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Evaluation of a Simple Automated Bioreactor for the Production of Pelargoniums Progress Report –March 1993

Date February 26, 1993

Title of Project Evaluation of a Simple Automated Bioreactor for the Production of Pelargoniums

Institution where work is being conducted The Pennsylvania State University

Amount of Endowment Grant $ 5, 000
Covering Period 6/92 to 5/93

Anticipated Date of Project Completion/Final Report 6/96

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

Progress Report to the American Floral Endowment, 2/26/93
A. Project Objectives:
Our major objectives for the second year of this project are to:
1. Manipulate environmental conditions, plant hormone amounts and ratios and othermedia supplements to promote maximum growth of callus and cell suspension growth;

2. Induce plantlet formation and maximize the number of plantlets formed withdifferent hormonal treatments in callus and cell suspension cultures;

3. Once we have maximized plantlet formation we will transfer these plantlets togrowth media containing different ratios of plant hormones in order to induce root

formation and then grow the plants in tissue culture to increase the size and number. We

will then begin work to acclimate these plants for transfer to the greenhouse where they will

be grown to flowering.

B. Summary of Work Conducted:
At the present time the results to date are preliminary and not suitable for dissemination.
C. Results to Date:
Our work over the past six months (September 92 to March 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 our

existing cell fines. We have been successful in maximizing growth rates of our suspension

cultures by modifying environmental conditions, plant hormone amounts and ratios and

other media supplements to promote maximum growth of the callus and suspension

cultures while still maintaining chromosome stability. All attempts thus far have been

unsuccessful in cryopreserving either callus or suspension culture lines, however, we are

still working on this. We have been successful in the induction of organogenic callus,

however, our major stumbling block to date has been that we are unable 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 the greenhouse. Our future goals will be to focus on overcoming these

problems so we may proceed with the project.

D. Future Plans Covered by the Endowment Grant:
Our goals for the next six months of this project are to manipulate environmentalconditions, plant hormone amounts and ratios and other media supplements to maximize

plantlets with roots per gram of callus and promote enlargement of these plants for transfer

to the greenhouse and for use in bioreactor studies.

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;
– induce plantlet production with treatments developed earlier and transfer plantlets tosoil and grow.

The last phase of this project will be to design a larger bioreactor and evaluate how initialstudies work on a commercial scale.

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 Pelargoniuma at a reduced cost which is a true

benefit to the floral industry.