Author's Note:

Plant Tissue Culture Propagation (1981) was created to give students, hobbyists and teachers, who were not able to attend a micropropagation Workshop, the next best thing... a teach-yourself ability through written instructions and a set of 872 colour photographs illustrating most aspects of tissue culture.

 

This book was extensively updated in 1993 but, shortly after this revision, the slide set of photographs went out of print and the widespread availability of computers and CDs meant that the slide selection was obsolescent, if not actually obsolete.

 

Now the photographs in this slide set have not only been magnificently restored but also have been placed on a CD along with the text of the book. Furthermore, an update as a 100-page addendum is included along with photographs of Dr.Ronald de Fossard's small commercial laboratory.

 

The author, who is of French-Irish-English descent, graduated from the University of Edinburgh in 1954 and did citrus research in Jamaica and South Africa before accepting a research position with the CSIRO's Division of Food Preservation in Sydney and, it was there in 1962, that he started using tissue culture techniques. But it was at the University of New England (1964-1979) that he evolved many new tissue culture techniques and this continued after his retirement (due to cataracts) in commercial laboratories (1979-2006) and is continuing.

 

Tissue Culture for Plant Propagators (1976) was written as a take-home manual for participants at his many 5-day-long Micropropagation Workshops. This book became the University of New England's bestseller worldwide. Later books included Micropropagation (1990), Nursery Costs (1992) and Commercial Micropropagation (2000).

 

Micropropagation was the earlier version of Commercial Micropropagation and included many spreadsheet files on a floppy disc. It was my intention that readers/users would be able to make their own calculations of chemicals, media, cost of production , inventory, projections and analysis of different media with these files. Unfortunately, these files were not only written with a program called LOTUS 123 but also a version that required the MS-DOS platform.

 

Now Commercial Micropropagation has been placed on a CD and the calculations, formulae, etc., used in this book have been transformed into EXCEL and is Windows-based. So users now have the opportunity to work their cost of production as well as many other calculations required in a tissue culture laboratory.

 

These two CDs, from a commercial scientist who is self-taught in this area of expertise, are likely to become the essential helper, companion and advise-giver to tissue culturists throughout the world. Also they will be very useful for the training of staff in techniques and of students in experimental design and implementation.

 

- Dr. Ronald A. de Fossard (2006)

 

Introduction to
"Case Histories"

In 1976, I wrote my first tissue culture book, Tissue Culture for Plant Propagators, specifically as a take-home manual for participants at my 5-day-long Workshops.

Since then, I have used part of that book in Commercial Micropropagation (2000) and also added new material in Plant Tissue Culture Propagation (1981), in Micropropagation (1990) and in Nursery Costs (1992).

In 2007, I incorporated most of the material in these books into three CDs, namely, Plant Tissue Culture Propagation, Commercial Micropropagation and Nursery Costs and the remaining material from the original book was incorporated in this, my 4th CD, namely Plant Tissue Culture Case Histories.

 

I must confess that the idea for this 4th CD came from the Plant Tissue Culture
e-mail group when several people urged members to write up their protocols as a joint endeavour bringing together the combined knowledge of all users. It was also proposed that "senior" members could be persuaded to "edit" these protocols and it was suggested that I might be one such person. Frankly, I turned that suggestion down because I knew, from experience writing Case Histories in Tissue Culture for Plant Propagators in 1976, how difficult this would be. I suggested instead that if someone could organize Wikipedia to receive individual contributions in this field that a practical assemblage of protocols could be obtained. Furthermore, I suggested that, as a model, individuals could use the Case History format that I used in my 1976 bookd and that, instead of anyone editing these contributions, each should stand as the individual author's statement, his/her responsibility. The main thing in my view was to agree on a suitable Case History format and to find out how to send these to Wikipedia.

 

The discussion "died" but it prompted me to resurrect these old Case Histories (from 30 years ago) in a new CD and my intention was then to make new Case Histories covering the period 1976 to 2008. It didn't take me long to realise the impossibility of this task quate apart from the limited access I have these days to the literature - the field is already simply too vast. So back the other idea. This time use the "old" Case Histories to show how interesting this earlier work was and then to demonstrate, in spreadsheet format, how relatively easy it would be for others to simply fill these in with thier own protocols. Maybe the idea of an unrefereed Wikipedia slot could become a reality and I like the idea of individual authors taking responsibility for their work/reports.

