Expression of a Bacillus thuringiensis cry1Ab gene in transgenic white spruce and its efficacy against the spruce budworm (Choristoneura fumiferana)

Lachance, Denis; Hamel, Louis-Philippe; Pelletier, F.; Valero, José R. Lerma; Bernier‐Cardou, Michèle; Chapman, Kent D.; Frankenhuyzen, Kees van; Séguin, Armand

doi:10.1007/s11295-006-0072-y

Cited by 36 publications

(18 citation statements)

References 41 publications

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“…Low copy of the integration of the transgene may be considered as good result, since several studies show the positive correlation between high copy number of transgenes and cytosine methylation mediated silencing (Kohli et al 1996;Lechtenberg et al 2003;Matzke and Matzke 1995). Western blotting also reconfirmed the proper integration and expression of cry1Ab gene in the transgenic plants, however, we found smaller fragments that reacted with Cry1Ab antibody in leaf extracts of transgenic plants could be due to protein degradation by intrinsic leaf proteases (Australia New Zealand Food Authority (ANZFA) 2001) or degradation of protein during sample preparation (Breitler et al 2004;Cheng et al 1998;Fujimoto et al 1993;Lachance et al 2007). CryIAb levels in transgenic plants was estimated to be about 0.1% of total soluble leaf proteins by comparing with Bt protoxin as standard.…”

Section: Discussionsupporting

confidence: 63%

Enhanced resistance to a lepidopteran pest in transgenic sugar beet plants expressing synthetic cry1Ab gene

Jafari

Norouzi²,

Malboobi

et al. 2008

Euphytica

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Two diploid sugar beet genotypes of agronomical importance were transformed using Agrobactrium tumefaciens harboring pBI35Scry containing a synthetic cry1Ab gene. Leaf blade with attached shoot bases, a highly regenerative tissue, were used as explant substratum for transformation. PCR screening with cry1Ab-specific primers showed the presence of transgene in more than 50% of the regenerated kanamycin-resistant plants after treatment with the antibiotic. A transformation rate of 8.8-12.2% (depending on genotype) was achieved as revealed by genomic DNA dot blotting. The intact integration of transgene cassette into the genome was furthermore confirmed by Southern blot analysis. The expression of the cry1Ab gene encoding a truncated endotoxin (67 kDa) at about 0.1% of total soluble protein was achieved in the leaves of transgenic plants as shown by Western blot analysis. Bioassays under in vitro conditions with Spodoptera littoralis, one of the most important pests in sugar beet fields, demonstrated enhanced resistance against this pest. The inheritance of the inserted transgene was confirmed in F 1 plants obtained through crossing of T 0 plants with a cytoplasmic male sterile line. Transgenic plants are currently grown in a greenhouse and will be subjected to further bioassay analyses against other lepidopteran pests of sugar beet.

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Section: Discussionsupporting

confidence: 63%

Enhanced resistance to a lepidopteran pest in transgenic sugar beet plants expressing synthetic cry1Ab gene

Jafari

Norouzi²,

Malboobi

et al. 2008

Euphytica

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“…Transgenic poplars carrying antisense CAD and COMT genes with altered lignin grown for 4 years under field conditions in France and England showed relatively stable growth and development (Pilate et al 2002;Halpin et al 2007). Transgenic white spruce (Picea glauca) carrying the Bt gene also showed a continued insecticidal activity in the needles against budworm under confined field trials for 5 years (Lachance et al 2007). Likewise, transgenic poplars have shown relatively stable expression of the herbicide resistance gene (BAR) up to 8 years under field trials (Li et al 2008a(Li et al , 2009).…”

Section: Transgene Stability In the Genomementioning

confidence: 99%

Fate of transgenes in the forest tree genome

Ahuja

2010

Tree Genetics & Genomes

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During the last two decades, genetic engineering (GE) has been progressing at a steady pace in the forest trees. Transgenic trees carrying a variety of different transgenes have been produced, and are undergoing confined field trials in the world. However, there are questions regarding stability of transgene expression, and transgene escape that need to be addressed in the long-lived forest trees. Although relatively stable transgene expression has been reported for several target traits, including herbicide resistance, insect resistance, and lignin reduction in the vegetative propagules of several forest tree species, there were still unintended unstable events in transgenic plants. Long-term stability of transgene expression involved in these traits and others affecting yield (impacting growth) would be desirable in the vegetative propagules, and also in the generative progeny of the forest trees. Transgene escape through pollen, seed, and vegetative propagules from GE trees to native forest populations, although inevitable, remains an important regulatory issue. However, it may be possible to manage/minimize the risk of transgene spread via isolation in confined areas, and use of incompatible genotypes of feral tree populations in the vicinity of transgenic forest trees. Therefore, it is desirable to produce genetically stable transgenic trees, and have biocontainment measures in place for testing and deployment of the GE forest trees. Toward these goals (transgene stability and containment), innovative biotech strategies are being actively pursued, with reasonable success, in forest trees.

