胁迫条件下碳酸酐酶参与了对光合作用的调节,并在15.00%土壤含水量时活性升至最高,使得番茄仍能维持较高的光合速率,以维持正常的生理机能;随着干旱胁迫程度的加剧(12.00%土壤含水量),碳酸酐酶活性与净光合速率都迅速下降。综上分析,当土壤含水量低于15.00%并高于12.00%时,对作物进行灌溉最为合适。抗氧化酶及碳酸酐酶活性可为作物最佳灌溉时间点的预测提供科学依据。     

Prior temperature exposure affects subsequent chilling sensitivity

The chilling sensitivity of small discs or segments of tissue excised from chillingsensitive species was significantly altered by prior temperature exposure subsequent to holding the tissue at chilling temperatures as measured by a number of physiological processes sensitive to chilling. This temperature conditioning was reversible by an additional temperature exposure before chilling, and mature-green and red-ripe tomato tissue exhibit similar chilling sensitivities. Exposing pericarp discs excised from tomato fruit (Lycopersicon esculentum Mill. cv. Castelmart), a chilling-sensitive species, to temperatures from 0 to 37°C for 6 h before chilling the discs at 2.5°C for 4 days significantly altered the rate of ion leakage from the discs, but had no effect on the rate of ion leakage before chilling and only a minimal effect on discs held at a non-chilling temperature of 12°C. Exposing chillingsensitive tissue to temperatures below that required to induce heat-shock proteins but above 20°C significantly increased chilling sensitivity as compared to tissue exposed to temperatures between 10 and 20°C. Rates of ion leakage after 4 days of chilling at 2.5°C were higher from fruit and vegetative tissue of chilling-sensitive species (Cucumis sativus L. cv. Poinsett 76, and Cucurbita pepo L. cv. Young Beauty) that were previously exposed for 6 h to 32°C than from similar tissue exposed to 12°C. Exposure to 32 and 12°C had no effect on the rate of ion leakage from fruit tissue of chilling tolerant species (Malus domestica Borkh. cv. Golden Delicious, Pyrus communis L. cv. Bartlett). Ethylene and CO₂ production were higher and lycopene synthesis was lower in chilled tomato pericarp discs that were previously exposed for 6 h to 32°C than the values from tissue exposed to 12°C for 6 h before chilling. Increased chilling sensitivity induced by a 6 h exposure to 32°C could be reversed by subsequent exposure to 12°C for 6 h.     

Nitrate reductase activity, nitrogenase activity and photosynthesis of black alder exposed to chilling temperatures

Actinorhizal ( Frankia -nodulated) black alder [ Alnus glutinosa (L.) Gaertn.] seedlings fertilized with 0.36 m M nitrate (low nitrate fertilizer treatment) or 7.14 m M nitrate (high nitrate fertilizer treatment) and acclimated in a growth chamber for 2 weeks were exposed to 2.5 h of night-time chilling temperatures of −1 to 4°C. Cold treatment decreased nitrogenase activity (acetylene reduction activity) 33% for low nitrate fertilized plants and 41% for high nitrate fertilized plants. Recovery of nitrogenase activity occurred within 7 days after chilling treatment. In contrast, in vivo nitrate reductase (NR) activities of leaves and fine roots increased immediately after chilling then decreased as nitrogenase activities recovered. Fine roots of alder seedlings exhibited NR activities proportional to the amounts of nitrate in the rooting medium. In contrast, the NR activities of leaves were independent of substrate and tissue nitrate levels and corresponded to nitrogenase activity in the root nodules. In a separate experiment, net photosynthesis (PS) of similarly treated black alder seedlings was measured before and after chilling treatments. Net PS declined in response to chilling by 17% for plants receiving low nitrate fertilizer and 19% for plants receiving high nitrate fertilizer. After chilling, stomatal conductance (g_s) decreased by 39% and internal CO₂ concentration (c_i) decreased by 5% in plants receiving the high nitrate fertilizer, whereas plants receiving the low nitrate fertilizer showed no change in g_s and a 13% increase in c_i. Results indicate that chilling stimulates stomatal closure only at the high nitrate level and that interference with biochemical functions is probably the major impact of chilling on PS.     

Water relations under root chilling in a sensitive and tolerant tomato species

The shoots of cultivated tomato (Lycopersicon esculentum cv. T5) wilt if their roots are exposed to chilling temperatures of around 5 °C. Under the same treatment, a chilling‐tolerant congener (Lycopersicon hirsutum LA 1778) maintains shoot turgor. To determine the physiological basis of this differential response, the effect of chilling on both excised roots and roots of intact plants in pressure chambers were investigated. In excised roots and intact plants, root hydraulic conductance declined with temperature to nearly twice the extent expected from the temperature dependence of the viscosity of water, but the response was similar in both species. The species differed markedly, however, in stomatal behaviour: in L. hirsutum, stomatal conductance declined as root temperatures were lowered, whereas the stomata of L. esculentum remained open until the roots reached 5 °C, and the plants became flaccid and suffered damage. Grafted plants with the shoots of one genotype and roots of another indicated that the differential stomatal behaviour during root chilling has distinct shoot and root components.     

Strigolactones positively regulate chilling tolerance in pea and in Arabidopsis

Strigolactones (SL) fulfil important roles in plant development and stress tolerance. Here, we characterized the role of SL in the dark chilling tolerance of pea and Arabidopsis by analysis of mutants that are defective in either SL synthesis or signalling. Pea mutants (rms3, rms4, and rms5) had significantly greater shoot branching with higher leaf chlorophyll a/b ratios and carotenoid contents than the wild type. Exposure to dark chilling significantly decreased shoot fresh weights but increased leaf numbers in all lines. Moreover, dark chilling treatments decreased biomass (dry weight) accumulation only in rms3 and rms5 shoots. Unlike the wild type plants, chilling‐induced inhibition of photosynthetic carbon assimilation was observed in the rms lines and also in the Arabidopsis max3‐9, max4‐1, and max2‐1 mutants that are defective in SL synthesis or signalling. When grown on agar plates, the max mutant rosettes accumulated less biomass than the wild type. The synthetic SL, GR24, decreased leaf area in the wild type, max3‐9, and max4‐1 mutants but not in max2‐1 in the absence of stress. In addition, a chilling‐induced decrease in leaf area was observed in all the lines in the presence of GR24. We conclude that SL plays an important role in the control of dark chilling tolerance.