 

The first part of this CD contains most of the 1976-book's Case Histories covering more than 60 species and 30 familes. This is followed by a section describing several well-known media and how they can be prepared using concentrated chemical stock solutions specifically prepared for each medium. The next section describes the same well-known culture media but this time prepared using the stock solutions used for the Case Histories. This leads logically into a section describing the mechanics of preparing Case Histories in which parts of well-know media are used. For example, one contributor to this Wikipedia "scheme" could have used 1/2-strength Murashige & Skoog's macronutrients with WPM- micronutrients. And so on. Simply by COPYing parts of perviously described culture media and PASTEing them on a new part of a spreadsheet we can put togehter any/all combinations of culture media

 

I have been using Broad Spectrum (BS) technology since it was evolved in the early 1970s and the next sections describes BS-media and the BS-concentrated chemical stock solutions. It includes a list of all BS-media prepared in my laboratory, for one purpose of another, over the last 40 years. One table lists all media in chronological order (BS-media 1 to 114) and a second table re-arranges these media so as to show their varying degrees of relationships between the major components in culture media. This is followed by the preparation details (component chemicals, stock concentrations, amounts per litre, ets...) of 28 BS-media ready for use by anyone wishing to prepare these media.

 

The Broad Spectrum part of this CD is followed by a description of our experimental methods and this leads ot the final section of this CD which describes the statistical analyses of experiments, using EXCEL. I am not averse to doing what many tissue culturists do and that is to making "look-see" experiments in which 3 or 4 well-known culture media or 3 or 4 variations of a single medium are used, explants placed on several replicates of these and, at various times, these are examined, notes taken and finally (hopefully) we are able to chose one of the experimental media for culturing our species. Maybe this is all that is necessary to get a satisfactory culture performance, reflecting on the luck of choice or the experience of the researcher. I have described how I do this in my CD Plant Tissue Culture Propagation, viz. if I have a new woody species, I test 3 or 4 media used successfully with other woody species. Same approach with herbaceous species. Many times one has no other option because of the scarcity of starting material and pressure of time or whatever may persuade one to leave it as that, whatever media is selected is used thenceforth. The fact is that most species can grow on a very wide range of media but the question I raise is: But what is the species capable of? We'll not know until and unless we experiment more extensively.

 

Rather that re-wording / re-writing a lot of my earlier advice on experimentation, I've decided to simply re-print some of my earlier writings. Thus, reades will find:-

 

1. (1) Chapter 5:- The Broad Spectrum approach to media.
   1. from my CD Commercial Micropropagation (2007)
      
2. (2) Step-by-Step Procedures to Achieve Clonal Propagation of a Cultivar.
   1. from my CD Plant Tissue Culture Propagation (2007)
3. (3) Appendix E:- Experimentation.
   1. from my CD Commercial Micropropagation (2007)
      
4. (4) Finding Suitable Culture Media
   1. from my CD Plant Tissue Culture Propagation (2007)

 

The latter includes the advice to include [-16]-Minerals in the earliest stages of experimentation. This is a mixture of Low, Medium and High concentration minerals first used in my Medium-16 for Eucalyptus cultures but later found to be useful for both Nepenthes and Sarracenia.

 

My BS-approach has allowed me to get many different species into culture fairly quickly but there is no guarantee that it will always lead to success. However, the more we learn from whatever source, the more likely it is that an ever-improving, successful Broad Spectrum technology will be evolved. I've included in this section a story, a Case History, of one of my "failures", namely the micropropagation of Capparis spinosa L. (capparidaceae), the caper bush.

 

I would like to think that my CDs are valuable and of interest to a wide range of tissue culturists from the newcomer learning about the chemicals we use and the equipment options availble, to the people thinking of a retirement hobby/commercial occupation, to large scale commercial laboratories and to students planning research for their thesis and to people just wanting to unravel a few of the secrets of plant life and behaviour.

 

Best Wishes

 

Ronald de Fossard

B.Sc. (Hons)(Edinburgh) M.Sc.(Witwatersrand) Ph.D.(New England)

 

March 2008