Cloning, Expression, and Characterization of (+)-δ-Cadinene Synthase: A Catalyst for Cotton Phytoalexin Biosynthesis

Chen, Xiaoya; Chen, Yuan; Heinstein, Peter; Davisson, V. Jo

doi:10.1006/abbi.1995.0038

Cited by 193 publications

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“…20 This isolated a cDNA clone that contained an insert of 1890 bp, with an open reading frame encoding a protein of 554 residues ( Figure 1), exactly the same size as the proteins encoded by cad1-C1 and cad1-C14. 11 Alignment of its nucleotide and de- duced amino acid sequences with members of the G. arboreum cad1 family revealed the highest identity with cad1-C1, 95.6% at nucleotide sequence level and 97.3% at amino acid sequence level, comparable to the identities between cad1-C1 and cad1-C14 (96.9% at amino acid sequence level). In the 3' noncoding region, its sequence identities with other members of the cad1-C subfamily were comparatively lower (86.4% with cad1-C1 and 85.8% with cad1-C14).…”

Section: Resultsmentioning

confidence: 90%

“…Nanking, as previously described. 11 The library was screened using a PCR-96-well plate method as described. 20 Briefly, the library of about 10 7 pfu was spread into 96 wells and mixed with E. coli strain XL1-Blue.…”

Section: Methodsmentioning

confidence: 99%

“…14 In G. arboreum cell suspension cultures, mRNA levels of both cad1-C and cad1-A subfamilies were increased by 10 times at 6-10 h posttreatment with V. dahliae cellular extracts, followed by increased levels of cyclase proteins. 11,12 Similarly, the expression of genes of a number of other plant terpene cyclases, such as tobacco 5-epi-aristolochene synthase (EAS), 15,16 Hyoscyamus muticus vetispiradiene synthase, 17 and Ricinus communis casbene synthase, 18 was induced in cell cultures or plant tissues by elicitation.Formation of pigment glands and their sesquiterpene content is genetically and developmentally controlled in cotton species but appears not to be related to environmental stimuli. 19 In developing cottonseeds, the young cotyledons are well protected by the seed-coat.…”

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Coordinated Accumulation of (+)-δ-Cadinene Synthase mRNAs and Gossypol in Developing Seeds of Gossypium hirsutum and a New Member of the cad1 Family from G. arboreum

et al. 1998

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A new member of the (+)-δ-cadinene synthase (CAD1) family was isolated from a Gossypium arboreum cDNA library. This cDNA encodes a protein that showed 97.3%, 96.9%, and 79.2% sequence identities with the proteins encoded by previously isolated cDNAs of cad1-C1, cad1-C14, and cad1-A, respectively. It may be grouped into the cad1-C subfamily as cad1-C2. Seeds of a glanded cotton cultivar, G. hirsutum cv. Sumian-6, were collected at different intervals during maturation, and the cad1 mRNA levels were analyzed by quantitative RT-PCR. The transcripts could be detected in seeds of 27 DPA (days postanthesis) and increased dramatically along with the seed maturation, which coordinated with an increase in sesquiterpene cyclase activities and subsequently the accumulation of gossypol. The transcription level detected with primers specific to cad1-C (including at least C1, C14, and cdn1) was higher than that detected with primers specific to cad1-A, and mRNA was detected also with cad1-C2-specific primers. This investigation indicates that, in developing seeds of the glanded cotton cultivar, genes of both the CAD1-C and CAD1-A subfamilies are expressed and there is an active biosynthesis of cadinene-type sesquiterpenes.Plant defense mechanisms involve the induced expression of a complex assortment of genes, including those coding for enzymes catalyzing the formation of antimicrobial secondary metabolites (phytoalexins). In cotton [Gossypium hirsutum L. and Gossypium arboreum L. (Malvaceae)], the production of cadinene-type sesquiterpenes was induced clearly in plants inoculated with the phytopathogenic bacterium Xanthomonas campestris 1,2 and in cell suspension cultures treated with elicitors prepared from the wilt disease-causing fungus Verticillium dahliae. 3 However, cotton sesquiterpenes, including the phenolic aldehyde gossypol, also are constitutive components present in subepidermal pigment glands. 4,5 The occurrence of pigment glands is a distinctive character in the tribe Gossypieae of the family Malvaceae. 4 The glands are found throughout the plant, and they are particularly dense in cotyledons of mature seeds of most cotton cultivars, and high levels of sesquiterpenes (mainly gossypol) are accumulated and deposited in seed glands. 6,7 Sesquiterpene cyclase catalyzes the cyclization of linear farnesyl diphosphate (FPP), the first step committed to the formation of various cyclic sesquiterpenes. [8][9][10] Previous investigations have characterized three cDNAs of the cotton sesquiterpene cyclase, (+)-δ-cadinene synthase (CAD1), from G. arboreum. On the basis of sequence similarities, they were grouped into two subfamilies, cad1-C (including C1 and C14) and cad1-A. 11,12 A cDNA from G. hirsutum, cdn1, which is over 95% identical to G. arboreum cad1-C1 and cad1-C14, has been isolated recently. 13 In addition, the enzyme was purified from bacterial-inoculated G. hirsutum folia tissues. 14 In G. arboreum cell suspension cultures, mRNA levels of both cad1-C and cad1-A subfamilies were increased by 10 times at 6-10 h p...

