Transformation of floral organs with GFP in Medicago truncatula

Kamaté, K.; Rodríguez‐Llorente, Ignacio D.; Scholte, Marije; Durand, P.; Ratet, Pascal; Kondorosi, Éva; Kondorosi, Ádám; Trinh, T. H.

doi:10.1007/s002999900168

Cited by 40 publications

(22 citation statements)

References 31 publications

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“…For study of symbiotic nitrogen fixation, mycorrhizal interactions and legume-pathogen interactions, Medicago truncatula is a good legume model species (Cook, 1999) because of its small genome size, self-fertile nature and rapid generation period. In addition, for Medicago truncatula protocols for regeneration from leaves and cotyledons (Trieu and Harrisson, 1996;Trinh et al, 1998;Iantcheva et al, 1999) and transformation (Chabaud et al, 1996;Kamate´et al, 2000) were developed.…”

Section: Introductionmentioning

confidence: 99%

Embryo induction and regeneration from root explants of Medicago truncatula after osmotic pre-treatment

Iantcheva

Slavov

Prinsen

et al. 2005

Plant Cell Tiss Organ Cult

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Embryo induction and regeneration from suspension culture of two Medicago truncatula cvs. (cv. R 108 1 and cv. Jemalong) have been studied. The influence of osmotic pre-treatment (1 M solution of sucrose for 48 h and 72 h) of roots as an initial explant, on embryogenic efficiency of the suspension culture was assessed. In comparison to the control, the level of abscisic acid (ABA) increased significantly after osmotic stress. The increased ABA level did not correlate with the induction of embryogenesis neither with the improved embryogenic potential of cv. R 108 1. The shortest regeneration period and the highest percent of conversion to plants were found in cv. R 108 1 after 72-h pre-treatment of roots. The efficiency of somatic embryo conversion was less after 48-h pre-treatment and much less for the untreated control. Osmotic stress did not positively affect the process of embryogenesis from root explants of cv. Jemalong, confirming its cultivar dependence. A single cell suspension fraction was produced in both Medicago trunacatula cvs. during the somatic embryo maturation stage. A higher embryogenic potential than the initial suspension culture was established only for the cell suspension originating from 72-h pre-treated roots of cv. R 108 1. The data confirms that the process of somatic embryo induction and embryo conversion from root explants of cv. R 108 1 could be promoted by osmotic stress pre-treatment.

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Section: Introductionmentioning

confidence: 99%

Embryo induction and regeneration from root explants of Medicago truncatula after osmotic pre-treatment

Iantcheva

Slavov

Prinsen

et al. 2005

Plant Cell Tiss Organ Cult

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“…One aspect of this effort has been the development of enabling methodologies, such as efficient transformation methods (Trinh et al 1998;Kamaté et al 2000;Zhou et al 2004), high-throughput systems for forward and reverse genetics, including insertional mutagenesis (d 'Erfurth et al 2003), RNAi (Limpens et al 2003(Limpens et al , 2004, and TILLING (VandenBosch and Stacey 2003), and an effective network among research groups (http://www.medicago.org). In parallel to these activities, national and international programs are collaborating to characterize the genome of M. truncatula at the transcript (Fedorova et al 2002;Journet et al 2002;Lamblin et al 2003), protein (Gallardo et al 2003;Watson et al 2003;Imin et al 2004), and whole genome sequence levels (Young et al 2005).…”

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confidence: 99%

Distribution of Microsatellites in the Genome ofMedicago truncatula: A Resource of Genetic Markers That Integrate Genetic and Physical Maps

et al. 2006

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Microsatellites are tandemly repeated short DNA sequences that are favored as molecular-genetic markers due to their high polymorphism index. Plant genomes characterized to date exhibit taxonspecific differences in frequency, genomic location, and motif structure of microsatellites, indicating that extant microsatellites originated recently and turn over quickly. With the goal of using microsatellite markers to integrate the physical and genetic maps of Medicago truncatula, we surveyed the frequency and distribution of perfect microsatellites in 77 Mbp of gene-rich BAC sequences, 27 Mbp of nonredundant transcript sequences, 20 Mbp of random whole genome shotgun sequences, and 49 Mbp of BAC-end sequences. Microsatellites are predominantly located in gene-rich regions of the genome, with a density of one long (i.e., $20 nt) microsatellite every 12 kbp, while the frequency of individual motifs varied according to the genome fraction under analysis. A total of 1,236 microsatellites were analyzed for polymorphism between parents of our reference intraspecific mapping population, revealing that motifs (AT) n , (AG) n , (AC) n , and (AAT) n exhibit the highest allelic diversity. A total of 378 genetic markers could be integrated with sequenced BAC clones, anchoring 274 physical contigs that represent 174 Mbp of the genome and composing an estimated 70% of the euchromatic gene space.

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“…Initially we tested vacuum inWltration for germinating Medicago seeds (Trieu et al 2000), which has been reported to be similar to the vacuum inWltration of Arabidopsis inXorescences (Ye et al 1999). This approach should have been a rapid method for generating transgenic plants because it omitted steps involved in conventional tissue culture during the transformation and regeneration process (Kamaté et al 2000). However, we were not able to obtain any transgenic plants with the method previously described (Trieu et al 2000).…”

Section: Resultsmentioning

confidence: 96%

Heterologous expression of IAP1, a seed protein from bean modified by indole-3-acetic acid, in Arabidopsis thaliana and Medicago truncatula

Walz

Rusak

Park

et al. 2007

Planta

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The seed protein IAP1 from bean (PvIAP1; Phaseolus vulgaris L.) that is modified by the phytohormone indole-3-acetic acid (IAA) was heterologously expressed in the two reference plant species Arabidopsis thaliana and Medicago truncatula. For the transformation of Medicago we devised a novel protocol using seedling infiltration. When PvIAP1 was overexpressed under the control of the constitutive 35SCaMV promoter in Arabidopsis, the plants showed signs of earlier bolting and enhanced branching. Expression of a fusion protein of PvIAP1 with both a green fluorescence protein (GFP) as reporter and 6x histidine (His) tag under the control of the native bean IAP1 promoter resulted in the accumulation of the protein in both plant species exclusively in seeds as shown by immunoblotting and by fluorescence microscopy. During seed development, PvIAP1 was first expressed in the vascular bundle of Arabidopsis, whereas in later stages GFP fluorescence was visible essentially in all tissues of the seed. Fluorescence decreased rapidly after imbibition in the seeds for both Arabidopsis and Medicago, although the fluorescence persisted longer in Arabidopsis. GFP fluorescence was distributed evenly between an organelle fraction, the microsomal membrane fraction, and the cytosol. This was also confirmed by immunoblot analysis. Clusters of higher GFP fluorescence were observed by confocal microscopy. Although PvIAP1 protein accumulated in seeds of both Arabidopsis and Medicago, neither species post-translationally modified the protein with an indoleacyl moiety as shown by quantitative GC-MS analysis after alkaline hydrolysis. These results indicate an apparent specificity for IAA attachment in different plant species.

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Transformation of floral organs with GFP in Medicago truncatula

Cited by 40 publications

References 31 publications

Embryo induction and regeneration from root explants of Medicago truncatula after osmotic pre-treatment

Embryo induction and regeneration from root explants of Medicago truncatula after osmotic pre-treatment

Distribution of Microsatellites in the Genome ofMedicago truncatula: A Resource of Genetic Markers That Integrate Genetic and Physical Maps

Heterologous expression of IAP1, a seed protein from bean modified by indole-3-acetic acid, in Arabidopsis thaliana and Medicago truncatula

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