Almost all research reports which i ever seen work on Early mature embryo for Agrobacterium mediated maize transformation. In other grass families like rice, we can use mature seed and what do you think this will work in maize?
Article Advances in Maize Transformation Technologies and Developmen...
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I think the embryo should be in between milk and dough stages! Check some literature.
I think it ma be possible with A. tumefaciens strain harbouring a tumor inducing plasmid, but this is for early only
Dear Temesgen!
In my opinion, success in Agrobacterium mediated transformation depends on access of the Agrobacterium to easily responding tissue. The major problem in transforming maize seed seems to me that the germ (embryo) in maize is positioned deep into the endosperm in maize while it is easily accessible by the Agrobacterium in other cereals like wheat and rice. In wheat, rice and other cereals, when you sock the seeds, the central endosperm loosens and the Agro can infect while in maize that is not easy. Therefore, immature embryos more preferably embryogenic calluses are the best materials. If you centrifuge the agro and soaked wheat seed you can get better results.
Open the attached file or look the following for further information
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Source: https://wheat.org/whole-grains/
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Source: https://www.frontiersin.org/articles/10.3389/fgene.2019.01392/full
Frontiers in Genetics | www.frontiersin.org 1 February 2020 | Volume 10 | Article 1392
Again, I would like to clarify that embryos of wheat and rice kernels are covered with a thin layer, much thinner than in maize kernel which is the embryo is covered by a thick layer of Lenin, son immature kernel or embryo is the best to use in transformation. Cheers.
Alan Gould ; this is to study the drought stress response of the transgenic and non transgenic maize. Mainly expression analysis and functional genomics.
Temesgen Assefa Gelaw But, how is this < this is to study the drought stress response of the transgenic and non transgenic maize. Mainly expression analysis and functional genomics. > link to use mature seeds for genetic transformation?
Temesgen Assefa Gelaw Here is a study: " Establishment of transformation systems using mature seeds of maize inbred lines. " (a dissertation I think)(https://lib.dr.iastate.edu/cgi/viewcontent.cgi?article=15908&context=rtd)
Alan Gould < Why would anyone want to do this? >
One of the reasons listed in the study I mentioned above is:
" The major problem for using the immature embryos is that it requires maize plants to be grown in the greenhouse year-round to meet the research demands. This practice requires large greenhouse spaces, quality growth conditions and experienced supporting staff. "
Note that this thesis uses embryogenic maize callus, not mature maize seed for transformation. My group published on field trials of transgenic Bt maize in 1993. I have not heard of anyone successfully transforming mature maize seed by direct gene insertion, biolistic or Agrobacterium, but would be very interested to hear if anyone has done this.
Thank you respected scientists for your interaction and the thesis (https://lib.dr.iastate.edu/cgi/viewcontent.cgi?article=15908&context=rtd) used mature seed also and i have't heard or read any paper which is ever successful in establishing mature seed transformation protocol. Working on embryo will take time and other extra ordinary aspects, how ever till now am on the way to use immature seed ( https://www.nature.com/articles/nprot.2007.241.pdf , doi: 10.1007/978-1-59745-494-0_4. But glad to hear from anyone who has done or going to do this.
Article Advances in Maize Transformation Technologies and Developmen...
Have you checked this before?
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