OVERVIEW OF COMMERCIALVEGETABLEBREEOING RESEARCH(VARI_-FYDEVELOPMEN!].)

L.R. BAKER, EXECUTIVEDIRECTOR,VEGETABLE R & DASGROW SEED COMPANY,KALAMAZOO,HI 49001

PREFACE

This presentation is not meant as a documentary of the seed trade. It

represents personal and professionalviews of the author who was at

Michigan State University as a public breeder in vegetable breeding

research for 13 years followed by his current position with Asgrow Seed

Company. The main thrust of this paper is to provide a discussion of

private vegetable seed company research for proprietaryvarieties.

INTRODUCTION

The advent of hybrid varieties to replace open-pollinated varieties

makes possible the protection of proprietarydevelopmentsand product.

Hybridization of vegetable speciesbegan in the 1920's with sweet corn,

followed by onions in the 1940's. Since that time, most species Of

vegetables have been changed into hybrid varieties for the marketplace.

Most recently, hand-pollinatedvarietieshave been developed, but costs

for hand-pollinated seed production are relatively expensive compared

to open-field production. However, the advantages of hybrid

performance related to uniformityand vigor have outweighedthe cost of

seed.

Most recently, plant variety protection (PVP) in the'U.S., and other

forms of protection throughout the developed countries of the world,

have afforded product security for species where hybridizationis not

possible(1). The latter would include the major vegetable speciesof

peas and beans. Such product protection has presented a business

incentive to corporations to invest in the seed industry (EXHIBITI).

Most seed companies were "family-owned" operations based upon

individual breeding successes in localized markets. For example, the

Clark family owned the Asgrow Companywhich excelled in the development

20 (over)

and production of high quality bean and pea varieties for processing.

Joseph Harris owned the Harris Seed Company in Rochester, New York, and

commanded a lead position throughout the northeastern U.S.in several

vegetable species con_only produced in that region. The introduction

of corporate business to the seed business permitted in major infusions

of capital. This supported a dramatic increase in private R & D

followed by strong competition in the marketplace between these major

seed companies.

The major worldwide vegetable species and estimated seed values are

presented below based on market surveys by Asgrow (EXHIBIT 2). The

estimated worldwide market value of these vegetable species is $I010MM.

A general estimate of R & D activities in the seed business approaches

10% of sales for R & D. Such an estimate would generate $100MM for

varietal development and other aspects of private seed research. Seed

companies have upgraded facilitiesand research staff over the last 10-

15 years. Plant breeders with Ph.D. degrees in genetics and plant

breeding with vegetable/horticulture backgrounds are commonplace

throughout the vegetable seed industry. The capability and

responsibility to develop proprietary varieties is commonplace

contrasted to previous public varieties. Prior to the 1970's, plant

breeders were more involved in seedstock work than varietal

development.

The transition for variety development from the public to the private

sector is well along. The cost for R & D to develop new varieties is

shifting from the public supported research programs to the customers

of the major seed companies. The majority of current varietiessold are

proprietary products developed by private R & D. These varieties are

more dependable in performance and available on a more reliable basis.

A significant consequence of the private increase in R & D for variety

development has been a reduction in public breeding work to develop

varieties. There is no question that the continued need for

fundamental breeding research is critical to support developmentof new

technology and expansion of the knowledge base which support varietal

development.

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Competition between proprietary varieties results in owner-companies

doing the best possible research to develop their own products and to

compete on genetic and physiologicalquality of seed in the marketplace

rather than price. Public varieties tend to be highly sensitive to

price; and therefore,cost saving measures are often taken in order to

compete on the basis of price. This often leads to a "running-out_ of

originally superiorvarieties. Reasonable profit margins are necessary

to pay-back the R & D costs to the owner and fund future research on

developing even better varieties to stay competitive in the

marketplace. There is considerable genetic variation within the

various vegetable specieswhich can be exploited in the developmentof

proprietary superior varieties. The consequences of this dynamic

situation will mean relativelyshort-livedvarietiesreplaced by either

the owner of _he variety or a competitor company. This intense

competition w_l mean constantly improved and more sophisticated

varieties for the vegetable industry.

