Isolation and characterization of tomato cDNA and genomic clones encoding the ubiquitin gene ubi3

Hoffman, Neil E.; Ko, Kenton; Milkowski, Deborah M.; Pichersky, Eran

doi:10.1007/bf00028735

Cited by 71 publications

(59 citation statements)

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“…Furthermore, and contrary to what happsns in some animal systems (Muller-Taubenberger et al, L989), plant extension proteins are rapidly cleaved to produce free ubiquitin (Callis et al, 1990). Their expressim patterns would also be different from that of UbiS "As, because extension proteins are usually expressed in higlily dividing cells and seldom up-regulated in response to stress (Callis et al, 1990;Hoffman et al, 1991). In contrast expression during the desiccation stages of embryogenesis of proteins in band IV (Fig.…”

Section: Discussionmentioning

confidence: 99%

“…This, together with observation of differential regulation of severa1 plant ubiquitin genes, has suggested that their multiplicity and diversity reflect an evolutionary divergence of their gene regulation to cope with different specific tasks, such as functions related to cell proliferation and dedifferentiation (Gausing and Barkardottir, 1986;Jamet et al, 1990;Genschik et al, 1992); stress, light, or hormone responses (Burke et al, 1988;Binet et al, 1991;Hoffman et al, 1991;Garbarino et al, 1992;Genschik et al, 1992;Reynolds and Hooley, 1992); and organ (flower, leaf, etc.) development (Gausing and Barkardottir, 1986;Burke et al, 1988;Hoffman et al, 1991;Garbarino et al, 1992). The differential regulation of tetraubiquitin mRNAs during seed maturation and detection of putative, seed-specific, protein-ubiquitin conjugates reported in this study suggest that distinct ubiquitin genes could perform specific functions during plant zygotic embryogenesis.…”

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confidence: 99%

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Differential Accumulation of Sunflower Tetraubiquitin mRNAs during Zygotic Embryogenesis and Developmental Regulation of Their Heat-Shock Response

Almoguera¹,

Coca²,

Jordano³

1995

Plant Physiol.

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We have isolated and sequenced Ha UbiS, a cDNA for a dryseed-stored mRNA that encodes tetraubiquitin. We have observed differential accumulation of tetraubiquitin mRNAs during sunflower (Helianthus annuus 1.) zygotic embryogenesis. These mRNAs were up-regulated during late embryogenesis and reached higher prevalence in the dry seed, where they were found to be associated mainly with provascular tissue. UbiS mRNA, as confirmed by RNase A protection experiments, accumulated also in response to heat shock, but only in leaves and later during postgerminative development. These novel observations demonstrate expression during seed maturation of specific plant polyubiquitin transcripts and developmental regulation of their heat-shock response. Using ubiquitin antibodies we also detected discrete, seed-specific proteins with distinct temporal expression patterns during zygotic embryogenesis. Some of these patterns were concurrent with UbiS mRNA accumulation in seeds. The most abundant ubiquitin-reacting proteins found in mature seeds were small (16-22 kD) and acidic (isoelectric points of 6.1-7.4). Possible functional implications for UbiS expression elicited from these observations are discussed.

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

Differential Accumulation of Sunflower Tetraubiquitin mRNAs during Zygotic Embryogenesis and Developmental Regulation of Their Heat-Shock Response

Almoguera¹,

Coca²,

Jordano³

1995

Plant Physiol.

