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1.
Photodynamic and photoprotective responses at different irradiances were investigated in transgenic rice (Oryza sativa) expressing Bradyrhizobium japonicum 5-aminolevulinic acid synthase (ALA-S). With high irradiance (HI) of 350 µmol m?2 s?1, transgenic lines P5 and P14 showed a decrease in contents of chlorophyll (Chl) and the chloroplast-encoded gene psbA mRNA, whereas a decrease in light-harvesting Chl-binding proteins was observed only in P14. These effects were not observed in the wild-type (WT) line treated with HI or all of the lines treated with low irradiance (LI) of 150 µmol m?2 s?1. HI resulted in a greater decrease in the quantum yield of photosystem 2 and a greater increase in non-photochemical quenching (NPQ) in the transgenic lines, particularly in P14, compared to WT. Photoprotective zeaxanthin contents increased at HI, even though carotenoid contents were lower in the transgenic lines compared to WT. When exposed to HI, superoxide dismutase greatly increased in transgenic lines P5 and P14, but peroxidase and glutathione reductase increased only in P14, in which more photodynamic damage occurred. Thus the greater expression of ALA-S in the transgenic plants developed the stronger protective functions, i.e. the increased values of NPQ and zeaxanthin, as well as more photodynamic reactions, i.e. decreased photosynthetic component and efficiency, in the photosynthetic complexes. However, the photodynamic reactions indicate that the antioxidant capacity was insufficient to cope with the severe stress triggered by photoactive porphyrins in the transgenic rice expressing ALA-S.  相似文献   

2.
Ferredoxins are the main electron shuttles in chloroplasts, accepting electrons from photosystem I and delivering them to essential oxido-reductive pathways in the stroma. Ferredoxin levels decrease under adverse environmental conditions in both plants and photosynthetic micro-organisms. In cyanobacteria and some algae, this decrease is compensated for by induction of flavodoxin, an isofunctional flavoprotein that can replace ferredoxin in many reactions. Flavodoxin is not present in plants, but tobacco lines expressing a plastid-targeted cyanobacterial flavodoxin developed increased tolerance to environmental stress. Chloroplast-located flavodoxin interacts productively with endogenous ferredoxin-dependent pathways, suggesting that its protective role results from replacement of stress-labile ferredoxin. We tested this hypothesis by using RNA antisense and interference techniques to decrease ferredoxin levels in transgenic tobacco. Ferredoxin-deficient lines showed growth arrest, leaf chlorosis and decreased CO(2) assimilation. Chlorophyll fluorescence measurements indicated impaired photochemistry, over-reduction of the photosynthetic electron transport chain and enhanced non-photochemical quenching. Expression of flavodoxin from the nuclear or plastid genome restored growth, pigment contents and photosynthetic capacity, and relieved the electron pressure on the electron transport chain. Tolerance to oxidative stress also recovered. In the absence of flavodoxin, ferredoxin could not be decreased below 45% of physiological content without fatally compromising plant survival, but in its presence, lines with only 12% remaining ferredoxin could grow autotrophically, with almost wild-type phenotypes. The results indicate that the stress tolerance conferred by flavodoxin expression in plants stems largely from functional complementation of endogenous ferredoxin by the cyanobacterial flavoprotein.  相似文献   