GERMPLASM

The term germplasm means different things to different people.

Generally, it has been defined as germ that from which the next

springs, and plasm formative material...the bearer of the

characteristic nature of the species and individual. Germplasm

connotates the science of heredity and genetics, but implies the

unknown mysteries of the spark of life. No wonder that developed

nations have collected and saved it over the last 50 years. The USA

has some 250M plant germplasm accessions stored in the National Seed

Storage Laboratory (NSSL) in Fort Collins, CO. The epidemic in field

corn of southern corn leaf blight in 1970 struck a devastatingblow to

corn production i_ the USA. The upshot of this was a NationalAcademy

of Science (NAS) study in 1972 which identified the genetic

vulnerabilityof our major food species. The result was to establisha

National Plant Germplasm System to collect, maintain, describe and

enhance germplasm.A Plant Introduction(PI) Service has been operated.° .

by the United States Department of Agriculture (USDA) since 1898, but

simply collected germplasm with limited follow-up research. This PI

system is now part of the upgraded NPGS which is being expanded each

year. Seed of the accessions is given free upon reasonable request.

22 (over)

Next, enter biotech and the mass media, patents and lawyers...germplasm

becomes a sacred national heritage. No wonder that the Food and

Agriculture Organization (FAO) of the United Nations passes Resolution

9/83 to monitor operations and arrangementson germplasmcollection. A

classical socio-politicalconfrontation between developed and lesser-

developed countries was created (2). We must collect germplasm,

conduct research to improve it and provide the improved varieties for

production of food to feed people. The best possible science and

technology is used to do this...single-gene traits to introgressive

hybridization;backcrossingto trdnsgenic plants; etc. We run the risk

of famine, war, etc. should progressive science and research be

compromised.

BREEDING METHODS

The pedigree method is widely used to develop arrays of "eliten inbreds

of superior homogeneousphenotypes. Inbreedingdepression is a problem

with most open-pollinated species, so genotypic homozygosity is

avoided. For hybrid programs, elite inbreds are then tested for

combining ability in the variety developmentwork. Outstanding hybrid

performance is usually from general combining ability, but specific

combining ability is also important.

An elite germplasm "pooln is developed by a breeder after 10-20 years

of research. The backcrossmethod is often used to selectively add new

, simply inheritedtraits to an otherwise desirable elite inbred. Limited

use of recurrent selection schemes has been made for quantitative

traits.

HYBRIDS

The use of hybrid vigor in vegetable variety developmentwas initiated

in 1924 with sweet corn (Zea mays) hybrids by the Connecticut

Agricultural ExperimentStation. The genetic term for hybrid vigor is

heterosis. Heterosis may be defined in a practical way as a hybrid

performance greater than either parent for uniformity and/or economic

( yield. Since the 1940's, essentially all cross-pollinatedvegetable_'..

species have been converted from open-pollinatedto hybrid cultivars.

Today, there is a strong trend towards hybrid cultivars in self-

pollinated species'such as tomato, pepper and eggplant (3).23

Cytoplasmic-geneticmale sterility (CMS7 is widely used to produce

hybrid seed of onions (Allium ceDaT, carrot (Daucus carota), and table

beet (Beta vulqaris) (4-6). Current research efforts are aimed at

developing CMS cabbage, cauliflowerand broccoli (Brassic_oleracea7to

replace the current complicatedself-incompatibilitysystem for hybrid

seed production. French workers have also developed a CMS system for

radish (Raphanissativus).

