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1.
Overexpression of the trehalose-6-phosphate synthase gene <Emphasis Type="Italic">OsTPS1</Emphasis> enhances abiotic stress tolerance in rice 总被引:1,自引:0,他引:1
Trehalose plays an important role in metabolic regulation and abiotic stress tolerance in a variety of organisms. In plants,
its biosynthesis is catalyzed by two key enzymes: trehalose-6-phosphate synthase (TPS) and trehalose-6-phosphate phosphatase
(TPP). The genome of rice (Oryza sativa) contains 11 OsTPS genes, and only OsTPS1 shows TPS activity. To demonstrate the physiological function of OsTPS1, we introduced it into rice
and found that OsTPS1 overexpression improved the tolerance of rice seedling to cold, high salinity and drought treatments without other significant
phenotypic changes. In transgenic lines overexpressing OsTPS1, trehalose and proline concentrations were higher than in the wild type and some stress-related genes were up-regulated,
including WSI18, RAB16C, HSP70, and ELIP. These results demonstrate that OsTPS1 may enhance the abiotic stress tolerance of plants by increasing the amount of trehalose
and proline, and regulating the expression of stress-related genes. Furthermore, we found that overexpression of some Class
II TPSs also enhanced plant tolerance of abiotic stress. This work will help to clarify the role of trehalose metabolism in abiotic
stress response in higher plants. 相似文献
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Analysis of trehalose-6-phosphate synthase (TPS) gene family suggests the formation of TPS complexes in rice 总被引:2,自引:0,他引:2
Trehalose-6-phosphate (T6P), an intermediate in the trehalose biosynthesis pathway, is emerging as an important regulator
of plant metabolism and development. T6P levels are potentially modulated by a group of trehalose-6-phosphate synthase (TPS)
and trehalose-6-phosphate phosphatase (TPP) homologues. In this study, we have isolated 11 TPS genes encoding proteins with both TPS and TPP domains, from rice. Functional complement assays performed in yeast tps1 and tps2 mutants, revealed that only OsTPS1 encodes an active TPS enzyme and no OsTPS protein possesses TPP activity. By using a yeast two-hybrid analysis, a complicated
interaction network occurred among OsTPS proteins, and the TPS domain might be essential for this interaction to occur. The
interaction between OsTPS1 and OsTPS8 in vivo was confirmed by bimolecular fluorescence complementation and coimmunoprecipitation
assays. Furthermore, our gel filtration assay showed that there may exist two forms of OsTPS1 (OsTPS1a and OsTPS1b) with different
elution profiles in rice. OsTPS1b was particularly cofractionated with OsTPS5 and OsTPS8 in the 360 kDa complex, while OsTPS1a
was predominantly incorporated into the complexes larger than 360 kDa. Collectively, these results suggest that OsTPS family members may form trehalose-6-phosphate synthase complexes and therefore potentially modify T6P levels to regulate
plant development. 相似文献
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Cloning and Characterization of Functional Trehalose-6-Phosphate Synthase Gene in Maize 总被引:1,自引:0,他引:1
Wei Jiang Feng-Ling Fu Su-Zhi Zhang Ling Wu Wan-Chen Li 《Journal of Plant Biology》2010,53(2):134-141
Trehalose is a non-reducing disaccharide of glucose that functions as a compatible solute in the stabilization of biological
structures under heat and desiccation stress in bacteria, fungi, and some “resurrection plants”. In the plant kingdom, trehalose
is biosynthesized by trehalose-6-phosphate synthase (TPS) and trehalose-6-phosphate phosphatase (TPP). Over-expression of
exogenous and endogenous genes encoding TPS and TPP is reported to be effective for improving abiotic stress tolerance in
tobacco, potato, tomato, rice, and Arabidopsis. On the basis of bioinformatics prediction, we cloned a fragment containing an open reading frame of 2,820 bp from maize,
which encodes a protein of 939 amino acids. Phylogenetic analysis showed that this gene belongs to the class I subfamily of
the TPS gene family. Analysis of conserved domains revealed the presence of a TPS domain and a TPP domain. Yeast complementation
with TPS and TPP mutants demonstrated that this protein has the activity of trehalose-6-phosphate synthase. Semi-quantitative RT-PCR and real-time
quantitative PCR indicated that the expression of this gene is upregulated in response to both salt and cold stress. 相似文献
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The rice terpene synthase gene OsTPS19 functions as an (S)‐limonene synthase in planta,and its overexpression leads to enhanced resistance to the blast fungus Magnaporthe oryzae
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Xujun Chen Joshua S. Yuan Tobias G. Köllner Yuying Chen Yufen Guo Xiaofeng Zhuang Xinlu Chen Yong‐jun Zhang Jianyu Fu Andreas Nebenführ Zejian Guo Feng Chen 《Plant biotechnology journal》2018,16(10):1778-1787
