IBGS 13,
Riga 03-07 July 2022
Session 1
Martin
Mascher
Inversion
in 2H close to CEN, found only in domesticated barleys, probably in Europe.
141 Mb
inversion in 7H in Planet. Goes back to
Diamant, mutagenesis, originated from Valticky. Also chrom block in 7H than
cannot be easily broken
Talks about
genepool concept of Harlan.
Expanding
the barley pangenome, to circa 80 cultivars
Latest
thing. Creation of genus-wide pan genomics, International barley pan-genome
consortium created,
HiFi technique
(2020) allows access to genomes easily, genus-wide pangenomics accessible
https://pangenome.github.io
graphical visualization of pangenomes
Single copy
pangenome, a hack for GWAS (ONLY all genes in single copy considered), single
copy kmer extracted from SV (PAVSs). Naked gene on 7H found with this technique
Monat et al
2019, frozen diversity
They have
resequenced about 1000 barleys, will be published over next 2 years. Also with
a genome browser (name?)
Einar B
Haraldsson
Hordeum
erectifolium (M von Korff group), perennial species
OBJ:
Decipher control of annual vs perennial life history in cereals
First,
generate genome of erectifolium, South-american species, Argentina. Unique leaf
morphology, leaf rolls up like a cinnamon roll under drought. Potential for
WUE. Field experiment in Dusseldorf, C13 discrimination, C:N ratio, large diffs
to barley.
Explains
sequence construction, 4.5 Gb (vs 5.4 Morex), more sequence statistics…
Explains
inversions, rearrangements between species, complete inversion in chrom 3
Transcriptomics
at 2 time points, 22 samples, several tissues (6 vegetative, 4 reproductive)
PacBio
isoseq, isoseq v3, minimao2? and cupcake….
CodAn, Helixer, etc
ISoSeq
Helixer plus annotations with MapMan/Mercator
38322 loci:
28446 protein coding, 140k potential isoforms, 10k non-coding genes
Produced a
hybrid with H intercedens (annual?)
Genome
assembly of 24 additional species ongoing (PanHordeum Project)
Postdoc
position open
Ale
Tondelli
Seed color
in barley.
Black seed,
phytomelanin accumulation, from early dough stage, gets darker as seed fills
and matures. Single dom gene in 1HL, BLP1?
Whealbi
GWAS, 450 accessions, 440k SNPs, gene found at end of 1H. HvPAP27 gene found.
From Chloroplast.
20
pangenome, gene duplicated in black seed accessions? HOR10350, 2 full copies,
same scaffold, HOR13821, 2 genes in diff scaffold, one of them partial.
Heterozygous
mappings found, same as disease resistance from Carlos, when mapping to Morex
Confirmed
by seq depth analysis
Description
at protein level
Promoter
has several landing sites for ABA, and other regulators
HvPAP27
expression analysis, CAPS assays to discriminate paralogs in semiquantitative
PCR.
BMC Plant
Biology paper.
Black seed
protection to high light or UV? Present in highlands.
Apparently,
no nutritional value
Martin
Kovacik (Olomouc,
Ales Pecinka)
Atlas on
gene expression. Focus on grain development, 4 tissues, 5 time points after
pollination
Syncytial
endosperm has a unique transcriptome.
Endosperm
transcriptome is extreme. Loss of transcription going on, many specific genes.
Nice bidimentsional
representation of DEG, up and down, along the dates of sampling (for Chesco?). DE
genes specific of each date found.
Finding
clusters, expression network analysis, summarizing modules of genes, and
relating modules to each other. Dimension of seed maturation found. Possible
relations, functions and regulators.
BAR bioanalytic resource…..search Nicholas
Provart
https://data.nal.usda.gov/dataset/bar-bio-analytic-resource-plant-biology
NF-YB TF
specific from aleurone DAP analysis, also DELLA protein GAI, contributes to
dormancy and regulates germination in Arabidopsis. Strong expression since
syncytial endosperm.
Session 2
Biotic stress, Robert Brueggeman
Brian
Steffenson
Optimum
temp for barley 18-27 degrees (2017 study)
Stem rust
of barley Puccinia graminis. Ug99 virulent on 95% of barleys, rpg4/Rpg5
effective against Ug99 race group.
Temperature
affects disease reaction; resistance at 27 degrees erodes rapidly, whereas at
19 is fine. In the field, up to 40% severities in hot summers (in material with
resistance)
Work with
wild species, mostly spontaneum. It is everywhere in Middle East (very common,
not like wild wheats, rather rare), WBDC wild barley diversity collection,
ecogeographically diverse, 314 accessions.
OBJ:
introducing wild resistance alleles through gene cassette transgenesis, big
five against rust disease (Sr45, Sr50, Sr35, Sr22, Sr55/Lr67/Yr46), from
different origins. Extreme resistance shown
Xu et al
2021, 2022, GWAS. But GWAS leaves low freq alleles undetected!. Rpg5 detected
though linkage mapping, not in GWAS.
AG-RenSeq
of Brande Wulff, tested with A tauschii, several genes discovered and cloned.
They applied to wild barley and rusts (10 pathotypes, the more the better): 144
seqs related to reactions to 10 races. Rph15 found on 2H, though known before,
high value gene.
AG-Renseq
shortcomings: dichotomous pattern rarely found (resistance-susceptibility);
also, little regulatory seq captures; Only R genes of NLR type found.
International Wild Barley Sequencing Consortium created during pandemics. June
2020, crowdfunding (150k donated), 61 researchers, 35 organizations, 14
countries.
285 wilds
sequenced, 10x coverage, 114 million SNps called, mean kmer counts per
accessions 1.98x109. Published
in September 2022. Data probably earlier. Ines Walde, Axel Himmelbach IPK,
involved in seq.
Wilds good
for 57 disease resistance datasets. Microelement content, cold hardiness (wilds
good for winter Minnesota!!), insect resistance, agronomic, morphologic,
everything.
SNP-based
vs kmer based GWAS, kmer offers new hits, fewer peaks but encouraging results.
Karl
Effert WSU, PhD
candidate, very good
Rpt5
cloning, netblotch res, reduces yield and seed quality
P. teres f.
maculate and f. teres (netblotch)
Effertz et
al 2021, nice graph of res genes in literature (book chapter), 6HL most
reported (Rpt5). Coincident with gene SPT1. Could be same or different.
World
minicore collection screened for Rpf5, receptor like protein
Transformation
at Sainsbury lab in Golden Promise, good results, but not total resistance.
