Plastid transformation in Arabidopsis thaliana

Sikdar, S. R.; Serino, Germán; Chaudhuri, S.K.; Maliga, Pál

doi:10.1007/s002990050525

Cited by 147 publications

(101 citation statements)

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“…In many cases, however, low transformation efficiencies have been reported for species other than tobacco (Sikdar et al 1998;Sidorov et al 1999;Ruf et al 2001;Hou et al 2003;Skarjinskaia et al 2003;Zubko et al 2004;Nguyen et al 2005;Nugent et al 2006;De Marchis et al 2009). Low recovery of transplastomic shoots has been attributed to multiple reasons, such as a relatively inefficient homologous recombination system, non-optimal homology and length of flanking regions, the promoter used for the expression of the selectable marker gene, the kind of explant, or the regeneration protocol.…”

Section: Introductionmentioning

confidence: 99%

High efficiency plastid transformation in potato and regulation of transgene expression in leaves and tubers by alternative 5′ and 3′ regulatory sequences

Valkov¹,

Gargano

Manna³

et al. 2010

Transgenic Res

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Transformation of potato plastids is limited by low transformation frequencies and low transgene expression in tubers. In order to improve the transformation efficiency, we modified the regeneration procedure and prepared novel vectors containing potato flanking sequences for transgene integration by homologous recombination in the Large Single Copy region of the plastome. Vector delivery was performed by the biolistic approach. By using the improved regeneration procedure and the potato flanking sequences, we regenerated about one shoot every bombardment. This efficiency corresponds to 15-18-fold improvement compared to previous results with potato and is comparable to that usually achieved with tobacco. Further, we tested five promoters and terminators, and four 5 0 -UTRs, to increase the expression of the gfp transgene in tubers. In leaves, accumulation of GFP to about 4% of total soluble protein (TSP) was obtained with the strong promoter of the rrn operon, a synthetic rbcL-derived 5 0 -UTR and the bacterial rrnB terminator. GFP protein was detected in tubers of plants transformed with only four constructs out of eleven. Best results (up to approximately 0.02% TSP) were achieved with the rrn promoter and rbcL 5 0 -UTR construct, described above, and another containing the same terminator, but with the promoter and 5 0 -UTR from the plastid clpP gene. The results obtained suggest the potential use of clpP as source of novel regulatory sequences in constructs aiming to express transgenes in amyloplasts and other non-green plastids. Furthermore, they represent a significant advancement of the plastid transformation technology in potato, of relevance to its implementation in potato breeding and biotechnology.

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Section: Introductionmentioning

confidence: 99%

High efficiency plastid transformation in potato and regulation of transgene expression in leaves and tubers by alternative 5′ and 3′ regulatory sequences

Valkov¹,

Gargano

Manna³

et al. 2010

Transgenic Res

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“…Transformations of the Arabidopsis, potato, and tomato chloroplast genomes were achieved using the tobacco chloroplast genome flanking sequences but efficiencies were much lower than in tobacco because of less than 100% sequence identities (Sikdar et al 1998;Sidorov et al 1999;Ruf et al 2001). In another study, Petunia flanking sequences were used to transform the tobacco plastid genome and transformation efficiency was lower than was observed with endogenous flanking sequences (DeGray et al 2001).…”

Section: Implications For Homologous Recombination Of Transgenes In Cmentioning

confidence: 99%

The complete nucleotide sequence of the cassava (Manihot esculenta) chloroplast genome and the evolution of atpF in Malpighiales: RNA editing and multiple losses of a group II intron

et al. 2008

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The complete sequence of the chloroplast genome of cassava (Manihot esculenta, Euphorbiaceae) has been determined. The genome is 161,453 bp in length and includes a pair of inverted repeats (IR) of 26,954 bp. The genome includes 128 genes; 96 are single copy and 16 are duplicated in the IR. There are four rRNA genes and 30 distinct tRNAs, seven of which are duplicated in the IR. The infA gene is absent; expansion of IRb has duplicated 62 amino acids at the 3′ end of rps19 and a number of coding regions have large insertions or deletions, including insertions within the 23S rRNA gene. There are 17 intron-containing genes in cassava, 15 of which have a single intron while two (clpP, ycf3) have two introns. The usually conserved atpF group II intron is absent and this is the first report of its loss from land plant chloroplast genomes. The phylogenetic distribution of the atpF intron loss was determined by a PCR survey of 251 taxa representing 34 families of Malpighiales and 16 taxa from closely related rosids. The atpF intron is not only missing in cassava but also from closely related Euphorbiaceae and other Malpighiales, suggesting that there have been at least seven independent losses. In cassava and all other sequenced Malphigiales, atpF gene sequences showed a strong association between C-to-T substitutions at nucleotide position 92 and the loss of the intron, suggesting that recombination between an edited mRNA and the atpF gene may be a possible mechanism for the intron loss.

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“…Plastid transformation of tobacco (Nicotiana tabacum) is routine in a number of laboratories, but there are only a limited number of isolated reports relating to genera outside Nicotiana. These include tomato (Ruf et al 2001), rice (Khan and Maliga 1999), potato (Sidorov et al 1999), carrot (Kumar et al 2004) and several Brassicaceae, including Arabidopsis (Sikdar et al 1998), Brassica napus (Hou et al 2002) and Lesquerella fendleri (Skarjinskaia et al 2003). All of these investigators used biolistics to deliver DNA into chloroplasts.…”

Section: Introductionmentioning

confidence: 99%

Plastid transformants of tomato selected using mutations affecting ribosome structure

Nugent

Have²,

Gulik³

et al. 2005

Plant Cell Rep

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Tomato plastid transformants were obtained using two vectors containing cloned plastid DNA of either Nicotiana tabacum or Solanum nigrum and including point mutations conferring resistance to spectinomycin and streptomycin. Transformants were recovered after PEGmediated direct DNA uptake into protoplasts, followed by selection on spectinomycin-containing medium. Sixteen lines contained the point mutation, as confirmed by mapping restriction enzyme sites. One line obtained with each vector was analysed in more detail, in comparison with a spontaneous spectinomycin-resistant mutant. Integration of the cloned Solanum or Nicotiana plastid DNA, by multiple recombination events, into the tomato plastome was confirmed by sequence analysis of the targeted region of plastid DNA in the inverted repeat region. Maternal inheritance of spectinomycin and streptomycin resistances or sensitivity in seedlings also confirmed the transplastomic status of the two transformants. The results demonstrate the efficacy in tomato of a selection strategy which avoids the integration of a dominant bacterial antibiotic resistance gene.Communicated by H. Lörz

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Plastid transformation in Arabidopsis thaliana

Cited by 147 publications

References 0 publications

High efficiency plastid transformation in potato and regulation of transgene expression in leaves and tubers by alternative 5′ and 3′ regulatory sequences

High efficiency plastid transformation in potato and regulation of transgene expression in leaves and tubers by alternative 5′ and 3′ regulatory sequences

The complete nucleotide sequence of the cassava (Manihot esculenta) chloroplast genome and the evolution of atpF in Malpighiales: RNA editing and multiple losses of a group II intron

Plastid transformants of tomato selected using mutations affecting ribosome structure

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