Study on nuclear and cytoplasmic genome expression in wheat by two-dimensional gel electrophoresis

Zivy, Michel; Thiellement, Hervé; Vienne, Dominique de; Hofmann, Jean-Paul

doi:10.1007/bf00281838

Cited by 98 publications

(21 citation statements)

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“…The gly II protein was purified as described (15) and used to raise polyclonal antibodies in rabbits (New Zealand White) as per standard procedures (16). For Western blotting, extraction of soluble proteins from tobacco was essentially carried out as reported (17). The amount of protein was estimated by the Bradford method (18).…”

Section: Methodsmentioning

confidence: 99%

Genetic engineering of the glyoxalase pathway in tobacco leads to enhanced salinity tolerance

Singla‐Pareek

Reddy²,

Sopory³

2003

Proc. Natl. Acad. Sci. U.S.A.

367

261

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The glyoxalase pathway involving glyoxalase I (gly I) and glyoxalase II (gly II) enzymes is required for glutathione-based detoxification of methylglyoxal. We had earlier indicated the potential of gly I as a probable candidate gene in conferring salinity tolerance. We report here that overexpression of gly I؉II together confers improved salinity tolerance, thus offering another effective strategy for manipulating stress tolerance in crop plants. We have overexpressed the gly II gene either alone in untransformed plants or with gly I transgenic background. Both types of these transgenic plants stably expressed the foreign protein, and the enzyme activity was also higher. Compared with nontransformants, several independent gly II transgenic lines showed improved capability for tolerating exposure to high methylglyoxal and NaCl concentration and were able to grow, flower, and set normal viable seeds under continuous salinity stress conditions. Importantly, the double transgenic lines always showed a better response than either of the single gene-transformed lines and WT plants under salinity stress. Ionic measurements revealed higher accumulation of Na ؉ and K ؉ in old leaves and negligible accumulation of Na ؉ in seeds of transgenic lines as compared with the WT plants. Comparison of various growth parameters and seed production demonstrated that there is hardly any yield penalty in the double transgenics under nonstress conditions and that these plants suffered only 5% loss in total productivity when grown in 200 mM NaCl. These findings establish the potential of manipulation of the glyoxalase pathway for increased salinity tolerance without affecting yield in crop plants.

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Section: Methodsmentioning

confidence: 99%

Genetic engineering of the glyoxalase pathway in tobacco leads to enhanced salinity tolerance

Singla‐Pareek

Reddy²,

Sopory³

2003

Proc. Natl. Acad. Sci. U.S.A.

367

261

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“…However, visualization of proteins with silver staining gave similar results (data not shown). Also, the amount of the protein can be reduced due to its proteolytic degradation [16,17]. Nevertheless, rubisco large fragments were not found on the 2-D gels.…”

Section: Resolution and Identification Of The Proteins From Maize Leavesmentioning

confidence: 98%

The proteome of maize leaves: Use of gene sequences and expressed sequence tag data for identification of proteins with peptide mass fingerprints

Porubleva

Velden²,

Kothari³

et al. 2001

Electrophoresis

140

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As a first step in establishing a proteome database for maize, we have embarked on the identification of the leaf proteins resolved on two-dimensional (2-D) gels. We detected nearly 900 spots on the gels with a pH 4-7 gradient and over 200 spots on the gels with a pH 6-11 gradient when the proteins were visualized with colloidal Coomassie blue. Peptide mass fingerprints for 300 protein spots were obtained with matrix assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometer and 149 protein spots were identified using the protein databases. We also searched the pdbEST databases to identify the leaf proteins and verified 66% of the protein spots that had been identified using the protein databases. Sixty-seven additional protein spots were identified from expressed sequence tags (ESTs). Many abundant leaf proteins are present in multiple spots. Functions of over 50% of the abundant leaf proteins are either unknown or hypothetical. Our results show that EST databases in conjunction with peptide mass fingerprints can be used for identifying proteins from organisms with incomplete genome sequence information.

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“…Zivy et al [9], in 1983, investigated the genetic variability of 18 wheat (Triticum aesti- vum) alloplasmic lines. Genetic differences between two wheat lines and two groups of cytoplasms were observed on proteins extracted from five developmental stages or organs.…”

Section: Genetic Diversitymentioning

confidence: 99%

“…2). However the same gene may be represented in 2-D gels by several spots resulting from post-translational modifications or degradation products [9,74]. PS are those cases where two spots with similar aspects and staining intensity are characterized by the following criteria: (i) They are located relatively close to each other on 2-D gels (i.e., they have close pI and/or apparent molecular masses); (ii) they present a monogenic and codominant Mendelian mode of inheritance in a segregating pedigree (1:1 segregation ratio in haploid and double haploid lines; 1:2:1 segregation ratio in F2 selfed families; see [8] for a review), and (iii) last but not least, they correspond to the same protein.…”

Section: Mutant Characterizationmentioning

confidence: 99%

Proteomics for genetic and physiological studies in plants

et al. 1999

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Proteomics is becoming a necessity in plant biology, as it is in medicine, zoology and microbiology, for deciphering the function and role of the genes that are or will be sequenced. In this review we focus on the various, mainly genetic, applications of the proteomic tools that have been developed in recent years: characterization of individuals or lines, estimation of genetic variability within and between populations, establishment of genetic distances that can be used in phylogenetic studies, characterization of mutants and localization of the genes encoding the revealed proteins. Improvements in specifically devoted software have permitted precise quantification of the variation in amounts of proteins, leading to the concept of “protein quantity loci” which, combined with the “quantitative trait loci” approach, results in testable hypotheses regarding the role of “candidate proteins” in the metabolism or phenotype under study. This new development is exemplified by the reaction of plants to drought, a trait of major agronomic interest. The accumulation of data regarding genomic and cDNA sequencing will be connected to the protein databases currently developed in plants.

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Study on nuclear and cytoplasmic genome expression in wheat by two-dimensional gel electrophoresis

Cited by 98 publications

References 0 publications

Genetic engineering of the glyoxalase pathway in tobacco leads to enhanced salinity tolerance

Genetic engineering of the glyoxalase pathway in tobacco leads to enhanced salinity tolerance

The proteome of maize leaves: Use of gene sequences and expressed sequence tag data for identification of proteins with peptide mass fingerprints

Proteomics for genetic and physiological studies in plants

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