3.
Glycine betaine (GB) is a compatible solute that accumulates rapidly to enhance heat tolerance in many plants grown under heat stress. In this study, a BADH gene (betaine aldehyde dehydrogenase) from spinach was introduced into tomato (Lycopersicon esculentum cv. ‘Moneymaker’) via Agrobacterium-mediated transformation. Transgenic tomato lines expressing BADH exhibited higher capabilities for GB accumulation. Chlorophyll fluorescence analysis of wild type (WT) and transgenic plants exposed to heat treatment (42 °C) showed that transgenic plants exhibited higher photosynthetic capacities than WT plants. This finding suggests that GB accumulation increases tolerance to heat-enhanced photoinhibition. This increased tolerance was associated with an improvement in D1 protein content, which accelerated the repair of photosystem II (PSII) following heat-enhanced photoinhibition. Significant accumulations of hydrogen peroxide (H2O2) and superoxide radical (O2 ?) were observed in WT plants under heat stress. However, these accumulations were much less for the transgenic plants. An important finding reported herein is that exogenous GB cannot directly reduce the content of reactive oxygen species (ROS). In accordance with a lower relative electrolyte conductivity and malondialdehyde content, the activities of antioxidant enzymes were higher in transgenic lines than in WT plants, indicating that the degree of membrane injury in the transgenic plants was lower compared to the WT plants. These results suggest that GB accumulation in vivo cannot directly eliminate ROS. Rather, higher antioxidant enzyme activities must be maintained to lessen the accumulation of ROS in transgenic plants and to decrease the degree of membrane injury.  相似文献   

4.
Phosphoenolpyruvate carboxylase (PEPC) is known to play a key role in the initial fixation of CO2 in C4 photosynthesis. The PEPC gene from sugarcane (a C4 plant) was introduced into indica rice (Hang2), a process mediated by Agrobacterium tumefaciens. Integration patterns and copy numbers of the gene was confirmed by DNA blot analysis. RT-PCR and western blotting results showed that the PEPC gene was expressed at both the mRNA and protein levels in the transgenic lines. Real-time PCR results indicated that expression of the sugarcane PEPC gene occurred mostly in green tissues and changed under high temperature and drought stress. All transgenic lines showed higher PEPC enzyme activities compared to the untransformed controls, with the highest activity (11.1 times higher than the controls) being observed in the transgenic line, T34. The transgenic lines also exhibited higher photosynthetic rates. The highest photosynthetic rate was observed in the transgenic line, T54 (22.3 μmol m?2 s?1; 24.6 % higher than that in non-transgenic plants) under high-temperature conditions. Furthermore, the filled grain and total grain numbers for transgenic lines were higher than those for non-transgenic plants, but the grain filling (%) and 1,000-grain weights of all transgenic lines remained unchanged. We concluded that over-expression of the PEPC gene from sugarcane in indica rice (Hang2) resulted in higher PEPC enzyme activities and higher photosynthesis rates under high-temperature conditions.  相似文献   

5.
Maintenance of energy balance under changeable light conditions is an essential function of photosynthetic organisms to achieve efficient photochemical reactions. Among the photosynthetic organisms, diatoms possess light-harvesting fucoxanthin chlorophyll (Chl) a/c-binding protein (FCP) as peripheral antennas. However, how diatoms regulate excitation-energy distribution between FCP and the two photosystem cores during light adaptation is poorly understood. In this study, we examined spectroscopic properties of a marine diatom Chaetoceros gracilis adapted in the dark and at photosynthetic photon flux density at 30 and 300?μmol?photons?m?2?s?1. Absorption spectra at 77?K showed significant changes in the Soret region, and 77-K steady-state fluorescence spectra showed significant differences in the spectral shape and relative fluorescence intensity originating from both PSII and PSI, among the cells grown under different light conditions. These results suggest alterations of pigment composition and their interactions under the different light conditions. These alterations affected the excitation-energy dynamics monitored by picosecond time-resolved fluorescence analyses at 77?K significantly. The contributions of Chls having lower energy levels than the reaction center Chls in the two photosystems to the energy dynamics were clearly identified in the three cells but with presumably different roles. These findings provide insights into the regulatory mechanism of excitation-energy balance in diatoms under various light conditions.  相似文献   

