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C O M P U T E R P R O G R A M FOR C O M P A R I S O N OF N U T R I E N T M E D I A FOR BIOLOGICAL C U L T U R E S G R O W N IN VITRO Stanley L. Nickel and Mary C. Grant

College of Pharmacy University of Toledo Toledo, Ohio 43606 SUMMARY: A computer program in BASIC computer language for use in storing and comparing the components of various biological culture media is described. The program contains 11 operations that deal with manipulation of data related to components, component quantities, and comparison of media. The program is capable of filing up to 50 different media lists each containing up to 75 components.

Key words: computer program; comparison; nutrient media; BASIC. I.

INTRODUCTION

II. MATERIALS

Over the years a variety of different nutrient media have been developed for support of in vitro cell, tissue, and organ culture growth. For example, there are at least 14 commonly used plant culture media, some claimed to be specific for growing woody plant cultures, some for flowering annuals, some for grasses, et cetera (1-4). As new media are reported in the literature, it is of interest to know if indeed the "new" medium is new or identical to media previously developed. The process of qualitative comparison by eye and quantitative comparison by calculator can become both tedious and error prone when several media lists must be compared. The computer program herein described is used to organize various media compilations for the purposes of comparison as well as efficient storage and retrieval. E n t r y of new media listings into the data base as well as retrieval and comparison runs are all accomplished interactively with the computer. The computer provides descriptive prompts for each operation, and the operator responds appropriately. Once entered into the computer system, the program requires no programming knowledge by laboratory personnel for operation. Although developed for plant tissue culture medium, the program may be used without modification for any other biological culture medium as weil.

A. Equipment Computer, PDP 11/70 Digital Equipment Corporation ~ Software, BASIC-PLUS 1 Terminal, Hazeltine 2000 with a thermal printer, Hazeltine Corporation ~

III. PROCEDURE A. Program: Initial entry 1. Sign on to the computer, type NEW COMPAR.BAS [ENTER], (Note: A word in brackets denotes a key with that function; for example, [ENTER] means push the ENTER key, do not type the word "enter"). Enter the sequence as given in Fig. 1. After each line push [ENTER]. Following the final statement, i.e. 9999 END, be sure to save the program as a permanent file by typing SAVE [ENTER]. The program is now ready to use. 2. To alter or rewrite the program for use in computer systems with BASIC software different from BASIC-PLUS, refer to the flow diagram (Fig. 2) and to the explanatory remarks interspersed throughout the program itself (Fig. 1). B. Use of theprogram 1. Type OLD COMPAR.BAS [ENTER] to call the program into operation. Start the program by typing RUN [ENTER]. The computer response will be:

' Digital Equipment Corporation, Marlboro, MA. 2 Hazeltine Corporation, Commack, NY. Journal of Tissue Culture Methods Vol. 7, No. 3, 1982

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N I C K E L AND G R A N T -- P R O C E D U R E NO. 99600

DO YOU W A N T TO: 1 ) E N T E R L I S T OF C O M P O N E N T S F O R T H E F I R S T TIME; 2) A D D TO T H E L I S T OF C O M P O N E N T S ; 3) C O R R E C T L I S T OF C O M P O N E N T S ; 4) P R I N T T H E L I S T OF C O M P O N E N T S ; 5) I N P U T A N E W M E D I U M ; 6) C O R R E C T A MEDIUM LIST; 7) D E L E T E A MEDIUM; 8)LIST A MEDIUM; 9)LIST THE NAMES OF THE MEDIA; 10) C O M P A R E TWO M E D I A ; 11) E N D THE PROGRAM? After the question mark, enter the number of the operation desired. 2. E n t r y of D a t a Base a. The first item is to E N T E R L I S T OF COMPONENTS FOR THE FIRST T I M E . This is accomplished b y typing 1 [ENTER] in response to the question mark. The computer responds with instructions that p r o m p t the operator to enter each chemical component of the medium. Either the chemical name or chemical formula m a y be entered. This list of components serves as a master list and will be used for all media listings. If necessary, this master list m a y be expanded at any subsequent time by Operation 2 {see below}. A maximum of 75 components m a y be entered. Operation 1 m a y only be done one time--the first time; all subsequent editorial changes to the component list m u s t be done b y operational commands 2 or 3. b. The second item is to enter the quantities of each component for a specific medium. This is accomplished b y typing 5 [ENTER] in response to the question mark indicated in section I I I B.1. The computer responds b y asking for a name or designation of the medium. The name m a y be 16 characters long. A sample name is, " H I L D E B R A N D T 1946", which designates the medium to be entered. The computer then lists each component followed b y a question mark. The operator enters the quantity of that component in milligrams or milliliters and presses the [ENTER] key. The next component is listed and the quantity entered; this process is repeated until quantities for all components have been entered. If a particular component is missing from the medium being input, a zero is entered for that component. On entry of the last quantity, the program automatically returns to the original listing of operations {see I I I B.I.}. 128

