Expression of alcohol dehydrogenase in rice embryos under anoxia

Ricard, Bérénice; Mocquot, Bernard; Fournier, Alain; Delseny, Michel; Pradet, A.

doi:10.1007/bf00032562

Cited by 47 publications

(44 citation statements)

References 22 publications

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“…Immunoprecipitation of in vivo and in vitro synthesized sucrose synthase was done by the protein A-Sepharose method described by Kopczynski and Scandalios (7). SDS or twodimensional PAGE of proteins, before or after immunoprecipitation, was performed as described previously (13,16). Gels were prepared for fluorography by impregnation with dimethyl sulfoxide/2,5-diphenyloxazole, dried, and exposed on Kodak X-Omat AR film.…”

Section: Immunoprecipitation and Pagementioning

confidence: 99%

“…For denaturing (SDS) gels, 100 ,ug of proteins were boiled for 5 min in 65 mm Tris-HCl, pH 6.8, 5% 2-mercaptoethanol, 2.3% SDS, and 10% glycerol, then subjected to electrophoresis on 15% polyacrylamide-SDS gels as described previously (16).…”

Section: Immunodetectionmentioning

confidence: 99%

“…Inbred rice (Oryza sativa L., var Cigalon) seeds were surface-sterilized with a commercial preparation of bleach, then germinated for 48 h in the dark at 25°C under distilled water with vigorous agitation ( 16 …”

Section: Plant Materialsmentioning

confidence: 99%

“…We have already shown that many of the glycolytic and fermentative enzymes increase in activity (19). ADH (16), cytosolic glyceraldehyde 3-phosphate dehydrogenase (18), and PDC (our unpublished data) are all induced at both the mRNA and protein levels. The rice seedlings referred to above are still attached to the seed that contains starchy reserves.…”

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confidence: 99%

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Anaerobic Stress Induces the Transcription and Translation of Sucrose Synthase in Rice

Ricard¹,

Rivoal²,

Spiteri³

et al. 1991

Plant Physiol.

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129

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Sucrose synthase activity increased in 2-day-old rice (Oryza sativa) seedlings submitted to anaerobic stress. Likewise The unusual behavior of maize sucrose synthase isozymes under anaerobiosis led us to study the anaerobic induction of the enzyme in rice. Rice shares the metabolic strategy of maize roots in response to anaerobic stress. However, in contrast to maize roots, rice is extremely resistant to anoxia. Rice seedlings that have germinated for 48 h aerobically survive more than 21 d and are capable of coleoptile elongation under complete oxygen deprivation. Their metabolism is high enough to permit DNA, RNA, as well as protein synthesis (1,12,13). We have already shown that many of the glycolytic and fermentative enzymes increase in activity (19). ADH (16), cytosolic glyceraldehyde 3-phosphate dehydrogenase (18), and PDC (our unpublished data) are all induced at both the mRNA and protein levels. The rice seedlings referred to above are still attached to the seed that contains starchy reserves. It seemed likely that the existence of these reserves played a role in the active metabolism seen in such seedlings under anoxia. Because starch mobilization is mediated by sucrose synthase, we hypothesized that in rice, sucrose synthase activity and protein levels should be induced by anaerobiosis coordinately with the other enzymes of sugar phosphate metabolism. In maize roots, however, lack ofstarch reserves means that sucrose synthase accumulation would be unnecessary.Our results were obtained using several independent techniques, including enzyme activity measurements, immunodetection on Western blots, SDS, and two-dimensional PAGE of in vivo-and in vitro-labeled total proteins and immunoprecipitates, as well as hybridization analyses of Northern blots and run-on transcripts. Sucrose synthase activity and protein both increased upon anaerobic stress. This was accompanied by an increase in the steady-state level of sucrose synthase mRNA and in the rate of transcription ofthe sucrose synthase gene. We conclude that sucrose synthase is a typical anaerobic polypeptide in rice and is induced at both the protein and mRNA levels. MATERIALS AND METHODS Plant MaterialInbred rice (Oryza sativa L., var Cigalon) seeds were surface-sterilized with a commercial preparation of bleach, then germinated for 48 h in the dark at 25°C under distilled water with vigorous agitation ( 16). Under such conditions, seedlings are under normoxia and have coleoptiles and primary roots about 0.5 cm in length. Anaerobic treatment consisted of www.plantphysiol.org on April 3, 2019 -Published by Downloaded from

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Section: Immunoprecipitation and Pagementioning

confidence: 99%

Section: Immunodetectionmentioning

confidence: 99%

Section: Plant Materialsmentioning

confidence: 99%

mentioning

confidence: 99%

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Anaerobic Stress Induces the Transcription and Translation of Sucrose Synthase in Rice

Ricard¹,

Rivoal²,

Spiteri³

et al. 1991

Plant Physiol.

