共查询到20条相似文献,搜索用时 31 毫秒
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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
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Wei Hu Jialu Feng Fan Zhang Lihong Chen Chao Huang Qingchen Luo Yanzhen He Guangxiao Yang Guangyuan He 《Physiologia plantarum》2013,149(3):367-377
Calcineurin B‐like protein‐interacting protein kinases (CIPKs) are components of Ca2+ signaling in responses to abiotic stresses. In this work, the full‐length cDNA of a novel CIPK gene (TaCIPK14) was isolated from wheat and was found to have significant sequence similarity to OsCIPK14/15. Subcellular localization assay revealed the presence of TaCIPK14 throughout the cell. qRT‐PCR analysis showed that TaCIPK14 was upregulated under cold conditions or when treated with salt, PEG or exogenous stresses related signaling molecules including ABA, ethylene and H2O2. Transgenic tobaccos overexpressing TaCIPK14 exhibited higher contents of chlorophyll and sugar, higher catalase activity, while decreased amounts of H2O2 and malondialdehyde, and lesser ion leakage under cold and salt stresses. In addition, overexpression also increased seed germination rate, root elongation and decreased Na+ content in the transgenic lines under salt stress. Higher expression of stress‐related genes was observed in lines overexpressing TaCIPK14 compared to controls under stress conditions. In summary, these results suggested that TaCIPK14 is an abiotic stress‐responsive gene in plants. 相似文献
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Transgenic cotton (Gossypium hirsutum L.) lines expressing the tobacco glutathione S-transferase (GST) Nt107 were evaluated for tolerance to chilling, salinity, and herbicides, antioxidant enzyme activity, antioxidant compound levels, and lipid peroxidation. Although transgenic seedlings exhibited ten-fold and five-fold higher GST activity under normal and salt-stress conditions, respectively, germinating seedlings did not show improved tolerance to salinity, chilling conditions, or herbicides. Glutathione peroxidase (GPX) activity in transgenic seedlings was 30% to 60% higher under normal conditions, but was not different than GPX activity in wild-type seedlings under salt-stress conditions. Glutathione reductase, superoxide dismutase, ascorbate peroxidase, and monodehydroascorbate reductase activities were not increased in transgenic seedlings under salt-stress conditions, while dehydroascorbate reductase activity was decreased in transgenic seedlings under salt-stress conditions. Transgenic seedlings had 50% more oxidized glutathione when exposed to salt stress. Ascorbate levels were not increased in transgenic seedlings under salt-stress conditions. Malondialdehyde content in transgenic seedlings was nearly double that of wild-type seedlings under normal conditions and did not increase under salt-stress conditions. These results show that expression of Nt107 in cotton does not provide adequate protection against oxidative stress and suggests that the endogenous antioxidant system in cotton may be disrupted by the expression of the tobacco GST. 相似文献
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Supaporn Hasthanasombut Valentine Ntui Kanyaratt Supaibulwatana Masahiro Mii Ikuo Nakamura 《Plant biotechnology reports》2010,4(1):75-83
Glycine betaine has been reported as an osmoprotectant compound conferring tolerance to salinity and osmotic stresses in plants.
We previously found that the expression of betaine aldehyde dehydrogenase 1 gene (OsBADH1), encoding a key enzyme for glycine betaine biosynthesis pathway, showed close correlation with salt tolerance of rice. In
this study, the expression of the OsBADH1 gene in transgenic tobacco was investigated in response to salt stress using a transgenic approach. Transgenic tobacco plants
expressing the OsBADH1 gene were generated under the control of a promoter from the maize ubiquitin gene. Three homozygous lines of T2 progenies with single transgene insert were chosen for gene expression analysis. RT-PCR and western blot analysis results
indicated that the OsBADH1 gene was effectively expressed in transgenic tobacco leading to the accumulation of glycine betaine. Transgenic lines demonstrated
normal seed germination and morphology, and normal growth rates of seedlings under salt stress conditions. These results suggest
that the OsBADH1 gene could be an excellent candidate for producing plants with osmotic stress tolerance. 相似文献
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Manaswini Das Harsh Chauhan Anju Chhibbar Qazi Mohd. Rizwanul Haq Paramjit Khurana 《Transgenic research》2011,20(2):231-246
Osmotin and osmotin-like proteins are stress proteins belonging to the plant PR-5 group of proteins induced in several plant
species in response to various types of biotic and abiotic stresses. We report here the overexpression of tobacco osmotin
in transgenic mulberry plants under the control of a constitutive promoter (CaMV 35S) as well as a stress-inducible rd29A promoter. Southern analysis of the transgenic plants revealed the stable integration of the introduced genes in the transformants.
