Functional expression of the <i>Erwinia uredovora</i> carotenoid biosynthesis gene <i>crtl</i> in transgenic plants showing an increase of β‐carotene biosynthesis activity and resistance to the bleaching herbicide norflurazon

Misawa, Norihiko; Yamano, Shigeyuki; Linden, Hartmut; Felipe, María Rosario de; Lucas, M. Mercedes; Ikenaga, Hiroshi; Sandmann, Gerhard

doi:10.1046/j.1365-313x.1993.04050833.x

Cited by 124 publications

(70 citation statements)

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“…The accumulation of astaxanthin in Ph-6 (and no additional hydroxylase expressed in the form of GlBCH) compared with Ph-7 (expressing Glbch), supports the proposed competition between the ketolase and hydroxylase activities in our transgenic lines (Fig. 3) in agreement with the previous in vitro (34,35) and in vivo (36) studies.…”

Section: Discussionsupporting

confidence: 91%

Combinatorial genetic transformation generates a library of metabolic phenotypes for the carotenoid pathway in maize

Zhu

Naqvi

Breitenbach

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

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331

263

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Combinatorial nuclear transformation is a novel method for the rapid production of multiplex-transgenic plants, which we have used to dissect and modify a complex metabolic pathway. To demonstrate the principle, we transferred 5 carotenogenic genes controlled by different endosperm-specific promoters into a white maize variety deficient for endosperm carotenoid synthesis. We recovered a diverse population of transgenic plants expressing different enzyme combinations and showing distinct metabolic phenotypes that allowed us to identify and complement rate-limiting steps in the pathway and to demonstrate competition between ␤-carotene hydroxylase and bacterial ␤-carotene ketolase for substrates in 4 sequential steps of the extended pathway. Importantly, this process allowed us to generate plants with extraordinary levels of ␤-carotene and other carotenoids, including complex mixtures of hydroxycarotenoids and ketocarotenoids. Combinatorial transformation is a versatile approach that could be used to modify any metabolic pathway and pathways controlling other biochemical, physiological, or developmental processes.induced mutation ͉ metabolic engineering ͉ transgenic plant ͉ provitamin A

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Section: Discussionsupporting

confidence: 91%

Combinatorial genetic transformation generates a library of metabolic phenotypes for the carotenoid pathway in maize

Zhu

Naqvi

Breitenbach

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

331

263

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“…PMI was used as the selectable marker gene. The construct combines the bacterial phytoene desaturase gene crtI, fused with the pea (Pisum sativum) RuBisCo small subunit transitencoding sequence (Misawa et al, 1993) and under the control of the constitutive 35S cauliflower mosaic virus (CaMV) promoter with the daffodil (Narcissus pseudonarcissus) psy, driven by the endospermspecific Gt1 promoter. To use a combination of only these two structural genes coding for carotenoid biosynthetic enzymes, omitting lcy, is the outcome of our previous findings showing that these were able to reconstitute the entire pathway, including the formation of ␣-and ␤-carotene and derived xanthophylls (Ye et al, 2000).…”

