Regulation of Sesquiterpene Cyclase Gene Expression (Characterization of an Elicitor- and Pathogen-Inducible Promoter)

Yin, Shaohui; Mei, Ling; Newman, Jeffrey D.; Back, Kyoungwhan; Chappell, Joseph

doi:10.1104/pp.115.2.437

Cited by 62 publications

(57 citation statements)

References 80 publications

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“…MJ elicitation has also been implicated in the activation of the defense related leucine aminopeptidase promoter in Nicotiana, the strictosidine synthase promoter in Catharanthus, and a sesquiterpene synthase promoter in Nicotiana are activated with MJ elicitation (17,36,40). In this investigation, we have shown that five paclitaxel biosynthetic pathway gene promoters are likewise activated by MJ elicitation.…”

Section: Withdrawn At Authors Requestmentioning

confidence: 61%

WITHDRAWN: TcJAMYC: A bHLH transcription factor that activates paclitaxel biosynthetic pathway genes in yew

Nims¹,

Vongpaseuth²,

Roberts³

et al. 2009

Journal of Biological Chemistry

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Abstract:Paclitaxel (Taxol ®) is a highly modified diterpene anti-cancer agent produced by the gymnosperm Taxus. Taxus suspension cell cultures have the potential to provide a sustainable source of paclitaxel, but the paclitaxel biosynthetic pathway is not fully characterized, making metabolic engineering efforts difficult. Methyl jasmonate (MJ) is used to elicit paclitaxel production in suspension cultures. Here we show that the promoters of five genes encoding enzymes of the paclitaxel biosynthetic pathway are activated by MJ elicitation.Thus, elicitation of paclitaxel production by MJ is regulated at least in part at the level of transcription. A transcription factor that positively activates the promoters of paclitaxel biosynthetic genes has been cloned. This transcription factor, TcJAMYC, possesses a high degree of similarity to AtMYC2 and JAMYC2, which are known to regulate the expression of jasmonate inducible genes in other systems. TcJAMYC binds to E-boxes found in the promoters of the paclitaxel pathway genes, and these promoters are transiently activated by TcJAMYC overexpression. Addition of MJ attenuates the effect of TcJAMYC on the pathway promoters, suggesting that TcJAMYC could be engineered into Taxus cells to avoid the negative regulation of MJ-induced genes that follows initial MJinduced positive regulation.This strategy could prolong the expression of paclitaxel pathway genes, and increase paclitaxel production.

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Section: Withdrawn At Authors Requestmentioning

confidence: 61%

WITHDRAWN: TcJAMYC: A bHLH transcription factor that activates paclitaxel biosynthetic pathway genes in yew

Nims¹,

Vongpaseuth²,

Roberts³

et al. 2009

Journal of Biological Chemistry

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“…In tobacco, formation of sesquiterpenoid phytoalexins is also catalyzed by multiple isozymes of EAS, and the tobacco genome is believed to contain 12-15 copies EAS genes. 15,16 Total RNAs were isolated from seeds of a glanded cotton cultivar G. hirsutum cv. Sumian-6, at four intervals ranging from 20, 27, 35, to 40 days postanthesis (DPA), approximately corresponding to stages of embryogenensis of the later cotyledon, early maturation, mid-maturation, and late maturation.…”

Section: Resultsmentioning

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.…”

mentioning

confidence: 99%

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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“…The coordinated responses suggest that GaWRKY1 and CAD1-A are components of the same pathway of cotton defense reaction, which involves induced biosynthesis of sesquiterpene phytoalexins. Searching of databases revealed the presence of W-box elements in the promoter region of sesquiterpene cyclase genes of other plants, such as 5-epi-aristolochene synthase gene EAS4 of tobacco (Yin et al, 1997), putative sesquiterpene cyclase genes of Arabidopsis (At1g31950), and rice (Oryza sativa; BAC99543.1). Therefore, WRKY-W-box interactions are likely operating also in other plants in regulation of sesquiterpene biosynthesis.…”

Section: Discussionmentioning

confidence: 99%

Characterization of GaWRKY1, a Cotton Transcription Factor That Regulates the Sesquiterpene Synthase Gene (+)-δ-Cadinene Synthase-A

et al. 2004

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The cotton (+)-δ-cadinene synthase (CAD1), a sesquiterpene cyclase, catalyzes a branch-point step leading to biosynthesis of sesquiterpene phytoalexins, including gossypol. CAD1-A is a member of CAD1 gene family, and its promoter contains a W-box palindrome with two reversely oriented TGAC repeats, which are the proposed binding sites of WRKY transcription factors. We isolated several WRKY cDNAs from Gossypium arboreum. One of them, GaWRKY1, encodes a protein containing a single WRKY domain and a putative N-terminal Leu zipper. Similar to genes encoding enzymes of cotton sesquiterpene pathway, GaWRKY1 was down-regulated in a glandless cotton cultivar that contained much less gossypol. GaWRKY1 showed a temporal and spatial pattern of expression comparable to that of CAD1-A in various aerial organs examined, including sepal, stigma, anther, and developing seeds. In suspension cells, expression of both GaWRKY1 and CAD1-A genes and biosynthesis of sesquiterpene aldehydes were strongly induced by a fungal elicitor preparation and methyl jasmonate. GaWRKY1 interacted with the 3× W-box derived from CAD1-A promoter in yeast (Saccharomyces cerevisiae) one-hybrid system and in vitro. Furthermore, in transgenic Arabidopsis plants, overexpression of GaWRKY1 highly activated the CAD1-A promoter, and transient assay in tobacco (Nicotiana tabacum) leaves demonstrated that W-box was required for this activation. These results suggest that GaWRKY1 participates in regulation of sesquiterpene biosynthesis in cotton, and CAD1-A is a target gene of this transcription factor.

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Regulation of Sesquiterpene Cyclase Gene Expression (Characterization of an Elicitor- and Pathogen-Inducible Promoter)

Cited by 62 publications

References 80 publications

WITHDRAWN: TcJAMYC: A bHLH transcription factor that activates paclitaxel biosynthetic pathway genes in yew

WITHDRAWN: TcJAMYC: A bHLH transcription factor that activates paclitaxel biosynthetic pathway genes in yew

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

Characterization of GaWRKY1, a Cotton Transcription Factor That Regulates the Sesquiterpene Synthase Gene (+)-δ-Cadinene Synthase-A

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