Molly
Bergum (Moscou lab)->
Moscou moving to Minnesota this year
Rps8 gene,
actually 2 genes. Tilling pop S. Salvi, (Holden et al 2022). Mutants found with
loss of resistance, complementation tests done. Resistance recovered when the 2
genes are transformed, only one does not work. What are their functions?
PUR1 and
ortholog Xa21, with kinase domains with some diffs.
Xo70
proteins involved in exocytosis, highly expanded family in plants. Involved in receptor-kinase-mediated
immunity. Other clades found in grasses (Exo70FX), but lack critical domains,
probably undergoing neofunctionalization. Hypothesis on this….
EXO70FX has
specialized role in immunity
Pin Yang (recorded)
Bymovirus
resistance in barley and wheat
Soil-borne
bymoviruses infecting cereals.1970-90s BYMV large effects in Europe and East
Asia
Rym4/rym5
(elF4E gene) Stein et al 2005, BYMV BaMMV, are bymoviruses. Also resistance in
rym1/rym11…. WYMV y WSSMV, not reported in Spain. Symptoms occur in young
leaves in early spring. Late sowing and crop rotation recommended.
Most resistance
genes are recessive. rym4/rym5 widely used in winter cereal breeding.
Rym1/11 are
allelic
HvelF4E
knock out results in resistance, but also in reduced grains per plant,
dispensable gene.
In wheat,
polyploidy probably buffers effects of recessive genes. They have done triple
editing of TaDIL5-1 in wheat (Kan et al New Phytol 2022). Triple mutant more
resistant than single or double. No yield penalty of triple mutant, resistant.
Triple
editing TaelF4E results in WYMV resistance. Pictures very impressive about
plant development in pots with or without virus. Triple mutant is taller and
delayed maturity.
PDIL6-1 and
elF4E are resistance genes for bymoviruses, and are dispensable.
Session
3……chair, no notes
Sergey
Shabala, Eyal Fridman, Agata Daszkowska-Golec, Maitry Paul
Session 4,
Wendy Harwood, Kevin Smith
Jochen
Kumlehn
Many
improvements in gene editing. Things are crystallizing.
Julián
García Maldonado
Genomic
assisted selection. ICARDA (master IAMZ).
MET
proposal, one location strategy first cycle, MET in next cycle. 3 MET strategies,
2 reps, augmented design, genomics assisted sparse testing (from more expensive
to cheaper). Last one turns out the best.
Reduced MET
with partial replication of genotypes.
Genomic
prediction enables MET in early generation, etc
1000 F6 + 6
checks, 96 markers from the 50k chip, well spaced. Includes several QTL.
Representative of 50k, gives similar distances as full 50k.
Sparse
trial testing 316 genot unrep per location, 24 genot replicated. Some entries
replicated within locations, besides the replicated across locations. It is the
strategy they are launching now.
4 environments,
rather different
Results
explained too fast, but apparently they are convinced of carrying on with the
genomics assisted sparse testing.
Wendy
Harwood
Lawrenson
et al Frontiers in Genome Editing. Updated review on gene editing
Agrobacterium
and hygromicin to introduce CRISPr in immature embryos, very genotypinc
dependent.
Transformation
improved with growth regulators (GRF-GIF). In wheat, they can edit almost any
genotype. In barley? It does not work the same. Calli did not regenerate. But,
using another gene, WOX5, things worked. Also other techniques (PLTP). Wox5 generates
too many changes in plants. PLTP plants look better.
KNockout
efficiency in 15 diff genes variable, but rather high. Improvement tried with
intron-mediated enhancement (Cas9 gene containing multiple introns). Seems to
work.
ttHsCas12a??
better than HvCas9 for gene editing.
Cas9 +
Cas12, with artificial introns, best results for editing up to 4 genes at a
time.
Alan
Schulman
Update on NBT
policies. Currently under reconsideration in EU, but no new decisions probably
until 2024.
Session
Barley breeding success stories
Birger
Eriksen (Sejet)
Quench in
the pedigree of many current important varieties (data from Paul Shaw), 6 of
the 8 major cvs in Denmark in the last years. Quench is Sebastian x Drum
Current
varieties highly related
Denmark, 4
years of approval for a malting barley, at least grown in 500 ha, stress tests
included, strict and regulated process, they must exceed in malting propertied
to a set of checks. Few cvs pass.
Success
rate in breeding, 0.001%, from 10000 to 10, and then 1 in 10 years succeed…
(1.5M€)
Winter
wheat and spring barley are the winning combination now in Denmark. Push for
cover crops, more yield winter wheat, less input spring barley.
20% barley
for malting and food, 80& for feed.
Malting
barley now is as productive as feed barley, while malting quality has shown
good improvement.
Grain
skinning (loose husks) is a problem.
Future:
Geen Deal, gools…. Phasing out important fungicides, etc. Yield loss due to no
fungicides, loss of circa 10% yield, and increase of CO2 for hectare harvested.
Disease resistance has to be improved.
CO2
footprint depends on yield per hectare. Labeling of Global warming potential
(GWP) (proposed or in place?). Opportunities for reduction of footprint by
improving protein and energy content of wheat and barley. Shows labeling
examples.
Voss Fels
Nat Plants, increase yield potential, carry over effects for all environments.
Winter
barley 2row and 6row are not big, but is part of the future
Alexander
Strube, Ackermann
One big
headache, IP. Golden or dark age for innovation.
Winter and
spring barley breeding (and soy, for crop rotation), part of Saaten Union
Results of
survey together with Bill Thomas. Breeder’s exemption most important.
Patenting, few people like it.
Fundamental
innovations only achievable by subsidized public research!!! It should stay in
public research
Patenting
prohibitive IP protection. Incompatible with breeder’s exemption. Heavy
transaction costs. Global standard for IP protection is good. Exclusivity
allows you to refinance innovations. Exemption allows pooling and socializing
of breeding progress. Patenting is opposite.
Patent
Licensing Platform, the solution? ACLP initiative, on the making. Agricultural
crop licensing platform. Simple legal framework. You could use materials
carrying patented IP, if there is a commercial use in the end, an standard
license agreement about royalties must be signed.
Aclp, eu….SEARCH
PlantBreedingRights+Patents+ACLP,
better system tan current. The actual technologies patented, not accessible,
though. Only material developed using the patent (for crossing).
