Improving transformation efficiency ofArabidopsis thaliana by modifying the floral dip method

Martı́nez-Trujillo, Miguel; Limones-Briones, Verónica; Cabrera-Ponce, José Luis; Herrera-Estrella, Luis

doi:10.1007/bf02773350

Cited by 133 publications

(84 citation statements)

References 15 publications

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“…SUC2:AcFT and 35S:AcFT binary vectors were transformed into Agrobacterium tumefaciens GV3101 by electroporation. Arabidopsis were transformed using the method of Martinez-Trujillo et al 56 as follows: Agrobacterium harbouring the binary vector were streaked onto LB media plates with selective antibiotics and incubated at 28°C for 2 days. The bacteria were then scraped off the plate and resuspended in 10 ml of infiltration media (0.5 Â MS (Duchefa), 0.05% Silwett L-77 (Lehle Seeds), 5% sucrose) to an OD 600 B2 and droplets of this solution were pipetted onto unopened flower buds of Arabidopsis thaliana ft-1 mutants 56 .…”

Section: Methodsmentioning

confidence: 99%

FLOWERING LOCUS T genes control onion bulb formation and flowering

et al. 2013

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Onion (Allium cepa L.) is a biennial crop that in temperate regions is planted in the spring and, after a juvenile stage, forms a bulb in response to the lengthening photoperiod of late spring/ summer. The bulb then overwinters and in the next season it flowers and sets seed. FLOWERING LOCUS T (FT) encodes a mobile signaling protein involved in regulating flowering, as well as other aspects of plant development. Here we show that in onions, different FT genes regulate flowering and bulb formation. Flowering is promoted by vernalization and correlates with the upregulation of AcFT2, whereas bulb formation is regulated by two antagonistic FT-like genes. AcFT1 promotes bulb formation, while AcFT4 prevents AcFT1 upregulation and inhibits bulbing in transgenic onions. Long-day photoperiods lead to the downregulation of AcFT4 and the upregulation of AcFT1, and this promotes bulbing. The observation that FT proteins can repress and promote different developmental transitions highlights the evolutionary versatility of FT.

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Section: Methodsmentioning

confidence: 99%

FLOWERING LOCUS T genes control onion bulb formation and flowering

et al. 2013

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“…For the PLDZ2 promoter fusion, a 1.3-kb region upstream of the ORF of PLDZ2 was amplified from genomic Arabidopsis DNA by PCR using the following primers: FWDAttB1pPLDZ2 (5Ј-GGGG-ACA-AGT-TTG-TAC-AAA-AAA-GCA-GGC-TAA-gga-gta-cca-ttc-aca-aac-3Ј) and REVAttB2p PLDZ2 (5Ј-GGGG-AC-CAC-TTT-GTA-CAA-GAA-AGC-TGG-GTAccg-tcg-aca-tcg-att-aac-3Ј). The PCR product was cloned in the vector pDONR221 and transferred to pKGWFS7 (30) by recombination using Gateway BP and LR Clonase enzyme mixes, respectively, to produce the pPLDZ2:GUS-GFP gene fusion, which was then introduced into the Arabidopsis genome via the Agrobacterium-mediated transformation system using the protocol reported by Martinez-Trujillo et al (31). GUS histochemical analysis was carried out as reported (6).…”

Section: Methodsmentioning

confidence: 99%

Phospholipase DZ2 plays an important role in extraplastidic galactolipid biosynthesis and phosphate recycling in Arabidopsis roots

Cruz‐Ramírez¹,

Oropeza-Aburto²,

Razo-Hernández³

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

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Low phosphate (Pi) availability is one of the major constraints for plant productivity in natural and agricultural ecosystems. Plants have evolved a myriad of developmental and biochemical mechanisms to increase internal Pi uptake and utilization efficiency. One important biochemical pathway leading to an increase in internal Pi availability is the hydrolysis of phospholipids. Hydrolyzed phospholipids are replaced by nonphosphorus lipids such as galactolipids and sulfolipids, which help to maintain the functionality and structure of membrane systems. Here we report that a member of the Arabidopsis phospholipase D gene family (PLDZ2) is gradually induced upon Pi starvation in both shoots and roots. From lipid content analysis we show that an Arabidopsis pldz2 mutant is defective in the hydrolysis of phospholipids and has a reduced capacity to accumulate galactolipids under limiting Pi conditions. Morphological analysis of the pldz2 root system shows a premature change in root architecture in response to Pi starvation. These results show that PLDZ2 is involved in the eukaryotic galactolipid biosynthesis pathway, specifically in hydrolyzing phosphatidylcholine and phosphatidylethanolamine to produce diacylglycerol for digalactosyldiacylglycerol synthesis and free Pi to sustain other Pi-requiring processes.phosphate starvation ͉ phospholipids ͉ root architecture ͉ sulfolipids P hosphate (Pi) influences virtually all developmental and biochemical processes in plants. Pi is not only a constituent of key cell molecules such as ATP, nucleic acids, and phospholipids, but it is also a pivotal metabolic regulator of many processes including energy transfer, protein activation, and carbon and nitrogen metabolism. However, Pi availability can be one of the major constraints for plant growth in both natural and agricultural ecosystems because of its low mobility and high absorption capacity in the soil. As a response to this limitation, plants have evolved a range of developmental, biochemical, and symbiotic adaptive strategies to cope with low Pi availability (1, 2). In Arabidopsis, a general, 3-fold strategy to cope with low Pi availability has been described. (i) The release and uptake of Pi from external sources that are not readily available for plant uptake. This mechanism includes the transcriptional activation of high-affinity Pi transporters and the excretion of RNases, acid phosphatases, and organic acids (3-5). (ii) Changes in the architecture of the root system that reflect alterations in cell length, root meristem activity, root hair elongation, and an increased number of lateral roots (6, 7). These changes presumably increase the exploratory capacity of the root and the absorptive surface area. (iii) Optimization of Pi utilization due to a wide range of metabolic alterations, and the mobilization of Pi from internal reserves by the hydrolysis of nucleic acids, proteins, and the recycling of Pi from membrane phospholipids.During Pi deprivation, the total content of diverse phospholipids such as phosphatidylcholin...

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“…The presence of phytohormones in the tissue culture during regeneration also increases the chance of somatic mutations (Bent, 2000;Bent, 2006;Tague and Mantis, 2006). In order to resolve these difficulties, several plant transformation methods have been established and improved (Feldmann and Marks, 1987;Bechtold et al, 1993;Chang et al, 1994;Katavic et al, 1994;Clough and Bent, 1998;Richardson et al, 1998;Martinez-Trujillo et al, 2004;Zhang et al, 2006). Feldmann and Marks (1987) first reported that imbibition of A. thaliana germinating seeds with an appropriate A. tumefaciens strain can generate transformed progeny in the next generation.…”

Section: Stable Transformation By Floral Dippingmentioning

confidence: 99%