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WP 9- FROST TOLERANCE IN OAT. P8 Agricultural Research Council CRA-GPG Fiorenzuola, Italy. Avena Genetic Resources for Quality in Human Consumption (AVEQ) Final project meeting Tuesday 19th – Thursday 21st October 2010 6th Meeting of the ECPGR Avena Working Group - PowerPoint PPT Presentation
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Avena Genetic Resources for Quality in Human Consumption (AVEQ) Final project meeting Tuesday 19th – Thursday 21st October 2010
6th Meeting of the ECPGR Avena Working GroupThursday 21st – Friday 22nd October 2010
Bucharest, Romania
P8 Agricultural Research Council CRA-GPG Fiorenzuola, Italy
WP 9- FROST TOLERANCE IN OAT
Among winter cereals, oat is the most frost sensitive and its insufficient level of winterhardiness is the main factor limiting winter sowing in many major oat growing regions in Europe
Fiorenzuola 1996-’97
-15°C without snow
Frost resistance not only improves winter survival but is also a fundamental component in high yielding winter
cultivars
IS THERE GENETIC VARIABILITY FOR THIS TRAIT?
Sixty-two genotypes with different origin (Europe, North America) were grown in Northern Italy (Fiorenzuola)
in 1996-’97 Frost damage was estimated by a visual score (0: no damage 9:
all plants killed)
A BROAD GENETIC VARIABILITY FOR
WINTERHARDINESSEXISTS IN OAT
Range between 3 and 9 (mean
5.8)
LARGE GENETIC VARIABILITY FOR BOTH TRAITS IN CEREALS (Ex. barley, diploid model for the Triticeae )
In winter-sown cereals the acquisition of frost tolerance is associated with the occurrence of a cold HARDENING adaptive process at low non-freezing temperature
FROST TOLERANCE, the main component of winter hardiness is a crucial factor for cereal cultivation, not only improves winter survival but is also a fundamental component in high yielding winter cultivars
WINTER HARDINESS is assessed in the field as ability to survive throughout the winter
AVEQ PROJECTWorkpackage 9:
PARTECIPANTSP8 Agricultural Research Council CRA-GPG Fiorenzuola, Italy (Valeria Terzi)P9 Sucaeva Genebank, Sucaeva, Romania (Danela Muraru)P10 IPGR “K.Malkow”, Sadovo, Bulgaria (Nadia Antonova)
OBJECTIVESField screening for cold tolerance for up to 500 accessionsGrowth chamber tests for cold tolerance for 100 accessions
hardening treatment (3/1°C) for 4 weeks
Plants at first leaf stage cold acclimated during 4 weeks at 3/1° C (day/night), 200E m-2 s-1. Hardened plants are subjected to a freezing treatment at temperature from -10 to –13°C
FROST TOLERANCE EVALUATION AT FIRST LEAF STAGE
Chlorophyll fluorescence responds to changes in PSII photochemistry and therefore represents a convenient and rapid tool to evaluate the capacity of the photosynthetic machinery at low temperature.
Fv Fm-F0
--- = ---- Fm Fm
The ratio of variable to maximal fluorescence (Fv/Fm) in dark-adapted state measures the maximum quantum yield for PSII photochemistry and represents a diagnostic probe for measuring low temperature stress-induced injury of photosynthesis
PAM-2000, Walz, Effeltrich, Germany
= 0.8in dark-adapted healthy plants
Rizza et al. (Plant Breed.120: 389-396, 2001).
one further reason of interest....
Compare the reliability of direct evaluations of FROST TOLERANCE and WINTER HARDINESSIntermediate-high tolerance
Optimize a simple and reliable protocol for PHENOTYPING LARGE POPULATIONS
Breeding
Biodiversity
Association mapping
Fr-H1
Fr-H2
Francia et al.,Theor Appl Genet (2004) 108:670–680
Two QTLs for frost tolerance were
mapped on barley chromosome 5H.