Genic control of sex expression has been manipulated in the cucurbits

to condition dioecious, monoecious, etc., sex expressions which are

conducive to hybrid seed production (77. The best commercial example

is cucumber (Cucumis sativus). Possibly some million kilos of hybrid

cucumber seed is produced annually using gynoecious, all-female seed

parents crossed with monoecious pollen patents. Three or more major

loci are thought to control sex expression in cucumber (87. The same

phenotypes have been identified in cantaloupe, but the genetics are

different from cucumber and not well understood (g).

Plant growth regulators have proven useful in certain cases. Ethephon

is widely used for production of hybrid squash seed. Pistillate

flowering is induced temporarily on monoecious squash (Cucurbita

and C. moschata) by several applicationsfrom the one-leaf to _everal-

leaves stage of growth (107. There is evidence to suggest cthephon

might prove useful for hybrid cucumber and cantaloupe seed pr_ction

similar to squash. However, these two speciesare not so responsiveto

ethephon as squash. Moreover, monoecious lines are necessary for

successful inducting of pistillate flowering; and, determinant plant

types are more easily controlled than indeterminant. Silver

thiosulfate is commonly used to induce staminate flowering on

gynoecious cucumber seed parents for seed stock increases. Prior to

silver, gibberellicacid (GA7 was used for the same purpose. The AgS203

also induces staminatefloweringon gynoeciouscantaloupe seed parents,

whereas GA is ineffective.

24 (over)

Spinach (Spinaci_ oleracea) breeding research has elucidated a method

for hybrid seed production called sex reversal (SR). Two major loci

are thought to condition three different sex phenotypes. One of the

genotypes is nearly gynoecious with pistillate flowering until near

senescence whereupon staminate flowering provides pollen for

maintenance of the gynoecious line. The SR lines are used as seed

parents for hybrid seed production.

Genic male sterility (MS) has been observed by researchers since the

outset of plant breeding. There are more than 60 different genes

recorded in tomato (Lycopersiconesculentum)which condition MS. Tomato

researchers have identifiedm__ss35as one of the more desirable loci to

use in seed-parent line development. A marker gene for seedling

hypocotyl color is linked to ms35 which permits roguing at the seedling

stage. Pepper (Capsicum annum) MS has been reported by several

workers, but the work by INRA in France appears most promisingregards

hybrid seed production. Two loci for MS in cantaloupe (Cucumismelo)

were reported in the 1960's by USDA researchers in California,but have

not proven practical (10). Recent promising work was reported on

watermelon (Citrulluslanatus) seed production using genic MS linked

with a seedling marker for glacous leaves. Recessive single gene MS's

have been reported in Brassica oleracea and lettuce (Lactuca sativa),

but have not been useful to date.

COMMERCIAL PLANT BREEDING

We are in the business of manipulating genes to improve plant variety

performance for a profit. If it doesn't make reasonable profit, the

research is worthless. The focus must be on research to improve

economic performance starting with the seed production costs, to the

grower-shipper/processor-enduser. If any link in this sequence of

events is weak or broken,the new variety will likely fail.

The most importantprincipleof variety development is the equation:

P = G + E + (G x E)

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Phenotype is the sum of genotype, environment and genotype x

environment effects. The environment and interaction effects are

extremely importantin variety performance. Accordingly,new varieties

must be tested for performance in all markets before sales.

Developmental breeding must also be done in major production areas

unique from others, or where competition is keen in order to maximize

performance.

A second principle of variety development is the nSOS" rule, i.e.,

SCALE-ORGANIZATION-SCIENCE.No practical breeding programwill succeed

without large numbers of lines (genotypes) to evaluate, select,

recombine and inbreed (fix genetically). This effort must be organized

so valid conclusionscan be reached and made (decisions). Scientists,

support people and facilities,budgets, management are requirementsto

assure success. Science must be state-of-the-art(SOTA) to maximize

use of scale and organizationsin a competitivebusiness environment.

