Doubled haploids from anther or pollen culture

1. S
Asian Seed Congress 2012, Bali, Indonesia, 5 – 9 November 2012
j
Breakthrough in Tomato Breeding Efficiency
Speaker: Bert van Duij n - CSO for Fytagoras BV, Leiden,
The Net herlands

2. Breakthrough in tomato
breeding efficiency
A joint project of Fytagoras, APSA and APSA members

3. AIM
S Establishment of efficient and robust
protocols for the production of doubled
haploids in tomato.
S Realize this in a joint project between
APSA members, APSA and Fytagoras

4. Why Doubled Haploids?
S Homozygous plants in only one step (generation)
S Real pure homozygous plants (haploids have 1 allele of
every gene).
S More lines in less time
S Doubled haploid technology higly accelerates
conventional plant breeding.

5. Why tomato?
S Tomato is the second largest vegatable crop in the world
S Breeding progress in tomato is important
S Tomato is a very recalcitrant crop for DH technology
S Tomato is an important crop for a large number of APSA
members
S Joint develoment of tomato DH protocols can be non-
competitive

6. What is the basis of DH
technology?

7. What is the basis of DH
technology?
Different systems making doubled haploids
DH technology starts with the use of haploid cells from the plant
in tissue culture
S Anther/microspore culture (male flower part)
S Ovule culture or megaspore culture (female flower part)
S Hordeum bulbosum technique

9. 1. Donorplant
3. Formation of plants
2. Induction of celldivision
and growth of multicellulair
structures

10. Critical steps in the DH technology
S Donor plants; stage of development
S Pretreatment; induction of cell division (many different
possibilities)
S Culture; formation of multi-cellular structures; embryos
S Formation of plants
S Implementation of the protocol
S Adaptations of the protocol for all varieties
What is the basis of DH
technology?

11. Why Fytagoras?
S Over 15 years experience in DH technology
S Protocol development, Protocol implementation, Consultancy
S Basic scientific research and scientific publications
S Worked for over 8 years on exclusive basis on DH of vegetable crops for one of
Holland's leading breeding companies. Ongoing development for different crops (also
ornamentals).
S Development of numerous successful protocols and commercial implementation.
S First steps for tomato explored and promising.
S Great experience in contract research and in-company instructions.
S On-going research program (in cooperation with Leiden University) on the
fundamentals of microspore regeneration: evolutionary aspects, metabolism aspects,
genes and transcriptions factor, cell signaling aspects.

12. How to approach?
PART 1
DEVELOPMENT OF INITIAL PROTOCOL
PHASE I
generation of multi cellular structures
PHASE II
embryo and
plant
formation
_____1____
preparation
and
sterilization
of
microspores
___ 2___
induction
of cell
division
___ 3____
formation
of
multi-cellular
structures
PART 2
OPTIMIZING
OF PROTOCOL
- - - - - - - - -
APPLICATION TO
OTHER VARIETIES

13. How to approach?
S PART 1: From microspores to doubled haploid plants
S Phase 1 Induction of multi-cellular structures
S Step 1 Selection of plant material, technical aspects concerning
the preparation of microspores, determination of developmental
stages, and characterization of microspores
S Step 2 Pretreatment and induction of cell division
S Step 3 Cultivation and formation of multi-cellular structures
S Phase 2 Embryo and plant formation
S PART 2 Optimizing of the procedure and
implementation

14. How to proceed?
S Set up joint project: APSA members-APSA-Fytagoras
S Share costs: general protocol
S Adapted protocol for your own varieties and implementation
S Broad company based knowledge input
S Joint results and progress meetings;
S Workshops
S Web-seminars
S Results restricted to participants
S APSA is contract partner
S Start of project January 2013

15. How to participate?
S Show your interest on the evaluation sheet.
S Send e-mail to: bert.vanduijn@fytagoras.nl
(cc. tom.burns@apsaseed.org)
S Come to Holland Pavilion (3rd floor)
(possibility to organize special meeting for potential
participants this week)
S Detailed technical and scientific back ground information
available upon request

16. Thank you for your attention!