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“…The qRT-PCR reaction was performed using miraculin-specific primers and ubiquitin3-specific primers. Ubiquitin3 has been described previously (Hoffman et al 1991) and was used as a control (Hackel et al 2006;Leclercq et al 2005). The primer sequences have also been described previously (Kim et al 2010).…”

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confidence: 99%

Gene dosage and genetic background affect miraculin accumulation in transgenic tomato fruits

Kim

Hirai

Kato

et al. 2010

Plant Biotechnology

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Gene dosage and genetic background are factors that influence transgene expression in transgenic plants. In our previous studies, we produced transgenic tomato plants that accumulate miraculin, a taste-modifying protein, in a genetically stable manner. To elucidate the effects of gene dosage and genetic background on miraculin accumulation in transgenic tomato fruits, we generated hybrid tomato lines between the homozygous transgenic line 56B (background cultivar 'Moneymaker') and the pure cultivars 'Micro-Tom,' 'Moneymaker,' 'Ailsa Craig,' 'M82,' 'Rutgers' and 'Aichi-first' and analyzed them for miraculin mRNA expression and miraculin protein accumulation. The hybrid lines exhibited variation in their fruit structures. Among the hybrid lines heterozygous for the miraculin gene, miraculin accumulation in the fruits varied from 111.0 mg g Ϫ1 fresh weight (FW) to 159.4 mg g Ϫ1 FW. Furthermore, the homozygous line 56B showed higher miraculin accumulation and miraculin mRNA expression than the heterozygous line 56Bϫ'Moneymaker.' These results demonstrate the profound effects of gene dosage and genetic background on miraculin accumulation in transgenic tomato fruits.Key words: Gene dosage, genetic background, miraculin, taste-modifying protein.Plant Biotechnology 27, 333-338 (2010) Original PaperThis article can be found at http://www.jspcmb.jp/ allelic composition and genetic background affect transgene expression and inheritance in white clover. Another study in rice showed that the expression pattern of Xa3/Xa26, which is a rice disease resistance gene, was affected by the genetic background (Cao et al. 2007). The full effects of gene dosage and genetic background on transgene expression and translation remain to be elucidated.In this study, we generated hybrid lines between transgenic line 56B (background cultivar 'Moneymaker'), which is homozygous for the miraculin gene, and the pure line cultivars 'Micro-Tom,' 'Moneymaker,' 'Ailsa Craig,' 'M82,' 'Rutgers' and 'Aichi-first.' Characterization of the hybrids' miraculin mRNA expression and miraculin protein accumulation, revealed the effects of gene dosage and genetic background on miraculin accumulation in transgenic tomato fruits, and in addition we discuss the significance of combining molecular breeding with cross-breeding to produce transgenic tomatoes that accumulate miraculin. Materials and methods Plant material and growth conditionsThe transgenic tomato line 56B was produced in our previous work (Sun et al. 2007). In brief miraculin encoding 660-bp DNA fragment was inserted into the XbaI/SacI sites of the plant transformation vectors pBI121 (Sun et al. 2006). Line 56B possesses a single copy of the miraculin gene driven by the CaMV 35S promoter.In the present study, the miraculin-accumulating transgenic tomato line 56B (background cultivars 'Moneymaker') was crossed with six other cultivars: 'Micro-Tom (accession number TOMJPF00001), ' 'Moneymaker (accession number TOMJPF00002), ' 'Aichi-first (accession number TOMJPF00003), ' 'Ailsa Cra...

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“…Expression of genes associated with phenylpropanoid and flavonoid biosynthesis was assessed using probes generously provided by Dr. Cathie Martin (John Innes Centre, Norwich, UK). A Datura stramonium ubiquitin sequence was generated by PCR from genomic DNA using primers derived from the tomato sequence (sense, 5Ј-ATGCAGATCTTCGTGAAAAC-3Ј; antisense, 5Ј-CTTAACCTTCTTCTTCTTCTGCTT-3Ј [Hoffman et al, 1991]). Tobacco ubiquitin and ␤-1,3-glucanase (PR2) coding sequences were amplified by PCR from tobacco genomic DNA using oligonucleotide primers derived from published sequences (ubiquitin sense, 5Ј-TGGCTCAGGATGAACGCTGGC-3Ј; antisense, 5Ј-CATCTT-TGAGACCTCAGTAGAC-3Ј [Karrer et al, 1998]; PR2 sense, 5Ј-CAT-GCAAACAATTTACCATCAGACC-3Ј; antisense, 5Ј-CCAGGTTTCTTT-GGAGTTCCTGCC-3Ј [Ward et al, 1991]).…”

