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1.
Hidenori Hayashi Alia Atsushi Sakamoto Hideko Nonaka Tony H. H. Chen Norio Murata 《Journal of plant research》1998,111(2):357-362
Arabidopsis thaliana was transformed previously with thecodA gene from the soil bacteriumArthrobacter globiformis. This gene encodes choline oxidase, the enzyme that converts choline to glycinebetaine. Transformation with thecodA gene significantly enhanced the tolerance of transgenic plants to low temperature and high-salt stress. We report here that
seeds of transgenic plants that expressed thecodA gene were also more tolerant to salt stress during germination than seeds of non-transformed wild-type plants. Seedlings
of transgenic plants grew more rapidly than those of wild-type plants under salt-stress conditions. Furthermore, exogenously
applied glycinebetaine was effective in alleviating the harmful effects of salt stress during germination of seeds and growth
of young seedlings, a result that suggests that it was glycinebetaine that had enhanced the tolerance of the transgenic plants.
These observations indicate that synthesis of glycinebetaine in transgenic plantsin vivo, as a result of the expression of thecodA gene, might be veryuseful in improving the ability of crop plants to tolerate salt stress.
The extended abstract of a paper presented at the 13th International Symposium in Conjugation with Award of the International
Prize for Biology “Frontier of Plant Biology” 相似文献
2.
Transformation of tomato with a bacterial codA gene enhances tolerance to salt and water stresses 总被引:1,自引:0,他引:1
Goel D Singh AK Yadav V Babbar SB Murata N Bansal KC 《Journal of plant physiology》2011,168(11):1286-1294
Genetically engineered tomato (Lycopersicon esculentum) with the ability to synthesize glycinebetaine was generated by introducing the codA gene encoding choline oxidase from Arthrobacter globiformis. Integration of the codA gene in transgenic tomato plants was verified by PCR analysis and DNA blot hybridization. Transgenic expression of gene was verified by RT-PCR analysis and RNA blot hybridization. The codA-transgenic plants showed higher tolerance to salt stress during seed germination, and subsequent growth of young seedlings than wild-type plants. The codA transgene enhanced the salt tolerance of whole plants and leaves. Mature leaves of codA-transgenic plants revealed higher levels of relative water content, chlorophyll content, and proline content than those of wild-type plants under salt and water stresses. Results from the current study suggest that the expression of the codA gene in transgenic tomato plants induces the synthesis of glycinebetaine and improves the tolerance of plants to salt and water stresses. 相似文献
3.
Transformation of Synechococcus with a gene for choline oxidase enhances tolerance to salt stress 总被引:12,自引:0,他引:12
Patcharaporn Deshnium Dmitry A. Los Hidenori Hayashi Laszlo Mustardy Norio Murata 《Plant molecular biology》1995,29(5):897-907
Choline oxidase, isolated from the soil bacterium Arthrobacter globiformis, converts choline to glycinebetaine (N-trimethylglycine) without a requirement for any cofactors. The gene for this enzyme, designated codA, was cloned and introduced into the cyanobacterium Synechococcus sp. PCC 7942. The codA gene was experssed under the control of a strong constitutive promoter, and the transformed cells accumulated glycinebetaine at intracellular levels of 60–80 mM. Consequently the cells acquired tolerance to salt stress, as evaluated in terms of growth, accumulation of chlorophyll and photosynthetic activity. 相似文献
4.