6.
Plant growth and crop production are limited by environmental stress. We used a large population of transgenic Arabidopsis expressing rice full-length cDNAs to isolate the rice genes that improve the tolerance of plants to environmental stress. By sowing T2 seeds of the transgenic lines under conditions of salinity stress, the salt-tolerant line R07047 was isolated. It expressed a rice gene, OsSMCP1, which encodes a small protein with a single C2 domain, a Ca2+-dependent membrane-targeting domain. Retransformation of wild-type Arabidopsis revealed that OsSMCP1 is responsible for conferring the salt tolerance. It is particularly interesting that R07047 and newly constructed OsSMCP1-overexpressing Arabidopsis showed enhanced tolerance not only to high salinity but also to osmotic, dehydrative, and oxidative stresses. Furthermore, R07047 showed improved resistance to Pseudomonas syringae. The OsSMCP1 expression in rice is constitutive. Particle-bombardment-mediated transient expression analysis revealed that OsSMCP1 is targeted to plastids in rice epidermal cells. It induced overexpression of several nuclear encoded genes, including the stress-associated genes, in transgenic Arabidopsis. No marked morphological change or growth retardation was observed in R07047 or retransformants. For molecular breeding to improve the tolerance of crops against environmental stress, OsSMCP1 is a promising candidate.  相似文献   

7.
Trifoliate orange (Poncirus trifoliata L. Raf.), a rootstock widely used for citrus species, is salt-sensitive. Worldwide, salinity is a major abiotic stress affecting citrus growth and yield. Glycinebetaine (GB) is an important osmoprotectant involved in responses to salt stress. However, current evidence regarding the effect of salt stress on GB accumulation in trifoliate orange is limited, and the GB synthesis gene has not yet been shown to confer enhanced salt stress tolerance to this species in a transgenic context. In the current study, we first examined the change in GB level of trifoliate orange seedlings exposed to salt stress, and found that salt increased endogenous GB level in a concentration-dependent manner. A betaine aldehyde dehydrogenase gene (AhBADH) cloned from Atriplex hortensis was introduced into the trifoliate orange by means of Agrobacterium-mediated transformation. RT-PCR analysis on three selected transgenic lines showed that the AhBADH gene was overexpressed in each of them. GB levels in these lines were also higher than those in untransformed wild-type (WT) plants. In the transgenic lines, exposure to 200 mM NaCl resulted in significantly less serious leaf burning and defoliation, lower MDA accumulation, and higher chlorophyll contents than those in the WT plants. Moreover, when exposed to salt, shoots of transgenic plants contained lower levels of Na+ and Cl, higher levels of K+, and a higher K/Na ratio, while the same was true for the roots in most cases. Taken together, the data suggest that overexpression of the AhBADH gene in transgenic trifoliate orange enhanced salt stress tolerance. This may be correlated with the low levels of lipid peroxidation, protection of the photosynthetic machinery, and increase in K+ uptake.  相似文献   

8.
In this study, chlorophyll fluorescence parameters (?F/F m′, F v/F m) and oxygen evolution of female vegetative tissues of Porphyra katadai var. hemiphylla in unisexual culture (FV) and in mixed culture with male vegetative tissues (FV-M) were followed at 5–20 °C, 10 and 80 μmol photons m?2 s?1. The formation of reproductive tissues was closely correlated with decreasing photosynthetic activities. At the same temperature the tissues cultured under 80 μmol photons m?2 s?1 showed a greater extent of maturation than those under 10 μmol photons m?2 s?1, and their decrease in photosynthesis was also larger. Under the same light intensity the extent of maturation increased with increasing temperature, and both cultures showed higher values of ?F/F m′ and F v/F m at 10 and 15 °C, while their oxygen evolution became negative at 15–20 °C during the later period. Under the same culture condition the maturation of FV-M culture was relatively faster than that of FV culture, while their photosynthetic activity, especially ?F/F m′, was lower.  相似文献   

9.