Section: Molecular Analysis Of Plantsmentioning

confidence: 99%

Rerouting the Plant Phenylpropanoid Pathway by Expression of a Novel Bacterial Enoyl-CoA Hydratase/Lyase Enzyme Function

Mayer¹,

Narbad²,

Parr³

et al. 2001

The Plant Cell

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The gene for a bacterial enoyl-CoA hydratase (crotonase) homolog (HCHL) previously shown to convert 4-coumaroylCoA, caffeoyl-CoA, and feruloyl-CoA to the corresponding hydroxybenzaldehydes in vitro provided an opportunity to subvert the plant phenylpropanoid pathway and channel carbon flux through 4-hydroxybenzaldehyde and the important flavor compound 4-hydroxy-3-methoxybenzaldehyde (vanillin). Expression of the Pseudomonas fluorescens AN103 HCHL gene in two generations of tobacco plants caused the development of phenotypic abnormalities, including stunting, interveinal chlorosis and senescence, curled leaf margins, low pollen production, and male sterility. In second generation progeny, the phenotype segregated with the transgene and transgenic siblings exhibited orange/red coloration of the vascular ring, distorted cells in the xylem and phloem bundles, and lignin modification/reduction. There was depletion of the principal phenolics concomitant with massive accumulation of novel metabolites, including the glucosides and glucose esters of 4-hydroxybenzoic acid and vanillic acid and the glucosides of 4-hydroxybenzyl alcohol and vanillyl alcohol. HCHL plants exhibited increased accumulation of transcripts for phenylalanine ammonia-lyase, cinnamate-4-hydroxylase, and 4-coumarate:CoA ligase, whereas ␤ -1,3-glucanase was suppressed. This study, exploiting the ability of a bacterial gene to divert plant secondary metabolism, provides insight into how plants modify inappropriately accumulated metabolites and reveals the consequences of depleting the major phenolic pools. INTRODUCTIONThe plant phenylpropanoid pathway is responsible for the synthesis of a wide variety of secondary metabolic compounds, including lignins, salicylates, coumarins, hydroxycinnamic amides, flavonoid phytoalexins, pigments, UV light protectants, and antioxidants (Dixon and Paiva, 1995). In addition to their roles in the structure and protection of the plant, phenylpropanoids have an important effect on plant qualities such as texture, flavor, color, and processing characteristics. Molecular engineering provides a tool to determine the complex biochemical pathways involved in the synthesis and regulation of phenylpropanoids and to manipulate pathways to increase or initiate the production of economically desirable traits or compounds (Dixon et al., 1996). Expression of plant genes in heterologous plant systems can lead to improved or novel synthesis of valuable compounds (Yun et al., 1992) and improved disease resistance (Hain et al., 1993). The overexpression or downregulation of enzymes involved in phenylpropanoid and lignin biosynthesis has demonstrated that it is possible to alter the content and properties of lignin and its associated phenolics (reviewed in Boudet, 1998). This finding has important implications for the manipulation of plant quality with respect to pulping, forage digestibility, texture, and defense responses (Campbell and Sederoff, 1996). The expression of bacterial genes in transgenic plants also has proven to be effec...

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Isolation and characterization of tomato cDNA and genomic clones encoding the ubiquitin gene ubi3

Cited by 71 publications

References 36 publications

Differential Accumulation of Sunflower Tetraubiquitin mRNAs during Zygotic Embryogenesis and Developmental Regulation of Their Heat-Shock Response

Differential Accumulation of Sunflower Tetraubiquitin mRNAs during Zygotic Embryogenesis and Developmental Regulation of Their Heat-Shock Response

Gene dosage and genetic background affect miraculin accumulation in transgenic tomato fruits

Rerouting the Plant Phenylpropanoid Pathway by Expression of a Novel Bacterial Enoyl-CoA Hydratase/Lyase Enzyme Function

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