Emmanuelle
Dyrszka
First
hybrid in 2016. Reliable seed production at reasonable cost is essential.
Trying to improve full male sterility of female and seed set. Using molecular
approaches. Remove partial fertile females by markers, removing lines with bad
GS data, etc. Several strategies combined.
Seed set
scoring visually in the field. One major focus found by GWAS of >500
females. Now introducing a QTL found into lines with poorer seed set.
Seed set is
a quantitative trait. Approach using genomic selection, to capture genes with
small effect, accuracy 0.61. Lines with lower seed set predicted than the
economic threshold they have calculated are removed.
Undesired
fertility restoration in cms found in chr 6H, Rfm1, Rfm3. Several haplotypes
found for Rfm3. Some haplotypes related
to full sterility, other to full fertility, several to partial fertility. These
last haplotypes can be removed by MAS.
Pernille
Merete Sarup
Genomic
selection in 2rowed spring barley. Nordic Seed
GS allows
that now they can select from 2000 lines, instead of 250!
9 years of
data, 40k yield plots, 3145 lines, 20 traits, 25 lines per trial, 634 envs.
15K array,
with imputation
Huge amount
of variance due to additive variation, much more than gxe (Denmark!!!), of
other genetic variation. Need high additive variation to be a good crossing
parent (no epistasis, for instance).
Predictive
ability, much larger with siblings in the TP(0.7), while lower (0.45) for lines
without siblings. Still usable, though, because of huge savings for no
phenotyping!
How many
lines for accurate GS? after 1500 lines in TP, accuracy does not increase
How many
lines/years to predict breeding cycles? Same, about 1500 lines
Adding
about 100 lines per year to the TP seems enough to keep up with accuracy
GS good for
early selection, even better for late predictions of deselected lines. Update
TP continuously.
Session
Genetics and breeding of biomass
Daniel
Miralles
Perspective
on barley production in Argentine. European varieties introduced in 90s
produced big improvement.
Field experiment,
2 years, 4 reps, with 11 cvs released 1982-2019. Detailed phenotypic
evaluation, intercepted radiation during cycle, biomass, RUE, phenology, Yield
and components, N in grains and shoots, malting parameters.
Breeding
history affects: days to anthesis, longer; gfp unchanged, in days and GDD.
Plant height reduced, stem diameter increased. Grain yield increased 69
kg/ha/year, 0.9%/year. Biomass, flat, harvest index increased (from 0.4 to
0.5). Cumulative intercepted radiation increased (due to longer cycle, mostly),
no change almost in RUE. Grain number and TKW increased, both. Spikes per m2,
no change, grains per spike increased, at least until 2000. Grain protein was
not affected, no dilution effect due to grain yield increase. N harvest index,
slightly increased.
Lana
Shabala, presented
by Sergey
KUE and K
uptake efficiency. QTLs very minor. Potash price went up from 150 USD per
ton in 2003 to 800 USD now
Experiment
with 4 K nutrition treatments, 30 genotypes, divided in 3 clusters according to
K response. Shoot biomass, bad predictor of KUE. Phloem transport critical.
Microelectrode
ion flux measuring (MIFE) technique, combined with FACS (fluorescence activated
cell sorting)
Paolo
Pesaresi
Happy under
the sun 1 (hus1), pale green crops.
Only 25% of
sunlight used for phot at top of canopy. Excess may damage plants. At the
bottom of canopy, on the contrary, the problem is too much shade (not enough
light).
More
uniform light penetration would be beneficial, reducing photo-oxidative damage
at top of canopy, higher production at high density cultivation, reducing temp
at canopy level, with higher reflected shortwave radiation (achievable with pale
plants, less chlorophyll).
AGRIVOLTAIC
plant REM-TEC, company that produces solar panels compatible with agricultural
production interested in this approach.
Mutants in
Hortillus and Tillmore, SPAD and Pocketpea used. Reduced chlo content, but high
radiation use efficiency and PHOT. hus1 from hortillus, reduced total chlo,
increased a/b ratio. PHOT and color maintained along the cycle.
F2 mapping
pop, dark and pale, excap, mutation found in 4H, stop codon. Monogenic
recessive allele. Protein like an Arabidopsis prot. Function is moving antenna
proteins into the chloroplast.
Rotasperti
et al 2022
Heterodimer
cannot be formed in the mutant, function loss. Plants cultivated in Poland and
Spain, very similar to controls. No penalty at standard density.
They then
isolated a mutant from TILLMORE. In principle, similar to the hortillus, but
sowing it in autumn, the mutant turned yellow and died, probably due to cold.
Chloroplasts collapsed. BSA RNA-seq approach to localize the gene. F2
population. Mutation in 7H. Candidate gene XANTHA-H, involved in chlo2
synthesis. The aa substitution was important for the protein-protein
interaction domain.
So, not all
pale green advantageous under field conditions (Wang et al 2018)
PHOT in
hus1 increases because there is less investment in photoprotection. Actually,
PHOT in dark adapted leaves is always higher, because there is no need for
photoprotection. So, less is more in this case.
Sonia
Negrao
Irish
whiskey industry booming in Ireland. Interest in impact of terroir, etc, to
differentiate their produce.
Waterlogging
induces hypoxia in the soil.
Heritage
germplasm (old cultivars) largely uncharacterized, may have stress-tolerance features.
Exhibit
collection, 365 accesions, 2rowed spring, 3 from Spain, 4 from Italy.
50k
genotyped, pop structure 5 groups. Some 6 rows were found, germplasm bank
mistakes. Year of release had an impact -> landrace, pre-90, post-90 formed
different groups. No geographical diffs.
From 365->230
lines core collection.
Field 2020:
0-2-5-10 days waterlogged. 5 days chosen as best. Very tricky to do and to
phenotype, muddy fields!
4 days of irrigation
at early tillering, 12h per day of irrigation
Waterlogged
flowered later, less biomass, reduced height. Grain protein and Bglucan reduced
in waterlogging. Machine learning and artificial intelligence to study lots of drone-image
data. Imaging is only possible with UAVs (due to mud), and irrigation elements
have to be removed soon, to avoid interference with machine learning.
Session on
barley end uses
Henrik
Brinch Pedersen
Herz
Dockter
Carlsberg
search for raw materials, new ingredients, etc
Specialty
barley, sorghum, wheatgrass for the future!
Targeted
breeding, NU-LOX3G and 4G barley, with variants of LOX genes, improves beer
quality and lowers climate impact (really?)
