How to introduce a mutagenesis in plant? - (Sep/06/2012 )
I want to make a mutagenesis of a seeds of a plant .What type of mutagen to use and how to make the mutagen?
Btw. if the question is little stupid that is because I am from the I.T. and I am totaly new to bio stuff.
there are many different methods that have been used throughout the years, I remember at some point there were a lot of radioactive induced mutations, there's also the use of EMS, this paper could probably be useful http://www.plantphysiol.org/content/131/1/139.full
or i just had a quick look at this paper which might get you started http://www.iaea.org/Publications/Magazines/Bulletin/Bull344/34405682533.pdf
Ok.And how to make EMS?Can I use DMS?If yes then is there a half-life of DMS?Can I use UV lamp?
hi gregre,
I'm sorry, but that's not really my area of expertise. Normally when you're working with radioactive stuff you need a special certification, which you need to renew every 5 years or so. I don't think using a UV lamp will do much to generate mutants. From the papers I've read previously the seed treatment is quite harsh and lots of embryos actually turn out defective and die. Plus, you'll need thousands of plants to detect a successful mutant.
This is a more recent publication of mutation in tomato to obtain resistance to a parasitic plant
http://www.springerlink.com/content/668j7880272x3352/fulltext.html
The method they use is irradiation by fast neutrons. Quoted from the paper, so you get a rough idea
"Seeds of M-82 were irradiated by 60 Gy fast neutrons. About 17,000 M2 plants were screened for resistance against Phelipanche under field conditions, and were allowed to self-pollinate. Approximately 133 of the screened plants were found resistant to Phelipanche, 99 of which had viable seeds."
try ems mutagenesis/activation tagging/x-ray mutagenesis rotocol in Arabidopsis protocol book by - Glazebrook and Weigel. Will give you some starting material. A population of around 100000 seeds that can be done in 2-3 50 ml tubes for arabidopsis (?) may b be good to start but then you have to design the screen based on what you are looking for.
Hope that helps