Transformation of Japanese persimmon (Diospyros kaki Thunb.) with a bacterial gene for choline oxidase 总被引:4,自引:0,他引:4
Gao Mei Sakamoto Atsushi Miura Keisuke Murata Norio Sugiura Akira Tao Ryutaro 《Molecular breeding : new strategies in plant improvement》2000,6(5):501-510
This report describes the first successful genetic engineering of tolerance to salt in an agriculturally important species of woody plants by Agrobacterium-mediated transformation with the codA gene of Arthrobacter globiformis. This gene encodes choline oxidase, which catalyzes the oxidation of choline to glycinebetaine. The binary plasmid vector pGC95.091, containing a kanamycin-resistance gene (nptII), a gene for -glucuronidase (gusA) and the codA gene in its T-DNA region, was used with a disarmed strain of Agrobacterium tumefaciens, EHA101, to transform Japanese persimmon (Diospyros kaki Thunb. `Jiro') by the leaf disk transformation method. The pRS95.101 plasmid that included only nptII and gusA in the T-DNA region was used as a control. We selected eight transgenic lines with one or two copies of the T-DNA after transformation with pGC95.091 (PC lines) and three lines after transformation with pRS95.101 (PR lines). The eight PC lines produced choline oxidase and glycinebetaine whereas neither was found in untransformed `Jiro' and in the control PR lines. Transgenic plants grew normally, resembling wild-type plants both in vitro and ex vitro. The activity of photosystem II in leaves of the transgenic Japanese persimmon plants under NaCl stress was determined in terms of the ratio of the variable (F
v) to the maximum (F
m) fluorescence of chlorophyll (F
v/F
m). The rate of decline in (F
v/F
m under NaCl stress was lower in the PC lines than in the control PR lines. These results demonstrated that genetic engineering of Japanese persimmon, which allowed it to accumulate glycinebetaine, enhanced the tolerance to salt stress of this plant. 相似文献
5.
Genetically engineered tobacco (Nicotiana tabacum L.) with the ability to synthesis glycinebetaine (GB) in chloroplasts was established by introducing the BADH gene for betaine
aldehyde dehydrogenase from spinach (Spinacia oleracea L.). The genetic engineering resulted in enhanced tolerance of growth of young seedlings to salt stress. This increased tolerance
was not due to improved water status, since there were no significant differences in accumulation of sodium and chloride,
leaf water potential, and relative water content between wild type and transgenic plants under salt stress. Salt stress resulted
in a decrease in CO2 assimilation and such a decrease was much greater in wild type plants than in transgenic plants. Though salt stress showed
no damage to PSII, there were a decrease in the maximal PSII electron transport rate in vivo and an increase in non-photochemical
quenching (NPQ) and these changes were greater in wild type plants than in transgenic plants. In addition, salt stress inhibited
the activities of ribulose 1,5-bisphosphate carboxylase/oxygenase, chloroplastic fructose-1,6-bisphosphatase, fructose-1,6-bisphosphate
aldolase, and phosphoribulokinase and such a decrease was also greater in wild type plants than in transgenic plants, suggesting
that GB protects these enzymes against salt stress. However, there were no significant changes in the activities of phosphoglycerate
kinase, triose phosphate isomerase, ribulose-5-phosphate isomerase, transketolase, and sedoheptulose-1,7-bisphosphatase in
both wild type and transgenic plants. The results in this study suggest that enhanced tolerance of CO2 assimilation to salt stress may be one of physiological bases for increased tolerance of growth of transgenic plants to salt
stress. 相似文献
6.
7.
An efficient gene transfer system without tissue culture steps was developed for kidney bean by using sonication and vacuum
infiltration assisted, Agrobacterium-mediated transformation. Transgenic kidney bean with a group 3 lea (late embryogenesis abundant) protein gene from Brassica napus was produced through this approach. Among 18 combinations of transformation methods, Agrobacterium-mediated transformation combined with 5 min sonication and 5 min vacuum infiltration turned to be optimal, resulting in the
highest transformation efficiency. Transgenic kidney bean plants demonstrated enhanced growth ability under salt and water
deficit stress conditions. The increased tolerance was also reflected by delayed development of damage symptoms caused by
drought stress. Transgenic lines with high level of lea gene expression showed higher stress tolerance than lines with lower expression level. Stress tolerance of transgenic kidney
bean correlated much better with lea gene expression levels than with gene integration results. There is no prior report on the production of transgenic kidney
bean using both ultrasonic and vacuum infiltration assisted, Agrobacterium-mediated transformation. 相似文献
8.
