Ectopic expression of a novel cold-resistance protein 1 from Brassica oleracea promotes tolerance to chilling stress in transgenic tomato

AbstractCold stress is considered as one of the major environmental factors that adversely affects the plant growth and distribution. Therefore, there arises an immediate need to cultivate effective strategies aimed at developing stress-tolerant crops that would boost the production and minimise the risks associated with cold stress. In this study, a novel cold-responsive protein1 (BoCRP1) isolated from Brassica oleracea was ectopically expressed in a cold susceptible tomato genotype Shalimar 1 and its function was investigated in response to chilling stress. BoCRP1 was constitutively expressed in all the tissues of B. oleracea including leaf, root and stem. However, its expression was found to be significantly increased in response to cold stress. Moreover, transgenic tomato plants expressing BoCRP1 exhibited increased tolerance to chilling stress (4 °C) with an overall improved rate of seed germination, increased root length, reduced membrane damage and increased accumulation of osmoprotectants. Furthermore, we observed increased transcript levels of stress responsive genes and enhanced accumulation of reactive oxygen species scavenging enzymes in transgenic plants on exposure to chilling stress. Taken together, these results strongly suggest that BoCRP1 is a promising candidate gene to improve the cold stress tolerance in tomato.

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Ectopic Expression of a Novel Cold-resistance Protein 1 (BoCRP1) from Brassica Oleracea Promotes Tolerance to Cold via Modulating Stress Associated Components

10.21203/rs.3.rs-168627/v1 ◽

2021 ◽

Author(s):

Umer Majeed Wani ◽

Tahir Majeed Sheikh ◽

Vaseem Raja ◽

Zubair Ahmad Wani ◽

Nelofer Jan ◽

...

Keyword(s):

Cold Stress ◽

Brassica Oleracea ◽

Chilling Stress ◽

Membrane Damage ◽

Ectopic Expression ◽

Ros Scavenging ◽

Effective Strategies ◽

Protective Shield ◽

Scavenging Enzymes ◽

Susceptible Tomato

Abstract Cold stress is considered as a major environmental factor that adversely affect the plant growth and distribution. Therefore, there arises an immediate need to cultivate effective strategies aimed at developing stress-tolerant crops that would boost the production and minimise the risks associated with cold stress. In this study, a novel cold-responsive protein1 isolated from Brassica oleracea (BoCRP1) was ectopically expressed in a cold susceptible tomato genotype Shalimar 1 and its function was investigated in response to chilling stress. BoCRP1 was constitutively expressed in all the tissues of B. oleracea including leaf, root and stem however, its expression was found to be significantly increased in response to cold stress. Moreover, transgenic tomato plants expressing BoCRP1 exhibited increased tolerance to chilling stress (4oC) with an overall improved rate of seed germination, increased root length, reduced membrane damage and increase in accumulation of osmoprotectants. Furthermore, we observed increased transcript levels of stress responsive genes and enhanced accumulation of ROS scavenging enzymes in transgenic on exposure to chilling stress. These results are therefore strongly in support of the role of BoCRP1 in offering the plant a protective shield and heightened resilience to chilling stress by maintaining osmotic balance, utilising the cellular antioxidant system and enhancing the transcription of cold responsive genes.

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Changes in SBPase activity influence photosynthetic capacity, growth, and tolerance to chilling stress in transgenic tomato plants

Scientific Reports ◽

10.1038/srep32741 ◽

2016 ◽

Vol 6 (1) ◽

Cited By ~ 45

Author(s):

Fei Ding ◽

Meiling Wang ◽

Shuoxin Zhang ◽

Xizhen Ai

Keyword(s):

Photosynthetic Capacity ◽

Chilling Stress ◽

Transgenic Tomato ◽

Tomato Plants ◽

Transgenic Tomato Plants

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SiFBA5, a cold-responsive factor from Saussurea involucrate promotes cold resilience and biomass increase in transgenic tomato plants under cold stress

10.21203/rs.3.rs-54573/v1 ◽

2020 ◽

Author(s):

Jianqiang Mu ◽

Yajuan Fu ◽

Bucang Liu ◽

Yao Zhang ◽

Aiying Wang ◽

...