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Section: Resultsmentioning

confidence: 90%

Section: Methodsmentioning

confidence: 99%

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confidence: 99%

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Coordinated Accumulation of (+)-δ-Cadinene Synthase mRNAs and Gossypol in Developing Seeds of Gossypium hirsutum and a New Member of the cad1 Family from G. arboreum

et al. 1998

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“…DNA was isolated from the leaves of A. tricolor according to the method described by Chen et al [17]. DNA was cleaved and denatured in the gel and transferred to Hybond-N + membrane (Amersham, UK).…”

Section: Dna Gel Blot Analysismentioning

confidence: 99%

Isolation of a choline monooxygenase cDNA clone from Amaranthus tricolor and its expressions under stress conditions

Meng

Zhang

et al. 2001

Cell Res

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Plants synthesize the osmoprotectant glycine betaine (GB) via choline betaine aldehyde glycine betaine [1]. Two enzymes are involved in the pathway, choline monooxygenase (CMO) and betaine aldehyde dehydrogenase (BADH). A full length CMO cDNA (1,643bp) was cloned from Amaranthus tricolor. The open reading frame encoded a 442-amino acid polypeptide, which showed 69% identity with CMOs in Spinacia oleracea L. and Beta vulgaris L. DNA gel blot analysis indicated the presence of one copy of CMO gene in the A. tricolor genome. The expressions of CMO and BADH proteins in A.tricolor leaves significantly increased under salinization, drought and heat stress (42 o C), as determined by immunoblot analysis, but did not respond to cold stress (4 o C), or exogenous ABA application. The increase of GB content in leaves was parallel to CMO and BADH contents.

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“…For example, in cotton, sesquiterpene phytoalexins are elicited in response to bacterial or fungal infection. Chen et al (1995) observed that Gossypium arboreum cell suspension culture showed an increase of transcripts of a (+)-delta-cadinene synthase when challenged by a preparation from Verticillium dahliae. The authors concluded that such observation was consistent with a role for this enzyme as the first step in the pathways leading to the biosynthesis of phytoalexins gossypol and lacinilene C in cotton.…”

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confidence: 99%

A genomic approach to characterization of the Citrus terpene synthase gene family

Dornelas

Mazzafera

2007

Genet. Mol. Biol.

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Terpenes are a very large and structurally diverse group of secondary metabolites which are abundant in many essential oils, resins and floral scents. Additionally, some terpenes have roles as phytoalexins in plant-pathogen relationships, allelopathic inhibitors in plant-plant interactions, or as airborne molecules of plant-herbivore multitrophic signaling. Thus the elucidation of the biochemistry and molecular genetics of terpenoid biosynthesis has paramount importance in any crop species. With this aim, we searched the CitEST database for clusters of expressed sequence tags (ESTs) coding for terpene synthases. Herein is a report on the identification and in silico characterization of 49 putative members of the terpene synthase family in diverse Citrus species. The expression patterns and the possible physiological roles of the identified sequences are also discussed.

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Cloning, Expression, and Characterization of (+)-δ-Cadinene Synthase: A Catalyst for Cotton Phytoalexin Biosynthesis

Cited by 193 publications

References 0 publications

Coordinated Accumulation of (+)-δ-Cadinene Synthase mRNAs and Gossypol in Developing Seeds of Gossypium hirsutum and a New Member of the cad1 Family from G. arboreum

Coordinated Accumulation of (+)-δ-Cadinene Synthase mRNAs and Gossypol in Developing Seeds of Gossypium hirsutum and a New Member of the cad1 Family from G. arboreum

Isolation of a choline monooxygenase cDNA clone from Amaranthus tricolor and its expressions under stress conditions

A genomic approach to characterization of the Citrus terpene synthase gene family

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