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1
Supplementary Material for
Speed breeding: a powerful tool to accelerate crop research and breeding
This file includes:
Supplementary Information
Supplementary Figures 1 to 12
Supplementary Tables 1 to 37
Supplementary References
Other supporting online material for this manuscript includes the following:
Supplementary Media File 1 (Time-lapse video)
2
Supplementary Information
Seed count
Supplementary Table 5 shows the average number of seeds per spike in T. aestivum cvs.
Chinese Spring and Paragon under speed breeding condition I and control conditions
(glasshouse with natural UK summer photoperiod and no supplementary light). Three different
spikes per plant were individually bagged to score manually the total number of seeds obtained
per spike in each treatment. Slight statistical differences (p = 0.0495) were observed between
treatments in T. aestivum cv. Chinese Spring (Supplementary Table 5). Control spikes from
plants grown in the glasshouse with natural UK summer photoperiod showed a higher average
of seed count than spikes from plants grown under speed breeding conditions (Supplementary
Table 5). The minimum and maximum number of seeds per treatment was 41 and 53,
respectively, in the control spikes and 30 and 51, respectively, in spikes from plants grown
with a photoperiod of 22 hours. On the other hand, no significant differences were found in T.
aestivum cv. Paragon between treatments (p = 0.0718) (Supplementary Table 5). The counting
of seed was the same as described above. Control plants grown in the glasshouse showed a
higher number of seeds per spike than plants grown in the chamber with a 22-hour photoperiod
(Supplementary Table 5). The minimum and maximum number of seeds per treatment was 41
and 76, respectively, in the control spikes and 39 and 72, respectively, in spikes from plants
grown with a photoperiod of 22 hours.
Analysis of meiosis
The results of the chromosome pairing at meiotic metaphase I (MI) of wheat carrying the Ph1
(Pairing Homoeologous I) locus and wheat-rye hybrids carrying and lacking the Ph1 locus
under speed breeding condition I (Supplementary Table 1: Speed Breeding I) and variable UK
summer as control are summarized in Supplementary Table 32. At MI in these plants, a variable
number of univalents, bivalents and multivalents were observed in both treatments
(Supplementary Fig. 4). In wheat carrying the Ph1 locus (Triticum aestivum cv. Chinese
Spring), the vast majority of chromosomal structures were bivalents (Supplementary Fig. 4a,b;
Supplementary Table 32). Univalents were rarely observed (Supplementary Fig. 4a,b;
Supplementary Table 32), at an average of 0.1 univalents under speed breeding and 0.2
univalents under control conditions, indicating a stable meiosis in wheat. Total number of
chiasmata/crossovers (COs) were not statistically different between treatments showing an
average of chiasmata/COs quite similar (40.1 under speed breeding conditions and 40.4 under
3
control conditions). In wheat-rye hybrids carrying the Ph1 locus, little variation was observed
between univalents and bivalents (Supplementary Fig. 4c,d). An average of 27.0 univalents
and 0.5 bivalents were observed in plants grown under speed breeding and 26.8 univalents and
0.6 bivalents were observed in the control plants, showing no significant differences (p > 0.05)
in both treatments. No multivalents were observed in the presence of the Ph1 locus, as expected
(Supplementary Fig. 4c,d). This wheat-rye hybrid genotype did not show significant
differences in the total number of chiasmata/COs between treatments resulting in a meiosis not
affected by the increase in the number of hours of light (Supplementary Table 32). In wheat-
rye hybrids lacking the Ph1 locus, the average number of univalents, bivalents and multivalents
was quite similar in both treatments, indicating also a stable meiosis (Supplementary Fig. 4e,f).
The number of univalents was higher than the number of bivalents and multivalents under both
treatment conditions, at an average of 13.6 under speed breeding and 13.8 under glasshouse
conditions. The frequency of multivalents in both treatments was quite low (0.1 both in 22-
hour light and in glasshouse). Moreover, the number total of chiasmata was not statistically
different in both treatments (8.3 under speed breeding and 8.7 under control conditions)
(Supplementary Table 32).
4
Supplementary Figures
Supplementary Fig. 1 | Development stages under speed breeding condition I (left) and
control conditions (right) of (a) M. truncatula (one branch of a plant in each condition pictured)
at 60 days post transfer of seedlings and (b) P. sativum JI 2822 at 37 days post sowing,
respectively. Scale bar = 20 cm. Control conditions for pea are described in Supplementary
Table H, while those for M. truncatula are given in Supplementary Table G.
Supplementary Fig. 2 | A set-up for speed breeding using LED lighting (Supplementary Table
1: Speed breeding III).
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Supplementary Fig. 3 | Representative chromosome pairing in wheat carrying Ph1,
wheat-rye hybrids carrying Ph1 and wheat-rye hybrids lacking Ph1 grown under speed
breeding condition I and control conditions. (a,b) Pollen mother cells (PMCs) of wheat with
42 chromosomes. (a) Wheat (T. aestivum cv. Chinese Spring) grown under speed breeding
condition I (20 bivalents (rings) + 1 bivalent (rod)) and (b) Wheat (T. aestivum cv. Chinese
Spring) grown under a natural UK summer photoperiod (19 bivalents (rings) + 2 bivalents
(rods)). (c,d) PMCs of wheat-rye hybrid in presence of Ph1 with 28 chromosomes. (c) Wheat-
rye hybrid grown under speed breeding condition I (28 univalents) and (d) wheat-rye hybrids
grown under a natural UK summer photoperiod (28 univalents). (e,f) PMCs of wheat-rye
hybrid in absence of Ph1 with 28 chromosomes. (e) Wheat-rye hybrid grown under speed
breeding condition I (4 bivalents (rings) + 6 bivalents (rods) + 8 univalents) and (f) wheat-rye
hybrid grown under a natural UK summer photoperiod (1 bivalent (ring) + 5 bivalents (rods)
+ 16 univalents). Scale Bar = 10 μm.
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Supplementary Fig. 4 | Development of callus on transition medium [S1] derived from
immature embryos of H. vulgare cv. Golden Promise grown under speed breeding condition I,
after 6 weeks of callus induction. Transformed green shoots are seen developing in 5 out of 12
immature embryo-derived calli.
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Supplementary Fig. 5 | Development of transformed explants of barley (H. vulgare cv. Golden
Promise) under speed breeding condition I (left) and 16-hour photoperiod control condition
(right) at 84 days post transfer of 14-week old explants to respective conditions. Scale bar = 40
cm.
SupplementaryFigure
8
Supplementary Fig. 6. Pendulum apparatus used for pod shattering resistance method (left).
Canola pod positioning during method (right).