Excessive heavy metal (HM) levels in soil have become a source of concern due to their adverse effects on human health and the agriculture industry. Soil contamination by HMs leads to an accumulation of reactive oxygen species (ROSs) within the plant cell and disruption of photosynthesis-related proteins. The response of tobacco lines overexpressing flavodoxin (Fld) and betaine aldehyde dehydrogenase (BADH) to cadmium (Cd) toxicity was investigated in this study. PCR results demonstrated the expected amplicon length of each gene in the transgenic lines. Absolute qRT-PCR demonstrates a single copy of T-DNA integration into each transgenic line. Relative qRT-PCR confirmed overexpression of Fld and BADH in transgenic lines. The maximum quantum yield of photosystem II (Fv/Fm) was measured under Cd toxicity stress and revealed that transgenic lines had a higher Fv/Fm than wild-type (WT) plants. Accumulation of proline, glycine betaine (GB), and higher activity of antioxidant enzymes alongside lower levels of malondialdehyde (MDA) and hydrogen peroxide (H2O2) was indicative of a robust antioxidant system in transgenic plants. Therefore, performing a loop in reducing the ROS produced in the photosynthesis electron transport chain and stimulating the ROS scavenger enzyme activity improved the plant tolerance to Cd stress.

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11.
Tocopherols (α-, β-, γ-, and δ-tocopherol) represent a group of lipophilic antioxidants that are synthesized only by photosynthetic organisms. It is widely believed that the main functions of tocopherols are protection of pigments and proteins of photosystem and polyunsaturated fatty acids from oxidative damage caused by reactive oxygen species. In the present study, we report on the cloning and characterization of NtTC, which is a tocopherol cyclase (TC) ortholog isolated from tobacco. To enhance tocopherol contents, we generated independent transgenic rice events in expressing NtTC or NtTC along with Perilla γ-tocopherol methyltransferase genes. The transgenic TC line significantly increased α-, total tocopherol, total glutathione, and total antioxidant status activity levels compared with the wild type. Furthermore, TC rice plants showed higher tolerance to drought than wild-type rice plants. On the basis of these studies, we concluded that overexpression of NtTC could increase the tolerance to drought stress and that the increase in tocopherol affects cellular signaling and antioxidant defense of plants in response to drought.  相似文献   

12.
The high light sensitivity of three submerged aquatic freshwater plant species, Egeria densa, Elodea nuttallii and Myriophyllum heterophyllum, which have been cultivated at a photosynthetically active radiation (PAR, 400-700 nm) of 70 μmol photons m−2 s−1, was studied by means of chlorophyll fluorescence and pigment analyses. Exposure of plants to 100, 300, 600 and 1000 μmol photons m−2 s−1 PAR for up to 360 min induced a strong reduction of the Fv/Fm ratio, indicating a pronounced inactivation of PSII even at the lowest PAR applied. These changes were accompanied by a reduction of the chlorophyll content to about 60-70% of control values at the highest PAR. Rapidly inducible photoprotective mechanisms were not affected, as derived from the rapid generation of pH-dependent energy dissipation under these conditions. At PAR higher than 100 μmol photons m−2 s−1, however, the primary quinone acceptor of photosystem II, QA, was reduced to about 80% and the effective quantum yield of photosystem II, ΦPSII, dropped to values of about 10%, indicating a high reduction state of the photosynthetic electron transport chain. These data support the notion that the three aquatic macrophytes have a very low capacity for the acclimation to higher light intensities.  相似文献   

13.
Salt stress has attracted increasing attention due to its toxic ability to restrict plant growth, and the photorespiration pathway has been shown to develop improved plant tolerance to abiotic stress. In this study, an Arabidopsis photorespiratory pathway gene serine: glyoxylate aminotransferase (SGAT), named as AtAGT1, was successfully overexpressed in duckweed (Lemna minor) to investigate the salinity defense capability in three transgenic overexpressed (OE) lines. Increased SGAT activity and decreased endogenous serine levels in these transgenic plant lines under salt stress resulted in enhanced protection against root abscission, higher maximum quantum yield of photosystem II (Fv/Fm), increased defense from cell damage as a result of improved cell membrane integrity, a decrease of reactive oxygen species (ROS) accumulation, and a strengthened antioxidant system. The salt tolerance in these transgenic OE lines indicates that the improvement of photorespiration stimulated the antioxidant system to scavenge ROS. The change of serine level also suggests the role of serine during salt stress. This transgenic engineering in duckweed not only introduced salt tolerance to this aquatic plant but also reveals a significant role of photorespiration during salinity stress.  相似文献   