Genetic engineering of the biosynthesis of glycinebetaine enhances thermotolerance of photosystem II in tobacco plants 总被引:1,自引:0,他引:1
Genetically engineered tobacco (Nicotiana tabacum L.) with the ability to accumulate glycinebetaine was established. The wild type and transgenic plants were exposed to heat
treatment (25–50°C) for 4 h in the dark and under growth light intensity (300 μmol m−2 s−1). The analyses of oxygen-evolving activity and chlorophyll fluorescence demonstrated that photosystem II (PSII) in transgenic
plants showed higher thermotolerance than in wild type plants in particular when heat stress was performed in the light, suggesting
that the accumulation of glycinebetaine leads to increased tolerance to heat-enhanced photoinhibition. This increased tolerance
was associated with an improvement on thermostability of the oxygen-evolving complex and the reaction center of PSII. The
enhanced tolerance was caused by acceleration of the repair of PSII from heat-enhanced photoinhibition. Under heat stress,
there was a significant accumulation of H2O2, O2− and catalytic Fe in wild type plants but this accumulation was much less in transgenic plants. Heat stress significantly
decreased the activities of catalase, ascorbate peroxidase, glutathione reductase, dehydroascorbate reductase, and monodehydroascorbate
reductase in wild type plants whereas the activities of these enzymes either decreased much less or maintained or even increased
in transgenic plants. In addition, heat stress increased the activity of superoxide dismutase in wild type plants but this
increase was much greater in transgenic plants. Furthermore, transgenic plants also showed higher content of ascorbate and
reduced glutathione than that of wild type plants under heat stress. The results suggest that the increased thermotolerance
induced by accumulation of glycinebetaine in vivo was associated with the enhancement of the repair of PSII from heat-enhanced
photo inhibition, which might be due to less accumulation of reactive oxygen species in transgenic plants. 相似文献
9.
Transformation of Arabidopsis thaliana with the codA gene for choline oxidase; accumulation of glycinebetaine and enhanced tolerance to salt and cold stress 总被引:12,自引:0,他引:12
Hidenori Hayashi Alia Laszlo Mustardy Patchraporn Deshnium Miki Ida Norio Murata 《The Plant journal : for cell and molecular biology》1997,12(1):133-142
Glycinebetaine is one of the compatible solutes that accumulate in the chloroplasts of certain halotolerant plants when these plants are exposed to salt or cold stress. The codA gene for choline oxidase, the enzyme that converts choline into glycinebetaine, has previously been cloned from a soil bacterium, Arthrobacter globiformis. Transformation of Arabidopsis thaliana with the cloned codA gene under the control of the 35S promoter of cauliflower mosaic virus enabled the plant to accumulate glycinebetaine and enhanced its tolerance to salt and cold stress. At 300 mM NaCl, considerable proportions of seeds of transformed plants germinated well, whereas seeds of wild-type plants failed to germinate. At 100 mM NaCl, transformed plants grew well whereas wild-type plants did not do so. The transformed plants tolerated 200 mM NaCl, which was lethal to wild-type plants. After plants had been incubated with 400 mM NaCl for two days, the photosystem II activity of wild-type plants had almost completely disappeared, whereas that of transformed plants remained at more than 50% of the original level. When exposed to a low temperature in the light, leaves of wild-type plants exhibited symptoms of chlorosis, whereas those of transformed plants did not. These observations demonstrate that the genetic modification of Arabidopsis thaliana that allowed it to accumulate glycinebetaine enhanced its ability to tolerate salt and cold stress. 相似文献
10.
Phytochelatins (PCs) are heavy metal binding peptides that play an important role in sequestration and detoxification of heavy
metals in plants. In this study, our goal was to develop transgenic plants with increased tolerance for and accumulation of
heavy metals from soil by expressing an Arabidopsis
thaliana
AtPCS1 gene, encoding phytochelatin synthase (PCS), in Indian mustard (Brassica juncea L.). A 35S promoter fused to a FLAG–tagged AtPCS1 cDNA was expressed in Indian mustard, and transgenic lines, designated pc lines, were evaluated for tolerance to and accumulation
of Cd and Zn. Transgenic plants with moderate AtPCS1 expression levels showed significantly higher tolerance to Cd and Zn
stress, but accumulated significantly less Cd and Zn than wild type plants in both shoot and root tissues. However, transgenic
plants with highest expression of the transgene did not exhibit enhanced Cd and Zn tolerance. Shoots of Cd-treated pc plants
had significantly higher levels of phytochelatins and thiols than wild-type plants. Significantly lower concentrations of
gluthatione in Cd-treated shoot and root tissues of transgenic plants were observed. Moderate expression levels of phytochelatin
synthase improved the ability of Indian mustard to tolerate certain levels of heavy metals, but at the same time did not increase
the accumulation potential for Cd and Zn. 相似文献
11.