Rice blast disease, caused by the fungus Magnaporthe oryzae, is the most devastating disease of rice. In our ongoing characterization of the defence mechanisms of rice plants against M. oryzae, a terpene synthase gene OsTPS19 was identified as a candidate defence gene. Here, we report the functional characterization of OsTPS19, which is up‐regulated by M. oryzae infection. Overexpression of OsTPS19 in rice plants enhanced resistance against M. oryzae, while OsTPS19 RNAi lines were more susceptible to the pathogen. Metabolic analysis revealed that the production of a monoterpene (S)‐limonene was increased and decreased in OsTPS19 overexpression and RNAi lines, respectively, suggesting that OsTPS19 functions as a limonene synthase in planta. This notion was further supported by in vitro enzyme assays with recombinant OsTPS19, in which OsTPS19 had both sesquiterpene activity and monoterpene synthase activity, with limonene as a major product. Furthermore, in a subcellular localization experiment, OsTPS19 was localized in plastids. OsTPS19 has a highly homologous paralog, OsTPS20, which likely resulted from a recent gene duplication event. We found that the variation in OsTPS19 and OsTPS20 enzyme activities was determined by a single amino acid in the active site cavity. The expression of OsTPS20 was not affected by M. oryzae infection. This indicates functional divergence of OsTPS19 and OsTPS20. Lastly, (S)‐limonene inhibited the germination of M. oryzae spores in vitro. OsTPS19 was determined to function as an (S)‐limonene synthase in rice and plays a role in defence against M. oryzae, at least partly, by inhibiting spore germination. 相似文献
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Hongxiu Gao Lin Zhu Tianqi Liu Xueyu Leng Zhenxing Zhu Wei Xie Haitao Lv Zhengxun Jin Ping Wu Zhongchen Zhang 《Phyton》2023,92(6):1743-1763
Cyclophilin (CYP) plays an important role in plant response to stress, and OsCYP2, one gene of cyclophlilin
family, is involved in auxin signal transduction and stress signaling in rice. However, the mechanism that OsCYP2
is involved in rice response to low temperature is still unclear. We identified a new OsCYP2 allelic mutant, lrl3,
with fewer lateral roots, and the differences in shoot height, primary root length and adventitious root length
increased with the growth process compared to the wild-type plant. Auxin signaling pathway was also affected
and became insensitive to gravity. The transgenic rice plants with over-expression of OsCYP2 were more tolerant
to low temperature than the wild-type plants, suggesting that OsCYP2 was involved in the low temperature
response in rice. In addition, OsCYP2 negatively regulated the expression of OsTPS38, a terpene synthase gene,
and was dependent on the OsCDPK7-mediated pathway in response to low temperature stress. OsTPS38-
overexpressed transgenic line ox-2 was more sensitive to low temperature. Therefore, OsCYP2 may negatively
regulate OsTPS38 through an OsCDPK7-dependent pathway to mediate the response to low temperature in rice.
These results provide a new basis for auxin signaling genes to regulate rice response to low temperature stress. 相似文献
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Sha Y Li S Pei Z Luo L Tian Y He C 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2004,108(2):306-314
Insertional mutagenesis provides a rapid way to clone a mutated gene. Transfer DNA (T-DNA) of Agrobacterium tumefaciens has been proven to be a successful tool for gene discovery in Arabidopsis and rice (Oryza sativa L. ssp. japonica). Here, we report the generation of 5,200 independent T-DNA tagged rice lines. The T-DNA insertion pattern in the rice genome was investigated, and an initial database was constructed based on T-DNA flanking sequences amplified from randomly selected T-DNA tagged rice lines using Thermal Asymmetric Interlaced PCR (TAIL-PCR). Of 361 T-DNA flanking sequences, 92 showed long T-DNA integration (T-DNA together with non-T-DNA). Another 55 sequences showed complex integration of T-DNA into the rice genome. Besides direct integration, filler sequences and microhomology (one to several nucleotides of homology) were observed between the T-DNA right border and other portions of the vector pCAMBIA1301 in transgenic rice. Preferential insertion of T-DNA into protein-coding regions of the rice genome was detected. Insertion sites mapped onto rice chromosomes were scattered in the genome. Some phenotypic mutants were observed in the T1 generation of the T-DNA tagged plants. Our mutant population will be useful for studying T-DNA integration patterns and for analyzing gene function in rice.Electronic Supplementary Material Supplementary material is available in the online version of this article at .Communicated by D. Mackill 相似文献
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We describe here the isolation and characterization of OsiSAP8, a member of stress Associated protein (SAP) gene family from rice characterized by the presence of A20 and AN1 type Zinc
finger domains. OsiSAP8 is a multiple stress inducible gene, induced by various stresses, namely heat, cold, salt, desiccation, submergence, wounding,
heavy metals as well as stress hormone Abscisic acid. OsiSAP8 protein fused to GFP was localized towards the periphery of
the cells in the epidermal cells of infiltrated Nicotiana benthamiana leaves. Yeast two hybrid analysis revealed that A20 and AN1 type zinc-finger domains of OsiSAP8 interact with each other.
Overexpression of the gene in both transgenic tobacco and rice conferred tolerance to salt, drought and cold stress at seed
germination/seedling stage as reflected by percentage of germination and gain in fresh weight after stress recovery. Transgenic
rice plants were tolerant to salt and drought during anthesis stage without any yield penalty as compared to unstressed transgenic
plants.