How to
accelerate the process, without GMO?
FIND IT, is
their new strategy, and accelerated and traditional trait breeding method, with
mutagenesis: Fast identification of nucleotide variants by droplet digital PCR,
to find the mutations you want in an efficient way (there is a publication)
TRaitomic
platform after Find-it
They have
isolated 100 knockout variants of interest, including GW2 (found in from rice)
DELLA
mutation in Paustian, earlier, taller, larger grain and spike
Climate
targets, increasing robustness of malting barley yield, drought and salt tolerance,
keeping quality, getting competitive yields
Crops for
the future project, accelerating domestication in sorghum, perfecting
wheatgrass for food and brewing; ancient rice and global food security
Leona Leisova-Svobodova
Sponsor presentation, ICARDA
ICARDA, Miguel Sánchez-García
CGIAR, 13
centres
257 barley
varieties released since 1979, 50 in the last 10 years
Excellence
in Breeding Platform, new in CGIAR: more focused, faster and more efficient
Identification
of product profiles, widely grown varieties in a particular area, which can be
improved for some characteristic. They co-develop varieties with National
Agriculture Research Services (NARS). Now targets grouped into “feed for arid
and semiarid”, “feed and forage for favorable”, “food and fodder”, “malt and
fodder barley”. Countries and regions grouped in the 4 categories. Same for
traits, essential traits identified per megaenvironment.
Crossing
block genotyped with 50k. Use PopVar to make simulation progenies. Used for
crossing strategies, can calculate yield expectations, but also correlation
with malt extract, for instance. Crosses are decided on a combination of traits
and correlations among them.
Speed
breeding up to F5, then multilocation in F6 (stage 1), using sparse genomic
selection approach. Stage 2 multilocation, using alpha lattice. Stage 3,
networks f field trials IBYT-ASA or OBYT-FFM.
Up to F5,
genomic selection and a little phenotypic. 4 gens/year of speed breeding
(capacity of 100.000). Includes disease resistance screening, with mixtures of
isolates, NB, scald, etc. They are usually more susceptible in the chamber than
in the field. Also NIRS measurements, for B-glucan and Fe content. Crosses with
spontaneum provide up to 5% more Fe, and 42% more B-glucan (measured by NIRS)
Multilocation
testing needed when 1 loc does not represent all envs. Identify locations that
represent target population if environments (TPE). Sparse genomic selection
ensures good accuracies compared to carrying out stage 1 in only one location
with all genotypes. Also, dispersion protects against catastrophic losses of
entire locations, which sometimes happen.
Collaborators
in stage 1 in 2021, includes Spain and Portugal (Andalucía).
Stage 2,
testing locations specific for some stresses, including disease hotspots.
Phenotyping with phenomobile. They have nice location for diseases. Poster 12,
disease resistance found, horizontal?
Stage 3.
Every year 2 yield trials to more than 25 countries, 22 IBYT-ASA (Arid and
semiarid), 22 IBYT-FFM (feed forage malt)
Also have
regional networks, like Maghreb-ICARDA barley initiative.
New lines
HB1963 and 1964 released in Ethiopia, 48% more grain, and 38% more straw
(highly valued by farmers), positive for resilience of crop-livestock farming
system. Circular economy incorporated in local value chains, 20% premium price
for malt barley grain. 44% more revenues at farm level.
Extensive
use of landraces and CWR in the program (31% of stage 1 has LR and CWR). Even
lines crossed with bulbosum.
Would it be
posible to test ICARDA materials as source of breeding? Registration problems? Previous
signed agreement needed? Apparently, they only ask for 1.5%, according to ITPGRFA.
lash&Dash
session
Alina
Klaus, poster 13
Drought
experiments with PEG, root tissue collected, split in regions. Weighted
coexpression analyses, combined with DE genes from RNAseq. Network construction
for the different root zones. Combination of the two approaches helps to
pinpoint candidates.
Kumsal
Ecem Colpan, poster 19
High
temperature effect on plant growth …. Isolines based on Bowman from Maria; PHYC
and PPD1. Both sets of isolines also crossed to each other. Floret fertility in
PPD1+PHYC-e, higher than others, because of production of double seeds. PPD1
and PHYC interact, possible epistasis.
Ewa
Sybilska, poster 14
Cbp20/cbp80
double mutant under drought, seedling. Part of cap binding complex, involved in
miRNA processing, splicing, RNA export, RNA decay. Mutant hypersensitive to ABA
during germination, and better tolerance to drought stress. Faster stomatal
closure, etc.
Eyal
Bdolach poster
B1K
collection, chloroplast diversity found with molecular markers. Reciprocal F1
hybrids made, diffs found between reciprocal crosses due to chloroplasts.
Cytonuclear multi parent population, 10 parents stress resistance index for
CMPP lines.
EA
mapping, Chang poster 50
Landraces
fdrom Europe, Asia, Africa. Geographical origin inference using deep learning,
GEA, PPdH1, FT4 signal in Manhattan plot. They use the geographic info for extracting
climate data. 10-fold cross validations for geographical inference, >0.9
accuracy. Associations with temperature seasonality (1626 LR georeferenced);
8338 accessions with geographical data inferred. Chrom 2H, Signal at CEN. Tiene
la SBCC.
Laura
Paire Naked barley,
organic…
Jian,
poster 77, IPK, flo.a
paired spikelet mutant, extra spikelet
and fused glumes, gene HvALOG1, responsible for flo.a, short arm of 6H, 477 kb
deletion region on short arm of 6H.
Poster
56. What makes
barley itchy? Awn roughness in barley, genes A (Raw1) and B (5H and 7H). The
two genes segregating in two populations, F1 crossed, and F2 segregating. Raw 1
controls the size; Raw7HS controls the frequency. Raw1 epistatic to Raw7HS.
Li Guo, U Bonn, poster 75, barley enhanced
gravitropic 2. Gravitropism
controls guidance of gravity. Shallow roots better for phosphorus, deep roots
better for N and water. Mutant egt2-1, enhanced gravitropism. DEG analysis of
RNAseq, and Y2H
Mingjiu
Li, Genes CMF7, CMF3,
causan white stripe y xantha phenotypes respectively. HvClpC1 causes a yellow
striped phenotype
Ilyse
Putz, HHU
Dusseldorf, Life history traits in
annual and perennial species of barley. Rains cause soil erosion in bare
fields. Perennial agriculture can help, Mosler et al 2021 Front Sust. But
breeding for perennial takes very long. Find key genes. H.chilense is perennial.