QTL for three distinct measures
of low-temperature resistance are
coincident
‘NURE’ x ’TREMOIS’ LINKAGE MAPThird leaf
First leaf
Fr-H1 is closely linked to Vrn-H1 vernalization response locus
THE SELECTED GENOTYPESgrowth chamber experiments 2008
104 oat genotypes (included 9 standards) out of 317 of the set 1
5
4
1
812
4
8
2
6
2 18
4
14
6
9
16 Holder countries
Holder
n.of genotyp
es moderncultivat
edstandar
d spring winterinterme
diate
origin from the
Holder country
origin from other
countries
germinability
n.of selecte
d genotyp
es*
AUT 11 11 - - 11 - - 11 - high 5
BGR 7 1 5 1 6 - 1(w) 4+? variable 4
BOR 1 1 - - 1(FIN) - high 1
CZE 25 13 11 1 13 1 19 6 variable 8
DEU 65 18 46 1 56 31 1? 33 variable 12
EST 12 2 9 1 9 12 - variable 4
FRA 37 15 21 1 25 1 1+3(w) 32 4 variable 8
GBR 3 3 - - 2 1 - 3 - high 2
ITA 8 6 - 2 8 - high 6
LTU 6 2 4 - 4 3 3 variable 2
LVA 2 - 2 - 1 2 - high 1
POL 38 16 22 - 23 24 14 variable 8
ROM 9 - 8 1 8 9 - variable 4
RUS 47 - 47 - 46 - 1(s) 6 41 variable 14
SVK 16 2 14 - 14 7 9 variable 6
SWE 29 15 13 1 13 20 9 high 9
USA 2 - 2 - 1 - 1 - 2 variable 1
THE SELECTED GENOTYPESgrowth chamber experiments 2008
FREEZING STRESS AT TWO MINIMUM TEMPERATURES
-10°C discrimination of the most frost susceptible genotypes
-12°C good discrimination of genotype with intermediate or superior FT
THE SELECTED GENOTYPES growth chamber experiments 2008
On the basis of the two experiments (ANOVA, LSD0.05):
6 genotypes were identified as the most susceptible
11 “ “ “ tolerant (4 with Fv/Fm close to 0.78)
The best genotypesFv/Fm 24h recovery
Mean 104 genotypes** 0.45
Range (min-max) 0.11 ÷ 0.77
Accession number
NAME FAO code - 12°C
A7BM0005 83/200-CR BGR001 0.77
200113379 Local 7 RUS001 0.75
200110905 Kinelskjj RUS001 0.73
605 Donata ITA037 0.73
1005 Vendelin SVK001 0.70
AVE 4659 Lueneburgher Kley Neue Zucht DEU146 0.68
19903 Gagybatory K Tajfajta SVK001 0.68
200107910 Local 3 RUS001 0.67
708 Novella Antonia ITA037 0.66
200111655 Local 5 RUS001 0.66
Millennium Millennium GBR016 0.65
breeding line
The same group (104) was tested in the field (P9)
-30.0
-20.0
-10.0
0.0
10.0
20.0
30.0
0 50 100 150 200
°C
maximum
minimum
9
87
6
Suceava meteorological Center, Romania 2009-2010
visual score
A7BM0005 83/200-CR BGR001 6
200113379 Local 7 RUS001 6
Millennium Millennium GBR016 7
10 Ava ITA037 7
605 Donata ITA037 7
set 1: correlation between Fv/Fm (24h recovery) with the field data of Bulgaria 2008-09 and Romania 2009-2010
ROMANIA field 2009-10 visual
score (0-9, Rizza et al.)