Real-time PCR analysis provided evidence for the expression of osmotin in the transgenic plants under both the constitutive
and stress-inducible promoters. Transgenic plants with the stress-inducible promoter were observed to better tolerate salt
and drought stress than those with the constitutive promoter. Transgenic plants when subjected to simulated salinity and drought
stress conditions showed better cellular membrane stability (CMS) and photosynthetic yield than non-transgenic plants under
conditions of both salinity and drought stress. Proline levels were very high in transgenic plants with the constitutive promoter
relative to those with the stress-inducible promoter. Fungal challenge undertaken with three fungal species known to cause
serious losses to mulberry cultivation, namely, Fusarium pallidoroseum, Colletotrichum gloeosporioides and Colletotrichum dematium, revealed that transgenic plants with osmotin under control of the constitutive promoter had a better resistance than those with osmotin under the control of the stress-inducible promoter. Evaluation in next generation was undertaken by studying bud break in
transgenic and non-transgenic plants under simulated drought (2% polyethylene glycol) and salt stress (200 mM NaCl) conditions.
The axillary buds of the selected transgenic lines had a better bud break percentage under stressed conditions than buds from
non-transgenic mulberry lines. A biotic assay with Bombyx mori indicated that osmotin protein had no undesirable effect on silkworm rearing and feeding. We therefore conclude that 35S
transgenic plants are better suited for both abiotic stress also biotic challenges (fungal), while the rd29A transgenic plants
are more responsive to drought. 相似文献
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Jing Qu Shuang Liu Peng Jiao Zhenzhong Jiang Jianbo Fei Shuyan Guan Yiyong Ma 《Phyton》2022,91(8):1709-1719
To develop cold-tolerant maize germplasms and identify the activation of INDUCER OF CRT/DRE-BINDING FACTOR EXPRESSION (ICE1) expression in response to cold stress, RT-PCR was used to amplify the complete open reading frame sequence of the ICE1 gene and construct the plant expression vector pCAMBIA3301-ICE1-Bar. Immature maize embryos and calli were transformed with the recombinant vector using Agrobacterium tumefaciens-mediated transformations. From the regenerated plantlets, three T1 lines were screened and identified by PCR. A Southern blot analysis showed that a single copy of the ICE1 gene was integrated into the maize (Zea mays L.) genomes of the three T1 generations. Under low temperature-stress conditions (4°C), the relative conductivity levels decreased by 27.51%–31.44%, the proline concentrations increased by 12.50%–17.50%, the malondialdehyde concentrations decreased by 16.78%–18.37%, and the peroxidase activities increased by 19.60%–22.89% in the T1 lines compared with those of the control. A real-time quantitative PCR analysis showed that the ICE1 gene was ectopically expressed in the roots, stems, and leaves of the T1 lines. ICE1 positively regulates the expression of the CBF genes in response to cold stress. Thus, this study showed the successful transformation of maize with the ICE1 gene, resulting in the generation of a new maize germplasm that had increased tolerance to cold stress. 相似文献
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Shunwu Yu Aona Huang Jia Li Lin Gao Yanni Feng Erinn Pemberton Chunli Chen 《Plant Growth Regulation》2018,84(3):519-531
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Z. -Y. Wang F. -B. Song H. Cai Y. -M. Zhu X. Bai W. Ji Y. Li Y. Hua 《Biologia Plantarum》2012,56(3):516-520
Glutathione-S-transferases (GSTs) are ubiquitous enzymes that play a key role in stress tolerance and cellular detoxification.
