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1.
Changes in gene expression during foliar senescence and fruit ripening in tomato (Lycopersicon esculentum Mill.) were examined using in-vitro translation of isolated RNA and hybridization against cDNA clones.During the period of chlorophyll loss in leaves, changes occurred in mRNA in-vitro translation products, with some being reduced in prevalence, whilst others increased. Some of the translation products which changed in abundance had similar molecular weights to those known to increase during tomato fruit ripening. By testing RNA from senescing leaves against a tomato fruit ripening-related cDNA library, seven cDNA clones were identified for mRNAs whose prevalence increased during both ripening and leaf senescence. Using dot hybridization, the pattern of expression of the mRNAs corresponding to the seven clones was examined. Maximal expression of the majority of the mRNAs coincided with the time of greatest ethylene production, in both leaves and fruit. Treatment of mature green leaves or unripe fruit with the ethylene antagonist silver thiosulphate prevented the onset of senescence or ripening, and the expression of five of the seven ripening- and senescence-related genes.The results indicate that senescence and ripening in tomato involve the expression of related genes, and that ethylene may be an important factor in controlling their expression.Abbreviations cDNA
copy-DNA
- MW
molecular weight
- PAGE
polyacrylamide gel electrophoresis
- SDS
sodium dodecyl sulphate 相似文献
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P. Hiscott H. A. Waller I. Grierson M. G. Butler D. Scott 《Cell and tissue research》1992,267(1):185-192
Summary The distribution of fibronectin mRNA and fibronectin in adult human retina and epiretinal membranes was investigated by in situ hybridisation and immunohistochemical techniques. The cells in normal adult retina contained little or no fibronectin mRNA and the retina only showed fibronectin immunoreactivity in retinal vessels. The cells in detached neuroretina did not contain fibronectin message but the vitreoretinal interface of the detached retina exhibited variable fibronectin immunoreactivity. Retinal glia, retinal pigment epithelium and fibroblast-like cells in membranes at the vitreoretinal juncture (epiretinal membranes) showed variable labelling with the fibronectin mRNA probe and all the membranes immunostained for fibronectin. No difference could be detected between membrane cell types in the intensity of labelling with the mRNA probe or for fibronectin immunoreactivity. The results indicate that cells in situ in attached and detached adult human retina do not produce fibronectin. Although fibronectin at the vitreoretinal juncture in retinal detachment is probably partly derived from plasma fibronectin resulting from breakdown of the blood-retinal barrier, ectopic retinal cells produce fibronectin and contribute to the glycoprotein in epiretinal membranes. 相似文献
5.
Methods were developed for the isolation of plastids from mature green and ripening tomatoes (Lycopersicon esculentum Mill.) and purification by sucrose or Percoll density-gradient centrifugation. Assessment of the purity of preparations involved phase-contrast and electron microscopy, assays for marker enzymes and RNA extraction and analysis. Proteins were extracted from isolated plastids at different ripening stages and separated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. The profiles obtained from chloroplasts and chromoplasts showed many qualitative and quantitative differences. Labelling of proteins with [35S]methionine in vivo showed that there was active protein synthesis throughout ripening, but there was a change in the plastid proteins made as ripening proceeded. The cellular location of synthesis of specific proteins has yet to be established.Abbreviations CS
citrate synthase
- EDTA
ethylenediaminetetraacetic acid,-acetate
- GAPDH
NADP+-glyceraldehyde-3-phosphate dehydrogenase
- rRNA
ribosomal RNA
- SDS
sodium dodecyl sulphate
- SDS-PAGE
SDS-polyacrylamide gel electrophoresis
- Tris
2-amino-2(hydroxymethyl)-1,3-propanediol 相似文献
6.
Sequencing and identification of a cDNA clone for tomato polygalacturonase. 总被引:28,自引:2,他引:26
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The 2a isoenzyme of tomato polygalacturonase was purified from ripe fruit and characterised. The N-terminal amino acid sequence of the protein was determined in order to identify polygalacturonase cDNA clones. The nucleotide sequence of a ripening-related cDNA (pTOM 6) was determined and found to encode the N-terminal sequence of mature polygalacturonase 2a. The complete open reading frame encodes a polypeptide of molecular weight 50,051, including a putative pre-sequence of 71 amino acids. 相似文献
7.
Linkage analysis of seven kindreds with the X-linked lymphoproliferative syndrome (XLP) confirms that the XLP locus is near DXS42 and DXS37 总被引:5,自引:0,他引:5
James C. Skare Helen L. Grierson John L. Sullivan Robert L. Nussbaum David T. Purtilo Bakary S. Sylla Gilbert M. Lenon Dorothy S. Reilly Bradley N. White Aubrey Milunsky 《Human genetics》1989,82(4):354-358
Summary Analysis of seven kindreds indicates that the XLP locus exhibits 1% recombination with DXS42 (lod = 17.5) and no recombination with DXS37 (lod = 13.3). 相似文献
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Ethylene receptor expression is regulated during fruit ripening, flower senescence and abscission 总被引:21,自引:0,他引:21
Sharon Payton Rupert G. Fray Stephen Brown Don Grierson 《Plant molecular biology》1996,31(6):1227-1231
Using theArabidopsis ethylene receptorETR1 as a probe, we have isolated a tomato homologue (tETR) from a ripening cDNA library. The predicted amino acid sequence is 70% identical toETR1 and homologous to a variety of bacterial two component response regulators over the histidine kinase domain. Sequencing of four separate cDNAs indicates that tETR lacks the carboxyl terminal response domain and is identical to that encoded by the tomatoNever ripe gene. Ribonuclease protection showed tETR mRNA was undetectable in unripe fruit or pre-senescent flowers, increased in abundance during the early stages of ripening, flower senescence, and in abscission zones, and was greatly reduced in fruit of ripening mutants deficient in ethylene synthesis or response. These results suggest that changes in ethylene sensitivity are mediated by modulation of receptor levels during development. 相似文献
10.
Nucleotide sequence and expression of a ripening and water stress-related cDNA from tomato with homology to the MIP class of membrane channel proteins 总被引:13,自引:0,他引:13
Rupert G. Fray Andrew Wallace Donald Grierson Grantley W. Lycett 《Plant molecular biology》1994,24(3):539-543
The nucleotide sequence and derived amino acid sequence were determined for a full-length version of the tomato cDNA clone, pTOM75, the mRNA for which has previously been shown to accumulate in roots, ripening fruit and senescing leaves. Computer analysis of the predicted protein product, which we have named tomato ripening-associated membrane protein (TRAMP) indicates strong homology to known transmembrane channel proteins from other organisms. Northern analysis showed that this gene was induced by waterstress and that this induction was unaffected in an ABA-deficient genetic back-ground. 相似文献