9
Supplementary Fig. 7 | Spectral measurement of light composition in the Conviron chamber
used for speed breeding condition I (Supplementary Table 1: Speed Breeding I), measured
using the MK350N handheld spectrometer from UPRtek. X-axis values are wavelength in
nanometres, Y-axis represents proportion (1 unit = 0.1 proportion).
Supplementary Fig. 8 | Growth curves tracking the development of Triticum aestivum cv.
Paragon under speed breeding condition I (“Speed breeding”) and control glasshouse
conditions (“control conditions”) in UK Summer with no supplementary light. Germinated
seedlings were sown in both treatments on 17 March, 2017. Calculations are based on
recordings made by the CropQuant workstation developed at the Norwich Research Park
(Supplementary Media File 1).
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Supplementary Fig. 9 | Set-up for speed breeding in a temperature-controlled glasshouse fitted
with high pressure sodium vapour lamps (Supplementary Table 1: Speed breeding II). View
from outside (left) and inside (right).
Supplementary Fig. 10 | Spectral measurement of light composition for Philips SON-T 400
W sodium vapour lamps used in QLD glasshouse for speed breeding condition II
(Supplementary Table 1: Speed Breeding II). Sourced from http://www.lighting.philips.com.
11
Supplementary Fig. 11 | Spectral measurement of light composition in the LED-based growth
room chamber at PBI (Supplementary Table 1: Speed breeding III), measured using the
Lighting Passport™ from AsenseTek.
12
Supplementary Fig. 12 | Ear and seed morphology of Triticum aestivum cv. Chinese
Spring grown in speed breeding condition I and control conditions. a,b Spike morphology
of T. aestivum cv. Chinese Spring grown under (a) speed breeding condition I and under (b)
glasshouse conditions in UK summer without any supplementary lights. c,d Seed morphology
of T. aestivum cv. Chinese Spring under (c) speed breeding condition I and (d) glasshouse
conditions in UK summer without any supplementary light.
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Supplementary Tables
Supplementary Table 1 | Summary of speed breeding experiments, conditions and species
used.
Experiment Speed breeding
protocol
Species and cultivars/accessions
Speed breeding I:
Demonstration of
generation
advancement using
growth chamber
Conviron
Chamber
Protocol (John
Innes Centre,
UK)
(i) T. aestivum cvs. Paragon, Cadenza, Kronos,
AvocetS, Chinese Spring
(ii) H. vulgare cvs. Braemar, Golden Promise
(iii) B. distachyon accessions Bd21, Bd3-1
(iv) M. truncatula genotype Jemalong A17
(v) P. sativum accession JI 2822
Speed breeding II:
Demonstration of
generation
advancement using
glasshouse
supplemented with
sodium vapour lamps
Harvest indices under
speed breeding
conditions
Generation
advancement through
SSD using speed
breeding
Glasshouse
Protocol
(University of
Queensland,
Australia)
(i) T. aestivum cvs. Drysdale, EGA Gregory, EGA
Wylie, Mace, Scout, SeriM82, Suntop,
Westonia, Dharwar Dry, Spitfire
(ii) H. vulgare cvs. Commander, Compass, Flagship,
La Trobe, Shepherd, Westmister, NRB090257,
NRB090885, ND24260, Grout
(iii) C. arietinum cvs. PBA Boundary, PBA
HatTrick, Jimbour, Kyabra
(iv) B. napus cvs. Taparoo, ATR Cobbler, ATR
Beacon, CB Argyle, Boomer, Westar, Skipton,
Bravo TT
Phenotype of awn
suppressor mutants
under speed breeding
conditions
Conviron
Chamber
Protocol (John
Innes Centre,
UK)
T. aestivum cv. Paragon (wild-type) and mutants
thereof in the awn suppressor B1 locus
Expression of the
allelic series of the Rht
genes
Conviron
Chamber
Protocol (John
Innes Centre,
UK)
T. aestivum cv. Maringá, Maringá with Rht-1
introgressed, Maringá with Rht-3 introgressed
Fusarium
graminearum infection
studies
Conviron
Chamber
Protocol (John
Innes Centre,
UK)
T. aestivum cvs. Sumai 3, Timstein
Phenotype flowering
time difference
between the parents
Conviron
Chamber
Protocol (John
T. aestivum cv. Paragon, W-352 (landrace), and an F6
hybrid of the two
14
(W-352 and Paragon)
and their F1 hybrid,
due to a deletion in
the FT-B1 locus
Innes Centre,
UK)
Phenotype reduced
glaucousness in
Eceriferum cqu
mutants
Conviron
Chamber
Protocol (John
Innes Centre,
UK)
H. vulgare cv. Bonus and mutants thereof in the
Eceriferum cqu locus
Studies on
chromosome
associations and
chiasmata frequency
in wheat and wheat-
rye hybrids
Conviron
Chamber
Protocol (John
Innes Centre,
UK)
(i) T. aestivum cv. Chinese Spring (+/- Ph1)
(ii) S. cereale cv. Petkus (+/- Ph1)
Transformation of
barley
Conviron
Chamber
Protocol (John
Innes Centre,
UK)
H. vulgare cv. Golden Promise
Demonstration of
growth and
development of
transformed barley
explants
Conviron
Chamber
Protocol (John
Innes Centre,
UK)
H. vulgare cv. Golden Promise
Phenotype method for
resistance to pod
shatter
Glasshouse
Protocol
(University of
Queensland,
Australia)
Pendulum
method
(Graham Centre,
NSW
Department of
Primary
Industries,
Wagga Wagga,
Australia)
(i) B. napus cvs. ATR-Cobbler, ATR-Beacon, CB-
Argyle, Skipton, Bravo TT
Speed breeding III:
Demonstration of
generation
enhancement using
LED lighting
LED Protocol
(University of
Sydney,
Australia)
(i) T. aestivum cvs. Morocco, AvocetR
(ii) H. vulgare cvs. Gus, Baudin
(iii) Avena sativa cv. Swan
(iv) Triticosecale cvs. Jackie, Coorong
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Supplementary Table 2 | Growth stages (GS) of wheat under speed breeding condition I (SB)
and glasshouse conditions in UK summer without any supplementary lights (sown on 29 March
2016). Values indicated are expressed as mean days after sowing (DAS)1 ± SD based on three
replicates.