The GST gene GsGST14 selected from the gene expression profiles of Glycine soja under alkaline stress was transformed into alfalfa (Medicago sativa L.). Transgenic alfalfa plants showed 1.73–1.99 times higher GST activity than wild-type plants. Transgenic alfalfa grew
well in the presence of 100 mM NaHCO3, while wild-type plants exhibited chlorosis and stunted growth, even death. There were marked changes in malondialdehyde
content and relative membrane permeability caused by alkaline stress in non-transgenic lines compared to transgenic lines.
The results indicate that the gene GsGST14 could enhance alkaline resistance in transgenic alfalfa. 相似文献
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E. S. Seong J. H. Yoo N. J. Kim J. H. Choi J. G. Lee B. K. Ghimire I. M. Chung C. Y. Yu 《Russian Journal of Plant Physiology》2016,63(1):124-131
The tomato bZIP2-encoding gene was inserted into the Nicotiana benthamiana genome using Agrobacterium-mediated transformation to characterize resistance to oxidative stress and two herbicides, glyphosate and paraquat. We produced transgenic tobacco plants using the LebZIP2 gene, which were then utilized to examine salt stress and herbicide resistance through oxidative mechanisms. Transgenic LebZIP2-overexpressing plants were examined using specific primers for selection marker genes (PCR using genomic DNA) and target genes (RT-PCR). Based on microscopic examination, we observed an increase in leaf thickness and cell number in transgenic plants. The electrolyte leakage of leaves suggested that LebZIP2-overexpressing lines were weak tolerant to NaCl stress and resistant to methyl viologen. During our analysis, transgenic lines were exposed to different herbicides. Transgenic plants showed an increased tolerance based on visual injury, as well as an increased biomass. Based on these results, the LebZIP2 gene may be involved in oxidative stress tolerance and cell development in plants. 相似文献
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The study was conducted to determine the effects of expression of a transgene encoding adenine isopentenyl transferase (ipt), which controls cytokinin synthesis, on growth and leaf senescence of creeping bentgrass (Agrostis stolonifera L.), subjected to heat stress. Creeping bentgrass (cv. Penncross) was transformed with ipt ligated to a senescence-activated promoter (SAG12). Eight SAG12-ipt transgenic lines exhibiting desirable turf quality and a transgenic control line (transformed with the empty vector) were
evaluated for morphological and physiological changes under normal growth temperature (20°C) and after 14 days of heat stress
(35°C) in growth chambers. Six of the SAG12-ipt lines developed more tillers than the control line during establishment under normal growth temperature of 20°C. Following
14 days of heat stress, four of the SAG12-ipt lines had increased 65–83% of roots and for all six SAG12-ipt lines root elongation continued, whereas root production ceased and total root length decreased for the control line. Root
isopentenyl adenine (iPA) content increased 2.5–3.5 times in five of the SAG12-ipt lines, whereas in the control line iPA decreased 20% after 14 days at 35°C. Total zeatin riboside (ZR) content was maintained
at the original level or increased in five of the SAG12-ipt lines, whereas in the control line ZR decreased under heat stress. Our results suggest expression of SAG12-ipt in creeping bentgrass stimulated tiller formation and root production, and delayed leaf senescence under heat stress, suggesting
a role for cytokinins in regulating cool-season grass tolerance to heat stress. 相似文献
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Yun-Hee Kim Soon Lim Kyoung-Sil Yang Cha Young Kim Suk-Yoon Kwon Haeng-Soon Lee Xin Wang Zhilin Zhou Daifu Ma Dae-Jin Yun Sang-Soo Kwak 《Molecular breeding : new strategies in plant improvement》2009,24(3):233-244
Transgenic sweetpotato (Ipomoea batatas L. cv. Yulmi) plants expressing the Arabidopsis nucleoside diphosphate kinase 2 (AtNDPK2) gene under the control of an oxidative stress–inducible peroxidase (SWPA2) promoter (referred to as SN plants) were developed and evaluated for enhanced tolerance of SN plants under various abiotic stress conditions. The level of AtNDPK2 expression and NDPK activity in SN plants following methyl viologen (MV) treatment was positively correlated with the plant’s tolerance to MV. Interestingly, we observed that antioxidant enzyme activities such as peroxidase, ascorbate peroxidase, and catalase increased in MV-treated SN plants. In addition, SN plants showed enhanced tolerance to cold, high salinity, and drought stresses by an increase in the activity of H2O2 scavenging enzymes. These results indicate that overexpression of AtNDPK2 in sweetpotato might efficiently modulate oxidative stress from various environmental stresses. 相似文献