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“…A number of economically important genes (Table 2) (for example, herbicide resistance, insect and disease resistance, reduced lignin, and growth traits) have been transferred to produce transgenic plants in several forest tree species. These include (1) herbicide resistance genes (aroA, BAR, CP4) in Populus (Fillatti et al 1987;Donahue et al 1994;Meilan et al 2002;Li et al 2008a, b), Eucalyptus (Harcourt et al 2000), Pinus radiata and Picea abies (Bishop-Hurley et al 2001;Charity et al 2005); (2) insect resistance gene (Bt) in Populus (Leple et al 1995;Wang et al 1996;Meilan et al 2000;Hu et al 1999;Yang et al 2003); Pinus radiata , Pinus taeda (Tang and Tian 2003) and Picea glauca (Lachance et al 2007); (3) bacterial and fungal resistance genes (D4E1, ChitIV, STS, ESF39A, ech42) in hybrid poplar (Populus tremula 9 P. alba) (Mentag et al 2003), in Betula (Pasonen et al 2004), in Populus , in Ulmus Americana (Newhouse et al 2007), and in Picea mariana and hybrid poplar (Populus nigra 9 P. maximowiczii) (Noël et al 2005); (4) stress tolerance gene (CaPF1) in Pinus strobus (Tang et al 2007) and salt tolerance gene (codA) in Eucalyptus (Yu et al 2009); and (5) lignin modification genes (CAD,4Cl,COMT,CAld5H) in Populus (Hu et al 1999;Pilate et al 2002;Baucher et al 2003;Li et al 2003;Halpin et al 2007;Hancock et al 2007) and Betula pendula (Tiimonen et al 2005). In most of these short-term studies there was a fairly stable and predicable expression of transgenes in the selected transgenic trees under greenhouse and confined field trials.…”

Section: Stability Of Transgene Expressionmentioning

confidence: 99%

“…Transgenic poplars have shown relatively stable expression of the herbicide resistance genes up to 8 years under confined field trials (Meilan et al 2000(Meilan et al , 2002Li et al 2008a, b). Transgenic white spruce (Picea glauca) carrying the Bt gene also showed a continued insecticidal activity in the needles against budworm under confined field trials for 5 years (Lachance et al 2007). Generally, stable expression was detected in those transgenic lines carrying one to fewer copies of the transgene Li et al 2008a, b).…”

Section: Stability Of Transgene Expressionmentioning

confidence: 99%

Transgene stability and dispersal in forest trees

Ahuja

2009

Trees

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Transgenics from several forest tree species, carrying a number of commercially important recombinant genes, have been produced, and are undergoing confined field trials in a number of countries. However, there are questions and issues regarding stability of transgene expression and transgene dispersal that need to be addressed in long-lived forest trees. Variation in transgene expression is not uncommon in the primary transformants in plants, and is undesirable as it requires screening a large number of transformants in order to select transgenic lines with acceptable levels of transgene expression. Therefore, the current focus of plant transformation is toward fine tuning of transgene expression and stability in the transgenic forest trees. Although a number of studies have reported a relatively stable transgene expression for several target traits, including herbicide resistance, insect resistance, and lignin modification, there was also some unintended transgene instability in the genetically modified (GM) forest trees. Transgene dispersal from GM trees to feral forest populations and their containment remain important biological and regulatory issues facing commercial release of GM trees. Containment of transgenes must be in place to effectively prevent escape of transgenic pollen, seed, and vegetative propagules in economically important GM forest trees before their commercialization. Therefore, it is important to devise innovative technologies in genetic engineering that lead to genetically stable transgenic trees not only for qualitative traits (herbicide resistance, insect resistance), but also for quantitative traits (accelerated growth, increased height, increased wood density), and also prevent escape of transgenes in the forest trees.

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Expression of a Bacillus thuringiensis cry1Ab gene in transgenic white spruce and its efficacy against the spruce budworm (Choristoneura fumiferana)

Cited by 36 publications

References 41 publications

Enhanced resistance to a lepidopteran pest in transgenic sugar beet plants expressing synthetic cry1Ab gene

Enhanced resistance to a lepidopteran pest in transgenic sugar beet plants expressing synthetic cry1Ab gene

Fate of transgenes in the forest tree genome

Transgene stability and dispersal in forest trees

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