Development of Bt-cry5 Insect-resistant Potato Lines 'Spunta-G2' and 'Spunta-G3'

Douches, David S.; Li, W.; Zarka, Kelly; Coombs, Joseph; Pett, W.; Grafius, E.; El-Nasr, T. H. S. A.

doi:10.21273/hortsci.37.7.1103

Cited by 37 publications

(22 citation statements)

References 12 publications

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“…The bioassay method used was based on the assumption that the level of Cry protein in excised leaves was similar to that in the foliage of intact plants. The same assumption has been made in similar studies involving PTM bioassays (Ebora et al, 1994;Jansens et al, 1995;Westedt et al, 1998;Beuning et al, 2001;Douches et al, 1998Douches et al, , 2002Li et al, 1999;Davidson et al, 2002Davidson et al, , 2004, and has been validated by the observed high correlation between excised leaves and intact plants for growth of PTM larvae in response to cry gene expression (Davidson et al, 2002).…”

Section: Discussionmentioning

confidence: 55%

“…In potato, it is often desirable to couple high-level expression of the transgene in foliage with no expression in tubers. Constitutive promoters have been used to drive cry gene expression in all previous studies targeting PTM resistance in transgenic potatoes (Douches et al, , 2002Jansens et al, 1995;Cañedo et al, 1999;Li et al, 1999;Chakrabarti et al, 2000;Mohammed et al, 2000;Davidson et al, 2002Davidson et al, , 2004. Since PTM generally infests foliage of potato crops prior to the tubers (Foot, 1979), preventing foliar infestation may be sufficient to avoid tuber infestation.…”

Section: Discussionmentioning

confidence: 99%

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Expression of cry1Ac9 and cry9Aa2 genes under a potato light-inducible Lhca3 promoter in transgenic potatoes for tuber moth resistance

et al. 2006

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In transgenic potato it is often desirable to couple high-level expression in foliage with no expression in the edible tubers, especially for resistance to pests that primarily infest foliage. To accomplish this we have investigated the use of a light inducible Lhca3 promoter for transcriptional control of cry1Ac9 and cry9Aa2 genes for resistance to potato tuber moth (PTM) (Phthorimaea operculella). Thirty-five and thirty-one independently derived transgenic lines of potato cultivar Iwa were regenerated for the cry1Ac9 and cry9Aa2 genes respectively. Significantly inhibited larval growth of PTM on excised greenhouse-grown leaves was observed in 51% of the cry1Ac9-trangenic lines and 84% of the cry9Aa2-transgenic lines. RT-PCR analysis identified several transgenic lines with high levels of cry gene mRNA in leaves and no to low levels in tubers. Southern and ELISA analyses on eight selected cry1Ac9-transgenic lines revealed that they contained 2 to 9 copies of the cry1Ac9 gene and the amount of Cry protein in leaves was less than 60 ng g −1 of fresh leaf tissue. Southern analysis for four selected cry9Aa2-transgenic lines revealed that they contained 2 to 6 copies of the cry9Aa2 gene. This study has established that the expression of either the cry1Ac9 gene or the cry9Aa2 gene in transgenic potato plants offers protection against PTM larval damage in foliage when expressed under the transcriptional control of a Lhca3 light-inducible promoter. Several transgenic lines were identified with high cry gene expression, high resistance to PTM larvae in the foliage, and no or minimal cry gene expression in tubers.

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Section: Discussionmentioning

confidence: 55%

Section: Discussionmentioning

confidence: 99%

Expression of cry1Ac9 and cry9Aa2 genes under a potato light-inducible Lhca3 promoter in transgenic potatoes for tuber moth resistance

et al. 2006

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“…Relatively high level of B.t. protein expression with better PTM control could be achieved by using codon modified and truncated cry1Ac9 (Beuning et al 2001;Davidson et al 2002), cry 1Ia1 (Mohammed et al 2000;Douches et al 2002), and a hybrid Bt toxin (SN19) gene consisting of domain I and III of cry1Ba and domain II of cry1Ia (Naimov et al 2003). Gleave et al cloned and sequenced a B.t.…”