5 species chosen, 3 ann, 2 perenn. All crossed to produce F1 and some BC;
RNAseq.
Maeva
Bicard,
understanding GEI in spring barley breeding. Unilasalle, Secobra, Kronenbourg,
Carlsberg. Determine ecoclimatic factors driving GEI. Build environmental
classification based on GEI drivers, and define high-throougput criteria to
select for GEI drivers (Understand-Predict-Control).
DSSAT, Ceres, 6 cvs 10 ens 2 years, and environ data. Then env classification.
Cvs with contrasted response to GEI drivers will be monitored by drones that
predict response of contrasting cvs. Poster 81
Barley
Genetic Resources workshop
NORDGEN,
Jan Svensson
17k barley
accessions
Bulbosum
introgressions, 176 accessions
Mutants,
9590 Scandinavian mutant collection, with many alleles for single genes
represented
991 Bowman
NILs
Chlorophyll
biosynthesis 373 mutants
Only stable
mutants kept (not het.)
Learn from
Arabidopsis community for organizing and sharing of resources
Community
infrastructure? It is a biological infrastructure
Andrea
Visioni, ICARDA
ICARDA new
genebank inaugurated in Rabat
30242
barley accessions, 2240 wild relatives, 455 are unique, many characterized
phenotypically.
Screening
of germplasm with leaf rust, for example, leaf rust in Marchouch. Also Bglucan.
FIGS to identify accessions probably carrying adaptive traits. FIGS for winter
survival in Minnesota, Stefensson, some lines survived, so FIGS strategy
succeeded. Pre-breeding, many crosses with spont to introduce disease
resistance. Examples of agronomic traits showing that spont derivated lines are
doing well, including yield. Bulbosum introgression lines outyielded the wild
line for yield at all locations. PHYSIOTRON facility, fully automated
lysimetric system. Also Phenobuggy (developed in connection with company hyphen).
AGENT, includes phenotyping genotyping and sharing.
Nils
Stein. IPK
Eurisco,
barley >21k accessions, largest genebank for crops, barley only second to
wheat
PCA with
76k SNPs (Milner)
The barley
core collection (Knupffer and van Hintum), 1995, not widely used. Was a vey
good compilation of germplasm. It is in IPK, you can order it. Pangenome on the
making, 3x 10x 50x (for 1000, 200, 50 accessions) prepublication stage. BPGv2,
50 accessions, wild and cultivated.
Not yet
official, Core1000, SSD out of Bridge, not available yet, BPGv1, BPGv2, not
available yet, some cannot be delivered by IPK
Gendibar
Icelandic
barley
One rainy
season: “all the time”. Long growing season. Barley Core Set (1000 genotypes). Ll
genotyped with 50k. Also IceBar (breeding material)
Chengdao
Li
For 100
years it was not posible to grow barley in Oz, it just did not work. Australian
Grains Genebank AGG (double skin freezer design), 2014. 20000 genotypes stored.
Project Ozbarley, 400 old varieties and breeding parents, gather datasets and
collate using FAIR principles. Cereal Rust Research Centre. Western Crop
Genetic Alliance (Joint Research Centre, Chengdao); 6000 accessions from 41
countries, 1000 with low coverage data. 5000 DH lines, 1000 JF Bowman BC lines,
RGT Planet mutant pop, Vlamingh mutant pop. Traits: phenology, heat tol, soil
abiotic stress tol.
Session
06-07-22, Genetic and database resources: harnessing diversity
Nils
Stein
Gap between
discovery and user friendly access, and sustainable use
About 70
genomes fully done, public over next months.
Problem for
opening genome data to public: generate all the information, annotation,
relation across genomes, etc. Not enough resources. TAIR has long term funding.
Barley has not.
Past:
Barley
Genomics Newsletter, past communication model, still alive in 2020
Next step,
Graingenes, main hub for map info
HarvEST,
CR-EST, next step, early 2000s
Next,
Barlex, to visualize genome, still accessible
Present:
Graingenes
still valid, Victoria C. Blake. USDA long term funding (how long?)
EnsemblPlants,
MorexV3 available, synteny research possible
IPK Galaxy
Blast Suite
JHI,
germinate, flapjack, etc, desktop applications
JHI, EoRNA,
transcription data
CROPPAL, Perth, proteomics, crop-pal.org ,
proteins, annotated, includes barley
Project related
databases, like BarleyVarDB, no longer available. Difficult to maintain. Send
data to permanent repositories
Eurisco,
Genesys, to access germplasm, Nordgen
Germinate,
for handling large datasets
BRIDGE IPK
BarleyDB in
Okayama, K Sato
T3/Barley,
triticeae toolbox barley.triticeaetoolbox.org
Databases
very distributed, often project-driven. Long-term resources not barley specific
Future:
GWAS
catalog, EBI, for humans, mandatory for human research. Graingenes has already included
GWAS results. Send GWAS chapter and
suppl info
Get used to
prepare research and data to be uploaded to databases
FAIR data,
findable, accessible, interoperable, reproducible.
The FAIR
cookbook, Beier et al….
Plan you
research as a resources project. DMP is important.
AGENT, precision collections of 500 barley and
500 wheat, evaluated for agronomic and biotic, a selection for abiotic stress.
Should we develop Graingenes into a new TAIR?
If yes, how?
Asis
Sherstha
(B Stitch)
HHU
Multiparent
pop, double round robin, 23 parents , 45 subpops using partial diallel, MARVIN
for grain traits, 50k, field trials, grain size characters. Huge genotypic
diversity among parents.
Grain
traits showed high h2.
mppR package for QTL analysis, special for multiparent. 62 consensus QTL, 13 new QTL, not
reported previously. Including proof of concept loci (vrs1, nud).
QTL
validation in Lakhan x Georgie, with 2 NILs, crossed, 924 F2 progenies,
validated in fine mapping
Comparison
of full QTL effect and GP model, GP with GBLUP was always better (30%). QTL
effects distributed across all parents, with many positive and negative alleles
in each one
Genetic
architecture of grain size is complex, more than previously thought. LR have
potential to improve these traits, not just for adaptation and stress
POSTER 64
Outmane
Bouhial
CGIAR
barley breeding toolbox
Assembly of
association panels. Specially for developing world, representative of CWANA
region
530 mostly
spring barleys, CGIAR, elite from all over, and LR.50k, flowering genes, VRN1,
PPD1, FT1, CEN, 16 allelic combinations in total.