Growth
Stage2
T. aestivum
cv. Paragon
T. aestivum
cv. Cadenza
T. aestivum
cv. AvocetS
T. aestivum
cv. Chinese
Spring
T. durum cv.
Kronos
SB GH SB GH SB GH SB GH SB GH
1st leaf
(GS11)
2.0 ±
0.0
4.0 ±
0.0
2.0 ±
0.0
4.0 ±
0.0
2.0 ±
0.0
4.0 ±
0.0
2.0 ±
0.0
4.0 ±
0.0
2.0 ±
0.0
4.0 ±
0.0
3rd leaf
(GS13)
7.0 ±
0.0
18.0
± 0.0
7.0 ±
0.0
18.0
± 0.0
7.7
± 0.9
18.0
± 0.0
6.0 ±
0.0
18.0
± 0.0
7.0 ±
0.0
18.0
± 0.0
Stem
extension
(GS32)
21.0
± 0.0
45.0
± 0.0
23.0
± 0.0
47.0
± 0.0
22.3
± 0.9
40.0
± 0.0
39.0
± 0.0
46.0
± 0.0
21.0
± 0.0
39.0
± 0.0
Boot
(GS45)
30
± 1.6
67.0
± 0.0
28.0
± 0.0
69.7
± 1.9
28.0
± 0.0
69.7
± 1.9
47.0
± 0.0
66.0
± 0.0
25.0
± 0.0
64.0
± 0.0
Head
(GS59)
36
± 1.6
76
± 1.4
34.0
± 0.0
74.7
± 1.9
34.3
± 0.9
77
± 1.4
57.7
± 0.9
74.0
± 0.0
30.7
± 0.9
70.0
± 0.0
Anthesis
(GS65)
39
± 0.8
82.7
± 0.9
38.0
± 0.0
80
± 1.6
37.3
± 0.9
82.7
± 1.9
63.0
± 0.0
78.0
± 0.0
35.7
± 0.5
74.0
± 0.0
Grain milk 47.0
± 0.0
95.0
± 0.0
47.0
± 0.0
95.0
± 0.0
47.0
± 0.0
95.0
± 0.0
72.0
± 0.0
95.0
± 0.0
44.0
± 0.0
95.0
± 0.0
Grain
dough (GS
87)3
56.0
± 0.0
105.0
± 0.0
56.0
± 0.0
105.0
± 0.0
56.0
± 0.0
105.0
± 0.0
80.0
± 0.0
105.0
± 0.0
56.0
± 0.0
102.0
± 0.0
1 DAS refers to the number of days (post transfer of germinated seedlings) to reach the indicated
developmental growth stages [S2]. Germination for all samples required 5 days. 2 All measurements are with respect to the main tiller. 3 Seeds for speed breeding were harvested on the 6th day following the GS87 reading. No water
was provided to the plants from Day 56 to Day 62 (harvest day).
NOTE: Post heading, plants were phenotyped every 2-4 days. This may cause whatever
differences there might be between replicates or varieties to even out at the time of
measurement, causing a net zero standard deviation.
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Supplementary Table 3 | Growth stages of barley under speed breeding condition I (SB) and
glasshouse conditions in UK summer without any supplementary lights (sown on 29 March
2016). Values indicated are expressed as mean days after sowing (DAS)1 ± SD based on three
replicates.
Development stage2 H. vulgare cv. Braemar
H. vulgare cv. Golden
Promise
SB GH SB GH
1st leaf 2.0 ± 0.0 4.0 ± 0.0 2.0 ± 0.0 4.0 ± 0.0
3rd leaf 7.0 ± 0.0 18.0 ± 0.0 7.0 ± 0.0 18.0 ± 0.0
Stem Extension (GS32) 24.0 ± 0.0 38.0 ± 0.0 26.0 ± 0.0 38.0 ± 0.0
Flag leaf 28.0 ± 0.0 55.0 ± 0.0 36.0 ± 0.0 55.0 ± 0.0
Head (GS51) 35.0 ± 0.0 71.0 ± 0.0 45.0 ± 0.0 74.0 ± 0.0
Anthesis 37.0 ± 0.0 76.0 ± 0.0 38.0 ± 0.0 80.0 ± 0.0
Grain milk 47.0 ± 0.0 85.0 ± 0.0 49.0 ± 0.0 90.0 ± 0.0
Grain dough
(viable seed collection) 55.0 ± 0.0 102.0 ± 0.0 60.0 ± 0.0 115.0 ± 0.0
1 DAS refers to the number of days (post transfer of germinated seedlings) to reach the indicated
developmental growth stages. Germination for all samples required 5 days. 2 All measurements are with respect to the main tiller.
NOTE: Post heading, plants were phenotyped every 2-4 days. This may cause any prior
differences among replicates and/or varieties to even out at the time of measurement, resulting
in a net zero standard deviation.
17
Supplementary Table 4 | Growth stages of two B. distachyon accessions under speed breeding
condition I (SB) and glasshouse conditions in UK summer without any supplementary lights
(sown on 29 April 2016). Values indicated are expressed as mean days after sowing (DAS)1 ±
SD based on three replicates.
Development stage1 Bd21 Bd3-1
SB GH SB GH
1st leaf 2.0 ± 0.0 4.0 ± 0.0 2.0 ± 0.0 4.0 ± 0.0
3rd leaf 5.0 ± 0.0 12.0 ± 0.0 5.0 ± 0.0 12.0 ± 0.0
Stem extension 14.0 ± 0.0 36.0 ± 0.0 14.0 ± 0.0 36.0 ± 0.0
Head 26.0 ± 0.0 54.0 ± 0.0 26.0 ± 0.0 54.0 ± 0.0
Viable seed collection 48.0 ± 0.0 73.0 ± 0.0 48.0 ± 0.0 73.0 ± 0.0 1 DAS (days after sowing) refers to the number of days (post transfer of germinated seedlings)
to reach the indicated developmental growth stages. Germination for all samples required 5
days. 2 All measurements are with respect to the tallest tiller.
Supplementary Table 5 | Average number of seeds per bagged spike from T. aestivum cvs.
Chinese Spring and Paragon grown under speed breeding condition I and glasshouse conditions
in UK summer without any supplementary lights. Values indicated are expressed as mean ±
SD based on three replicates and three spikes per replicate.
T. aestivum cultivar Speed breeding Glasshouse p value1
Chinese Spring 40.6 ± 9.3 45.8 ± 3.7 0.0495
Paragon 53.8 ± 9.6 64.4 ± 12.6 0.0718 1 LSD test.
Supplementary Table 6 | Spike counts of wheat and barley cultivars grown under speed
breeding condition I and glasshouse conditions in UK Summer with no supplementary light.
Sown on 29 March 2016. Values indicated are expressed as mean ± SD.
Spike number under
speed breeding1
Spike number in
glasshouse2
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. AvocetS 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 1 Grown in 900 ml of JIC Cereal Compost Mix (Supplementary Table 36). 2 Grown in 600 ml of JIC Cereal Compost Mix (Supplementary Table 36).
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Supplementary Table 7 | Germination percentage per day following chilling at hard dough
stage in wheat seeds harvested under speed breeding condition I. All seeds were stratified for
3 days at 4 C.
