Genetics and expression of two pectinesterase genes in Valencia orange
Identifieur interne : 000153 ( Istex/Corpus ); précédent : 000152; suivant : 000154Genetics and expression of two pectinesterase genes in Valencia orange
Auteurs : C. Joseph Nairn ; Dennis J. Lewandowski ; Jacqueline K. BurnsSource :
- Physiologia Plantarum [ 0031-9317 ] ; 1998-02.
English descriptors
- KwdEn :
Abstract
The genetics and expression of pectinesterase (PE) genes were examined in Valencia orange. Degenerate primers based on partial amino acid sequence of a 36 kDa PE protein isolated from juice vesicles were used to amplify a 350 bp DNA fragment from cDNA prepared from juice vesicle total RNA. Two groups of 350 bp PE clones with 66% sequence identity were isolated. A clone from each group was used to screen a Valencia orange genomic DNA λ library. Two different lambda clones that contained complete PE coding sequence (CsPME1 and CsPME3) and a third lambda clone that contained partial PE sequence (CsPME2) were characterized. The CsPME1 gene contained two exons (1063 and 689 bp) interrupted by a 1452 bp intron, whereas the CsPME3 gene had two exons (844 and 686 bp) interrupted by a 771 bp intron. CsPME1 shared significant sequence similarity with the partial clone CsPME2, including the entire cloned region of the first exon, a large region in the 5′ portion of the intron and the 3′ portion of the second exon, but the 3′ portion of the intron and the 5′ portion of the second exon were dissimilar. Southern blots suggested that Valencia orange has two genes within each PE group. Full‐length cDNA clones that shared 99% sequence identity with CsPME1 and CsPME3 were isolated. Both groups of PE genes were differentially expressed in tissues of Valencia orange, and in addition CsPME3 appeared to be ethylene‐regulated. The deduced proteins of PE cDNA clones CsPME1 (63.5 kDa) and CsPME3 (56.3 kDa) were considerably larger than the PE protein we isolated from Valencia orange juice vesicles and also other mature plant PE proteins. The estimated size of group I (2.2 kb) and group II (2.0 kb) PE mRNAs also predicted a larger protein than was isolated from juice vesicles. Alignment of the mature tomato and mung bean PE proteins, the most N‐terminal sequence we obtained from polypeptides derived from the 36 kDa PE isolated from juice vesicles and the deduced amino acid sequences of plant PE cDNA clones suggest that a post‐translational cleavage event separates the variable N‐terminus from the more conserved C‐terminal domain of the mature PE protein.
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DOI: 10.1034/j.1399-3054.1998.1020210.x
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), Univ. of Florida, Citrus Research and Education Center, 700 Experiment Station Road, Lake Alfred, FL 33850, USA.</unparsedAffiliation>
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<keywordGroup xml:lang="en"><keyword xml:id="k1">Abscission</keyword>
<keyword xml:id="k2"><i>Citrus sinensis</i>
</keyword>
<keyword xml:id="k3">gene expression</keyword>
<keyword xml:id="k4">molecular cloning</keyword>
<keyword xml:id="k5">pectinesterase</keyword>
<keyword xml:id="k6">post‐translational modification</keyword>
<keyword xml:id="k7">Valencia orange</keyword>
</keywordGroup>
<abstractGroup><abstract type="main" xml:lang="en"><p>The genetics and expression of pectinesterase (PE) genes were examined in Valencia orange. Degenerate primers based on partial amino acid sequence of a 36 kDa PE protein isolated from juice vesicles were used to amplify a 350 bp DNA fragment from cDNA prepared from juice vesicle total RNA. Two groups of 350 bp PE clones with 66% sequence identity were isolated. A clone from each group was used to screen a Valencia orange genomic DNA λ library. Two different lambda clones that contained complete PE coding sequence (<i>CsPME1</i>
and <i>CsPME3</i>
) and a third lambda clone that contained partial PE sequence (<i>CsPME2</i>
) were characterized. The <i>CsPME1</i>
gene contained two exons (1063 and 689 bp) interrupted by a 1452 bp intron, whereas the <i>CsPME3</i>
gene had two exons (844 and 686 bp) interrupted by a 771 bp intron. <i>CsPME1</i>
shared significant sequence similarity with the partial clone <i>CsPME2</i>
, including the entire cloned region of the first exon, a large region in the 5′ portion of the intron and the 3′ portion of the second exon, but the 3′ portion of the intron and the 5′ portion of the second exon were dissimilar. Southern blots suggested that Valencia orange has two genes within each PE group. Full‐length cDNA clones that shared 99% sequence identity with <i>CsPME1</i>
and <i>CsPME3</i>
were isolated. Both groups of PE genes were differentially expressed in tissues of Valencia orange, and in addition <i>CsPME3</i>
appeared to be ethylene‐regulated. The deduced proteins of PE cDNA clones <i>CsPME1</i>
(63.5 kDa) and <i>CsPME3</i>
(56.3 kDa) were considerably larger than the PE protein we isolated from Valencia orange juice vesicles and also other mature plant PE proteins. The estimated size of group I (2.2 kb) and group II (2.0 kb) PE mRNAs also predicted a larger protein than was isolated from juice vesicles. Alignment of the mature tomato and mung bean PE proteins, the most N‐terminal sequence we obtained from polypeptides derived from the 36 kDa PE isolated from juice vesicles and the deduced amino acid sequences of plant PE cDNA clones suggest that a post‐translational cleavage event separates the variable N‐terminus from the more conserved C‐terminal domain of the mature PE protein.</p>