W. Sawahel 《Biologia Plantarum》2003,47(1):39-44
Plasmid DNA (pChlCOD), containing the selectable hygromycin phosphotransferase hpt gene for hygromycin B resistance and the Arthrobacter globiformis codA gene for choline oxidase which catalyzes the direct conversion of choline to glycinebetaine, was delivered into rice plants
using Agrobacterium-mediated gene transfer via scutellum-derived calli. Southern, Northern and Western blot analyses demonstrated that the foreign gene had been transferred,
integrated into rice chromosomal DNA and expressed. Drought test indicated that glycinebetaine acts as an osmoprotectant and
its production in transgenic rice plant helped the cells to maintain osmotic potential and increased root growth, and thus
enhanced the ability of the plants to tolerate water deficit
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
12.
Phytochelatins (PCs) are post-translationally synthesized thiol reactive peptides that play important roles in detoxification
of heavy metal and metalloids in plants and other living organisms. The overall goal of this study is to develop transgenic
plants with increased tolerance for and accumulation of heavy metals and metalloids from soil by expressing an Arabidopsis
thaliana
AtPCS1 gene, encoding phytochelatin synthase (PCS), in Indian mustard (Brassica juncea L.). A FLAG-tagged AtPCS1 gDNA, under its native promoter, is expressed in Indian mustard, and transgenic pcs lines have been compared with wild-type
plants for tolerance to and accumulation of cadmium (Cd) and arsenic (As). Compared to wild type plants, transgenic plants
exhibit significantly higher tolerance to Cd and As. Shoots of Cd-treated pcs plants have significantly higher concentrations
of PCs and thiols than those of wild-type plants. Shoots of wild-type plants accumulated significantly more Cd than those
of transgenic plants, while accumulation of As in transgenic plants was similar to that in wild type plants. Although phytochelatin
synthase improves the ability of Indian mustard to tolerate higher levels of the heavy metal Cd and the metalloid As, it does
not increase the accumulation potential of these metals in the above ground tissues of Indian mustard plants. 相似文献
13.
14.
V. Prabhavathi J.S. Yadav P.A. Kumar M.V. Rajam 《Molecular breeding : new strategies in plant improvement》2002,9(2):137-147
In the present work, the bacterial mannitol-1-phosphodehydrogenase(mtlD) gene was introduced into eggplant(Solanummelongena L.) by Agrobacteriumtumefaciens-mediated transformation. Several transformants weregenerated and the transgene integration was confirmed by PCR, dot blot andSouthern blot analysis. Transgenic lines of T0 and T1generations were examined for tolerance to NaCl-induced salt stress,polyethylene glycol-mediated drought and chilling stress under bothinvitro and in vivo growth conditions. Aconsiderable proportions of transgenic seeds germinated and seedlings grew wellon 200 mM salt-amended MS basal medium, whereas seeds ofuntransformed control plants failed to germinate. Further, leaf explants fromthe transgenics could grow and showed signs of shoot regeneration onsalt-amended MS regeneration medium, whereas wild type did not respond, and infact the explants showed necrosis and loss of chlorophyll after about one week.The transgenic leaves could also withstand desiccation, and transgenics couldgrow well under chilling stress, and hydroponic conditions with salt stress ascompared to wild type plants. Thus, the transgenic lines were found to betolerant against osmotic stress induced by salt, drought and chilling stress.The morphology of the transgenic plants was normal as controls, but thechlorophyll content was higher in some of the lines. These observations suggestthat mtlD gene can impart abiotic stress tolerance ineggplant. 相似文献
15.