BIOTECH AND THE FUTUR_

Biotechnology is a new, and potentially powerful, tool to add to plant

breeding research programs (12). It can augment and/or accelerate

conventional variety development programs through (I) time saved, (2)

better products, (3) more genetic uniformity or (4) achieveresults not

possible by conventionalmeans.

The following concepts are being tested by private companies. This

list is not all-inclusive.

I. Cybridization - The hybridizing of cytoplasms from different

sources. The obvious application is related to cyto-male sterile

systems. This falls into either time saved or impossible by

conventionalmeans.

2. Cellular Screens - Methods to screen cells in a specific medium

for a specific reaction. The immediate applications are for

disease toxin resistanceand herbicide resistance (13).

26 (over)

3. Somaclonal Variatioq - By regenerating plants from single cells

through cell culture, new genetic variation is created. This

"new" genetic variation can be identified through either

conventional or biotech methods for incorporation into the

breeding research program. There are numerous examples of

somaclonal variation in the literature and in practice (14).

Current applications relate to disease resistance, processing

quality and seed producibility.

4. Anther Culture The cell culture of microspores to regenerate

haploid plants which are redoubledto form dihaploids. This

creates an "instant" inbred and saves the inbreeding time;

applicable to hybrid programs. In practice, this must be coupled

with several screens to eliminate undesirable genotypes/-

phenotypes. Otherwise, the plant breeder is quickly inundated

with hundreds to thousands of inbred lines to evaluate in the

conventional program. In practice, anther culture is used in

Brassica breeding with major research efforts in tomato. There

are many necessary pre-requisites to meet before it can be

commonly used in other species.

5. Transqenic Plants - this is the most difficult research area with

many pre-requisites to be satisfied prior to the actual gene

transfer. Gene identification,isolation,purification,transfer,

expression and regulation, all must be done successfully (15).

Recent reports on expression of the "bt" gene from Bacillus

thurinqensis in tomato plants are quite exciting. Other labs have

reported transfer of a virus gene to several plants species which

confers resistanceto that virus.

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SUMMARY

The area of commercial vegetablebreeding is dynamic and exciting (16).

There is no immediate "yield plateau" in sight. The area of biotech

promises to create even more opportunities for breeders to improve

varieties. We have seen resistance to certain diseases, insects and

herbicides via biotech in 1986/87. More and more "Hi-Tech" will be

used in the development of higher performing varieties. Research

developmentswhich have only been thought or dreamed about in the past

will become commonplace in the future. The proprietaryvegetable seed

business is coming of age in the USA and world. It is research-driven

which means vegetable breeder lead. Combined with the possibilities

from biotech, these are exciting times for commercial vegetable

breeding researchwith business opportunitiesfor profit.

NATLBRDS.PRS

MAY 1987

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SELECTED REFERENCES

I. Batcha, J.A. and J.A. Studebaker. 1983. A chronicle of plantvariety protection. Asgrow Seed Company. Kalamazoo, MI 49009. 26p.

2. Perlas, N. and J. Rifkin. 1986. Default at the gene banks. Garden10(3). NY Botanical Gardens, Bronx, NY.

3. Quagliotti, L. (Ed.). 1981. Symposium on vegetable and flower seedproduction.Acta Hort. 111. Castrocaro, Italy.

4. Jones, H.A. and A.E. Clarke. 1943. Inheritanceof male sterility inthe onion and the production of hybrid seed. Proc. Am. Soc. Hort.Science 43:189-194.

5. Hansche, P.E. and W.H. Gabelman. 1963. Digenic control of malesterility in carrots, Daucus carota L. Crop Science 3:383-386.

6. Bliss, F.A. and W.H. Gabelman. 1965. Inheritanceof male sterilityin beets, Beta vulqaris L. Crop Science 5:403-406.

7. Peterson, C.E. and J.L. Weigle. 1958. A new method of producinghybrid cucumber seed. MI Agr. Expt. Sta. Quart. Bull. 40:960-965.

