11
LupiBreed
Improving narrow-leafed lupin – Novel genetic resources for
higher yield and yield stability
Helge Flüß, Kristin Fischer, Steffen Roux, Regine Dieterich, Hans-Ulrich Jürgens
and Brigitte Ruge-Wehling
22ILC 2019 Cochabamba, Bolivia
LupiBreed – Consortium (2015-2018)
Brigitte Ruge-Wehling
Kristin Fischer
Steffen Roux
Peter Wehling
Gisela Jansen
Hans-Ulrich Jürgens
Regine Dieterich
Anna Beyer
Ulrike Lohwasser
Matthias Kotter
Herwart Böhm
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Part I
Improving yield…
– Identification of high-yield prebreeding lines based on a mutant population by
performance testing in different locations over 3 years (grain yield, protein
content, alkaloid content)
Improving yield stability…
– Early shattering resistance: field testing and molecular markers
– Anthracnose resistance: greenhouse and field testing, genetic mapping of
resistance loci LanrBo and Lanr1
Part II (Anne Zaar, JKI Institute for Resistance Research and Stress Tolerance)
Analysis of the range of variation for protein and alkaloid content in plant
genetic resources and novel mutant collection of narrow-leafed lupin
Aims of the consortium
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Improving yield
Increase genetic variation – EMS population
Field testing yield performance
5
M0
EMS-treatment
M1 M2 M3 M4
Selection new
growth types
Propagation of
new growth types
selfing
Selection homogenous
growth types
selfing selfing
seed plant plant plant plant
Development mutant lines
Terminal
wild type
New growth types
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‚Boruta‘
‚Probor‘
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Material
cv. ‘Boruta‘, ‘Boregine‘, ‘Probor‘
22 early M-lines A (Donor: ‘Boruta’)
(22 late M-lines B (Donor: ‘Boruta’, ‘Probor’))
3 locationsGroß Lüsewitz JKI I (conv) und II (eco), Bocksee
SZS
3 years2015, 2016, 2017
Sowed5m2 – plots, 100 grains/m2
Traits
flowering, ripening, plant height, grain yield
(dt/ha), TKG, protein content, protein yield
(dt/ha), alkaloid content
Yield performance field testing
4.6.2016
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Means yield and protein, early M-lines, 2015 – 2017
LocationGrain yield
(dt/ha)
Protein
content (%)
Protein yield
(dt/ha)
GL konv Min. 33.1 28.8 9.2
BP40-47 Max. 51.1 33.0 15.2
Mean 42.5 30.2 12.5
Boruta 43.7 30.6 13.1
Probor 52.4 31.9 16.3
Boregine 57.1 28.6 15.9
GL öko Min. 20.5 24.8 5.5
BP40-47 Max. 36.4 30.5 9.8
Mean 29.6 26.8 7.8
Boruta 29.6 25.7 7.4
Probor 35.1 29.8 10.2
Boregine 41.5 27.8 11.2
Bocksee Min. 10.3 24.6 2.4
BP18-20 Max. 18.7 28.5 7.6
Mean 15.1 26.4 4.1
Boruta 15.4 26.2 3.9
Probor 17.6 28.5 5.2
Boregine 18.2 26.5 6.0
gesamt Min. 21.4 26.1 5.8
Max. 35.4 29.3 10.0
Mean 29.1 27.6 8.0
Boruta 29.6 27.5 8.2
Probor 35.0 30.0 10.5
Boregine 38.9 27.6 10.7
Good soil compostition,
some M-lines better than
wild type
No herbicides (competition)
water accessability
plant diseases, -pests
Low soil quality, water
accessability
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M-line Grain yield (dt/ha)
A4Bor 35.4 *
A15Bor 33.8 *
A3Bor 33.3 *
A18Bor 32.9 *
A5Bor 32.6 *
A19Bor 32.2
A17Bor 31.3
A12Bor 30.9
A7Bor 30.5
A9Bor 29.2
A16Bor 29.1
A14Bor 29.1
A10Bor 28.4
Boruta 29.5
Probor 35.0
Boregine 38.9
M-lineProtein content
(%)
A9Bor 29.3 *
A4Bor 28.5 *
A11Bor 28.5 *
A17Bor 28.3 *
A5Bor 28.3 *
A13Bor 28.2
A8Bor 28.1
A3Bor 28.1
A18Bor 28.0
A15Bor 27.9
A22Bor 27.6
A12Bor 27.5
A19Bor 27.5
Boruta 27.5
Probor 30.0
Boregine 27.6
M-lineAlkaloid content
(%)
A8Bor 0.010 *
A20Bor 0.011 *
A2Bor 0.014 *
A13Bor 0.015 *
A14Bor 0.015 *
A1Bor 0.016 *
A6Bor 0.018 *
A15Bor 0.018 *
A16Bor 0.018 *
A19Bor 0.019 *
A9Bor 0.020 *
A12Bor 0.020 *
A22Bor 0.021 *
Boruta 0.026
Probor 0.027
Boregine 0.039
Yield performance early M-lines, A
9 environments (3 locations, 2015-2017)
Variant analysis LSD-Test, =0.05A Bor Mutant line based on cv ‚Boruta‘ Comparison to wild type ‚Boruta‘
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Early shattering resistance
Increasing yield stability
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Material
VB Antaniai -> strongly shattering
Tanjil, Uniwhite, Unicrop, Lila Baer -> high
resistance
22 early M-lines A + 10 breeding lines
(22 late M-lines B + 10 breeding lines)
2 locationsGroß Lüsewitz JKI, Bocksee SZS
