Arabidopsis Thaliana

Gracias, pero una vez que tienes el Agar líquido, ¿Cómo haces para lavarlo y no perderlo? lo pasas por una columna, o por un filtro cuando le añades el detergente o el ácido? O se podría hacer antes de disolverlo, cuando todavía está en suspensión?

use less dosage of silwet..0.01%

Have you tried SPME and GC/MS?

Finally, I got an answer from Lehle seeds. I am passing on the information I received for everyone who is interested in mutagenesis in Arabidopsis: "I apologize for your inconvenience, but LEHLE SEEDS is not providing the requested service at the present time. Our current staffing is not considered sufficient to initiate M1 productions at this time. Unfortunately, I am not aware of another commercial company which we can recommend to you. If and when our service is resumed, a notice will be posted on our web site at www.arabidopsis.com. However, we have no scheduled plans to resume this service in 2013. Chapter 2 entitled "Obtaining Mutants" in the CSHL Press book Arabidopsis A Laboratory Manual 2002 by Detlef Weigel and Jane Glazebrook contains a procedure for EMS mutation of Arabidopsis seeds. In addition, the EMS entry in the appendix includes important information, not found elsewhere to our knowledge, for how to chemically detoxify the chemical wastes generated by the procedure. I presume, that contacting the authors directly would yield additional details about their protocol. While EMS mutation is perhaps the most efficient approach, I bring to your attention the following references of the use of X-rays and Gamma-rays to mutate Arabidopsis pollen directly. Some early work in this area may also be found in the Arabidopsis Information Service (AIS, 1964-1990) see http://www.arabidopsis.org/ais/newaishint.jsp. Naito, K. et al. 2005 Transmissible and nontransmissible mutations induced by irradiating Arabidopsis thalian pollen with Gamma-rays and Carbon Ions. Genetics 169(2) 881-889 Dawson J Wilson ZA et al. 1993 Microspore and pollen development in six male-sterile mutants of Arabidopsis thaliana. Can J Botany 71(4) 629-638 Elsevier Ma T-H 1979 Micronuclei induced by X-rays and chemical mutagens in meiotic pollen mother cells of tradescantia: A promising mutagen test system. Mutation Research/ Environmental Mutagenesis and Related Subjects 64(5) pp. 307-315 And I remind you that it was was Dr. Laibach's PhD student Erna Reinholz's thesis work on the use of ionizing irradiation (X-rays) to mutate Arabidopsis flowers in the mid-1040's that first alerted plant genetists internationally to the relative ease by which mutations could be obtained from Arabidopsis. Reinholz E (1945) Auslösung von Röntgenmutationen bei Arabidopsis thaliana (L.) Heynh. und ihre Bedeutung für die Pflanzenzüchtung und Evolutionstheorie. Thesis, Univ. Frankfurt. Frankfurt am Main, Germany Disclaimer of intent to make recommendation. Providing this reference is for informational purposes only and is not and should not be considered a recommendation by LEHLE SEEDS that you or anyone else attempt or otherwise perform this procedure. LEHLE SEEDS does not make recommendations concerning chemical mutagenesis of plant seeds and is not doing so here."

Plant morphology is dramatically influenced by environmental signals. The growth and development of the root system is an excellent example of this developmental plasticity. Both the number and placement of lateral roots are highly responsive to nutritional cues. This indicates that there must be a signal transduction pathway that interprets complex environmental conditions and makes the “decision” to form a lateral root at a particular time and place. Lateral roots originate from differentiated cells in adult tissues. These cells must reenter the cell cycle, proliferate, and redifferentiate to produce all of the cell types that make up a new organ. Almost nothing is known about how lateral root initiation is regulated or coordinated with growth conditions. Here, a report of growth assay that allows this regulatory mechanism to be dissected in Arabidopsis (from American Society of Plant Physiologists, Environmental Regulation of Lateral Root Initiation in Arabidopsis1 by Jocelyn E. Malamy * and Katherine S. Ryan). When Arabidopsis seedlings are grown on nutrient media with a high sucrose to nitrogen ratio, lateral root initiation is dramatically repressed. Auxin localization appears to be a key factor in this nutrient-mediated repression of lateral root initiation. We have isolated a mutant, lateral root initiation 1 (lin1), that overcomes the repressive conditions. This mutant produces a highly branched root system on media with high sucrose to nitrogen ratios. The lin1 phenotype is specific to these growth conditions, suggesting that thelin1 gene is involved in coordinating lateral root initiation with nutritional cues. Therefore, these studies provide novel insights into the mechanisms that regulate the earliest steps in lateral root initiation and that coordinate plant development with the environment.

yes you can get those transgenic plants from ABRC or NASC website

You might want to look at this website http://botserv1.uzh.ch/home/grossnik/curtisvector/index_2.html, pMDC7 is a good candidate if you want to use gateway. But you will need to sign a MTA

Please also check the following: Alexandersson E, Saalbach G, Larsson C, Kjellbom P.Arabidopsis plasma membrane proteomics identifies components of transport, signal transduction and membrane trafficking. Plant Cell Physiol. 2004 Nov;45(11):1543-56 Nühse TS, Stensballe A, Jensen ON, Peck SC.Phosphoproteomics of the Arabidopsis plasma membrane and a new phosphorylation site database. Plant Cell. 2004 Sep;16(9):2394-405 Marmagne A, Rouet MA, Ferro M, Rolland N, Alcon C, Joyard J, Garin J, Barbier-Brygoo H, Ephritikhine G.Identification of new intrinsic proteins in Arabidopsis plasma membrane proteome. Mol Cell Proteomics. 2004 Jul;3(7):675-91

Thanks to all for the contribution. Although in the Arabidopsis gene nomenclature page no indication is specifically provided for mutants homozygous for a gene and hetrozygous for another (mostly embryo lethal mutants), I agree that gene/GENE is the best option

...thanks a lot for the comments! i will first try the GA treatment because we don´t have any 24h light chamber!

If you have the genomic DNA, you can perform a linker ligation-mediated PCR (maybe need nested) with dSpam-transposon end-specific primer(s) and follow by sequencing.

If anyone has problems germinating seed from either stock centre then please do tell us. Normally we will then perform a germination test to see what the germination frequency is in our hands (under the protocols specific to the lines if there are any documented). We also usually send replacement seeds even if the seeds are OK in our lab - but tell you how they performed for us and add some avuncular advice on how to grow arabidopsis (including our comedy cartoon at http://arabidopsis.info/information/kill.pdf and a link to our growing advice pages at http://arabidopsis.info/InfoPages?template=newgrow;web_section=arabidopsis ). Some ecotypes do have poor germination and have to be molly-coddled a little (to be technical :) - all the ebst, Sean