Potato Transformation with Modified Nucleotide Sequences of the cry9Aa2 Gene Improves Resistance to Potato Tuber Moth

Jacobs, Jeanne M. E.; Takla, Michael; Docherty, Lisa C.; Frater, Christina M.; Markwick, Ngaire P.; Meiyalaghan, Sathiyamoorthy; Conner, A. J.

doi:10.1007/s11540-009-9140-7

Cited by 12 publications

(14 citation statements)

References 32 publications

(57 reference statements)

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“…4). However, this level of resistance was no greater than that previously observed from the use of only the cry1Ac9 gene (Davidson et al 2002, 2004, Meiyalaghan et al 2006a or only the cry9Aa2 gene (Meiyalaghan et al 2005(Meiyalaghan et al , 2006aJacobs et al 2009). The failure to observe an enhanced level of resistance to PTM larvae is surprising given the previous experimental results from simulating the pyramiding of several cry genes (Meiyalaghan et al 2006c).…”

Section: Re-transformation Simultaneous Transformationcontrasting

confidence: 54%

“…Considerable variation in transgene expression among independently derived insect-resistant transgenic plants has been commonly reported in other studies (e.g. Peferoen et al 1990;Van Rie et al 1994;Beuning et al 2001), including Iwa potato lines transgenic for either the cry1Ac9 gene (Davidson et al 2002(Davidson et al , 2004Meiyalaghan et al 2006a) or the cry9Aa2 gene (Meiyalaghan et al 2005(Meiyalaghan et al , 2006aJacobs et al 2009). Such variation is usually attributed to unpredictable levels of transgene expression as a consequence of position effects resulting from differences Plant Biotechnol Rep (2010) 4:293-301 299 in the integration site of the transgenes within the plant genome and/or differences in T-DNA copy number (Conner and Christey 1994).…”

Section: Re-transformation Simultaneous Transformationsupporting

confidence: 52%

“…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%

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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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Section: Re-transformation Simultaneous Transformationcontrasting

confidence: 54%

Section: Re-transformation Simultaneous Transformationsupporting

confidence: 52%

Section: Introductionmentioning

confidence: 99%

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Pyramiding transgenes for potato tuber moth resistance in potato

Meiyalaghan

Pringle

Barrell

et al. 2010

Plant Biotechnol Rep

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“…One of the main aspects to be considered in this regard are position effects, which are a result of the random integration of transgenes into a genome (Barrell et al, 2013). Position effects frequently are reported in potato and can override the regulatory control of the pro moter associated with the transgene, such that the tran script level, as well as the temporal and spatial expression patterns of the transgene are altered (Meiyalaghan et al, 2006;Barrell & Conner, 2009;Jacobs et al, 2009;Bar rell et a l, 2013). It has been shown that in A. thaliana methylation of the 35S promoter can cause epigenetic si lencing of transgenic RNAi constructs, leading to strongly varying levels of small RNAs between lines (Kyndt et al, 2013).…”

Section: Meloidogyne Chitwoodi Rnai Resistance In Potatomentioning

confidence: 99%

Plant-mediated RNA interference of effector gene Mc16D10L confers resistance against Meloidogyne chitwoodi in diverse genetic backgrounds of potato and reduces pathogenicity of nematode offspring

Dinh

Zhang

Brown

et al. 2014

Nematol

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Meloidogyne chitwoodi is a major problem for potato production in the Pacific Northwest of the USA. In spite of long term breeding efforts no commercial potato cultivars with resistance to M. chitwoodi exist to date. The RMcUblb) resistance gene against M. chitwoodi has been introgressed from Solanum bulbocastanum into cultivated potato (S. tuberosum), but M. chitwoodi pathotypes are able to overcome this resistance. In this study, an RNA interference (RNAi) transgene targeting the M. chitwoodi effector gene Mcl6D10L was introduced into potato cvs Russet Burbank and Desiree, and the advanced breeding line PA99N82-4, which carries the RMcl(blb) gene. Stable transgenic lines were generated for glasshouse infection assays. At 35 days after inoculation (DAI) with M. chitwoodi race 1 the number of egg masses (g root)-1 formed on RNAi lines of cvs Russet Burbank and Desiree was reduced significantly by up to 68% compared to empty vector control plants. At 55 DAI, the number of eggs was reduced significantly by up to 65%. In addition, RNAi of Mcl6D10L significantly reduced the development of egg masses and eggs formed by the RMcl(blb) resistance breaking M. chitwoodi pathotype Roza on PA99N82-4 by up to 47 and 44%, respectively. Importantly, the plant-mediated silencing effect of Mcl6D10L was transmitted to M. chitwoodi offspring and significantly reduced pathogenicity in the absence of selection pressure on empty vector control plants. This finding suggests that the RNAi effect is stable and nematode infection decreases regardless of the genotype of the host once the RNAi process has been initiated in the nematode through a transgenic plant. In summary, plantmediated down-regulation of effector gene Mcl6D10L provides a promising new tool for molecular breeding against M. chitwoodi.

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“…When developing large populations of independently derived transgenic lines, only the first shoot to regenerate from each transformation event is usually selected [14,17]. This is based on a widely held view that minimum time in tissue culture is favourable to avoid undesired somaclonal variation [32], and the assumption that a shoot taking a longer time to regenerate has a greater chance of producing off-types.…”

Section: Discussionmentioning

confidence: 99%

Regeneration of multiple shoots from transgenic potato events facilitates the recovery of phenotypically normal lines: assessing a cry9Aa2 gene conferring insect resistance

et al. 2011

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BackgroundThe recovery of high performing transgenic lines in clonal crops is limited by the occurrence of somaclonal variation during the tissue culture phase of transformation. This is usually circumvented by developing large populations of transgenic lines, each derived from the first shoot to regenerate from each transformation event. This study investigates a new strategy of assessing multiple shoots independently regenerated from different transformed cell colonies of potato (Solanum tuberosum L.).ResultsA modified cry9Aa2 gene, under the transcriptional control of the CaMV 35S promoter, was transformed into four potato cultivars using Agrobacterium-mediated gene transfer using a nptII gene conferring kanamycin resistance as a selectable marker gene. Following gene transfer, 291 transgenic lines were grown in greenhouse experiments to assess somaclonal variation and resistance to potato tuber moth (PTM), Phthorimaea operculella (Zeller). Independently regenerated lines were recovered from many transformed cell colonies and Southern analysis confirmed whether they were derived from the same transformed cell. Multiple lines regenerated from the same transformed cell exhibited a similar response to PTM, but frequently exhibited a markedly different spectrum of somaclonal variation.ConclusionsA new strategy for the genetic improvement of clonal crops involves the regeneration and evaluation of multiple shoots from each transformation event to facilitate the recovery of phenotypically normal transgenic lines. Most importantly, regenerated lines exhibiting the phenotypic appearance most similar to the parental cultivar are not necessarily derived from the first shoot regenerated from a transformed cell colony, but can frequently be a later regeneration event.

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Potato Transformation with Modified Nucleotide Sequences of the cry9Aa2 Gene Improves Resistance to Potato Tuber Moth

Cited by 12 publications

References 32 publications

Pyramiding transgenes for potato tuber moth resistance in potato

Pyramiding transgenes for potato tuber moth resistance in potato

Plant-mediated RNA interference of effector gene Mc16D10L confers resistance against Meloidogyne chitwoodi in diverse genetic backgrounds of potato and reduces pathogenicity of nematode offspring

Regeneration of multiple shoots from transgenic potato events facilitates the recovery of phenotypically normal lines: assessing a cry9Aa2 gene conferring insect resistance

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