Expression of insect (Microdera puntipennis dzungarica) antifreeze protein MpAFP149 confers the cold tolerance to transgenic tobacco

Wang, Yan; Qiu, Liming; Dai, Chunying; Wang, Jing; Luo, Jian‐Ping; Zhang, Fuchun; Ma, Junjun

doi:10.1007/s00299-008-0562-5

Cited by 28 publications

(7 citation statements)

References 44 publications

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“…The comparative rate of lipid peroxidation was assayed from the leaves of transgenic and non-transformed plants by determining the level of malondialdehyde (MDA) production after exposure to cold stress (4°C) for 48 h, using the thiobarbituric acid (TBA) method as described by Wang et al (2008) with slight modifications of our own. Leaf samples (0.1 g) were homogenized with a mortar and pestle in 5 ml of 0.5% (v/v) TBA solution in 10% trichloroacetic acid (TCA) and centrifuged at 12,000 g for 15 min at 4°C.…”

Section: Malondialdehyde Estimationmentioning

confidence: 99%

Transgenic tomato cv. Pusa Uphar expressing a bacterial mannitol-1-phosphate dehydrogenase gene confers abiotic stress tolerance

Khare

Goyary

Singh

et al. 2010

Plant Cell Tiss Organ Cult

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A bacterial mannitol-1-phosphate dehydrogenase (mtlD) gene driven by the constitutive cauliflower mosaic virus (CaMV) 35S promoter was transferred into tomato plants using an Agrobacterium tumefaciens-mediated transformation protocol in an attempt to improve abiotic stress tolerance in the transformed plants. Transgene integration was confirmed by PCR analysis and Southern blot analysis, and transgene expression was confirmed by reverse transcription (RT)-PCR and direct mtlD (EC 1.1.1.17) activity. Upon exposure to low temperature stress (4°C) in a cold chamber, transgenic plants survived up to 48 h, while nontransformed plants were unable to survive and gradually died. Transgenic plants subjected to the chilling stress showed a significant decrease in electrolyte leakage and increased lipid peroxidation, as assessed by measuring malondialdehyde (MDA) content. Under the cold condition, transgenic plants also showed a significant increase in the activities of antioxidant enzymes (superoxide dismutase and catalase) and in relative water content (RWC) in comparison to non-transformed plants. Drought (polyethylene glycol in medium) and salinity (sodium chloride in medium) tolerance tests revealed that transgenic lines exhibited a higher tolerance for abiotic stresses than non-transformed plants. These findings indicate that the introduction of a bacterial mtlD gene into tomato conferred tolerance to abiotic stresses to the transformed tomato plants.

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Section: Malondialdehyde Estimationmentioning

confidence: 99%

Transgenic tomato cv. Pusa Uphar expressing a bacterial mannitol-1-phosphate dehydrogenase gene confers abiotic stress tolerance

Khare

Goyary

Singh

et al. 2010

Plant Cell Tiss Organ Cult

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“…Introduction of the hyperactive antifreeze proteins of insects into the genome of plants has the potential to achieve the latter. AFPs and/or IAPs from fish, insects, and plants have been successfully expressed in tomato, tobacco, potato, and Arabidopsis thaliana (Hightower et al 1991;Kenward et al 1993;Wallis et al 1997;Kenward et al 1999;Meyer et al 1999;Holmberg et al 2001;Wang et al 2008;Zhu et al 2010). In some systems, AFP expression in transgenic plants produced plants whose extracts exhibited low levels of thermal hysteresis (0.37°C/mg apoplastic protein by nanoliter osmometer, Holmberg et al 2001) and recrystallization inhibition in vitro (Worrall et al 1998), as well as reduced electrolyte leakage in leaves after freezing (Wallis et al 1997;Zhu et al 2010).…”

Section: Introductionmentioning

confidence: 97%

“…), and the freezing temperature of the whole plant without roots was lowered by 1-3°C depending on the freezing treatment method used. In a recent study, Wang et al (2008) expressed a beetle (Microdera puntipennis dzungarica) antifreeze protein in tobacco and showed decreased ion leakage, lower malondialdehyde content in the leaves, and improved cold tolerance at −1°C for 3 days.…”