3 years2015, 2016, 2017
Sowing2-row micro plots, 5 x 5 grids
Scoring1 – 9 (shattering resisant – highly shattering)
Shattering resistance field testing
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Shattering resistance, early M-lines, 2015 – 2017, A
highly shattering (9)
No shattering (1)
VB Antaniani
Tanjil
Uniwhite
Unicrop
Breeding lines
Mutant lines
Specimens
Sco
res
Boruta (1 Jahr)
Lila Baer (1 Jahr)
Kruskal –Wallis-Test
(c2 = 155.7, df = 35, p = < 0.05)
PostHoc: Dunn-Bonferroni-Test 22.03.2019 ILC 2019 Cochabamba, Bolivia 11
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Molecular characterization
Example: tardus
Marker: TaLi (tardus, Li et al., 2010)
+- + + + -- - -
Breeding lines
Mutant-lines
+ +-/ - -
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Anthracnose resistance
Increasing yield stability
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Resistance resources
Resource Resistance-
level
Inheritance Loucs LG Selection-
marker
cv. ‚Tanjil‘
(Australia)
High
(some spots on
pods)
monogenic
dominant
Lanr 1* NLL-11
(distal)
Lanr1_1
Lanr1_2
Zuchtlinie
Bo7212
(SZS)
High
(almost no
symptoms)
monogenic
dominant
LanrBo** NLL-11 BoSeq196
LanM_19
Greenhouse testing
Field testing*Yang et al. 2004
**Fischer et al. 2015
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Allelic association test
hor hor
x
her
Lanr1 LanrBo
Lanr1+ LanrBo
F1
S
her her sushor
F2-PopulationPhenotype – Resistance
0
50
100
150
200
250
300
resistant susceptible
Nu
mb
er
of
pla
nts
exp
obs
21
0
20
40
60
80
100
120
140
160
180
LanrBo +Lanr1
Lanr1 LanrBo -
Nu
mb
er
of
pla
nts
obs
exp
Genotype – Marker
15
c2 15:1= 0.66
1
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1
‚Tanjil‘ Bo7212
hor = homozygous resistant for Lanr1 and LanrBo
her = heterozygous resistant für Lanr1 and/or LanrBo
sus. = susceptible
N= 283
N= 283
262
151 49 62 21
c2 9:3:3:1 = 2.9
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Mapping of resistance locus
www.lupinexpress.com
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BoSeq37DAFWA3348ArSeq423LaIND_096BoSeq67BoSeq62ArSeq105BoSeq41 BoSeq61BoSeq720LaIND_085
LJM7_1Lanr1_1Lanr1_2DAFWA7361AntjM1LanM_02Lup015155LanM_03
BoSeq196
LanrBoLanM_16LanM_19BoSeq1108Lup011620BoSeq83BoSeq5Lup007149
LaIND_138
DAFWA2850
2.81.28.22.10.90.21.00.10.3
14.5
0.91.21.32.25.10.62.9
17.6
0.40.91.65.23.31.44.70.9
9.2
4.5
1
Lanr1
LanrBo
~ 30 cM
6.5 Mbp
9.5 Mbp
4.1 Mbp
15.5 Mbp
Genetic map (cM) Physical map (Mbp)
NLL-11
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Sum up yield performance and stability
Selection of mutant-lines with multiple improved traits pre-
breeding
Marker assisted breedingc approaches possible for shattering and anthracnose
resistance
Mutant lines:
Significantly higher grain yields compared to donor ‚Boruta‘ (9 environments)
Significantly higher protein content compared to high yield cultivar ‚Boregine‘ positive
effect on protein yield
22.03.2019 ILC 2019 Cochabamba, Bolivia
Significantly higher grain yield compared to high yield cultivar ‚Boregine‘ in poor soil
location
High heritability breeding progress
Significantly lower alkaloid contents in early types
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Madlen Christoffer
Juliane Böttcher
Kerstin Kadzban
Birgit Schütt
ILC 2019 Cochabamba, Bolivia
Thanks to...
Nicolas Krezdorn, GenXPro
Björn Rotter, GenXPro
Matt Nelson, UWA
Lars Kamphuis, Karam Singh, CSIRO
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19ILC 2019 Cochabamba, Bolivia
Vielen Dank für Ihr Interesse!
Thank you for your attention!
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2020
Part I
Improving yield…
– Identification of high-yield prebreeding lines based on a mutant population by
performance testing in different locations over 3 years (grain yield, protein
content, alkaloid content)
Improving yield stability…
– Early shattering resistance: field testing and molecular markers
– Anthracnose resistance: greenhouse and field testing, genetic mapping of
resistance loci LanrBo and Lanr1
Part II (Anne Zaar, JKI Institute for Resistance Research and Stress Tolerance)
Analysis of the range of variation for protein and alkaloid content in plant
genetic resources novel mutant collection of narrow-leafed lupin
Aims of the consortium
22.03.2019 ILC 2019 Cochabamba, Bolivia 20