Keyword(s):

Low Temperature ◽

Cold Stress ◽

Cold Tolerance ◽

Stress Responses ◽

Transgenic Tomato ◽

Dihydroxyacetone Phosphate ◽

Tomato Plants ◽

Transgenic Tomato Plants ◽

Photosynthesis Efficiency ◽

Fructose 1,6 Bisphosphate

Abstract Background Saussurea involucrate survives in extreme arctic conditions with strong cold resistant ability. The species occurs in rocky, mountainous habitats at elevations of approximately 2400-4100m with year-round snowfall and freezing temperatures providing a magnificent model and abundant gene pool for dissecting plant stress responses. Fructose-1,6-Bisphosphate Aldolase (FBA) mediates the reverse conversion of fructose 1,6-bisphosphate (FBP) into dihydroxyacetone phosphate (DHAP) and glycerol triphosphate (GAP) during glycolysis or gluconeogenesis, which is the key enzyme in this reaction. Saussurea involucrate can survive in extremely low temperature environment, which shows that it has extremely high photosynthesis efficiency. Significantly, the underlying mechanism of its cold tolerance is yet to be unveiled. Therefore, our work aims to explore potential molecular mechanisms. Results In this study, we identified a cold-responsive gene SiFBA5 that based on a preliminary low-temperature genome-wide transcriptional profiling in S. involucrata. Expression analysis showed that cold temperature rapidly induced transcriptional expression of SiFBA5, suggesting that SiFBA5 participates in initial stress responses. Subcellular localization showed that SiFBA5 is localized to the chloroplast. We then generated transgenic tomato plants overexpressing SiFBA5 derived by a CaMV 35S promoter. Phenotypic observation suggested that the transgenic overexpress plants displayed increased cold tolerance, photosynthesis efficiency, and carbohydrate accumulation in comparison with wild-type plants. Conclusion Collectively, our results demonstrated that SiFBA5 positively regulates plant response to cold stress, making it a promising candidate for improving cold tolerance in crops.

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SiFBA5, a cold-responsive factor from Saussurea involucrata promotes cold resilience and biomass increase in transgenic tomato plants under cold stress

BMC Plant Biology ◽

10.1186/s12870-021-02851-8 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Jianqiang Mu ◽

Yajuan Fu ◽

Bucang Liu ◽

Yao Zhang ◽

Aiying Wang ◽

...

Keyword(s):

Stress Response ◽

Low Temperature ◽

Cold Stress ◽

Cold Tolerance ◽

Crop Yield ◽

Photosynthetic Efficiency ◽

Transgenic Tomato ◽

Saussurea Involucrata ◽

Tomato Plants ◽

Transgenic Tomato Plants

Abstract Background Saussurea involucrata survives in extreme arctic conditions and is very cold-resistant. This species grows in rocky, mountainous areas with elevations of 2400–4100 m, which are snow-covered year-round and are subject to freezing temperatures. S. involucrata’s ability to survive in an extreme low-temperature environment suggests that it has particularly high photosynthetic efficiency, providing a magnificent model, and rich gene pool, for the analysis of plant cold stress response. Fructose-1, 6-bisphosphate aldolase (FBA) is a key enzyme in the photosynthesis process and also mediates the conversion of fructose 1, 6-bisphosphate (FBP) into dihydroxyacetone phosphate (DHAP) and glycerol triphosphate (GAP) during glycolysis and gluconeogenesis. The molecular mechanisms underlying S. involucrata’s cold tolerance are still unclear; therefore, our work aims to investigate the role of FBA in plant cold-stress response. Results In this study, we identified a cold-responsive gene, SiFBA5, based on a preliminary low-temperature, genome-wide transcriptional profiling of S. involucrata. Expression analysis indicated that cold temperatures rapidly induced transcriptional expression of SiFBA5, suggesting that SiFBA5 participates in the initial stress response. Subcellular localization analysis revealed that SiFBA5 is localized to the chloroplast. Transgenic tomato plants that overexpressed SiFBA5 were generated using a CaMV 35S promoter. Phenotypic observation suggested that the transgenic plants displayed increased cold tolerance and photosynthetic efficiency in comparison with wild-type plants. Conclusion Cold stress has a detrimental impact on crop yield. Our results demonstrated that SiFBA5 positively regulates plant response to cold stress, which is of great significance for increasing crop yield under cold stress conditions.