Self Cite

129

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“…Tissue Imprinting A tissue-printing technique (1) was combined with the histochemical staining procedure (19) to localize ADH activity in maize and rice seedlings. Untreated and cold-treated maize and rice seedlings were carefully removed from vermiculite, and, because of high endogenous levels of ADH activity and the difficulty of pressing, the seeds were removed.…”

Section: Rna Analysismentioning

confidence: 99%

Low-Temperature Accumulation of Alcohol Dehydrogenase-1 mRNA and Protein Activity in Maize and Rice Seedlings

Christie¹,

Hahn²,

Walbot³

1991

Plant Physiol.

112

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Low-temperature stress was shown to cause a rapid increase in steady-state levels of alcohol dehydrogenase-1 message (Adhl) and protein activity (ADHI) in maize (Zea mays) (B37N, A188) and rice (Oryza sativa) (Taipei 309, Calmochi 101) seedlings. Maize roots and rice shoots and roots from 7-day seedlings shifted to low temperature (100C) contained as much as 15-fold more Adhl mRNA and 8-fold more ADHI protein activity than the corresponding tissues from untreated seedlings. Time-course studies showed that these tissues accumulated Adhl mRNA and ADHI activity severalfold within 4-to 8-hour, levels plateaued within 20 to 24 hours, and remained elevated at 4 days of cold treatment. Within 24 hours of returning cold-stressed seedlings to ambient temperature, Adh1 mRNA and ADH1 activity decreased to pretreatment levels. Histochemical staining of maize and rice tissue imprints showed that ADH activity was enhanced along the lengths of cold-stressed maize primary roots and rice roots, and along the stems and leaves of rice shoots. Our observations suggest that short-term cold stress induces Adhl gene expression in certain plant tissues, which, reminiscient of the anaerobic response, may reflect a fundamental shift in energy metabolism to ensure tissue survival during the stress period.Cold temperature, a common environmental stress in the temperate zone, affects numerous biochemical, physiological, and metabolic functions in plants (8,13,29 roplast genes (psaB, psbB, rbsL, atpE) and by nuclear genes coding for chloroplast proteins (RbcS, Cab). A more detailed analysis of ribulose 1,5-bisphosphate carboxylase transcript and protein levels demonstrated strong suppression at the level of transcription and/or transcript stability, and the apparent loss of coordinate synthesis of the large and small subunits, suggestive of an effect of cold on nuclear-organellar signaling (9). Numerous physiological studies have demonstrated that cold impairs photosynthetic activity of chillingsensitive plants (13,18,29). Our findings indicate that a corresponding molecular consequence of cold stress is a reduced expression of chloroplast-related genes.Cold stress also disrupts respiratory activities of chillingsensitive plants (12)(13)(14)16). It has been proposed that chillingsensitive plants compensate for impaired mitochondrial function by shifting their metabolism from aerobic to anaerobic respiration (see ref. 13). This strategy is an important component of the response to oxygen deprivation, another environmental stress that prevents mitochondrial activity. When certain plant tissues-most notably the primary roots of maize-become anoxic, they rapidly shift to ethanolic fermentation to maintain energy production. Early genetic studies established the importance of ethanolic fermentation for maize tissue survival during anoxia, i.e. during flooding (25). More recent studies have shown several members of the glycolytic pathway, including Adh1,3 to be transcriptionally activated during anaerobic stress (10,11,(21)(22)(23). At le...

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The Anaerobic Stress Response and its Use for Studying Gene Expression in Conifers

Harry

Kinlaw

Sederoff

1988

Genetic Manipulation of Woody Plants

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Expression of alcohol dehydrogenase in rice embryos under anoxia

Cited by 47 publications

References 22 publications

Anaerobic Stress Induces the Transcription and Translation of Sucrose Synthase in Rice

Anaerobic Stress Induces the Transcription and Translation of Sucrose Synthase in Rice

Low-Temperature Accumulation of Alcohol Dehydrogenase-1 mRNA and Protein Activity in Maize and Rice Seedlings

The Anaerobic Stress Response and its Use for Studying Gene Expression in Conifers

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