Seed Type Cultivars/
Accessions
Seed
total
Day 1
%
Day 2
%
Day 3
%
Day 4
%
Day 5
%
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
T. aestivum (cross) Cadenza (♀) x
AvocetS (♂)
20 20 35 40 55 95
T. aestivum (cross) AvocetS (♀) x
Paragon (♂)
13 23 54 69 69 85
T. aestivum (cross) Cadenza (♀) x
Paragon (♂)
13 62 69 69 77 92
T. aestivum (cross) Paragon (♀) x
AvocetS (♂)
15 80 93 93 93 93
Inter-species
hybrid: T. durum x
T. aestivum
Kronos (♀) x
AvocetS (♂)
7 14 57 57 57 57
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Supplementary Table 8 | Efficiency rates for wheat crosses under speed breeding condition
I.
Pollen Recipient Pollen Donor
Efficiency
(% of pollinated ovaries that set
seed)
T. aestivum cv. Cadenza T. aestivum cv. AvocetS 87.5
T. aestivum cv. AvocetS T. aestivum cv. Paragon 62.0
T. aestivum cv. Cadenza T. aestivum cv. Paragon 65.0
T. durum cv. Kronos T. aestivum cv. AvocetS 70.0
T. aestivum cv. Paragon T. aestivum cv. AvocetS 82.0
Supplementary Table 9 | Medicago truncatula A17 growth rates under speed breeding
condition I and control conditions. Values indicated are expressed as mean ± SD.
Speed Breeding1,3 Control1,2,3
Days to flowering 35.3 0.5 70.0 1.0
Days to harvest 89.0 1.4 131.0 3.0
Number of pods 227.3 39.4 152.5 14.5
Number of seeds per pod 8.1 1.7 8.1 0.2
Harvested seed germination rates (%) 80.0 0.05 78.0 2.0 1 Stratified for 3 days, then placed in each condition on 24 June 2016. 2 Control conditions were 16-hour photoperiod with 22 C day and 20 C night temperatures,
80% relative humidity and light levels of 140-150 µmol m-2 s-1 at the bench level 335-350 µmol
m-2 s-1 at adult plant height. Light was provided by metal halide lamps (HQI) supplemented
with tungsten bulbs. 3 Calculations are based on three replicates under speed breeding conditions and two under
control conditions.
20
Supplementary Table 10 | Pea (Pisum sativum JI 2822) growth rates under speed breeding
condition I and control conditions. Values for traits are expressed as mean (± SD) based on
five replicates.
Speed breeding1 Control1,2
Days to flowering 26.4 ± 1.9 38.4 ± 1.3
Node number at flowering 8.2 ± 1.0 5.8 ± 0.4
Node number at harvest 12.4 ± 0.8 10.4 ± 0.9
Plant height (mm) 120.2 ± 12.6 295 ± 48.2
Number of pods 3.8 ± 0.7 7.2 ± 1.3
Number of side shoots 1.6 ± 0.5 0.8 ± 0.4
Number of seeds 11.2 ± 0.4 19.6 ± 3.4
Water stopped 37 DAS3 63 DAS
Harvest 51 DAS 84 DAS
% Germination 98.2 ± 4.0 100 ± 0 1 Seeds were chipped and sown into compost and placed either in speed breeding or control
conditions. Calculations based on five plants in each condition. 2 Control conditions in a glasshouse without supplementary lighting, natural UK spring
photoperiod with a minimum of 12 C. 3 DAS, days after sowing. Seeds under both conditions were sown on 3 April 2017.
Supplementary Table 11 | Development stages1 (GS) of wheat under speed breeding
condition II (SB) and glasshouse (GH) conditions in Queensland, Australia, with no
supplementary light (sown on 18 June 2016). Values indicated are expressed as mean days
after sowing (DAS)2 ± SD based on three replicates.
Development stage
GS13 GS65
T. aestivum cultivar SB GH SB GH
Dharwar Dry 15.7 ± 0.6 18.0 ± 1.0 43.0 ± 0.0 71.0 ± 3.5
Drysdale 14.0 ± 0.0 17.0 ± 0.0 40.0 ± 1.0 55.0 ± 1.7
EGA Gregory 14.0 ± 0.0 16.0 ± 0.0 45.3 ± 0.6 70.0 ± 2.6
EGA Wylie 14.0 ± 2.8 14.7 ± 0.6 42.0 ± 0.0 75.7 ± 1.2
Mace 13.3 ± 0.6 16.7 ± 1.2 44.0 ± 2.6 68.3 ± 1.5
Scout 13.5 ± 0.7 16.3 ± 0.6 41.0 ± 1.7 57.0 ± 2.6
SeriM82 15.5 ± 0.7 16.0 ± 2.6 42.3 ± 6.0 65.3 ± 4.2
Spitfire 13.7 ± 1.5 15.7 ± 0.6 41.3 ± 3.8 58.7 ± 7.4
Suntop 14.0 ± 1.0 18.0 ± 1.0 37.3 ± 0.6 54.3 ± 2.5
Westonia 12.7 ± 1.5 17.7 ± 0.6 37.7 ± 0.6 56.0 ± 1.0 1 GS13 = 3rd leaf emerged, GS65 = anthesis. 2 DAS refers to the number of days (post transfer of germinated seedlings) to reach the
indicated developmental growth stages. Germination for all samples required 5 days.
21
Supplementary Table 12 | Development stages1 (GS) of barley under speed breeding
condition II (SB) and glasshouse (GH) conditions in Queensland, Australia, with no
supplementary light (sown on 18 June 2016). Values indicated are expressed as mean days
after sowing (DAS)2 ± SD based on three replicates.
Development stage
GS13 GS49
H. vulgare cultivar SB GH SB GH
Commander 10.0 ± 1.7 17.0 ± 0.0 22.7 ± 1.5 101.5 ± 19.1
Compass 12.0 ± 1.7 17.7 ± 0.6 39.7 ± 2.9 105.0 ± 23.9
Flagship 10.7 ± 0.6 15.7 ± 0.6 29.3 ± 0.6 115.7 ± 11.7
La Trobe 10.0 ± 0.0 15.7 ± 0.6 23.0 ± 1.0 93.3 ± 11.5
Shepherd 13.5 ± 0.7 17.0 ± 1.0 38.0 ± 1.0 88.7 ± 21.8
Westminster 14.0 ± 1.0 18.3 ± 1.2 39.0 ± 3.5 94.7 ± 19.1
NRB090257 9.3 ± 0.6 16.7 ± 0.6 28.3 ± 1.2 89.3 ± 14.2
NRB090885 13.3 ± 1.2 18.0 ± 0.0 30.0 ± 0.0 78.7 ± 5.5
ND24260 13.3 ± 1.2 16.7 ± 0.6 33.7 ± 1.5 79.7 ± 5.9
Grout 9.5 ± 0.7 17.7 ± 0.6 21.0 ± 1.0 93.3 ± 11.5 1 GS13 = 3rd leaf emerged, GS49 = awn peep. 2 DAS refers to the number of days (post transfer of germinated seedlings) to reach the indicated
developmental growth stages. Germination for all samples required 5 days.