14.
The key step in the fermentation of glutamate by Acidaminococcus fermentans is a reversible syn-elimination of water from (R)-2-hydroxyglutaryl-CoA to (E)-glutaconyl-CoA catalyzed by 2-hydroxyglutaryl-CoA dehydratase, a two-component enzyme system. The actual dehydration is mediated by component D, which contains 1.0 [4Fe-4S]2+ cluster, 1.0 reduced riboflavin-5′-phosphate and about 0.1 molybdenum (VI) per heterodimer. The enzyme has to be activated by the extremely oxygen-sensitive [4Fe-4S]1+/2+-cluster-containing homodimeric component A, which generates Mo(V) by an ATP/Mg2+-induced one-electron transfer. Previous experiments established that the hydroquinone state of a flavodoxin (m=14.6 kDa) isolated from A. fermentans served as one-electron donor of component A, whereby the blue semiquinone is formed. Here we describe the isolation and characterization of an alternative electron donor from the same organism, a two [4Fe-4S]1+/2+-cluster-containing ferredoxin (m=5.6 kDa) closely related to that from Clostridium acidiurici. The protein was purified to homogeneity and almost completely sequenced; the magnetically interacting [4Fe-4S] clusters were characterized by EPR and Mössbauer spectroscopy. The redox potentials of the ferredoxin were determined as ?405 mV and ?340 mV. Growth experiments with A. fermentans in the presence of different iron concentrations in the medium (7–45 μM) showed that flavodoxin is the dominant electron donor protein under iron-limiting conditions. Its concentration continuously decreased from 3.5 μmol/g protein at 7 μM Fe to 0.02 μmol/g at 45 μM Fe. In contrast, the concentration of ferredoxin increased stepwise from about 0.2 μmol/g at 7–13 μM Fe to 1.1±0.1 μmol/g at 17–45 μM Fe.  相似文献   

15.
Genome methylation plays a key role in regulating gene expression, but limited knowledge exists concerning the link between DNA methylation and economic traits in forest trees. We measured photosynthetic characteristics and growth traits in 130 intraspecific hybrids of Chinese white poplar (Populus tomentosa Carr.) and detected their genome methylation. The phenotypic data were normally distributed, and each trait had a significant difference among the hybrids. The net photosynthetic rate (Pn, 14.83?±?3.76???mol?m?2?s?1), stomatal conductance (Gs, 0.29?±?0.09?mol?m?2?s?1), and intercellular CO2 concentration (Ci, 264.50?±?30.94???mol?mol?1) showed similar trends. Positive correlations were found between Pn and height (H, 133.59?±?50.44?cm) and basal diameter (D, 16.29?±?5.20?mm), respectively. Using methylation-sensitive amplification polymorphism (MSAP) analysis, 32 primer-pair combinations generated 715 polymorphic markers. Positive correlations between photosynthetic characteristics, such as Pn and Gs, and total relative methylation level and relative hemimethylation (CNG methylation) level were investigated. Eighty-one candidate markers were associated with Pn, Gs, or Ci, 13 of which were also associated with growth traits using single MSAP molecular marker association. Sequencing and BLAST analysis showed that candidate markers were linked to genes encoding protochlorophyllide reductase and proteins of cytochrome P450 CYP4/CYP19/CYP26 subfamilies, and linked to genes taking part in, e.g., photosystem II. Therefore, the regions defined by the MSAP candidate markers are linked to genes that are essential for photosynthetic characteristics that respond to DNA methylation and subsequently affect growth traits.  相似文献   