Genetic
subsetting….. 312 entries in public domain, mostly CGIAR and LR.
Cluster,
PCA, AMOVA, 6r more polymorphic than 2r, ICARDA most diverse AMOVA of flowering
genes indicated mot variation for flowering genes within pops. CEN the most
important among pops. All flowering genes associated with pop structure. 16
flowering combinatons AC1-AC16, would be interesting knowing them.
Pop
structure with sNMF. Identified pops have common origin and breeding history.
MTK (mean of transformed kinship) measures of
kinship pf genotypes in a subset and identifies the most dissimilar set of
genotypes
312 (CBBT)
do not include some of the more modern, negative side of PC1 (seen in the
picture of principal component analysis). But have most subpops and all 16
earliness combinations flowering. They offer it as a collaborative tool.
Available in 2023.
Ronja Wonneberger
Comprehensive
multi-omics database. OBJ: genomics and transcriptomics 2r springs. 209
selected. UMinn, IPK, JHI, 2 years, field trials, many traits, fertility
traits, etc
Available
not yet, it will be after the paper (soon)
Transcriptome
all 209 accessions, seedling crown and root, developing inflo, preduncle,
central spikelet, developing grain 5 days post anthesis.
50k SNP,
and RNAse SNP, WGS done, 40M SNPs, after filtering, LD pruning, imputation,
1509447 SNPs. ENA archive
Search preprint
Structure
of European 2-rowed spring, PCA
differentiates release period. FsT in sliding windows, 5H>0.7, old vs
new shift, most of new accessions carry the Barke alleles. Region of 3Mbp shown
50-300Mbp approx. Exactly the same as
region in our Molec Ecol paper.
Expression
of 800 genes in that region. PCA of expression divides the 2 groups clearly.
PCA of genes also done, 2 groups consistently showed diffs, across all tissues.
Strongly differentiated across clusters of genotypes. Off expression in old, on
expression in new. About 45 genes
Haploblock in a pedigree scheme. Very nice. Six cvs with strange patterns, 3 of
them, with diff expression pattern, come from laevigatum.
Global
distribution of NEW haplotype, checked in the IPK collection (hap2), mostly in
LRs from Mediterranean, including Spain, and Cvs also in Spain (and Europe).
Also found in winter and 6row barleys.
New
haplotype comprising 5% of barley genome under strong selection in the past 60
years, associated with upregulation of gene expression. Most prevalent in North
Africa and Southern Central Europe. Still unclear the reason (disease, leaf
rust?). It is definitely the same as the
one in Molecular Ecology. Check in databases if it may come from an spontaneum
introgression. Article sent to Ronja.
It will be
possible to publish papers in Journal of Agronomy, before September 1st.
Agatha
Walla (von Korff)
HHU
Quest for
elusive developmental gene in barley
Shoot
architecture, balance between traits, tillers, seeds…negative correlations
hamper yield progress.
What are
genetic key regulators?
Forward
genetics, Nodgen collection of mutants, mnd8,mnd6, mnd1, dwarfs, high
tillering. Genes cloned Hibara et al 2021, Mascher et al 2014, Walla et
al 2020 (mnd1)
RNAseq in
the right tissue and developmental stage. Bowman introgression lines tested. 2
major introgressions, 2H and 7H. In 7H the candidate mutation was found, null
mutant due to frameshift. Probably related to histone function and expression
downstream. Regulator of meristem transition. Tillers in mnd1 appear along the
whole shoot, not just in the two lower nodes, as usual (WT). They found
bracteae at the base of spike, and underneath primordia that formed spikelets. Gene
expressed in shoot apical meristem.
Further mnd
mutants mining found 5 mutants which were not in the 3 main genes. Mnd-like1,
mnd-like2. One is Xray, the other natural.
Phenotype not as severe as mnd1, tillers only in base of plants.
They found
mnd-like mutants were in the same place as for mnd1. But no candidates based on
RNA study. WGS for the mutants also did not work. Then biparental mapping in an
F2 population. There were 2 introgressions in the region, and a region with no
recombination. They found an inversion of 3Mbp in one of the mutants, just
upstream of mnd1. In the mnd-like2 they found an insertion in the upstream
region of mnd1. Expression of mnd-likes did not go up with time, like the wild.
The mutants did not complement
Other
mutant mnd7h also had no expression, no diff in coding. Probably also cis
variation.
Mnd1
represses vegetative growth, key regulator of many traits. Many cis-regulatory
mutations affecting expression. Dosage or spatio-temporal expression patterns
probable underlie the nuanced phenotypic traits.
C. Jiang
(recorded)
Reference
guided tilling by amplicon. Most tilling in EU material
Hatiexi?,
landrace NE China. 2row, black, grains and leaves, etc. It is far from East
Asia accession in PC Milner. Origin in Middle East. Susceptible to BaYMV,
BaMMV. Good for animal feed, high protein.
Mutated
with EMS. , 2.8% good germination. They used 2 levels of mutant agent. 8500 M3
seeds. Many phenotypes available.
WGS made to
build a reference genome. TILLING platform developed, 3D amplicon-seq.
5184 M2
included in TILLING service. 21 mutants per 1.5 kb !!!
Plant Communications July 2022, publication
ready
Session Morphology,
phenology, and development, Laura Rossini
Maria
von Korff
This is the
Udda Lundqvist memorial sponsored talk
Inflorescence
development and floral abortion under stress (high T)
Reproductive
level particularly sensitive to high temp, reducing seeds per spike
Reference
collection of 300 accessions in ICARDA, time ago, many from hot and dry places.
100 accessions LR and elite genotyped 50K. 20-16 degrees control, 28-24 degress
stress. Many traits measured in the chamber.
Days to
heading, LR from hot env accelerate flowering; elite strongly delay flowering.
Shows correlations
of diff vegetative traits with seed number, all positive under control
conditions; except fertility. Reproductive traits, positive in control with
spikelet number, in high T, fertility was the variable clearly correlated with
seed number.
PpdH1 is
repressed by ELF3, and PHYC is also involved in response to flowering and hight
T. VRN1 also involved, under high T is down regulated, then delayed flowering
through FT1.
PPDH1
variant Gly-Trp, effect of the two alleles. Recessive gene, lines flower later
in control, in high T they are further delayed (in days); the wild allele
actually speeds up development under high T.