22
Supplementary Table 13 | Development stages1 of canola flowering under speed breeding
condition II (SB) and glasshouse (GH) conditions in Queensland, Australia, with no
supplementary light (sown on 18 June 2016). Values indicated are expressed as mean days
after sowing (DAS)2 ± SD based on three replicates.
Development stage
Stage 3.3 Stage 3.6 Stage 4.1
B. napus cultivar SB GH SB GH SB GH
Taparoo 36.7 ±
1.2
119.0 ±
0.0
38.3 ±
1.0
147.5 ±
7.8
44.0 ±
1.0
158 ±
15.6
ATR Cobbler 39.0 ±
0.0
103.7 ±
19.0
41.0 ±
0.0
108.3 ±
18.5
46.3 ±
1.2
110.7 ±
16.6
ATR Beacon 37.7 ±
3.1
92.0 ±
22.0
40.3 ±
2.0
96.3 ±
19.9
45.0 ±
2.0
104.3 ±
21.9
CB Argyle 42.3 ±
1.5
118.3 ±
1.0
43.7 ±
1.0
130.5 ±
6.4
49.7 ±
1.2
136.7 ±
11.6
Westar 41.0 ±
0.0
87.7 ±
6.0
42.7 ±
1.0
90.3 ±
6.7
47.7 ±
0.6
97.7 ±
5.9
Skipton 37.0 ±
5.7
114.3 ±
23.0
40.0 ±
6.0
126.7 ±
28.6
44.0 ±
5.2
136.0 ±
30.0
Bravo TT 38.5 ±
4.9
75.0 ±
2.0
40.5 ±
5.0
81.7 ±
4.6
47.0 ±
4.2
90.0 ±
5.2 1 Stage 3.3 = green buds visible; Stage 3.6 = flower stalk extends; Stage 4.1 = first flower opens
[S3]. 2 DAS refers to the number of days (post transfer of germinated seedlings) to reach the indicated
developmental growth stages. Germination for all samples required 5 days.
Supplementary Table 14 | Number of days after sowing for which chickpea (C. arietinum)
cultivars reached early bloom1 under speed breeding condition II (SB) and glasshouse (GH)
conditions in Queensland, Australia, with no supplementary light (sown on 7 June 2016).
Values indicated are expressed as mean ± SD days after sowing (DAS)2 based on five
replicates.
Development
stage
PBA Boundary PBA HatTrick Jimbour Kyabra
SB GH SB GH SB GH SB GH
Early bloom 31.4 ±
1.1
63.4 ±
4.6
28.6 ±
0.5
67.8 ±
7.4
29.4 ±
0.5
60.3 ±
1.3
30.8 ±
0.8
58.8 ±
1.3 1 Early bloom = first flower opens.
2 DAS refers to the number of days (post transfer of germinated seedlings) to reach the
indicated developmental growth stages. Germination for all samples required 5 days.
23
Supplementary Table 15 | Growth stage1 (GS) of wheat grown under glasshouse conditions
in Queensland, Australia, with no supplementary light at the time when plants under speed
breeding condition II reached anthesis. Both treatments were sown on 18 June 2016.
T. aestivum cultivar GS65
Dharwar Dry GS13
Drysdale GS14
EGA Gregory GS14
EGA Wylie GS14 - GS15
Mace GS13 - GS14
SeriM82 GS13 - GS14
Spitfire GS14 - GS15
Suntop GS13 - GS14
Westonia GS13 1 GS65 = anthesis, GS13 = 3rd leaf emerged, GS14 = 4th leaf emerged, GS15 = 5th leaf emerged.
Supplementary Table 16 | Growth stage1 (GS) of barley grown under glasshouse (GH)
conditions in Queensland, Australia, with no supplementary light at the time when plants under
speed breeding condition II reached awn peep. Both treatments were sown on 18 June 2016.
H. vulgare cultivar SB GH
Commander GS491 GS132-GS143
La Trobe GS49 GS14
Grout GS49 GS13 1 GS49 = awn peep, GS13 = 3rd leaf emerged, GS14 = 4th leaf emerged.
Supplementary Table 17 | Development stages1 of canola grown under glasshouse (GH)
conditions in Queensland, Australia, with no supplementary light at the time when plants under
speed breeding condition II reached stage 3.3, 3.6 and 4.1. Both treatments were sown on 18
June 2016.
Development stages
SB at Stage 3.3 SB at Stage 3.6 SB at Stage 4.1
B. napus cultivars GH GH GH
Taparoo 4th leaf2 - 5th leaf 5th leaf 6th leaf – 7th leaf
ATR Cobbler 4th leaf - 5th leaf 5th leaf - 6th leaf 6th leaf
ATR Beacon 4th leaf - 5th leaf 6th leaf - 7th leaf 7th leaf – 9th leaf
Skipton 5th leaf 5th leaf 8th leaf – 10th leaf
Bravo TT 4th leaf - 5th leaf 6th leaf 8th leaf - 9th leaf 1 Stage 3.3 = green buds visible, Stage 3.6 = flower stalk extends, Stage 4.1 = first flower opens. 2 Leaf exposed.
24
Supplementary Table 18 | Development stage1 of chickpea (C. arietinum) grown under
glasshouse (GH) conditions in Queensland, Australia, with no supplementary light at the time
when plants under speed breeding condition II reached anthesis. Both treatments were sown
on 7 June 2016.
Development
stage
PBA Boundary PBA HatTrick Jimbour Kyabra
GH GH GH GH
SB at anthesis 13th leaf 12th leaf 9th leaf 10th leaf 1 Refers to unfolding of multifoliate leaves.
Supplementary Table 19 | Number of spikes per plant and grains per spike on self-pollinated
mature wheat grown under speed breeding condition II (SB) and glasshouse (GH) conditions
in Queensland, Australia, with no supplementary light (sown on 18 June 2016). Values
indicated are expressed as mean ± SD.
