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Re: HYB:embryo rescue


Response from Dennis Hager, Society For Japanese Irises

As for embryo culture, it is a sterile process. It has been written up in 
the AIS Bulletin at least 3 times, dating back to the early 1940's. Randolph 
wrote the article. For over a decade, it was one of the most cited articles 
published in the Bulletin. 
If you would like to try embryo culture, I would recommend that you take a 
course in plant tissue culture--or at least read a few books. Most 
universities and many community colleges offer courses. If you insist on educating 
yourself through the internet, there is a yahoo group for home tissue culture. 
However, it is a marketing tool for the owner to promote weekend seminars on 
home tissue culture. 
   
  I would make a distinction between tissue culture and embryo culture (or embryo rescue).  The technics and theory do overlap a lot, but there are diffeerences.
  I make this distinction because different recipes for the nutrient medium are needed, and some different proceedures, too.
  Tissue culture starts from plant tissue not limited to embryoes.  Typically it is used to multiply a given variety.  In iris, an immature bloomstalk is the only tissue I've read working.  One such stalk could give many more plants of a variety per year than natural divission.  This sounds like the method mentioned at first.  It has noting to do with rescuing hybrid embryoes.
  Embryo rescue, or embryo culture, starts with an immature embryo which is not expected to mature into a viable seed.  The immature seed is removed from a surface-sterilized pod, then the seed is surface-sterilized.  The seedcoat is opened and the embryo is removed, under sterile conditions.  The embryo is placed right side up, on the nutrient agar, and the proceedure is continued like tissue culture from then on.
  I leaned embryo rescue in grad school, while studying crosses between wheat, rye, an barley.  It isn't too hard to do at home, if you are willing to sanitize a room, preferibly a small one.  It needs to be operating room clean.  You need to be able to prepare or buy sterile nutient agar OF THE RIGHT  KIND.  Each recipe is good for certain plants.  Needs may even change with the maturity of the embryo.  Need for cold treatment changes with maturity of the seeds in some genera.  I don't know about iris.
  When I taught high-school biology and 7th grade life science, I had the class do tissue culture of roses and african violets.  The sterile meium in petri dishes was already in the store-room before I was hired, so why not?  Results were poor, due mostly to the difficulty of getting high-school students to clean a lab to operating room clean.
  As for using store-bought nutrient agar, It is by far the easiest way to go, IF you can get the right kind.  
  Aril Society International has pulbished recipes to make the medium from scratch, and how-to articals.  While a class in doing all this would be helpful, many people do without such classes with good results.  And many people fail even with such classes.  Attention to detail is essential doing this.  There are more ways to fail than there are ways to succeed.
  Walter

 		
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