ROMANIA field 2009-10 number
survived plant (total row 1,2)
BULGARIA field 2008-09 visual
score (0-9, Rizza et al.) Mean
replications I,II
BULGARIA field 2008-09 visual
score (0-9 IBPGR)
Fv/Fm means two laboratory experiments
0.447*** 0.432*** -0.224* -0.121ns
ROMANIA field 2009-10 visual score (0-9, Rizza et al.) -0.951*** 0.267** 0.287**
ROMANIA field 2009-10 number survived plant (total
row 1,2) -0.271*** -0.257** BULGARIA field 2008-09
visual score (0-9, Rizza et al.) Mean replications I,II 0.340***
n=104 for laboratory experimentsn=318 field experiments
- -
THE SELECTED GENOTYPESgrowth chamber experiments 2010
106 oat genotypes (included 11 standards) out of 309 of the set 2
Holdern.of
genotypes*
origin from the Holder
countryn.of selected genotypes*
AUT 6 3 3BGR 17 5 7CZE 40 5 10DEU 79 12 20ESP 10 3 3EST 1 1 1FRA 28 8? 8GBR 37 6 9HUN 36 8 9IRL 5 4 4ITA 1 1 1NDL 1 1 1POL 67 6 11PRT 1 1 1ROM 1 1 1RUS 2 2 2SWE 4 4 4
335 95
*standard excluded 11 STANDARDS
3
7
1020
1
8
9
9
4
1
1 11
1
1
4
17 Holder countries3
2
FREEZING STRESS AT TWO MINIMUM TEMPERATURES:
-11°C ,-12°Cgood discrimination between intermediate and
tolerant genotypes
THE SELECTED GENOTYPESgrowth chamber experiments 2010
Accession number status FAO CODE
after stress
24h recovery
Fv/Fm Fv/FmA7BM0002 BREEDING LINE cultivated BGR001 0.79 0.81A7BM0006 BREEDING LINE cultivated BGR002 0.78 0.8019632 Cimarron cultivated FRA040 0.78 0.7837 Luilbreg (YUG) cultivated GBR011 0.78 0.7836 Belyski (YUG) cultivated GBR011 0.77 0.77CPVO19981528 EVORA standard FRA261 0.77 0.75A7BM0003 BREEDING LINE cultivated BGR001 0.79 0.75BGR 24983 K06-08 local BGR002 0.78 0.758 ARGENTINA standard ITA037 0.76 0.751126 AJ 145/114 cultivated GBR011 0.77 0.74RCAT012421 Kulsovati B cultivated HUN003 0.78 0.7419620 La Gaillarde cultivated FRA040 0.76 0.74AVE 1994 grisea DEU146 0.76 0.72
Means 106 genotypes 0.75 0.52Range (min-max) 0.66 - 0.79 0.16 - 0.81
The 13 best genotypes -11°C
THE SELECTED GENOTYPESgrowth chamber experiments
2010
n=106 Fv/Fm 24h -11°C Fv/Fm 24h -12°Cmean 2
experiments
Fv/Fm 24h -11°C
Fv/Fm 24h -12°C 0.523***
Bulgaria field 2009-‘10 scale 0-9 -0.476*** -0.369*** -0.493***
Bulgaria field 2009-‘10 scale 0-7 (IBPGR) -0.400*** -0.248*(*) -0.385***
THE SELECTED GENOTYPESgrowth chamber experiments
2010
Correlation between the laboratory and field xperimentsSodovo, Bulgaria 2009-2010
Fv/Fm after stress
-11°C
Fv/Fm after
stress -12°C
mean 2 experime
nts
Fv/Fm after stress -
11°C
Fv/Fm after stress -
12°C -0.601***
Bulgaria field 2009-‘10 scale 0-9 -0.396*** -0.305** -0.396***
Bulgaria field 2009-‘10 scale 0-7 (IBPGR) -0.352*** -0.205* -0.298**
Romania prova campo 2010-11
To which extent the evaluation of the plant response in EARLY GROWTH STAGE or EARLY ACCLIMATION state plays a critical role in
determining the potential (maximum) frost hardiness?
Can we further simplify our evaluation of frost tolerance?
in progress............some more questions ...
CAN WE CONTRIBUTE IN UNDERSTANDING THE BASIS OF THE FROST TOLERANCE CAPACITY?
Is there genetic variability for the plant capacity for “early acclimation?”
The ability to induce early hardening is of advantage under field conditions, where temperature falls progressively, because it
enables resistant plants to prepare for cold before the susceptible ones.