Spikes per plant Grain per spike
T. aestivum cultivar SB GH SB GH
Dharwar Dry 3.3 ± 0.6 1.0 ± 0.0 32.0 ± 5.1 42.4 ± 9.4
Drysdale 4.0 ± 1.0 1.3 ± 0.6 35.8 ± 5.0 35.0 ± 9.6
EGA Gregory 5.7 ± 0.6 2.3 ± 0.6 33.8 ± 5.0 37.6 ± 9.4
EGA Wylie 6.0 ± 1.7 3.7 ± 0.6 25.9 ± 5.0 30.3 ± 9.6
Mace 5.0 ± 1.0 2.0 ± 0.0 36.7 ± 5.0 35.1 ± 9.6
Scout 4.3 ± 0.6 3.0 ± 1.0 36.3 ± 5.0 27.5 ± 9.4
SeriM82 4.7 ± 2.2 2.0 ± 1.0 37.2 ± 5.0 33.5 ± 9.4
Spitfire 6.0 ± 1.0 1.7 ± 1.2 27.7 ± 5.1 24.6 ± 9.4
Suntop 2.0 ± 1.0 1.7 ± 0.6 34.7 ± 5.1 35.5 ± 9.6
Westonia 2.3 ± 1.5 1.7 ± 1.2 30.9 ± 5.0 37.7 ± 9.4
25
Supplementary Table 20 | Number of spikes per plant and grains per spike on self-pollinated
mature barley grown under speed breeding conditions II (SB) and glasshouse (GH) conditions
in Queensland, Australia, with no supplementary light (sown on 18 June 2016). Values
indicated are expressed as mean ± SD.
H. vulgare cultivar Spikes per plant Grain per spike
SB GH SB GH
Compass 11.3 ± 2.1 17.0 ± 0.0 18.4 ± 1.8 21.6 ± 0.0
Flagship 12.0 ± 3.0 15.0 ± 4.6 14.0 ± 0.6 12.1 ± 3.9
La Trobe 14.0 ± 6.2 20.0 ± 9.0 11.5 ± 0.3 13.9 ± 1.3
Shepherd 11.2 ± 2.5 13.0 ± 8.2 21.0 ± 2.7 12.7 ± 2.7
Westminster 10.7 ± 0.6 11.3 ± 9.1 21.5 ± 2.1 15.5 ± 1.0
NRB090257 6.7 ± 1.2 19.3 ± 6.8 20.3 ± 2.1 17.4 ± 1.1
NRB090885 9.3 ± 1.5 15.0 ± 3.0 21.8 ± 1.0 19.4 ± 2.5
ND24260 7.7 ± 1.2 13.3 ± 2.9 21.2 ± 1.6 18.4 ± 3.1
Grout 9.3 ± 2.3 11.7 ± 5.7 12.7 ± 2.9 17.8 ± 8.5
Supplementary Table 21 | Germination percentage of 30 wheat (T. aestivum) seeds harvested
early (14 days post-anthesis) and at maturity from speed breeding condition II (SB) and
glasshouse (GH) conditions in Queensland, Australia, with no supplementary light (sown on
18 June 2016). All seeds were kept at 4 °C for 4 days prior to germination. Values indicated
are expressed as mean ± SD.
Cultivar
Germination percentage
Early Mature
SB GH SB GH
Dharwar Dry 95.0 ± 8.7 88.0 ± 10.4 100.0 ± 0.0 80.0 ± 24.1
Drysdale 90.7 ± 16.2 96.6 ± 3.8 100.0 ± 0.0 98.9 ± 1.9
EGA Gregory 87.0 ± 22.5 79.9 ± 14.8 96.7 ± 5.8 97.7 ± 4.0
EGA Wylie 80.7 ± 17.2 55.2 ± 3.2 96.7 ± 5.8 96.7 ± 0.0
Mace 91.3 ± 15.0 94.3 ± 7.3 100.0 ± 0.0 100.0 ± 0.0
Scout 100.0 ± 0.0 87.8 ± 17.2 100.0 ± 0.0 100.0 ± 0.0
SeriM82 100.0 ± 0.0 67.8 ± 34.4 100.0 ± 0.0 97.8 ± 3.9
Spitfire 77.7 ± 38.7 86.7 ± 20.5 100.0 ± 0.0 98.7 ± 2.2
Suntop 100.0 ± 0.0 97.8 ± 3.7 97.7 ± 4.0 97.8 ± 1.9
Westonia 100.0 ± 0.0 99.0 ± 1.6 99.0 ± 1.7 100.0 ± 0.0
26
Supplementary Table 22 | Germination percentage of 30 barley (H. vulgare) seeds harvested
at maturity (plants senesced) from speed breeding condition II (SB) and glasshouse (GH)
conditions in Queensland, Australia, with no supplementary light (sown on 18 June 2016). All
seeds were kept at 4 °C for 4 days prior to germination. Values indicated are expressed as mean
± SD.
H. vulgare cultivar Germination percentage (%)
SB GH
Commander 100.0 ± 0.0 30.00 ± 28.3
Compass 98.5 ± 2.1 60.0 ± 0.0
Flagship 100.0 ± 0.0 70.0 ± 28.3
Shepherd 100.0 ± 0.0 86.7± 11.5
Westminster 100.0 ± 0.0 75.0 ± 23.2
NRB090257 100.0 ± 0.0 78.3.0 ± 33.3
NRB090885 100.0 ± 0.0 100.0 ± 0.0
ND24260 100.0 ± 0.0 97.5 ± 3.5
Grout 83.5 ± 23.3 80.0 ± 34.6
Supplementary Table 23 | Germination percentage of canola seed (seed from three pods)
harvested at maturity (plants senesced) from speed breeding condition II (SB) and glasshouse
(GH) conditions in Queensland, Australia, with no supplementary light (sown on 18 June
2016). All seeds were kept at 4 °C for 4 days prior to germination. Values indicated are
expressed as mean ± SD.
B. napus cultivar Germination percentage (%)
SB GH
Taparoo 55.0 ± 21.2 80.0 ± 0.0
ATR Cobbler 90.0 ± 10.0 90.0 ± 14.1
ATR Beacon 96.7 ± 5.7 98.3 ± 2.9
CB Argyle 90.0 ± 10.0 91.7 ± 2.9
Westar 70.0 ± 42.4 100.0 ± 0.0
Skipton 100.0 ± 0.0 81.7 ± 27.5
Bravo TT 100.0 ± 0.0 100.0 ± 0.0
27
Supplementary Table 24 | Germination percentage of chickpea pods harvested at maturity
(plants senesced) from speed breeding condition II (SB) and glasshouse (GH) conditions in
Queensland, Australia, with no supplementary light (sown on 7 June 2016). All seeds were
kept at 4 °C for 4 days prior to germination. Values indicated are expressed as mean ± SD.
C. arietinum cultivar Germination percentage (%)
SB GH
PBA Boundary 96.9 ± 6.3 92.2 ± 7.5
PBA HatTrick 100.0 ± 0.0 93.6 ± 9.8
Jimbour 100.0 ± 0.0 100.0 ± 0.0
Kyabra 81.0 ± 12.6 70.0 ± 0.0
Supplementary Table 25 | Pod number per plant and total pod weight per plant (g) of mature
canola grown under speed breeding condition II (SB) and glasshouse (GH) conditions in
Queensland, Australia, with no supplementary light. Pods were dried at 35 °C for 10 days prior
to weighing. Values indicated are expressed as mean ± SD.