Section: Resultsmentioning

confidence: 99%

Golden Indica and Japonica Rice Lines Amenable to Deregulation

Hoa

Al‐Babili²,

Schaub³

et al. 2003

Plant Physiology

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As an important step toward free access and, thus, impact of GoldenRice, a freedom-to-operate situation has been achieved for developing countries for the technology involved. Specifically, to carry the invention beyond its initial "proof-ofconcept" status in a Japonica rice (Oryza sativa) cultivar, we report here on two transformed elite Indica varieties (IR64 and MTL250) plus one Japonica variety Taipei 309. Indica varieties are predominantly consumed in the areas with vitamin A deficiency. To conform with regulatory constraints, we changed the vector backbone, investigated the absence of beyondborder transfer, and relied on Agrobacterium tumefaciens-mediated transformation to obtain defined integration patterns. To avoid an antibiotic selection system, we now rely exclusively on phosphomannose isomerase as the selectable marker. Single integrations were given a preference to minimize potential epigenetic effects in subsequent generations. These novel lines, now in the T 3 generation, are highly valuable because they are expected to more readily receive approval for follow-up studies such as nutritional and risk assessments and for breeding approaches leading to locally adapted variety development.We reported previously on a genetically modified rice line, frequently termed GoldenRice, engineered to synthesize and accumulate pro-vitamin A (␤-carotene) in the endosperm (Ye et al., 2000). Since then, the concept of genetic engineering-based nutritional enhancement of rice to contribute to a sustained reduction of vitamin A deficiency (VAD) has evoked strong expectations to develop in the public sector and to deliver a safe product in a short period of time. However, to carry the project from the scientific discovery to impact, i.e. to actually provide grain to subsistence farmers and urban poor in developing countries free of charge for the technology used, a large number of obstacles, not only scientific in nature, need to be dealt with. These are contractual and regulatory aspects.Contractual issues have largely been solved to date, including a "freedom-to-operate" intellectual property situation for GoldenRice, the inventors, and their licensees. Free licenses have been granted by our license collaborators for patents used in this research designed to benefit resource-poor farmers in developing countries. The next step to be taken is to find regulatory acceptance in these countries, the prerequisite for most of the tasks ahead such as to allow grain production enabling feeding trials and to begin with diversified variety development.GoldenRice as published (Ye et al., 2000) demonstrates the feasibility of the scientific approach but does not yet represent a product. Its development was based on the finding that the precursor geranylgeranyldiphosphate, which wild-type rice endosperm is capable of synthesizing, can be used by the subsequent enzyme phytoene synthase (PSY) when the latter is supplemented by transformation (Burkhardt et al., 1997). Providing the full supplement of all necessary biosynthetic genes ...

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“…For construction of the binary vector pCas-Phyt, the TP-CrtB gene (Diretto et al, 2007) encoding the PSY CrtB from Pantoea ananatis fused with the pea (Pisum sativum) RbcS transit peptide (TP; Misawa et al, 1993) was isolated from pTrio1, a pBluescript KS derivative, using SacI and NcoI. The fragment thus obtained was ligated into the accordingly digested pCas-CrtI, a pBluescipt derivative harboring a TP-CrtI expression cassette driven by the cassava CP1 promoter (Beltrá n et al, 2010), to yield pCas-CrtB.…”

Section: Dna Gel Blot Analysismentioning

confidence: 99%

Provitamin A Accumulation in Cassava (Manihot esculenta) Roots Driven by a Single Nucleotide Polymorphism in a Phytoene Synthase Gene

et al. 2010

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Cassava (Manihot esculenta) is an important staple crop, especially in the arid tropics. Because roots of commercial cassava cultivars contain a limited amount of provitamin A carotenoids, both conventional breeding and genetic modification are being applied to increase their production and accumulation to fight vitamin A deficiency disorders. We show here that an allelic polymorphism in one of the two expressed phytoene synthase (PSY) genes is capable of enhancing the flux of carbon through carotenogenesis, thus leading to the accumulation of colored provitamin A carotenoids in storage roots. A single nucleotide polymorphism present only in yellow-rooted cultivars cosegregates with colored roots in a breeding pedigree. The resulting amino acid exchange in a highly conserved region of PSY provides increased catalytic activity in vitro and is able to increase carotenoid production in recombinant yeast and Escherichia coli cells. Consequently, cassava plants overexpressing a PSY transgene produce yellow-fleshed, high-carotenoid roots. This newly characterized PSY allele provides means to improve cassava provitamin A content in cassava roots through both breeding and genetic modification.

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Functional expression of the Erwinia uredovora carotenoid biosynthesis gene crtl in transgenic plants showing an increase of β‐carotene biosynthesis activity and resistance to the bleaching herbicide norflurazon

Cited by 124 publications

References 0 publications

Combinatorial genetic transformation generates a library of metabolic phenotypes for the carotenoid pathway in maize

Combinatorial genetic transformation generates a library of metabolic phenotypes for the carotenoid pathway in maize

Golden Indica and Japonica Rice Lines Amenable to Deregulation

Provitamin A Accumulation in Cassava (Manihot esculenta) Roots Driven by a Single Nucleotide Polymorphism in a Phytoene Synthase Gene

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