Matsunaga E Nanto K Oishi M Ebinuma H Morishita Y Sakurai N Suzuki H Shibata D Shimada T 《Plant cell reports》2012,31(1):225-235
Eucalyptus globulus is one of the most economically important plantation hardwoods for paper making. However, its low transformation frequency
has prevented genetic engineering of this species with useful genes. We found the hypocotyl section with a shoot apex has
the highest regeneration ability among another hypocotyl sections, and have developed an efficient Agrobacterium-mediated transformation method using these materials. We then introduced a salt tolerance gene, namely a bacterial choline
oxidase gene (codA) with a GUS reporter gene, into E. globulus. The highest frequency of transgenic shoot regeneration from hypocotyls with shoot apex was 7.4% and the average frequency
in four experiments was 4.0%, 12-fold higher than that from hypocotyls without shoot apex. Using about 10,000 explants, over
250 regenerated buds were confirmed as transformants by GUS analysis. Southern blot analysis of 100 elongated shoots confirmed
successful generation of stable transformants. Accumulation of glycinebetaine was investigated in 44 selected transgenic lines,
which showed 1- to 12-fold higher glycinebetaine levels than non-transgenic controls. Rooting of 16 transgenic lines was successful
using a photoautotrophic method under enrichment with 1,000 ppm CO2. The transgenic whole plantlets were transplanted into potting soil and grown normally in a growth room. They showed salt
tolerance to 300 mM NaCl. The points of our system are using explants with shoot apex as materials, inhibiting the elongation
of the apex on the selection medium, and regenerating transgenic buds from the side opposite to the apex. This approach may
also solve transformation problems in other important plants. 相似文献
16.
Tocopherols (vitamin E) are lipid soluble antioxidants synthesized by plants and some cyanobacteria. We have earlier reported that overexpression of the γ-tocopherol methyl transferase (γ-TMT) gene from Arabidopsis thaliana in transgenic Brassica juncea plants resulted in an over six-fold increase in the level of α-tocopherol, the most active form of all the tocopherols. Tocopherol levels have been shown to increase in response to a variety of abiotic stresses. In the present study on Brassica juncea, we found that salt, heavy metal and osmotic stress induced an increase in the total tocopherol levels. Measurements of seed germination, shoot growth and leaf disc senescence showed that transgenic Brassica juncea plants overexpressing the γ-TMT gene had enhanced tolerance to the induced stresses. Analysis of the chlorophyll a fluorescence rise kinetics, from the initial “O” level to the “P” (the peak) level, showed that there were differential effects of the applied stresses on different sites of the photosynthetic machinery; further, these effects were alleviated in the transgenic (line 16.1) Brassica juncea plants. We show that α-tocopherol plays an important role in the alleviation of stress induced by salt, heavy metal and osmoticum in Brassica juncea. 相似文献
17.
Brini F Hanin M Lumbreras V Amara I Khoudi H Hassairi A Pagès M Masmoudi K 《Plant cell reports》2007,26(11):2017-2026
Late Embryogenesis Abundant (LEA) proteins are associated with tolerance to water-related stress. A wheat (Triticum durum) group 2 LEA proteins, known also as dehydrin (DHN-5), has been previously shown to be induced by salt and abscisic acid
(ABA). In this report, we analyze the effect of ectopic expression of Dhn-5 cDNA in Arabidopsis thaliana plants and their response to salt and osmotic stress. When compared to wild type plants, the Dhn-5 transgenic plants exhibited stronger growth under high concentrations of NaCl or under water deprivation, and showed a faster
recovery from mannitol treatment. Leaf area and seed germination rate decreased much more in wild type than in transgenic
plants subjected to salt stress. Moreover, the water potential was more negative in transgenic than in wild type plants. In
addition, the transgenic plants have higher proline contents and lower water loss rate under water stress. Also, Na+ and K+ accumulate to higher contents in the leaves of the transgenic plants. Our data strongly support the hypothesis that Dhn-5, by its protective role, contributes to an improved tolerance to salt and drought stress through osmotic adjustment. 相似文献
18.