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<mods version="3.6"><titleInfo lang="en"><title>Genetics and expression of two pectinesterase genes in Valencia orange</title>
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<titleInfo type="alternative" contentType="CDATA" lang="en"><title>Genetics and expression of two pectinesterase genes in Valencia orange</title>
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<name type="personal"><namePart type="given">C.</namePart>
<namePart type="family">Joseph Nairn</namePart>
<affiliation>C. J. Nairn, Univ. of Georgia, Dept of Botany, 2502 Plant Sciences, Athens, GA 30602, USA; D. J. Lewandowski and J. K. Burns (corresponding author, e‐mail jkbu@tangelo.lal.ufl.edu), Univ. of Florida, Citrus Research and Education Center, 700 Experiment Station Road, Lake Alfred, FL 33850, USA.</affiliation>
<role><roleTerm type="text">author</roleTerm>
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<name type="personal"><namePart type="given">Dennis J.</namePart>
<namePart type="family">Lewandowski</namePart>
<affiliation>C. J. Nairn, Univ. of Georgia, Dept of Botany, 2502 Plant Sciences, Athens, GA 30602, USA; D. J. Lewandowski and J. K. Burns (corresponding author, e‐mail jkbu@tangelo.lal.ufl.edu), Univ. of Florida, Citrus Research and Education Center, 700 Experiment Station Road, Lake Alfred, FL 33850, USA.</affiliation>
<role><roleTerm type="text">author</roleTerm>
</role>
</name>
<name type="personal"><namePart type="given">Jacqueline K.</namePart>
<namePart type="family">Burns</namePart>
<affiliation>C. J. Nairn, Univ. of Georgia, Dept of Botany, 2502 Plant Sciences, Athens, GA 30602, USA; D. J. Lewandowski and J. K. Burns (corresponding author, e‐mail jkbu@tangelo.lal.ufl.edu), Univ. of Florida, Citrus Research and Education Center, 700 Experiment Station Road, Lake Alfred, FL 33850, USA.</affiliation>
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<originInfo><publisher>Munksgaard International Publishers</publisher>
<place><placeTerm type="text">Copenhagen</placeTerm>
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<dateIssued encoding="w3cdtf">1998-02</dateIssued>
<edition>Received 21 May, 1997; revised 12 November, 1997</edition>
<copyrightDate encoding="w3cdtf">1998</copyrightDate>
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<abstract lang="en">The genetics and expression of pectinesterase (PE) genes were examined in Valencia orange. Degenerate primers based on partial amino acid sequence of a 36 kDa PE protein isolated from juice vesicles were used to amplify a 350 bp DNA fragment from cDNA prepared from juice vesicle total RNA. Two groups of 350 bp PE clones with 66% sequence identity were isolated. A clone from each group was used to screen a Valencia orange genomic DNA λ library. Two different lambda clones that contained complete PE coding sequence (CsPME1 and CsPME3) and a third lambda clone that contained partial PE sequence (CsPME2) were characterized. The CsPME1 gene contained two exons (1063 and 689 bp) interrupted by a 1452 bp intron, whereas the CsPME3 gene had two exons (844 and 686 bp) interrupted by a 771 bp intron. CsPME1 shared significant sequence similarity with the partial clone CsPME2, including the entire cloned region of the first exon, a large region in the 5′ portion of the intron and the 3′ portion of the second exon, but the 3′ portion of the intron and the 5′ portion of the second exon were dissimilar. Southern blots suggested that Valencia orange has two genes within each PE group. Full‐length cDNA clones that shared 99% sequence identity with CsPME1 and CsPME3 were isolated. Both groups of PE genes were differentially expressed in tissues of Valencia orange, and in addition CsPME3 appeared to be ethylene‐regulated. The deduced proteins of PE cDNA clones CsPME1 (63.5 kDa) and CsPME3 (56.3 kDa) were considerably larger than the PE protein we isolated from Valencia orange juice vesicles and also other mature plant PE proteins. The estimated size of group I (2.2 kb) and group II (2.0 kb) PE mRNAs also predicted a larger protein than was isolated from juice vesicles. Alignment of the mature tomato and mung bean PE proteins, the most N‐terminal sequence we obtained from polypeptides derived from the 36 kDa PE isolated from juice vesicles and the deduced amino acid sequences of plant PE cDNA clones suggest that a post‐translational cleavage event separates the variable N‐terminus from the more conserved C‐terminal domain of the mature PE protein.</abstract>
<subject lang="en"><genre>keywords</genre>
<topic>Abscission</topic>
<topic>Citrus sinensis</topic>
<topic>gene expression</topic>
<topic>molecular cloning</topic>
<topic>pectinesterase</topic>
<topic>post‐translational modification</topic>
<topic>Valencia orange</topic>
</subject>
<relatedItem type="host"><titleInfo><title>Physiologia Plantarum</title>
</titleInfo>
<genre type="journal">journal</genre>
<identifier type="ISSN">0031-9317</identifier>
<identifier type="eISSN">1399-3054</identifier>
<identifier type="DOI">10.1111/(ISSN)1399-3054</identifier>
<identifier type="PublisherID">PPL</identifier>
<part><date>1998</date>
<detail type="volume"><caption>vol.</caption>
<number>102</number>
</detail>
<detail type="issue"><caption>no.</caption>
<number>2</number>
</detail>
<extent unit="pages"><start>226</start>
<end>235</end>
<total>10</total>
</extent>
</part>
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<identifier type="ArticleID">PPL1020210</identifier>
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