Master’s Seminar - I

“SPEED BREEDING-A Powerful

Tool To Accelerate Crop Research
And Breeding”

Shreekant T. Pugati
PGS17AGR7568 1
 Flow of seminar

Introduction

Earlier approaches to hasten breeding cycles

Speed breeding: As a powerful tool in crop

research and breeding

Case studies

Conclusion

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 Introduction
 To feed a global population of 9-10 billion by 2050
 Great concern-Current slow improvement rate of
several important crops
 Long generation time of crop plants
 Earlier efforts were made to advance generation
rapidly.

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 Earlier approaches
D
to hasten breeding
C cycles
B
A. Shuttle breeding
A B. Double haploid technology
C. Biotron breeding system
D. RGA
Just imagine you are a
plant breeder…..
First cross to
commercial release
of variety is only six
to seven years

Development of
improved F4 derived
lines within 12
months
Now possible to grow
as many as 6
generations of wheat
every year

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 Speed breeding: As a concept

 Inspiration – NASA

 Prolonged photoperiods to accelerate development rate

of plants and the harvesting and germination of immature
seeds, there by reducing generation time. (Watson et al.,
2018)

 Uses a glasshouse or an artificial environment with

enhanced lighting to create intense long day regimes. (Jhon
Innes centre)
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Current development pipeline for new
cultivars

Line Field Release

development testing (1-3
(5-10 years) (3-5 years) years)

Speed breeding targets here

Line
development Field testing Release
(2 years) (3-5 years) (1-3 years)

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 Drivers

Dr. Lee Hickey

Dr. Brande wulff
Research Scholar
Project leader, Crop genetics
University of Queensland
John innes centre, UK

http://www.abc.net.au/news/rural/2018-01-02/nasa-inspires-plant-speed-breeding-program 8
Protocols in high demand

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Speed breeding accelerates generation time!!!

Watson et al., 2018 10

 Speed breeding I Speed breeding II Speed breeding III

Speed breeding I- Controlled

environment chamber

 22-hour photoperiod and 2-hour dark

period
 Temperature: 22 oC during photoperiod
and 17 oC during 2 hours of dark period
 Humidity: 70 %
 Lighting: Mixture of white LED bars, far-
red LED lamps
 Light intensity -360-380 µmol m-2s-1 Watson et al., 2018 11
Anthesis in approximately half the time than
those in greenhouse

Watson et al., 2018 12

Plants produced healthy number of spikes per plant,
despite of rapid growth
Spike number under Spike number in
Wheat cultivars  speed breeding glasshouse
T. aestivum cv. Paragon 7.3 ± 0.5 3.7 ± 0.9
T. aestivum cv. Cadenza 7.0 ± 0.8 4.3 ± 0.5
T. aestivum cv. Avocet 9.0 ± 1.4 3.7 ± 0.9
T. aestivum cv. Chinese Spring 11.0 ± 0.8 5.3 ± 0.5
T. durum cv. Kronos 8.0 ± 2.5 8.7 ± 2.1
H. vulgare cv. Braemar 14.0 ± 2.9 9.0 ± 1.0

Viability of harvested seeds unaffected by speed breeding

Cultivars/ Seed Day 1 Day 2 Day 3 Day 4 Day 5
Seed Type Accessions total % % % % %
T. aestivum (self) Cadenza 20 90 95 95 95 95
T. aestivum (self) Paragon 20 65 85 90 90 90
T. aestivum (self) Chinese Spring 20 100 100 100 100 100
H. vulgare (self) Braemar 16 100 100 100 100 100
B. distachyon (self) Bd21 20 90 95 95 95 95
B. distachyon (self) Bd3-1 16 56 75 81 81 81
Watson et al., 2018 13
 Speed breeding
Speed breeding I Speed breeding III
II

Speed breeding II- Glasshouse speed

breeding conditions

 A temperature controlled glasshouse fitted with high

pressure sodium vapour lamps (22 hours photoperiod)

 Temperature: 17/22 oC regime with 12 hours turnover

 Light intensity -440-650 µmol m-2s-1

Watson et al., 2018 14

Significant reduction in time to anthesis

Canola (50 DAS) Chickpea (35 DAS)

Watson et al., 2018 15

 Generation time and yield measurements of wheat,
barley, canola and chickpea

Days to Total Mean yield per

 Crop anthesis DPA generation plant Gen./year
time (g/plant)

SB 41.4 ± 1.7 14 65.4 3.5 ± 1.2 5.6

Wheat
GH 63.1 ± 2.8 14 87.1 2.5 ± 1.0 4.2
SB 30.4 ± 1.4 28 68.4 5.9 ± 1.4 5.3
Barley
GH 94.0 ± 14.4 28 132.0 9.4 ± 4.8 2.8
SB 46.2 ± 2.2 42 98.2 8.3 ± 2.2 3.7
Canola
GH 119.1 ± 15.3 42 171.1 9.2 ± 2.4 2.1
SB 30.1 ± 0.7 42 82.1 1.9 ± 1.3 4.5
Chickpea
GH 62.6 ± 3.7 42 114.6 2.4 ± 1.7 3.2

Watson et al., 2018 16

 Speed breeding
Speed breeding I Speed breeding III
II

Speed breeding III- Homemade growth room

design for low cost speed breeding

 A room of 3m × 3m × 3m with insulated sandwich panelling

fitted with seven LED light boxes
 12- hours photoperiod for 4 weeks and then increased to an 18
hours
 1.5 horsepower inverter split system domestic air conditioner
(18 oC in darkness and 21 oC when LED lights on)
 Automatic watering was achieved with irrigation controller