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Masaru Tanaka Yasuhiro Takahata Hiroki Nakayama Makoto Nakatani Makoto Tahara 《Planta》2009,230(4):737-746
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Shu Chen Mingliang Chai Yufang Jia Zhongshan Gao Li Zhang Minxia Gu 《Plant Cell, Tissue and Organ Culture》2011,107(3):493-500
To study growth in the presence of NaCl, in vitro plantlets regenerated from callus of manilagrass (Zoysia matrella [L.] Merr.) were cultured on regeneration medium supplemented with or without 0.3 M NaCl. The results indicated that growth
was significantly inhibited by NaCl, with the leaves becoming relatively shorter and thicker. The differences of in vitro
plantlets grown under NaCl stress provided specific criteria for the selection of salt tolerant variants. The 6-year maintained
calli were treated with different doses (0, 5, 10, 20, 40, 80, 100, 150, 200, 250, and 300 Gy) of 60Co γ rays. Regeneration rate and regeneration capacity of the calli were highest after treatment with 20 Gy 60Co γ rays, 27.08 and 91.67% respectively. When the irradiation dose was increased to 100 Gy, 10.42% of the calli developed
shoots, but at 150 Gy, both regeneration capacity and regeneration rate declined significantly, and no shoot was observed
after 6 weeks of regeneration. Therefore, 100–150 Gy is the most appropriate irradiation span for inducing somaclonal variation.
The irradiated calli were selected in vitro for NaCl tolerance. Five NaCl tolerant variant lines, Ze1, Fv1, Te1, Tw1, Fr1,
were selected on subculture medium supplemented with 0.35 M NaCl, then transferred to regeneration medium containing 0.25 M
NaCl, and grown in a greenhouse. The dark green colour index (DGCI) was used to identify the amount of injury caused by NaCl
treatment. This was significantly higher in four of the lines, Ze1, Fv1, Te1, Fr1 (30.88, 31.17, 30.45 and 37.70%, respectively)
compared to the control line (CK), which was regenerated from calli subcultured monthly (27.39%), indicating that watering
with NaCl caused less injury in these four lines. These lines had lower proline contents than CK under salt stress. The superoxide
dismutase (SOD) activity was higher in Ze1 under control condition and its peroxidase (POD) activity increased significantly
under salt stress. With Fr1 catalase (CAT) activity was higher under salt stress. The higher activity of these antioxidant
enzymes may contribute to the enhanced salt tolerance of the four plant lines. 相似文献
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Young-Mi Shin Goh Choe Byoungchul Shin Gibum Yi Pil-Yong Yun Kiyoung Yang Joon Seol Lee Sang-Soo Kwak Kyung-Moon Kim 《Journal of Plant Biology》2007,50(2):206-212
We have used two aminoglycosides, G418 and paromomycin, to develop a reliable selection system fornptll transgenic sweet-potato (Ipomoea batatas (L.) Lam.). Embryogenic calli derived from shoot apical meristems were bombarded with gold particles coated with pCAMBIA2301,
which contained thenptll andgusA genes. When compared on a kill curve that was based on calli proliferation and cell viability, G413-selection proved to be more efficient and had fewer escapes than kanamycin. These bombarded expiants were then selected on
G418-containing media. The total time required from bombardment to plant establishment in soil was seven to nine months. Multiple
copies of the transgene were integrated into the sweetpotato genome. Northern analysis confirmed transgene expression in the
regenerated plants, and a paromomycin assay demonstrated that thenptll gene was functionally expressed in transformed sweetpotato. These molecular analyses and assays all showed that selection
with G418 and paromomycin is reliable. So far, we have produced 69 transgenic events with this system, at a transformation frequency
of approx. 1.1%. That efficiency is based on the number of transgenic plants obtained and the amount of calli bombarded. Thus,
this selection method that combines G418 with paromomycin is now available for selectingnptll transgenic sweetpotato. 相似文献