16.
Ascorbate (AsA) is an important antioxidant that can scavenge reactive oxygen species to protect plant cells against oxidative stress. Guanosine 5'-diphosphate (GDP)-L-galactose phosphorylase (GGP) is a key enzyme in the AsA biosynthetic pathway. To investigate the functions of GGP in AsA synthesis and oxidative stress tolerance in tomato, antisense lines with a reduced expression of SlGGP were obtained. Photobleaching after treatment of leaf disks with methyl viologen was more severe in transgenic lines compared to wild type (WT) plants. Moreover, compared with the WT plants, the transgenic plants showed a higher content of hydrogen peroxide, superoxide anion, malondialdehyde, as well as ion leakage, but a lower content of AsA and chlorophylls, ascorbate peroxidase activity, net photosynthetic rate, and maximal photochemical efficiency of photosystem II. Results of real-time quantitative polymerase chain reaction show that suppression of the SlGGP gene in the transgenic plants reduced their oxidative stress tolerance.  相似文献   

17.
Oxidative stress is one of the major causative factors for injury to plants exposed to environmental stresses. Plants have developed diverse defense mechanisms for scavenging oxidative stress-inducing molecules. The antioxidative enzyme 2-cysteine peroxiredoxin (2-Cys Prx) removes peroxides and protects the photosynthetic membrane from oxidative damage. In this study, transgenic potato (Solanum tuberosum L. cv. Atlantic) expressing At2-Cys Prx under control of the oxidative stress-inducible SWPA2 promoter or enhanced CaMV 35S promoter (referred to as SP and EP plants, respectively) was generated using Agrobacterium-mediated transformation. The transgenic plants were tested for tolerance to stress. Following treatment with 3 μM methyl viologen (MV), leaf discs from SP and EP plants showed approximately 33 and 15% less damage than non-transformed (NT) plants. When 300 μM MV was sprayed onto whole plants, the photosynthetic activity of SP plants decreased by 25%, whereas that of NT plants decreased by 60%. In addition, SP plants showed enhanced tolerance to high temperature at 42 °C. After treatment at high temperature, the photosynthetic activity of SP plants decreased by about 7% compared to plants grown at 25 °C, whereas it declined by 31% in NT plants. These results indicate that transgenic potato can efficiently regulate oxidative stress from various environmental stresses via overexpression of At2-Cys Prx under control of the stress-inducible SWPA2 promoter.  相似文献   

18.
Soil salinity is a serious worldwide problem. To improve the salt tolerance of plants, an increasing number of genes related to abiotic stress have been recently expressed by genetic engineers. In the present study, the successful introduction into tobacco of isopentenyl transferase (IPT) from Agrobacterium tumefaciens via Agrobacterium-mediated transformation is reported. A stress-inducible genetic construct was cloned using IPT under the control of the stress-inducible promoter rd29A from Arabidopsis thaliana. A total of 40 putative transgenic plant lines were obtained from independent Kan-resistant shoots. IPT integration into the tobacco genome was confirmed by polymerase chain reaction (PCR) and Southern blot analyses. Four positive transgenic lines each with a single T-DNA insertion were obtained. Real-time PCR confirmed a marked increase in IPT expression in young tobacco plants harboring rd29A-IPT after short-term exposure to salt. Ectopic IPT overexpression IPT under the control of the stress-inducible rd29A promoter resulted in significantly enhanced tolerance to salt stress. No obvious adverse effect on growth and development was observed in transgenic plants. Two IPT transgenic lines, T10 and T25, were chosen for further physiological analyses. The leaves of transgenic tobacco plants showed significantly prolonged chlorophyll retention times under a 2-week salt-stress treatment (150?mmol?L?1). In contrast, the leaves of the non-transformed plants (wild type) gradually senesced under the same condition. After re-watering for 2?weeks, chlorophyll in transgenic plants increased to a level comparable with that in the unstressed plants. On the other hand, the level in the non-transgenic control still remained low. Malondialdehyde (MDA) levels increased in both transgenic plants and the control after salt stress. However, the MDA levels only mildly increased in transgenic plants, and dramatically increased in the control. After re-watering for 7?days, MDA in transgenic plants returned to normal, whereas the level in the control remained high. Superoxide dismutase activity also similarly increased in transgenic plants during salt stress, and returned to normal after re-watering. These results indicate that enhanced reactive oxygen species scavenging capability may play a significant role in acquiring tolerance to abiotic stress.  相似文献   

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