Section: Introductionmentioning

confidence: 99%

Expression of the cry9Aa2 B.t. gene in tobacco chloroplasts confers resistance to potato tuber moth

et al. 2006

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We report here the control of potato tuber moth (Phthorimaea operculella) by incorporating a truncated Bacillus thuringiensis cry9Aa2 gene in the plastid genome. Plasmids pSKC84 and pSKC85 are derivatives of a new polycistronic plastid transformation vector, pPRV312L, that carries spectinomycin resistance (aadA) as a selective marker and targets insertions in the trnI-trnA intergenic region. The Cry9Aa2 N-terminal region (82.1 kDa; 734 amino acids) was expressed in a cassette, which consists of 49 nucleotides of the cry9Aa2 leader and the 3'-untranslated region of the plastid rbcL gene (TrbcL), and relies on readthrough transcription from the plastid rRNA operon. In a tobacco leaf bioassay, expression of Cry9Aa2 conferred resistance to potato tuber moth. In accordance, the Cry9Aa2 insecticidal protein accumulated to high levels, approximately 10% of the total soluble cellular protein and approximately 20% in the membrane fraction. However, high-level Cry9Aa2 expression significantly delayed plant development. Thus, a practical system to control potato tuber moth by Cry9Aa2 expression calls for down-regulation of its expression.

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“…However, with the advent of genetic engineering, genes for insect resistance can be transferred into plants to develop insectresistant plants. Transgenic potatoes that express cry genes derived from various strains of the soil bacterium Bacillus thuringiensis (Bt) have exhibited resistance to PTM (Jansens et al 1995;Cañedo et al 1999;Li et al 1999;Chakrabarti et al 2000, Douches et al 2002Davidson et al 2002Davidson et al , 2004Meiyalaghan et al 2005Meiyalaghan et al , 2006aHagh et al 2009;Jacobs et al 2009;Kumar et al 2010).…”

Section: Introductionmentioning

confidence: 99%

Pyramiding transgenes for potato tuber moth resistance in potato

Meiyalaghan

Pringle

Barrell

et al. 2010

Plant Biotechnol Rep

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The feasibility of two strategies for transgene pyramiding using Agrobacterium-mediated transformation was investigated to develop a transgenic potato (Solanum tuberosum L. cv. Iwa) with resistance to potato tuber moth (PTM) (Phthorimaea operculella (Zeller)). In the first approach, cry1Ac9 and cry9Aa2 genes were introduced simultaneously using a kanamycin (nptII) selectable marker gene. The second approach involved the sequential introduction (re-transformation) of a cry1Ac9 gene, using a hygromycin resistance (hpt) selectable marker gene, into an existing line transgenic for a cry9Aa2 gene and a kanamycin resistance (nptII) selectable marker gene. Multiplex polymerase chain reaction (PCR) confirmed the presence of the specific selectable marker gene and both cry genes in all regenerated lines. The relative steady-state level of the cry gene transcripts in leaves was quantified in all regenerated lines by real-time PCR analysis. Re-transformation proved to be a flexible approach to effectively pyramid genes for PTM resistance in potato, since it allowed the second gene to be added to a line that was previously identified as having a high level of resistance. Larval growth of PTM was significantly inhibited on excised greenhouse-grown leaves in all transgenic lines, although no lines expressing both cry genes exhibited any greater resistance to PTM larvae over that previously observed for the individual genes. It is anticipated that these lines will permit more durable resistance by delaying the opportunities for PTM adaptation to the individual cry genes.

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Development of Bt-cry5 Insect-resistant Potato Lines 'Spunta-G2' and 'Spunta-G3'

Cited by 37 publications

References 12 publications

Expression of cry1Ac9 and cry9Aa2 genes under a potato light-inducible Lhca3 promoter in transgenic potatoes for tuber moth resistance

Expression of cry1Ac9 and cry9Aa2 genes under a potato light-inducible Lhca3 promoter in transgenic potatoes for tuber moth resistance

Expression of the cry9Aa2 B.t. gene in tobacco chloroplasts confers resistance to potato tuber moth

Pyramiding transgenes for potato tuber moth resistance in potato

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