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Enhanced Methods for Doubled Haploid Production in Triticum

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Tyler Suelter

on 7 December 2016

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Transcript of Enhanced Methods for Doubled Haploid Production in Triticum

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Heartland Plant Innovations and the Kansas Wheat Innovation Center
Process Overview
Creating DH lines is a multi-stage procedure that takes an average of 52 weeks to compete

Process begins with growing clients' seeds to flowering stage
The Importance of Wheat Haploids
Image credit: E. Holbrook, RM Magazine
Enhanced Methods
for Doubled Haploid Production in
Triticum aestivum

T. SUELTER, F. Chumley, R. Berard, and H.N. Trick
In vitro

D
H Challenges
Current growth media is not optimized for embryo growth and development.

Germination Rate
Callus Formation

Chromosome doubling with colchicine treatments are a major stress to plant that can be difficult to recover from.

Timing
Chemistry
Recovery
A Closer Look at Rescue
Normal seeds compared to haploid seeds 2 weeks into development
Emasculation:

Glumes and awns are clipped from the spike and the anthers are stripped off each flower
Pollination:

Maize pollen is dusted onto the wheat stigma, inducing haploid embryogenesis
Sterilization:

After two weeks of development, haploid seeds are threshed from the flower spike and sterilized
Embryo Rescue:

Embryos are excised from the haploid seeds and cultured on MS medium
Embryo culture:

Haploid plantlets are allowed to grow to the three leaf stage, and vernalized for eight weeks
Haploid development:

Haploid plantlets are transplanted to the greenhouse to grow until they begin tillering
Chromosome doubling:

Plants are removed from potting medium, and soaked in a solution of colchicine and repotted
Recovery:

Plants recover in growth chambers before returning to the greenhouse to grow to maturity
Harvest:

After reaching maturity, DH lines are threshed and shipped to clients for field testing.
Image credit: J. McGowan, HPI
Seed coat opened on normal and halpoid seeds.
Excised normal embryo compared to haploid embryo on scalpel tip
Kansas
State University
WGRC
Kansas Wheat
Heartland
Wheat accounts for 20% of all the calories consumed by humans worldwide

World trade of wheat is larger than all other crops combined

Food production must to increase by over 70% by 2050 to feed 9 billion

Haploids reduce number of selfing generations before selection

Improve chances of selecting desired recombinant

Select and confirm in the same season

Combined time savings can dramatically cut time for releasing new wheat varieties

Accelerated gene discovery
Doubled haploid production in wheat and other food crops (triticale, sorghum, millet, etc.) must grow to meet the demand of breeding programs.

Overcoming the biological constraints that reduce efficiency of embryo culture and chromosome doubling is integral to achieving that growth in production.

Questions?

tsuelter@heartlandinnovations.com
General Mills
Kansas
Wheat Alliance
Image credit: Kansas Wheat
The Takeaway
Image credit: Kansas Wheat
Crossover and
Gamete Formation
F Generation

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Repeated Self Crossing
F Generation

6-8
6+ Years

Doubled Haploid

1 Year

F Hybrid

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Parents

Conversion Rates at HPI, a Three Year Average:


For every head emasculated we return, on average, 1.9 lines.

For every embryo rescued about 37% will return a developed line.

Turn around times for our orders has gone from 15.2 to 13.7 to 13.0 months

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Full transcript