c. Additional media listings for the data base m a y be entered b y repeating b above. 3. Operations -- a brief description of each a. Operation 1 -- E N T E R L I S T OF COMPONENTS FOR THE FIRST T I M E . See section III.B.2.a. above. b. Operation 2 -- A D D TO T H E L I S T OF C O M P O N E N T S . This editorial operation allows for the expansion of the list of components in case a new medium happens to have a component not given in the initial listing. c. Operation 3 -- C O R R E C T L I S T OF C O M P O N E N T S . This editorial operation allows for correction of a component name in case of misspelling or erroneous formula entry. d. Operation 4 -- P R I N T T H E L I S T OF C O M P O N E N T S . Allows the list of components to be viewed. e. Operation 5 -INPUT A NEW M E D I U M . See section III.B.2.b. f. Operation 6 -- C O R R E C T A M E D I U M LIST. This editorial operation allows for correction of quantitites in case an error was made in the original entry. g. Operation 7 -- D E L E T E A M E D I U M . If a particular medium is no longer needed in the data base, this operation allows for deletion of that list. h. Operation 8 -- L I S T A M E D I U M . A n y single medium m a y be listed b y this operation. i. Operation 9 -- L I S T T H E N A M E S OF T H E M E D I A . Names for all the media in the data base can be listed b y this command. j. Operation 10 -- C O M P A R E TWO M E D I A . This operation prompts the user to indicate which media are to be compared; the user responds with the numbers assigned to the desired media and percents of the component quantities in each medium in terms of the quantities in both Mediums A and B. k. Operation 11 -- E N D T H E P R O G R A M . E n t r y of Operation 11 allows exit from the program. All files and medium listings will be saved. IV. DISCUSSION

This program was originally developed for comparing various rhododendron media compilations with some of the traditional plant culJournal of Tissue Culture M e t h o d s Vol. 7, No. 3, 1982

N I C K E L AND G R A N T - - P R O C E D U R E NO. 99600

COMPONENT

(NH4)2S04 MGSO4-TH20 CACL2-2H20 KNO3 NH4N03 NAH2PO4-H20 KH2P04 FESO4-7H20 NA2-EDTA MNSO4-4H20 ZNSO4-TH20 CUSO4-SH20 COCL-6H20 FE(SEQUESTRENE) K1 H3B03 NA2M004-2H20 SUCROSE MYO-INOSITOL NICOTINIC ACID PYRIDOXINE HCL THIAMINE HCL CA PANTOTHENATE BIOTIN NICOTINAMIDE CYANOCOBALAMIN FOLIC ACID PABA RIBOFLAVIN CHOLINE CHLORIDE

NOTE:

MEDIUM A 6AMBORG 1968

B AS % OF A

MEDIUM B RT MEDIUM

134.000 500.000 150.000 3000.000

370.000 440.000 1900.000 1650.000

150.000

I0.000 2.000 0.025 0.025 28.000 3.000 0.025 20000.000 100.000 1.000 i0.000

170.000 27.850 37.250 16.900 0.011 0.025 0.025 0.850 6.200 0.250 30000.000 2000.000 2.000 1.000 1.000 1.000 2.000 0.002 0.500 0.500 0.500 1.000

0.00 74.00 293.33 63.33 XXX 0.00 XXX XXX XXX 169.00 0.53 I00.00 i00.00 0.00 XXX 206.67 1000.00 150.00 2000.00 0.00 XXX I0.00 XXX XXX XXX XXX XXX XXX XXX XXX

A AS % OF B

XXX 135.14 34.09 157.89 0.00 XXX 0.00 0.00 0.00 59.17 18867.92 I00.00 i00.00 XXX 0.00 48.39 10.00 66.67 5.00 XXX 0.00 i000.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00

XXX -- MEDIUM USED FOR STANDARD HAS 0 FOR THIS COMPONENT FIG. 3. Comparison of G a m b o r g 1968 and RT medium.

ture media. It was found that on visual qualitative comparison, one or several medium components would be overlooked. The computer program presents the data in tabular form {Fig. 3) in which it is very difficult to overlook components appearing in one medium and not another. In addition, the computer can very quickly and easily calculate exact proportions of medium components of one medium in terms of the other. The tabular listing of these data provides for instant understanding of quantitative relationships. An additional value of the program is its ability to store and retrieve media listings for laboratory use efficiently. The exact quantities Journal of Tissue Culture Methods Vol. 7, No. 3, 1982

of each component for any stored medium listing are instantly available from the computer {Operation 8) and precludes a manual search through old files. The program is generously endowed with R E M A R K statements to assist programmers who need to alter certain features to fit software other than B A S I C - P L U S in order to install the program on a non-PDP 11 system. Additionally, the program once installed is completely self-prompting to assist individuals without programming experience. This seilprompt feature also assists individuals who m a y not use the program frequently to run it without review of extraneous instructions. 129

NICKEL AND GRANT -- PROCEDURE NO. 99600

V.

REFERENCES

1. Anderson, W. C. Propagationofrhododendronbytissuecultures: Part 1. Development of a culture medium for multiplication of shoots. Proceedings of International Plant Propagators' Society 25: 129-134; 1975. 2. Gamborg, O.L.; Miller, R.A.; Ojima, K. Nutrient requirements of suspension cultures of soybean root ceUs. Exp. CeU Res. 50: 151-158; 1968.

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3. Narayanaswamy, S. Regeneration of plants from tissue cultures. Reinert, J.; Bajaj, Y. P. S. eds. Plant cell, tissue, and organ culture. Berlin: Springer-Verlag; 179-248; 1977.

4. Vieitez, A.M.; Vieitez, E. Plantlet formation from embryonic tissue of chestnut grown in vitro. Physiol. Plant. 50: 127-130; 1980.

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