Introgression
lines to study this. Golden Promise-derived, with introgression of wild PpdH1
from Igri; also Scarlett based, and Bowman based. Same results in the 3 sets.
Confirmed results from panel of genotypes, a crossover interaction effect. They
studied apex, Waddington scale, because seed number is determined by spikelet
initiation and floret abortion.
GP, delay
in floret development in high T, GP-fast, acceleration of floret development
under high T over control. 1.7 vs 0.9 spikelet primordia per day in GP; 1.6 vs
2.0 in GP-fast (control-high T).
Hight temp
affects Inflorescence meristem (IM) size, microscopy, nut not in GP-fast.
Fertility
score hugely different between GP and GP-fast. Female or male organs
susceptible? Experiments to test this.
Crosses done
between ovaries at 28 degress by pollen at 20 degrees, and the opposite. Found
that ovary fertility dropped, but pollen dropped much more. But not in GP-fast
(GP with Igri dominant PpdH1). Pollen looked shrunk in GP, not in GP-fast.
Molecular
changes? RNAseq profiling, at 4 stages: 1.0, 2.0, 3.5, 6.0 Waddington. Still
under analysis.
>90%
reads mapped to Barke. DE genes found for every genotype and stage. Many more
DE genes in GP than in GP-fast. 408 genes related to development AND
temperature, 222 annotated, 186 not. Stages drove the clustering of the genes,
except GP under heat in 2.0 and 3.5, found cluster of upregulated genes under
heat.
Zhong and Kong 2022, SEARCH
FT1 in the
leaf TPM of GP-fast much higher AFTER W1.0 than in GP. Downregulated by heat,
but not that much.
FT2 is
expressed in meristem, down regulated in GP as well, and by heat.
Balance of inducing and repressing genes, FT vs
CEN, if you have
more CEN you keep the meristem alive. Ratio FT1 in the leave to CEN. Ratio very
diff in GP and GP-fast, under heat ratio goes up in GP-fast, down in GP Could
explain crossover interaction.
Inflorescence
meristem affected by ITN-M/AP2, they analyzed, down regulated by time.
Scarecrow TF and trehalose-6-phosphate synthase,
important for sugar status. Both down regulated in GP heat.
MADS34/TaSEP5
and MADS15/BM3 also affected…..
Auxin
related genes also tested. Auxin response factor down regulated in GP heat.
Validation of hypothesis , HvpPIN1….. expression in IM, Fluorescence reporter lines being developed by Raffaella and Laura.
Hormone
homeostasis may be altered. Need to dissect the meristem tissue to search for
specific molecular networks.
Started
Edgar Damesa-Arevalo, scRNAseq. Molecular
Cartography company helps to locate the genes by color in the developing inflorescence,
easier than with other methods.
POSTER 18,
67
Higher T
effect in experiments with shift, with actually shorter high T treatments.
Ravi
Koppolu
Spikelet
determinacy for improving grain yield potential.
Rachilla
differs between barley-wheat. There is the potential in barley to form more
than 1 floret, of having an indeterminate rachilla
Mutants,
com1, com2, Int1, Vrs4 central spikeletes transform into a spike like inflor.
Rachilla developmental mutants.
Qgene,
APETALA2 TF regulated by miRNA172, responsible in wheat for rachilla mutants.
Mutants of barley also found (publication of 2021)
Mul2.b
mutant, lateral spikelet rachilla elongates to produce up to 3 florets (in a
row)
Shows
beautiful pictures showing early rachilla development
Data from
flo.a x mul2.b cross, indicates they are not allelic
The trait
can be treated as quantitative. Studied in pop Morex x mul2.b F2. QTL analysis
in F2, 2 large QTLs in 2H and 6H, minor in 3H from Morex! 6H and 2H QTL
interact for lateral supernumerary grains, not for central. They are
mendelizing the QTLs in lines, to continue with map-based cloning.
New mutant
1 found, nice pictures of rachilla elongation forming new spikes. Rachilla
would be like a secondary rachis. Also mutant 2….
Luke
Ramsay
Recombination
in barley is much lower than in Brachy. Why?
Resource:
desynaptic mutants in barley, 15 loci , 24 in Bowman, 3 biparental pops for 15
desynaptic mutants.
Des10
reduces recombination, does not help for breeding. Built a suppressor screen of
BW230 desynaptic10, 18000 seeds mutagenized. M2 families in polytunnel and
increased for increased fertility. 10 M4 plants with restored fertility
Line with
HvRECOQL4 C/T point mutation had highest fertility, in the coding seq. Crossed
to WT->FT1, alleles at Mlh3 and RecQL4 tracked, F3 lines with wild type, one
mutant or double mutant, 4 classes.
Arrieta et
al 2021.
Recomb of
recql4 F3 lines was twice than in WT. Double mutant similar to recql4. Mlh3 has
much reduced recombination.
They will
backcross recql4 into other backgrounds, to see if effect is maintained.
Ivan
Acosta
Auxin in
production of starch… in pollen maturation.
Pollen
matures accumulating starch. Mitosis during maturation forms vegetative cell
(energy) and the two sperm cells. Mutant msg39 anthers look normal, pollen
seems alive with staining, but seems much lower in starch. Starch accumulation
starting around W9 does not occur in the mutant, whereas mitosis occurs
normally. The mutant is sterile, has to be maintained with a maintainer line,
and inheritance is not Mendelian (16 instead of 25%).
Msg38
encodes an enzyme for auxin biosynthesis (YUCCA). Confirmed by actual
measurements of IAA in plants along the time, whereas precursors are
accumulated in the mutant. Curiously, at its peak, IAA is quickly degraded into
ox-IAA
Protein
found only in pollen, after W9. RNAseq, auxin involved in expression of
thousands of genes. Studies of diffs clusters, auxin required to boost energy
production at W9.25 and 9.35. So, starch was not made in the mutant because it
had no energy to do so. Co-expression of genes for energy production and msg38.
Auxin is required to increase flux through glycolysis and TCA cycle. The sugars
actually reach pollen during its growth, but pollen does not know what to do
with them, no energy in the mutant.
Sugar
accumulates at W8.75 and W9, inducing a transient boost of auxin synthesis, and
a boost of energy genes, High ATP, then starch synthesis.
The msg38
is quite normal. It belongs to a specific subfamily of YUCCAs, and has a
specific effect in pollen. Could be a mechanism for sterility in breeding!