Ectopic expression of wheat expansin gene TaEXPA2 improved the salt tolerance of transgenic tobacco by regulating Na+/K+ and antioxidant competence
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Yangyang Han Xiangzhu Kong Hanhan Kang Yuanqing Ren Wei Wang 《Physiologia plantarum》2017,159(2):161-177
High salinity is one of the most serious environmental stresses that limit crop growth. Expansins are cell wall proteins that regulate plant development and abiotic stress tolerance by mediating cell wall expansion. We studied the function of a wheat expansin gene, TaEXPA2, in salt stress tolerance by overexpressing it in tobacco. Overexpression of TaEXPA2 enhanced the salt stress tolerance of transgenic tobacco plants as indicated by the presence of higher germination rates, longer root length, more lateral roots, higher survival rates and more green leaves under salt stress than in the wild type (WT). Further, when leaf disks of WT plants were incubated in cell wall protein extracts from the transgenic tobacco plants, their chlorophyll content was higher under salt stress, and this improvement from TaEXPA2 overexpression in transgenic tobacco was inhibited by TaEXPA2 protein antibody. The water status of transgenic tobacco plants was improved, perhaps by the accumulation of osmolytes such as proline and soluble sugar. TaEXPA2‐overexpressing tobacco lines exhibited lower Na+ but higher K+ accumulation than WT plants. Antioxidant competence increased in the transgenic plants because of the increased activity of antioxidant enzymes. TaEXPA2 protein abundance in wheat was induced by NaCl, and ABA signaling was involved. Gene expression regulation was involved in the enhanced salt stress tolerance of the TaEXPA2 transgenic plants. Our results suggest that TaEXPA2 overexpression confers salt stress tolerance on the transgenic plants, and this is associated with improved water status, Na+/K+ homeostasis, and antioxidant competence. ABA signaling participates in TaEXPA2‐regulated salt stress tolerance. 相似文献
19.
该研究在实验室前期研究的基础上,将受脱水、盐胁迫和ABA诱导的柠条锦鸡儿CkLEA4基因转入野生型拟南芥,并利用实时荧光定量PCR从8株纯合体中筛选出3个表达量不同的株系,比较野生型和转CkLEA4基因过表达拟南芥种子在不同胁迫处理下的萌发率,以探讨CkLEA4基因在植物抵抗逆境胁迫中的功能。结果发现:(1)在不同浓度NaCl、甘露醇及ABA处理下,转CkLEA4基因过表达拟南芥种子的萌发率均高于野生型,随着NaCl、甘露醇及ABA浓度增加,各株系萌发率均降低,但野生型的萌发率下降幅度均高于3个过表达株系,并且在200mmol/L NaCl和400mmol/L甘露醇处理下,过表达株系子叶绿化率均显著高于野生型。(2)在低浓度ABA处理下,CkLEA4过表达植株子叶的绿化率也高于野生型。研究表明,柠条锦鸡儿CkLEA4基因提高了拟南芥种子萌发阶段对盐、ABA及渗透胁迫的耐受性。 相似文献
20.
In order to investigate the function of chloroplast ascorbate peroxidase under temperature stress, the thylakoid-bound ascorbate
peroxidase gene from tomato leaf (TtAPX) was introduced into tobacco. Transformants were selected for their ability to grow on medium containing kanamycin. RNA gel
blot analysis confirmed that TtAPX in tomato was induced by chilling or heat stress. Over-expression of TtAPX in tobacco improved seed germination under temperature stress. Two transgenic tobacco lines showed higher ascorbate peroxidase
activity, accumulated less hydrogen peroxide and malondialdehyde than wild type plants under stress condition. The photochemical
efficiency of photosystem 2 in the transgenic lines was distinctly higher than that of wild type plants under chilling and
heat stresses. Results indicated that the over-expression of TtAPX enhanced tolerance to temperature stress in transgenic tobacco plants. 相似文献