Section: Introductionmentioning

confidence: 98%

Expression of Two Self-enhancing Antifreeze Proteins from the Beetle Dendroides canadensis in Arabidopsis thaliana

2011

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Antifreeze proteins depress the non-equilibrium freezing point of aqueous solutions, but only have a small effect on the equilibrium melting point. This difference between the freezing and melting points has been termed thermal hysteresis activity (THA). THA identifies the presence and relative activity of antifreeze proteins. Two antifreeze protein cDNAs, dafp-1 and dafp-4, encoding two self-enhancing (have a synergistic effect on THA) antifreeze proteins (DAFPs) from the beetle Dendroides canadensis, were introduced into the genome of Arabidopsis thaliana via Agrobacterium-mediated floral dip transformation. Southern blot analysis indicated multiple insertions of transgenes. Both DAFP-1 and/or DAFP-4 were expressed in transgenic A. thaliana as shown by RT-PCR and Western blot. Apoplastic fluid from T 3 DAFP-1+DAFP-4-producing transgenic A. thaliana exhibited THA in the range of 1.2-1.35°C (using the capillary method to determine THA), demonstrating the presence of functioning antifreeze proteins (with signal peptides for extracellular secretion). The freezing temperature of DAFP-1+DAFP-4-producing transgenic A. thaliana was lowered by approximately 2-3°C compared with the wild type.

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“…Holmberg et al (2001) also reported antifreeze activity in applications with very low concentrations of AFPs. Wang et al (2008) reported that in order to transfer antifreeze protection to a plant, it is crucial to introduce AFPs into the apoplast space to confer an optimal antifreeze effect. This phenomenon occurs since ice forms preferentially in the apoplast where the solute concentration is the lowest.…”

Section: Introductionmentioning

confidence: 99%

The response of watercress (Nasturtium officinale) to vacuum impregnation: Effect of an antifreeze protein type I

Cruz

Vieira

Silva

2009

Journal of Food Engineering

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a b s t r a c tThe setting up of methodologies that reduce the size of ice crystals and reduce or inhibit the recrystallisation phenomena could have an extraordinary significance in the final quality of frozen products and consequently bring out new market opportunities. In this work, the effect of an antifreeze protein type I (AFP-I), by vacuum impregnation (VI), on frozen watercress was studied. The VI pressure, samples' weight, Hunter Lab colour, scanning electron microscopy (SEM), and a wilting test were analysed in this work.The water intake of watercress samples augmented with vacuum pressure increase. The results also showed that, independently from the vacuum pressure used, the Lab colour parameters between raw and impregnated samples were maintained, showing no significant differences (P > 0.05).A VI of 58 kPa, during 5 min, allowed impregnating the AFP-I solution (0.01 mg ml À1 ) into the watercress samples. The scanning electron microscopy (SEM) analysis showed the AFP-I impregnated frozen samples with better cell wall definition and rounded cell shape with smaller ice crystals compared with the control samples.The wilting test results corroborated that AFP-I is a valuable additive, since the leaves impregnated with AFP-I showed higher turgidity compared to the control samples.The present findings will help to better understand the effect of AFP-I, particularly, on frozen watercress microstructure and its importance as valuable food additive in frozen foods and mainly in leafy vegetables.

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Expression of insect (Microdera puntipennis dzungarica) antifreeze protein MpAFP149 confers the cold tolerance to transgenic tobacco

Cited by 28 publications

References 44 publications

Transgenic tomato cv. Pusa Uphar expressing a bacterial mannitol-1-phosphate dehydrogenase gene confers abiotic stress tolerance

Transgenic tomato cv. Pusa Uphar expressing a bacterial mannitol-1-phosphate dehydrogenase gene confers abiotic stress tolerance

Expression of Two Self-enhancing Antifreeze Proteins from the Beetle Dendroides canadensis in Arabidopsis thaliana

The response of watercress (Nasturtium officinale) to vacuum impregnation: Effect of an antifreeze protein type I

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