Cannot be restored by external auxin application, probably because it cannot
reach pollen cells.
Vanda B
Marosi
Wheat
barley transcriptomes comparison. Same growth stage.
Weighted
gene correlation network analysis
Mapping to
a single transcriptome for each species. Not much diff!
Ortholog
analysis: 36k genes vs 108k genes. Orthofinder to find25k orthologs; 11908
single copy orthologs, 1 copy per genome. Used for further analysis
34
coexpression modules in barley, clearly diff by tissues
Looks at
overlap of wheat and barley modules, similarities and differences (many)
CONCL:
tissue selection was instrumental
Silvio
Salvi
Roots.
Mentions Wallace
et al, On the road to breeding 4.0. Ann Rev Genet 2018
3 types of
roots. Seminal roots (primary or secondary), nodal roots starting from crown,
lateral roots starting from other roots
Traits:
root/shoot biomass rate. Anchorage to soil, root lodging. RSA (numbers, angles,
distribution, etc); anatomical traits (aerenchyma, etc); root hairs; root
exudates, Rhizosphere microbiome; nutrient and water absorptions.
fer1, the
only barley root architecture mutant in BGNesletter!!!! (fewer root 1), only
seminal root available, but seed source was lost. More mutants needed-> TILLING
TILLMore,
3600 lines. POSTER 72, example of forward genetics.
Screening
for root phenotypes, rolls, rhizoslides, dirt-rhizotrons….34 mutants found,
hypergravitropic, short, wavy, etc.
Enhanced
gravitropism 2, PNAS 2021, angle affected in barley and wheat, confirmed with
rotation test to ensure ACTIVE gravitropism. Bulked segregation analysis with
F2 plants, 5H, only one of the 6 mutations found with a stop codon. Gene edited
the gene to have further confirmation. Done. Controls gravitropism of ALL root
types. RNAseq of root sections placed it
in the meristematic zone, 67 DEG, related to plant cell functions (expansin and
peroxudases). EGT2 is close homolog of WEEP2 in peach (shoot gravitropism)
EGT1, PNAS,
just accepted. Phenotype similar to EGT2
Possible look for them in OxP????
6H, ask for
preprint!!!!!!!!!!!!!!!!!!!!!!!!!!!!!
Natural
variation found in Whealbi collection. Haplotypes II and IV showed diff in root
angle. Probably affects adaptation. Is independent from auxin.
RNA
hybridization indicated location of expression (diff to EGT2)
Type of
protein and genes DE related mainly to cell walls.
Are they
involved in cellular stiffness? Atomic Force Spectroscopy reveals egt1 cell
walls are less stiff.
Konstantinova et al 2021 int J Mol Sci
It seems
that the antigravitropic mechanism is cut by the mutants.
Kishner et al 2021 PNAS, Fusi et al 2022 PNAS.
Confirmed
in wheat, TILLING in Cadenza, triple mutants for both genes.
Now, it
needs to be tested in the field. Also Uauy and Ober (bread and durum wheat)
GWAS,
POSTER 51
Rescreen of
TILLMORE with CD cases!!! Interesting method. Easy to implement?
Session Baltic
and Nordic barley: a regional perspective
Therese
Bengtsson
Pre-breeding,
N-S, Cont-Maritime, climate constrasts, very long days
Public-Private
partnership, why? Climate changes, new pests, food security, small market!
Number of programs have been reduced. They depend from the Big 4 companies,
which have Nordic and Baltics outside their target. No one else will do the
job. Private companies are small.
Pre-breeding
is pre-competitive, companies can collaborate. Resources are not enough in companies
Germplasm
evaluation and introduction of traits in elite background, HT phenotyping
Funding
50/50 ministry (norden)/private partners. Project leader, Ahmed Jahoor. Boreal,
Sejet, Lantmannen, Nordic Seed, Graminor are the private partners; Arric. Univ
Iceland. SLU, Luke, and other University as public.
2012-14
screening of adapted material to get linked markers for MAS; materials
deposited at Nordgen; screening and crosses using adapted an hidstoric material
(MAGIC and biparental pops); phase III, delivery?
Building
trust in the beginning was necessary. Each breeder provided 30 lines or cvs,
180 genotypes. Screened for 7 diseases and development traits and yield and
straw strength. 9k genotyping and GWAS
KASP
markers prepared for the companies to use.
Example:
cereal oat cyst nematode, screened by a company. Two markers found on 2H,
converted to KASP, already used.
Bengtsson et al 2017, 2 publications. Goransson 2019. QTL for many traits identified.
Phase II
2015-17. 200 genotypes adapted and historic materials, screened at 2 locations
for disease and agronomics. MAGICs developed, balanced for row-type, some with
4 some with 8 parents, multiplication in New Zealand. 9 MAGIC and 2 biparental,
83-200 (303 max) lines per pop, DH or SSD.
Phase III.
Continue screening, 50k genotyping, disease screening massive, some agronomics
(heading, maturity, etc)
Hautsalo et al 2021, TAG 2021, Pops 1-4. Pyramiding of scald resistance.
Studies of
early vigor in adapted and historic material, at IPK Lemnatec, EPPN2020. Multitrait
GWAS in progress, 250 genotypes (180 from phase I).
Also
screened for powdery mildew. Scald and mildew resistance together found.
From Phase
III retrieving markers linked to traits, which can be used for MAS.
Many more
advantages of project, intangibles and tangibles (picture)
All info shared
through INTERNAL website
Alge
Leistrumaite
Barley in
Lithuania
Mara
Bleidere
Barley
breeding in Latvia. Interest in roots. Increasing perspective for cultivation
of winter barley.
Magnus
Goransson
Long days,
strong winds, heat sum, only 75% of Scandinavian. Sown end of April.
Traits:
maturity in low temp, early heading, straw stability. Icelandic barley is very
early. Does it occur at other envs? 17 trials in the PPP for pre-breeding
project, and found that earliness is stable over envs. But the 3 early
genotypes were too little for GWAS (169 genotypes). Tested in controlled envs.
84 genotpes, 30 hours day, 5 degrees at night, 15 degrees during day, and 15-20
(warm treatment). 9k chip. Heat sum to maturity same in LDC and LDW. Most
genotypes severely stressed in SDW. Three early genotypes were very tall, and
much more vigorous than LDC and LDW. Most lines delayed heading under short
days. Reseq PPD1, ELF3, FT1, CEN, 30 genotypes. Winter CEN gives earliness and
yield.