The TRESHOLD TEMPERATURE for
COR14 accumulation is higher in the resistant
cv
Crosatti et al. Planta 196:458-463, 1995
Winter, Resistant
Spring, Susceptible
SUBOPTIMAL HARDENING TEMPERATURE
THE SELECTED GENOTYPESgrowth chamber experiments
2010
104 oat genotypes (included 9 standards) out of 317 of the set 1
Plants at first leaf stage wereexposed 3 weeks at suboptimal hardening temperature12/7°C.
Freezing stress at -6°C and -7°C
Accession namber variety name FAO code after stress 24h recovery
Millennium Millennium GBR016 0.783 0.749A7BM0005 83/200-CR BGR001 0.775 0.747200113379 Local 7 RUS001 0.782 0.746605 Donata ITA037 0.755 0.742St. 4 Evora EVORA FRA040 0.782 0.72710 Ava ITA037 0.759 0.716200105652 Local 1 RUS001 0.745 0.673200110905 Kinelskjj RUS001 0.706 0.61419303 Neu Grosse FRA040 0.745 0.594Breton Breton POL040 0.725 0.590200111655 Local 5 RUS001 0.723 0.5908-ARGENTINA ARGENTINA ITA037 0.736 0.589AINTREE Aintree FRA261 0.743 0.583200107022 Local 2 RUS001 0.74 0.558200107910 Local 3 RUS001 0.735 0.531
Fv/Fm
Freezing -7°C after acclimation at suboptimal hardening temperature (12/7°C, day/night). SET 1
Mean 104 genotypes** 0.70 0.29Range (min ÷ max) 0.66 ÷ 0.78 0.02 ÷ 0.75
Comparison between laboratory tests (“early” or “standard” hardening) and with the field data for set 1
n=104-106 Fv/Fm 24h -6°C Fv/Fm 24h -7°C
mean 2 experiments -
6 -7°C
Fv/Fm 24h -7°C 0.656***
Fv/Fm 24h -12° e-10°C
means 0.477*** 0.453*** 0.512***
Fv/Fm 24h -12° 0.446*** 0.422*** 0.477***
ROMANIA field 2009-10 visual score (0-9) -0.435*** -0.623*** -0.588***
BULGARIA field 2009-10 scale 0-9 -0.234* -0.291** -0.291**
BULGARIA field 2009-10 scale 0-7 (IBPGR) -0.305** -0.361*** -0.369***
- one more stress temperature to test frost tolerance under “standard”hardening conditions (common publication)
WP9. The experiments planned in the AVEQ project are concluded, with the only exception of the second field in Romania for the evalaution of winter hardiness in the second set of 106 accessions (set 2)
This is due to a modified organization of the laboratory tests (200 accessions instead of 100) and a consequent oganization of the field experiments (environment subjected to severe frost)
about the AVEQ project......
beyond the objectives.....- effect of hardening at suboptimal hardening conditions (12/7°) for the set 2
- interest for analysing wild species, landraces...(adaptability, breeding)
.........and beyond? POSSIBLE QUESTIONS
frost tolerance and winter hardiness: how to dissect the components of WINTER HARDINESS (development traits. vernalization, photoperiod, earliness per se) in a large number of genotypes? GEOGRAPHYCAL ORIGINBREEDING EFFECTS
use of SELECTED MOLECULAR MARKERS for traits affecting winter hardiness (vernalization and photoperiod).The best approach?
What plants need for adaptability to the PRESENT and FUTURE environmental conditions?
AB
TR
EM
OIS
A B
lad
de
r
NU
RE
NU
RE
Phenotypelaboratory tests
Genotype. Selection ofadequate molecular markers
Field validation
genes for vernalization/photoperiod response
In barley
Growth at 12/7°C, associated to higher light intensity (700E) implied a morphological adaptation in frost
resistant and frost sensitive cultivars
Frost resistant
Frost sensitive
P9 Sucaeva Genebank, Sucaeva, Romania (Danela Muraru)P10 IPGR “K.Malkow”, Sadovo, Bulgaria (Nadia Antonova)
P8 Agricultural Research Council CRA-GPG Fiorenzuola, Italy
Valeria TerziMichele StancaLuigi CattivelliDonata PaganiFlavio Astesano
Thank You!
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