Pod number per plant Total pod weight per
plant (g)
B. napus cultivar SB GH SB GH
Taparoo 91.0 ± 9.6 85.0 ± 39.6 9.9 ± 0.9 8.4 ± 2.3
ATR Cobbler 94.0 ± 2.0 90.7 ± 8.5 7.6 ± 2.8 8.4 ± 1.5
ATR Beacon 97.7 ± 18.8 118.7 ± 20.8 7.0 ± 1.1 9.8 ± 1.4
CB Argyle 102.7 ± 29.7 75.7 ± 9.0 9.5 ± 2.3 6.3 ± 0.5
Westar 80.3 ± 4.0 161.3 ± 21.2 7.8 ± 0.5 12.9 ± 0.4
Skipton 97.0 ± 12.1 96.0 ± 38.7 9.6 ± 2.3 10.2 ± 3.8
Bravo TT 92.7 ± 15.3 113.3 ± 23.7 9.1 ± 1.7 9.2 ± 1.2
28
Supplementary Table 26 | Pod number per plant and total pod weight per plant (g) of mature
chickpea grown under speed breeding condition II (SB) and glasshouse (GH) conditions in
Queensland, Australia, with no supplementary light. Pods were dried at 35 °C for 10 days prior
to weighing. Values indicated are expressed as mean ± SD.
Pod number per plant Total pod weight per
plant (g)
C. arietinum cultivar SB GH SB GH
PBA Boundary 12.2 ± 6.7 14.0 ± 6.3 1.5 ± 1.0 2.0 ± 1.3
PBA HatTrick 14.6 ± 7.8 16.0 ± 5.2 1.5 ± 0.7 2.1 ± 1.0
Jimbour 14.4 ± 7.7 11.4 ± 5.8 2.2 ± 1.9 1.0 ± 0.7
Kyabra 15.6 ± 7.7 23.4 ± 6.2 2.5 ± 1.5 4.5 ± 1.8
Supplementary Table 27 | Summary of generation time and yield measurements of wheat,
barley, canola and chickpea grown in individual pots under speed breeding condition II (SB)
and glasshouse (GH) conditions in Queensland, Australia, with no supplementary light. Values
indicated are expressed as mean ± SD.
Days to
anthesis
Days to
seed
sampling1
Total
generation
time2
Mean yield
per plant
(g/plant)
Gen./year
Wheat SB 41.4 ± 1.7 14 65.4 3.5 ± 1.2 5.6
GH 63.1 ± 2.8 14 87.1 2.5 ± 1.0 4.2
Barley SB 30.4 ± 1.4 28 68.4 5.9 ± 1.4 5.3
GH 94.0 ± 14.4 28 132.0 9.4 ± 4.8 2.8
Canola SB 46.2 ± 2.2 42 98.2 8.3 ± 2.2 3.7
GH 119.1 ± 15.3 42 171.1 9.2 ± 2.4 2.1
Chickpea SB 30.1 ± 0.7 42 82.1 1.9 ± 1.3 4.5
GH 62.6 ± 3.7 42 114.6 2.4 ± 1.7 3.2 1 Number of days post-anthesis (or awn-peep for barley). 2 Total generation time calculated according to: mean days to anthesis of all cultivars + mean
days to sample seed + 5 days drying at 35 °C + 1 day imbibing seed at room temperature + 4
days chilling at 4 °C.
29
Supplementary Table 28 | Generation time and yield measurements of wheat (cv. Westonia)
grown in 100-cell trays under speed breeding condition II (SB) and glasshouse (GH) conditions
in Queensland, Australia, with no supplementary light (sown on 8 June 2016). Values indicated
for GS651 are expressed as days after sowing (DAS2).
SB GH
GS65 36 55
Percentage germination (early)3 80 100
Percentage germination (mature)4 100 100
Seed number per spike (Mean ± SD) 19.0 ± 1.3 18.0 ± 2.2
Seed weight per tray (g) 41.2 41.0
Complete generations per year5 6 4 1 GS65 = anthesis. 2 DAS refers to the number of days following direct sowing of ungerminated seeds into soil. 3 Early harvest was 14 days post-anthesis. 4 Mature harvest was 28 days post-anthesis. 5 Each generation time includes days to anthesis + 14 days maturing + 5 days drying at 35 °C
and 4 days chilling at 4 °C. Compete generations per year = 365/generation time.
Supplementary Table 29 | Generation time and yield measurements of wheat (cv.
Commander) grown in 100-cell trays under speed breeding condition II (SB) and glasshouse
(GH) conditions in Queensland, Australia, with no supplementary light (sown on 8 June 2016).
Values indicated for GS491 are expressed as days after sowing (DAS2).
SB GH
GS49 22 82
Percentage germination (early)3 0 20
Percentage germination (mature)4 100 90
Seed number per spike (Mean ± SD) 18.6 ± 1.8 12.0 ± 5.4
Seed weight per tray (g) 27.3 20.1
Complete generations per year5 6 3 1 GS49 = awn peep. 2 DAS refers to the number of days following direct sowing of ungerminated seeds into soil. 3 Early harvest was 14 days post-anthesis. 4 Mature harvest was 28 days post-anthesis. 5 Each generation time includes days to anthesis + 14 days maturing + 5 days drying at 35 °C
and 4 days chilling at 4 °C. Complete generations per year = 365/generation time.
30
Supplementary Table 30 | Fusarium head blight infection of wheat (T. aestivum cvs. Timstein
and Sumai 3) under speed breeding condition I.
cv. Timstein (susceptible) cv. Sumai 3 (resistant)
Replicate Inoculated
spikes
Susceptible
spikes
Inoculated
spikes
Resistant
Spikes
Plant 1 2 2 3 3
Plant 2 2 2 2 2
Plant 3 2 2 3 3
Supplementary Table 31 | Flowering time and leaf number on main tiller for observation of
the flowering time difference phenotype in wheat lines W352, Paragon and W352 x Paragon
F6 RIL grown under speed breeding condition I (SB) and glasshouse conditions in UK Summer
with no supplementary light. Sown 29 March 2016. Values indicated are expressed as mean ±
SD.
Leaf counts to flowering Days to flowering
Genotype SB GH SB GH
W352 7.3 0.5 8.3 0.5 34.3 2.4 78.0 0.0
Paragon 7.3 0.5 7.7 0.5 36.0 1.6 76.0 1.4
W352 x Paragon F6 9.0 0.0 9.3 0.5 50.3 2.4 86.0 1.4
31
Supplementary Table 32 | Chromosome associations during meiotic metaphase I in wheat in
presence of Ph1, and wheat-rye hybrids in presence of Ph1, and in absence of Ph1 per pollen
mother cells in speed breeding condition I and control conditions (GH) (UK summer
glasshouse with no supplementary light). Eighty pollen mother cells were analysed in three
plants. Values indicated are expressed as mean ± SD.