OsiSAP8 is deposited in the Genbank with the Accession number AY345599. 相似文献
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The bZIP transcription factor OsABF1 is an ABA responsive element binding factor that enhances abiotic stress signaling in rice 总被引:5,自引:0,他引:5
Md. Amir Hossain Yongjoo Lee Jung-Il Cho Chul-Hyun Ahn Sang-Kyu Lee Jong-Seong Jeon Hun Kang Choon-Hwan Lee Gynheung An Phun Bum Park 《Plant molecular biology》2010,72(4-5):557-566
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J Sook Chung 《Aquatic biosystems》2008,4(1):1-8
Trehalose in ectoderms functions in energy metabolism and protection in extreme environmental conditions. We structurally characterized trehalose 6-phosphate synthase (TPS) from hemocytes of the blue crab, Callinectes sapidus. C. sapidus Hemo TPS (CasHemoTPS), like insect TPS, encodes both TPS and trehalose phosphate phosphatase domains. Trehalose seems to be a major sugar, as it shows higher levels than does glucose in hemocytes and hemolymph. Increases in HemoTPS expression, TPS enzyme activity in hemocytes, and hemolymph trehalose levels were determined 24 h after lipopolysaccharide challenge, suggesting that both TPS and TPP domains of CasHemoTPS are active and functional. The TPS gene has a wide tissue distribution in C. sapidus, suggesting multiple biosynthetic sites. A correlation between TPS activity in hemocytes and hemolymph trehalose levels was found during the molt cycle. The current study provides the first evidence of presence of trehalose in hemocytes and TPS in tissues of C. sapidus and implicates its functional role in energy metabolism and physiological adaptation. 相似文献
16.
Zhiguo Zhang Quian Zhang Jinxia Wu Xia Zheng Sheng Zheng Xuehui Sun Quansheng Qiu Tiegang Lu 《PloS one》2013,8(2)
Plant ascorbate peroxidases (APXs), enzymes catalyzing the dismutation of H2O2 into H2O and O2, play an important role in reactive oxygen species homeostasis in plants. The rice genome has eight OsAPXs, but their physiological functions remain to be determined. In this report, we studied the function of OsAPX2 gene using a T-DNA knockout mutant under the treatment of drought, salt and cold stresses. The Osapx2 knockout mutant was isolated by a genetic screening of a rice T-DNA insertion library under 20% PEG-2000 treatment. Loss of function in OsAPX2 affected the growth and development of rice seedlings, resulting in semi-dwarf seedlings, yellow-green leaves, leaf lesion mimic and seed sterility. OsAPX2 expression was developmental- and spatial-regulated, and was induced by drought, salt, and cold stresses. Osapx2 mutants had lower APX activity and were sensitive to abiotic stresses; overexpression of OsAPX2 increased APX activity and enhanced stress tolerance. H2O2 and MDA levels were high in Osapx2 mutants but low in OsAPX2-OX transgenic lines relative to wild-type plants after stress treatments. Taken together, the cytosolic ascorbate peroxidase OsAPX2 plays an important role in rice growth and development by protecting the seedlings from abiotic stresses through scavenging reactive oxygen species. 相似文献
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Xiao B Huang Y Tang N Xiong L 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2007,115(1):35-46
Late embryogenesis abundant (LEA) proteins have been implicated in many stress responses of plants. In this report, a LEA
protein gene OsLEA3-1 was identified and over-expressed in rice to test the drought resistance of transgenic lines under the field conditions.
OsLEA3-1 is induced by drought, salt and abscisic acid (ABA), but not by cold stress. The promoter of OsLEA3-1 isolated from the upland rice IRAT109 exhibits strong activity under drought- and salt-stress conditions. Three expression
constructs consisting of the full-length cDNA driven by the drought-inducible promoter of OsLEA3-1 (OsLEA3-H), the CaMV 35S promoter (OsLEA3-S), and the rice Actin1 promoter (OsLEA3-A) were transformed into the drought-sensitive japonica rice Zhonghua 11. Drought resistance pre-screening of T1 families at anthesis stage revealed that the over-expressing families with OsLEA3-S and OsLEA3-H constructs had significantly
higher relative yield (yield under drought stress treatment/yield under normal growth conditions) than the wild type under
drought stress conditions, although a yield penalty existed in T1 families under normal growth conditions. Nine homozygous families, exhibiting over-expression of a single-copy of the transgene
and relatively low yield penalty in the T1 generation, were tested in the field for drought resistance in the T2 and T3 generations and in the PVC pipes for drought tolerance in the T2 generation. Except for two families (transformed with OsLEA3-A), all the other families (transformed with OsLEA3-S and OsLEA3-H
constructs) had higher grain yield than the wild type under drought stress in both the field and the PVC pipes conditions.
No significant yield penalty was detected for these T2 and T3 families. These results indicate that transgenic rice with significantly enhanced drought resistance and without yield penalty
can be generated by over-expressing OsLEA3-1 gene with appropriate promoters and following a bipartite (stress and non-stress) in-field screening protocol. 相似文献