8. Scott, J.W. and L.R. Baker. 1975. Inheritance of sex expressionfrom crosses of dioecious cucumber. J. Am. Soc. Hort. Science100:457-461.

9. Poole, C. and P. Grimball. 1939. Inheritanceof new sex forms inCucumis melo L. J. Hered. 30:21-25.

10. Coyne, D.P. 1970. Effect of 2-chloroethylphosphonicacid on sexexpression and yield in squash and its usefulness in producinghybrid squash. Hort. Science 5:227-228.

11. Bohn, G.W. and J.A. Principe.1964. A second male-sterilitygene inthe muskmelon. J. Hered. 55:211-215.

12. Brown, W.L. 1984. Genetic engineering of plants. National ResearchCouncil, Board on Agriculture.National Academy Press. Washington.p. 83.

13. Sun, M. 1986. Engineeringcrops to resist weed killers. Science231:1360-1361.

14. Evans, D.A., W.R. Sharp and H.P. Medina Filho. 1984, Somaclonaland gametoclonalvariation. Am. J. Bot. 71:759-774.

15. Goodman, R.M., H. Hauptli, A. Crossway and V.C. Knauf. 1987. Genetransfer in crop improvement. Science 236:48-54.

16. Innes, N.L. 1982. The potential from plant breeding for vegetablecrops. Sci. Horot. 33:60-66.

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EXHIBIT 1. Major Worldwide Vegetable Seed Companies

Asgrow(Upjohn)

- FerryMorse (LimaGraines)

- PetoSeed (GeorgeBall)

- NorthrupKing-Sluis& Groot(Sandoz)

- Harris-Moran-FMC(LaFarge)

- Arigenetics(Lubrizol)

- Sun Seeds-Dessert-Castle(U & F Seeds,Inc.)

- RoyalSluis

- IPB (Shell)

- Clause

- Takii

- Sakata

- 90% of WorldwideVegetableSeed Business.

30 (over)

EXHIBIT 2. Estimated Value Major Vegetable Species Worldwide (HI( =

millions).

- Onion $160/4/4 - Watermelon $ 60/4/4

- Tomato 135MM - Carrot 44MM

- Bean 125/4/4 - Cabbage 4OHM

Pea 110/4/4 Cantaloupe 30MM

- Cucumber gIMM - Lettuce 24MM

- Broccoli gOMM - SweetCorn 18MM

- Pepper 75MM - Squash 8MM

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ASGROWVEGETABLER & D

NOWFOR1995

I. INI'ROOUCTION

* Hybrids* PVP* CorporateInvestment/Participation.* MajorChanges

- Familyto Corp.Business.- PublicVarietiesto Proprietary.- ProprietaryR & D Investments.- Consolidation

* Germplasm- NPGS- FAO- LDC's

* BiotechHype.* Patentsand Lawyers.

II. COMMI_RCIALPLANTBREEDING

* Breedingfor a Profit.* P-G+E +GE* "SOS"Rule* AsgrowStatus

- People- Facilities- Budget- Results

* MajorPlayerin MajorMarkets.

III. BIOTECH

* UpjohnBiotechCommitment.* New Hi-TechTool.* IntegralPartof VarietyDevelopment.* Accelerate/AugmentMajorBreedingPrograms.* 1985for 1995,Lead-Time.* ExamplesCurrentResearch.

- Cybrids- DiseaseScreens- Dihaploids- Transgenics- ClonalSeeds

32 (over)

IV. FUIIJRE(Dynamic)

* GeneticVariation& Yield Plateaus.* More Hi-Tech.

* Asgrow's Place 1995.- Market Share Projections.- Now Research is 1995 Sales.

* Integration,People Chemistry...How?

V. CONCLUSION

Asgrow is positionedto be one of the "BIG 3".

LRBAKER5 JUNE 1987SAN DIEGO, CA

WRSCONF.INT

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