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Partial Resistance to Bacterial Wilt in Transgenic Tomato Plants Expressing Antibacterial Lactoferrin Gene

Journal of the American Society for Horticultural Science ◽

10.21273/jashs.127.2.158 ◽

2002 ◽

Vol 127 (2) ◽

pp. 158-164 ◽

Cited By ~ 23

Author(s):

Tae-Jin Lee ◽

Dermot P. Coyne ◽

Thomas E. Clemente ◽

Amitava Mitra

Keyword(s):

Southern Blot ◽

Bacterial Wilt ◽

Transgenic Tomato ◽

Mendelian Segregation ◽

Western Blots ◽

Tomato Plants ◽

Tomato Line ◽

Transgenic Tomato Plants ◽

Transgenic Tomatoes ◽

Susceptible Tomato

Expression of lactoferrin (LF) gene, a cationic iron-binding glycoprotein, was investigated in transgenic tomato plants (Lycopersicon esculentum Mill.). Resistance of the transgenic tomato plants to the pathogen (Ralstonia solanacearum Smith) causing bacterial wilt was also determined. Tomato line F7926-96, susceptible to bacterial wilt, was transformed with Agrobacterium strain C58C1 containing a plasmid construction carrying a modified LF cDNA. The introgression of LF cDNA into the susceptible tomato line was confirmed by Southern blot and the expression of full-length lactoferrin transcript and protein was also detected by northern and western blots, respectively. Based on resistance to kanamycin, a Mendelian segregation for a single locus insertion was observed in the T1 and T2 generations and all T1 and T2 plants resistant to kanamycin showed the single corresponding band of LF cDNA in Southern blot analysis. Two transgenic tomato lines inoculated with 1 × 107 and 1 × 108 colony-forming units (CFU)/mL with Rs isolate NC251 (K60, race 1) exhibited early resistance and subsequent susceptibility, while 44% to 55% of plants survived until maturity (fruit ripening) when inoculated with 1 × 105 CFU/mL in comparison with the fully susceptible tomato line. The latter resistance to bacterial wilt in transgenic tomatoes with the stable Mendelian segregation patterns for the LF gene suggests a potential new approach to consider for control of bacterial wilt of tomato. The possible value of this gene along with other plant genes to control bacterial pathogens is discussed.

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GRAS-domain transcription factor PAT1 regulates jasmonic acid biosynthesis in grape cold stress response

Plant Physiology ◽

10.1093/plphys/kiab142 ◽

2021 ◽

Author(s):

Zemin Wang ◽

Darren Chern Jan Wong ◽

Yi Wang ◽

Guangzhao Xu ◽

Chong Ren ◽

...

Keyword(s):

Stress Response ◽

Cold Stress ◽

Cold Tolerance ◽

Cold Treatment ◽

Ectopic Expression ◽

Bimolecular Fluorescence Complementation ◽

Luciferase Reporter ◽

Vitis Amurensis ◽

Mobility Shift

Abstract Cultivated grapevine (Vitis) is a highly valued horticultural crop, and cold stress affects its growth and productivity. Wild Amur grape (Vitis amurensis) PAT1 (Phytochrome A signal transduction 1, VaPAT1) is induced by low temperature, and ectopic expression of VaPAT1 enhances cold tolerance in Arabidopsis (Arabidopsis thaliana). However, little is known about the molecular mechanism of VaPAT1 during the cold stress response in grapevine. Here, we confirmed the overexpression of VaPAT1 in transformed grape calli enhanced cold tolerance. Yeast two-hybrid and bimolecular fluorescence complementation assays highlighted an interaction between VaPAT1 with INDETERMINATE-DOMAIN 3 (VaIDD3). A role of VaIDD3 in cold tolerance was also indicated. Transcriptome analysis revealed VaPAT1 and VaIDD3 overexpression and cold treatment coordinately modulate the expression of stress-related genes including lipoxygenase 3 (LOX3), a gene encoding a key jasmonate biosynthesis enzyme. Co-expression network analysis indicated LOX3 might be a downstream target of VaPAT1. Both electrophoretic mobility shift and dual luciferase reporter assays showed the VaPAT1-IDD3 complex binds to the IDD-box (AGACAAA) in the VaLOX3 promoter to activate its expression. Overexpression of both VaPAT1 and VaIDD3 increased the transcription of VaLOX3 and JA levels in transgenic grape calli. Conversely, VaPAT1-SRDX (dominant repression) and CRISPR/Cas9-mediated mutagenesis of PAT1-ED causing the loss of the C-terminus in grape calli dramatically prohibited the accumulation of VaLOX3 and JA levels during cold treatment. Together, these findings point to a pivotal role of VaPAT1 in the cold stress response in grape by regulating JA biosynthesis.

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