Watson et al., 2018 17

“Recipe” of SB for local purpose and
resource

A set-up for speed breeding using LED lighting

Watson et al., 2018 18
 Integrating speed breeding and SSD
technique

 Plants
Wheat are
(43 DAS) grown at higher density in 100
Barley (34 DAS) cell tray under

speed breeding and glasshouse condition

 The equated density of approximately 900 plants per m2
 Generation time was shorter than plants grown at lower
density in previous speed breeding experiments

Watson et al., 2018 19

Adult plant phenotypes under SB conditions

a. EMS induced mutation of the awn suppressor B1 locus (mutant on

left, wild type on right)

b. Green revolution reduced height (Rht) genes in wheat (from left: Wild,
Rht-1 and Rht-3)

Watson et al., 2018 20

 44 days 67 days

C. Fusarium head blight (Resistant: right, Susceptible: left)

d. Loss of function of FLOWERING LOCUS T-B1 in F6 RILs (F6 – Right

on both panels, Paragon-left, Land race W352- centre)

Watson et al., 2018 21

 Speed breeding in conjugation with genetic
transformation
Time of embryo Number of Number of Transformation
harvesting (days immature transformed efficiency
post sowing) embryos plants

SB Control SB Control SB Control SB Control

60 92 25 75 7 19 28% 25%

Transformation data of barley (H. vulgare) cv. Golden Promise under

speed breeding condition and control conditions
Speed Control
breeding

Let’s keep lights on

Watson et al., 2018 22

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Objectives:
 Assessing the potential use of speed breeding techniques in
Peanut breeding program.

Material and methods:

 F2 and F3 of cross between (Farnsfield×D147-p3-115)

 The experiment was carried out in a greenhouse speed breeding

conditions (PAR 450 watt)

O’ Cooner et al., 2013 24

  F2 - 113 DAS

 F3 - 89 DAS

F2 plants in the greenhouse under

SB condition

O’ Cooner et al., 2013 25

Reproductive success rate of F2 and F3 generations

Emergence Plants with Plants with Seed recovery

Generation Seeds sown
rate mature pods viable seed rate

F2 400 94% 281 270 68%

F3 270 91% 205 201 74%

Strategy 1: 17 months

Strategy 2: 23 months
Strategy 3: 42 months
O’ Cooner et al., 2013 26
Objective:
 Rapidly transfer multiple disease resistance into the Scarlett
genetic background.

Materials and methods:

 Four elite lines from Australia NRB090683-1 , NRB091033,
NRB091087 and NRB091092 used as donors
 F1 and BC1F1 generations were grown under speed breeding
conditions
 Scrabble and Explorer as standards for preliminary yield trial.

Hickey et al., 2017 27

 Disease screening in F2 and BC1F2

16 DAS inoculation and 25 DAS scoring

• Plants were inoculated with net form of net
blotch (NFNB) and spot form of net blotch
(SFNB) and incubated in a dew chamber for 24 h

27 DAS inoculation and 37 DAS scoring

• Plants were inoculated with spot blotch (SB)
and incubated in a dew chamber for 24 h

47 DAS inoculation and 57 DAS scoring

• Plants were inoculated with leaf rust and
incubated in a dew chamber for 16 h

Hickey et al., 2017 28

Rapid generation advance and
environmental control

2 Years

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Hickey et al., 2017
Summary of selection performed in segregating
generations
F2 BC1F2 BC1F3
Recurrent
Donor source parent Screened Selected Screened Selected Screened Selected

NRB090683-1 Scarlett 1091 20 (2) 1091 20 274 10

NRB091033 Scarlett 1004 53 (2) 1004 47 637 45

NRB091087 Scarlett 1340 36 (1) 1340 31 117 7

NRB091092 Scarlett 1565 65 (2) 1565 64 722 25

Total   5000 129 (7) 5000 162 1751 87

Hickey et al., 2017 30

Yield performance of elite introgression lines

Balcarce

Tres arroyos

Mean yield for the 12 elite introgression lines

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Hickey et al., 2017
Objective:
 Selection for traits like seminal root angle (RA), seminal root
number (RN), tolerance to crown rot (CR), resistance to leaf
rust (LR) and plant height (PH)

Material and Methods:

 F2 and F3 populations derived from the parents Outrob4 and
Caparoi
 Mace, Scout, Thatcher, Thatcher + Lr34, Sunguard and Yawa as
standards.
 Clear pot technique 32
The breeding strategy for applying selection
in early segregating generations

Alahmad et al., 2017 33

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Frequency distribution for the
F2 segregating population

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Density distribution of the weighted selection
index values for selected, unselected and the
entire F2 generation

Alahmad et al., 2017 36

 Density distribution and comparison of
population means for selected and unselected F3
population sets

Alahmad et al., 2017 37

Alahmad et al., 2017 38
• Rapid generation homozygosity • Cost involved
• Rapid production of cultivars • Maintenance of CEnvC
• Ideally suited to backcross • Breeding populations
breeding strategy • Intense multi trait selection
• Plant – pathogen interaction
studies
• Crop genetics

Pros Cons

Species responding to extended photoperiod

Protocols to short-day species

Integration with other advance technologies

Future Advances in LED light technologies

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Conclusion..
. Speed riding is “Danger” but……

Speed breeding is “Future”

Thank you 40