T. aestivum cv. Chinese
Spring
Wheat-Rye hybrid
(carrying Ph1)
Wheat-Rye hybrid
(lacking Ph1)
SB GH SB GH SB GH
Univalent 0.1a1 ± 0.5
(0-2)2
0.2a ± 0.6
(0-2)
27.0a ± 1.5
(22-28)
26.8a ± 1.6
(22-28)
13.6a ± 1.6
(10-18)
13.8a ± 2.3
(8-18)
Bivalent
20.9a ± 0.2
(20-21)
20.9a ± 0.3
(20-21)
0.5a ± 0.8
(0-3)
0.6a ± 0.8
(0-3)
7.1a ± 0.9
(4-9)
7.0a ± 1.2
(4-10)
Multivalent
- - - -
0.1a ± 0.3
(0-1)
0.1a ± 0.4
(0-2)
Number of
chiasmata
40.1a ± 1.6
(34-42)
40.4a ± 1.4
(37-42)
0.5a ± 0.8
(0-3)
0.6a ± 0.8
(0-3)
8.3a ± 1.6
(7-13)
8.7a ± 1.7
(6-12)
1 Different letters indicate significant differences according to LSD test (p < 0.05). 2 Values in parenthesis indicate the range of variation between cells.
Supplementary Table 33 | Transformation data of barley (H. vulgare) cv. Golden Promise
under speed breeding condition I (SB) and control conditions. Constructs used were from the
pBRACT series as described by Smedley et al. [S1]. All constructs contained the hygromycin
resistance gene under the control of a 35S promoter.
Experiment
Number
Time of embryo
harvesting (days
post sowing)
Number of
immature
embryos
Number of
transformed
plants
Transformation
efficiency
SB Control1 SB Control SB Control SB Control
1 60 92 25 75 7 19 28% 25%
2 66 92 50 50 13 18 26% 36% 1 Control conditions were 16-hour photoperiod with 15 C day and 12 C night temperatures,
80% relative humidity and light levels of 500 µmol m-2 s-1 at the mature plant canopy level.
Light was provided by metal halide lamps (HQI) supplemented with tungsten bulbs.
32
Supplementary Table 34 | Development of transformed barley (H. vulgare cv. Golden
Promise) explants under speed breeding condition I and control conditions.
Speed Breeding Control1,2
Rep. 1 Rep. 3 Rep. 23 Rep. 1 Rep. 2 Rep. 3
Days to heading 60 52 - 64 64 61
Days to harvest viable
seed 87 87 - 131 131 131
No. of tillers 25 72 6 32 20 22
Total number of seeds
produced 259 58 - 280 215 359
Percentage germination
(sample of 20 seeds) 95% 55% - 50% 90% 70%
114-week old explants generated from transformed barley tissue culture were transferred to the
respective conditions (3 explants per condition) to monitor growth and development. 2 Control conditions were 16-hour photoperiod with 15 C day and 12 C night temperatures,
80% relative humidity and light levels of 500 µmol m-2 s-1 at the mature plant canopy level.
Light was provided by metal halide lamps (HQI) supplemented with tungsten bulbs. 3 Flowering was severely delayed in this plant most likely due to variation imposed by tissue
culture conditions.
Supplementary Table 35 | Shatter resistance (RELSQ - rupture energy adjusted for pod
length), as measured with the pendulum test, of mature, dry canola pods grown under speed
breeding condition II and the field (Wagga Wagga, NSW, Australia). Higher value indicates
higher shatter resistance. Mean of ten replicate pods per plant and three replicates per cultivar
for speed breeding (values indicated are expressed as mean ± SD). Mean of ten mature pods
harvested from cultivars grown in field plots.
B. napus cultivar Speed breeding Field
Skipton 1.1 ± 0.2 1.08
CB Argyle 1.0 ± 0.1 1.31
ATR Cobbler 1.7 ± 0.9 1.35
ATR Beacon 1.4 ± 0.4 1.08
Bravo TT 2.0 ± 1.5 1.44
33
Supplementary Table 36 | JIC Cereal Compost Mix supplied by Petersfield Growing
(Leicester, UK).
Component Measure
Medium Grade Peat 40%
Sterilised Soil 40%
Horticultural Grit 20%
Fertilizer
PG Mix™ 14-16-18 + Trace Elements (TE) Base Fertiliser 1.3 kg/m³
Osmocote® Exact Mini 16-8-11+ 2MgO + TE 0.02% Boron 1 kg/m³
H2Gro® (Wetting Agent) from ICL Specialty Fertilizers
(Ipswich, UK)
Maglime 3 kg/m³
Insecticide
Exemptor® from ICL Specialty Fertilizers
(Ipswich, UK)
300 g/m³
Supplementary Table 37 | CGS20 Compost mix components and fertilisers designed by Mr
K. Hayes, Central Glasshouse Services, University of Queensland, Australia. The pH is
balanced with either FeSO (pH is high) or Dolomite (pH is low).
Component Measure
Composted pine bark (0-5mm) 70%
Coco peat 30%
Fertilizer
Yates Flowtrace® (Yates, Padstow, NSW, Australia) 1 kg/m3
Iron sulphate heptahydrate 1 kg/m3
Superphosphate 0.4 kg/m3
Copper sulphate 0.03 kg/m3
Gypsum 1 kg/m3
Supplementary Media File 1 | Timelapse video recording comparing plant growth under
speed breeding condition I and glasshouse conditions in UK Summer without any
supplementary light. Video depicts three replicates of Triticum aestivum cv. Paragon sown and
recorded under each treatment, with two replicates removed after stem extension stage was
reached in each condition. Recording were made using the CropQuant workstation developed
by Ji Zhou and colleagues at the John Innes Centre [S4]. Germinated seedlings of Paragon were
sown on 17 March, 2017. Growth curves are illustrated in Supplementary Fig. J.
https://drive.google.com/open?id=0B_nFDDKXi2jwdzhQWDQ2QW1oU2s
34
Supplementary References:
[S1] Smedley, M. A. & Harwood, W. A. Gateway®-Compatible Plant Transformation Vectors.
Agrobacterium Protocols: Volume 1, 3-16 (2015).
[S2] Tottman, D. The decimal code for the growth stages of cereals, with illustrations. Annals
of applied biology 110, 441-454 (1987).
[S3] Sylvester-Bradley, R. & Makepeace, R. code for stages of development in oilseed rape
(Brassica napus L.). Aspects of Applied Biology (1984).
[S4] Zhou, J., Reynolds, D. CropQuant: next-generation crop monitoring for precision
agriculture, <http://www.earlham.ac.uk/cropquant-next-generation-crop-monitoring-
precision-agriculture#Detail-1> (2016).