Evaluation of a Simple Automated Bioreactor for the Production of Pelargoniums Progress Report –September 1993
Date August 1993
Title of Project Evaluation of a Simple Automated Bioreactor for the Production of Pelargoniums
Institution where work is being conducted Department of Horticulture, Penn State University
Amount of Endowment Grant $ 5000
Covering Period Dec. 1993 to Nov. 1993
Anticipated Date of Project Completion/Final Report November 1996
Individual(s) Conducting Project:
(List Project Leader First)
Richard N. Arteca - Title Professor
Telephone Number (814) 863-2252
Evaluation of a Simple Automated Bioreactor for the Production of Pelargoniums
Richard N. Arteca
Pennsylvania State University
- A. Project Objectives:
- Our major objectives for the second year of this project were to:
- 1. Manipulate environmental conditions, plant hormone amounts/ratios and othermedia supplements to promote maximum growth of callus and cell suspensions;
- 2. Induce plantlet formation and maximize the number of plantlets formed withdifferent hormone treatments in callus and cell suspension cultures;
- 3. To induce root formation on plantlets with different hormone treatments and growplants in tissue culture to increase the size prior to transfer to the greenhouse where they
will first be acclimated and then grown to flowering;
- 4. To evaluate different cryopreservation and low temperature storage methods for ourdifferent cell lines.
- B. Summary of Work Conducted:
- The results reported within this progress report are suitable for dissemination.
- C. Results to Date:
- Our work over the past six months (March 93 to September 93) has been to manipulateconditions to maximize the growth rates, shoot and root formation in our callus and
suspension culture lines and to use different cryoprotectants in order to cryopreserve
existing cell lines. We have been successful in maximizing growth rates of our suspension
cultures by modifying environmental conditions, plant hormone amounts/ratios and other
media supplements to promote maximum growth of the callus and suspension cultures
while still maintaining chromosome stability. AR attempts thus far have been unsuccessful
in cryopreserving either callus or suspension culture lines, however, we are still working
on this. A temporary solution to this problem has been to place the callus or suspension
cultures at reduced temperatures to suppress their growth. We have been successful in
keeping the cultures at 12′C which dramatically reduces their growth rates. When the
cultures are transferred to 20′C they win grow at the same rate as prior to low temperature
treatments and maintain chromosome stability. We are presently evaluating lower
temperatures and the stability of these cultures over longer periods of time. We have been
successful in the induction of organogenic callus, however, our major stumbling block to
date has been our inability to grow adequate numbers of rooted plants per gram of callus in
order to proceed to the greenhouse and to start bioreactor work. While we have been
successful in obtaining some small plantlets only a very small percentage of these will
develop roots and enlarge to sufficient size for transfer to the greenhouse. The plants
which do grow to an adequate size will readily acclimate to greenhouse conditions.
- D. Future Plans Covered by the Endowment Grant:
- Our goals for the remainder of the 1992 to 1993 research year are to manipulateconditions to maximize the number of shoots per gram of callus and to induce root
formation in these shoots. To use different cryoprotectants in order to cryopreserve our
existing cell lines and to continue to evaluate the potential of low temperature storage of our
cultures. The proposed experiments for the 1993-1994 research year are to do the
following:
- 1. Maximize the number of plantlets per gram of tissue with different hormonetreatments in callus and cell suspension cultures;
- 2. To induce root formation on plantlets with different hormone treatments and growplants in tissue culture to increase the size prior to transfer to the greenhouse where they
will first be acclimated and then grown to flowering;
- 3. To evaluate different cryopreservation and low temperature storage methods for ourdifferent cell lines.
- After these goals are achieved we will evaluate bioreactor technology for the productionof Pelargonium plants. This will be accomplished by seeding an airlift bioreactor with cells
grown in suspension culture and do the following:
- - treat with hormones to promote maximum growth of callus;
- soil and grow.
- The last phase of this project will be to design a larger bioreactor and evaluate how initialstudies work on a commercial scale.
- At the present time the major problem which we are encountering is that the number ofshoots formed per gram of callus is very low and before scaling up to a bioreactor this
problem must be solved. It may appear to be a trivial problem, however, this is the reason
why Pelargoniums now propagated by cuttings are not commercially grown in tissue
culture. We are confident that we can overcome this problem, in fact, the entire project
should be successfully completed within the proposed four year period, since at the present
rate of progress we are keeping up with the schedule initially proposed.
- E. Anticipated Benefits for Floral Industry:
- The overall goal of the proposed research is to develop an automated bioreactor whichcan be used to increase the commercial production of selected Pelargoniums with desired
characteristics such as new genotypes or disease-free plants rapidly and efficiently. The
availability of such methods will also lead to increased breeding and selection efforts which
in turn will promote increased quality of Pelargoniums at a reduced cost which is a true
benefit to the floral industry.
