A simple and efficient method for testing Lettuce mosaic virus resistance in in vitro cultivated lettuce

Mazier, Marianne; German-Retana, Sylvie; Flamain, Fabrice; Dubois, Vincent; Botton, Emmanuel; Sarnette, Verane; Gall, Olivier Le; Candresse, Thierry; Maisonneuve, Brigitte

doi:10.1016/j.jviromet.2003.11.011

Cited by 18 publications

(16 citation statements)

References 18 publications

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“…Mazier et al [16] have described a simple and efficient system for in vitro inoculation of lettuce plants with Lettuce mosaic virus (LMV). The principle of their method relies on the mechanical inoculation of in vitro -grown lettuce plantlets using latex fingers dipped in sap extract from greenhouse-grown infected plants.…”

Section: Discussionmentioning

confidence: 99%

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An efficient in vitro-inoculation method for Tomato yellow leaf curl virus

Al‐Abdallat

Debei

Asmar

et al. 2010

Virol J

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BackgroundTomato yellow leaf curl virus (TYLCV) is a member of the family Geminiviridae, genus Begomovirus. To test the infectivity of TYLCV in tomato plants, an improved protocol for inoculation of in vitro-cultured tomato plants was developed.ResultsA TYLCV isolate was cloned, sequenced and used to construct a 1.8-mer infectious clone. Three weeks old microshoots of TYLCV-susceptible tomato plants were inoculated with Agrobacterium tumefaciens harboring the infectious clone for the TYLCV isolate. After two weeks, the TYLCV symptoms started to appear on the in vitro-inoculated plants and the symptoms became more severe and pronounced eight weeks post-inoculation. The method was used efficiently to uncover the resistance mechanism against TYLCV in Solanum habrochaites accession LA 1777, a wild tomato known for its high resistance to whitefly and TYLCV.ConclusionsThe reported in vitro-inoculation method can be used to screen tomato genotypes for their responses to TYLCV under controlled conditions and it will be a useful tool for better understanding of the TYLCV biology in tomato plants.

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Section: Discussionmentioning

confidence: 99%

“…Two previous reports have described the development of viral inoculation systems suitable for in vitro plants [16,17]. With the described systems, it has been possible to inoculate successfully plants grown in vitro using a mechanical approach.…”

Section: Introductionmentioning

confidence: 99%

An efficient in vitro-inoculation method for Tomato yellow leaf curl virus

Al‐Abdallat

Debei

Asmar

et al. 2010

Virol J

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“…Lettuce genetic transformation procedures are described by Dinant et al (1997) and the regeneration medium used is described by Mazier et al (2004). Leaves excised from 10-d-old seedlings (cultivated aseptically) were inoculated with A. tumefaciens strain C58 pGV2260 carrying the constructs to be introduced.…”

Section: In Vitro Culture Regeneration and Genetic Transformationmentioning

confidence: 99%

“…Newly in vitro-regenerated plantlets (without transformation) were produced from true leaves taken from sterile seedlings grown in vitro or from selected plants growing in the greenhouse, surface sterilized by immersing in a disinfecting solution for 30 min (Mazier et al, 2004), followed by three rinses with sterilized deionized water. In vitro regeneration procedures were performed as described by Mazier et al (2004).…”

Section: In Vitro Culture Regeneration and Genetic Transformationmentioning

confidence: 99%

“…It is the largest genome tested so far for retrotransposon mutagenesis. Lettuce is amenable to most in vitro culture techniques and is easy to transform via Agrobacterium tumefaciens (Michelmore et al, 1987;Chupeau et al, 1994;Mazier et al, 2004;Lelivelt et al, 2005). Interestingly, Yang et al (1993aYang et al ( , 1993b already tested transposon mutagenesis using the well-known DNA transposable activator/dissociation (Ac/Ds) elements from maize (Zea mays) in lettuce.…”

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Successful Gene Tagging in Lettuce Using the Tnt1 Retrotransposon from Tobacco

Mazier¹,

Botton²,

Flamain³

et al. 2007

Plant Physiol.

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The tobacco (Nicotiana tabacum) element Tnt1 is one of the few identified active retrotransposons in plants. These elements possess unique properties that make them ideal genetic tools for gene tagging. Here, we demonstrate the feasibility of gene tagging using the retrotransposon Tnt1 in lettuce (Lactuca sativa), which is the largest genome tested for retrotransposon mutagenesis so far. Of 10 different transgenic bushes carrying a complete Tnt1 containing T-DNA, eight contained multiple transposed copies of Tnt1. The number of transposed copies of the element per plant was particularly high, the smallest number being 28. Tnt1 transposition in lettuce can be induced by a very simple in vitro culture protocol. Tnt1 insertions were stable in the progeny of the primary transformants and could be segregated genetically. Characterization of the sequences flanking some insertion sites revealed that Tnt1 often inserted into genes. The progeny of some primary transformants showed phenotypic alterations due to recessive mutations. One of these mutations was due to Tnt1 insertion in the gibberellin 3b-hydroxylase gene. Taken together, these results indicate that Tnt1 is a powerful tool for insertion mutagenesis especially in plants with a large genome.

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Leafy Vegetables

Davey

Anthony

Power

et al. 2008

Compendium of Transgenic Crop Plants

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Leafy vegetables constitute an important part of a well‐balanced diet. Lettuce, chicory, and spinach are grown extensively as salad crops, with a range of cultivars being generated by conventional plant breeding. Somatic cell technologies, based on the regeneration of shoots from cultured cells and tissues by organogenesis and/or somatic embryogenesis, underpin conventional breeding through procedures such as in vitro pollination, microspore culture, embryo rescue, and exposure of somaclonal variation. Gene transfer techniques are most advanced in lettuce. Somatic hybridization through the fusion of isolated protoplasts has been exploited to circumvent sexual incompatibility barriers to introduce genes from wild species into cultivated lettuce. Transformation, based primarily on Agrobacterium ‐mediated DNA delivery, has been used to introduce specific traits into leafy vegetables, particularly in lettuce. Such genes include those for resistance to pests, and tolerance to herbicides and environmental conditions including drought, cold, and salinity. Improving the nutritional value and extending shelf life are other prime targets for genetic manipulation. The use of leafy vegetables as “plant factories” to synthesize high value recombinant proteins is an immediate target for transformation at the nuclear and organelle levels in leafy vegetables.

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A simple and efficient method for testing Lettuce mosaic virus resistance in in vitro cultivated lettuce

Cited by 18 publications

References 18 publications

An efficient in vitro-inoculation method for Tomato yellow leaf curl virus

An efficient in vitro-inoculation method for Tomato yellow leaf curl virus

Successful Gene Tagging in Lettuce Using the Tnt1 Retrotransposon from Tobacco

Leafy Vegetables

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