Workpackage 06 Analysis for protein and fat and minerals

AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ) March Quedlinburg Main topics Protein content (P5 – P9 – P11) - NIRS Calibration: Kjeldahl method (P9 – P11) sampling 10% of the whole harvest - NIRS Analyses (P5 – P9 – P11) on the whole harvest Oil content (P5 – P9 – P11) - NIRS Calibration: Soxhlet method (P9 – P11) - NIRS Analyses (P5 – P9 – P11) Avenins (P5) 600 accessions Micronutriments (P5 - External assistance by ENITA Clermont-Fd) 200 accessions

AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ) March Quedlinburg WP 6 FIRST AVENIN RESULTS Audrey Didier, Jean Koenig, Gerard Branlard, Roland Chavany, François Balfourier

AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ) March Quedlinburg 35 countries represented 5 to to to to 40 1 to accessions The oat collection

AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ) March Quedlinburg Modern oatOld cultivar Oat Landraces Varieties struck off catalogue Avena sativa 35 %65 % AACCDD Avena sativa 88 % AACCDD Avena strigosa 7 % Avena byzantina 4 % Avena abyssinica 1 % AsAs AACCDD AABB The collection

AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ) March Quedlinburg Evaluation of Agro- morphologic descriptors WP3 Put in place of evaluation plots Descriptors observation Agro-morphologic diversity of the collection Data analysis Method Avenin identification WP6 Avenin’s extraction protocol Allele identification at three loci Ave1, Ave2 et Ave3 Acid page gliadin pattern analysis Avenin allelic diversity Data analysis Method

AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ) March Quedlinburg Proteins block from 17 to 23 kDa 25 genes in average by haplöid genome Extraction in alcohol solution Avenins 3 loci: Ave1, Ave2 et Ave3 Prolamins

AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ) March Quedlinburg A? c Ave1 B?c Ave 2 C?c Ave 3 Main locus involved in synthesis of avenin endosperm storage proteins Avenin storage proteins are encoded by clusters of genes

AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ) March Quedlinburg Gliadins protocol - INRA Avenins protocol - Portyanko Condition A: Incubation at 39°C without shaking in a 70% alcohol solution Condition B: Incubation at 25°C with shaking in a 70% alcohol solution Condition A Condition B Elaboration of avenins extraction protocol

AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ) March Quedlinburg Three concentrations of gels: T = 8.5% C= 4.76%T = 12% C= 4.76%T = 14% C= 4.76% Acid page gliadin pattern analysis T=(A+B)/V C=B/(A+B)

AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ) March Quedlinburg 16 indicators of allelic pattern NameOriginSpecies NameOriginSpecies AstorNLD A. sat. var. aristata ManodGBR A. sat. var. mutica Avoine de versaillesFRA A. sat. var. brunnea MargotBEL A. sat. var. mutica BardseyNLD A. sat. var. mutica Maris tabardGBR A. sat. var. mutica Black MesdagFRA A. sativa var montana PelsoFIN A. sat. var. brunnea Blanche de WattinesFRA A. sat. var. aristata RheaFRA A. sat. var. inermis BoogBYS A. sativa SanoSWE A. sat. var. aristata HedeDNK A. sativa StarblondeFRA A. sat. var. mutica KytoFIN A. sat. var. aurea Svalof HellgeberSWE A. sat. var. mutica

AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ) March Quedlinburg Identification of allelic pattern at the locus Ave1 5 alleles were found against 7 in Portyanko’s analysis 4 alleles were indentical to Portyanko’s analysis (a, b, d, g) 1 new allele (o)

AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ) March Quedlinburg Identification of allelic pattern at the locus Ave2 6 alleles were found against 10 in Portyanko’s analysis 3 alleles were indentical (a, d, g) 3 new alleles (m, o, n)

AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ) March Quedlinburg Identification of allelic pattern at the locus Ave3 10 alleles were found against 9 in Portyanko’s analysis 6 alleles were indentical (a, b, c, e, g) 4 new alleles (k, o, p, q)

AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ) March Quedlinburg Accessions: correspond to the 323 accessions used in WP3 Sample seeds origin: multiplication 2007 in Clermont-Ferrand, other multiplication site Quedlinburg and Radzikow breeders Two analysis had been realised: migration of 5 seeds manually grind together migration of 1 manually grind seed Migration indicators : equal mixed of Margot and Mulga lay down three times on each gel Material used

AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ) March Quedlinburg Allelic pattern identification Identify locusNumber of accessionPresence percentage Ave A. byzantina subsp. byzantina (2) A. sativa (16) A. sativa var.nigra (2) A. sativa subsp. sativa (8) A. strigosa (1) Ave A. abyssinica (1) A. sativa (9) A. sativa subsp. nudisativa (2) A. sativa subsp. sativa (3) Ave A. byzantina (1) A. sativa (8) A. sativa var. nigra (2) A. sativa subsp. nudisativa (3) A. sativa subsp. sativa (6) Ave1 + Ave A. sativa (9)A. sativa subsp. nudisativa (2)A. sativa subsp. sativa (7) Ave1 + Ave A. sativa (39) A. sativa var. nigra (1) A. sativa subsp. nudisativa (1) A. sativa subsp. sativa (22)

AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ) March Quedlinburg Allelic pattern identification Identify locusNumber of accessionPresence percentage Ave2 + Ave A. abyssinica (1) A. byzantina subsp. byzantina (2) A. sativa (19) A. sativa var. macrantha (1) A. sativa subsp. nudisativa (4) A. sativa subsp. sativa (17) Ave1 + Ave2 + Ave A. byzantina subsp. byzantina (1) A. sativa (39) A. sativa subsp. nudisativa (6) A. sativa subsp. sativa (43) A. sativa var. brachytricha (1) A. strigosa subsp. brevis (1) None A. byzantina (1) A. byzantina subsp. byzantina(1) A. sativa (13) A. sativa subsp.nudisativa (1) A. sativa subsp.sativa (11) A. strigosa (5) A. strigosa subsp.brevis (1) A. strigosa var.nigra (1) A. strigosa var.typica (3) A. strigosa subsp.strigosa var.gilva (3)

AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ) March Quedlinburg 18% of accessions shown difference between one seed and five seeds pattern. This 18% are composed by 80% landraces and 20% modern cultivars. Hypothesis: genetic heterogeneity mix of accessions Results

AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ) March Quedlinburg Allele repartitions to the 3 locus (in %) Ave 1Ave2Ave3 a02111 b55-38 c--34 d1810- e--0 g2129 j--2 k--1 m- - n-10- o25354 p--0 q accessions

AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ) March Quedlinburg Difficulty to identify the allelic pattern at the three loci Different technical conditions Variation of accession in function of donor origin Analysis of avenin, on a 324 accessions set, did not allow an individually differentiation of the accessions Further studies on relation between agro-morphological characters and avenins Conclusions and Perspectives

AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ) March Quedlinburg What’s next? NIRS analysis (P5 – P9 – P11) Dehulling in process  Who sent to who?  What delay expected Start of NIRS analysis  2009 /2010 Calibration for protein and oil content  2009/2010 Avenins (P5)  2010 ? Micronutriments  2010

AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ) March Quedlinburg Evaluation of Agro- morphologic descriptors Put in place of evaluation plots Descriptors observation Agro-morphologic diversity of the collection Data analysis Method Avenin identification Avenin’s extraction protocol Allele identification at three loci Ave1, Ave2 et Ave3 Acid page gliadin pattern analysis Avenin allelic diversity Data analysis Method

AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ) March Quedlinburg Observations in evaluation plots (2008) Panicles descriptors (scale 1 to 3) Disease and lodging descriptors (scale 1 to 9) Plant height (cm) Heading date (day) april june jullyaugust may Sowing Harvest Yield (q/ha) TGW (g) Glume color (scale 1 to 6)

AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ) March Quedlinburg Principal Components Analysis 2 first axis explain 51% of diversity variation Correlation: Positive between lodging and plant height Negative between (lodging plus plant height) and sensibility to black rust Negative between yield and heading date Descriptors projection on factorial plan (1 x 2) Active and Additional Variables

AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ) March Quedlinburg Less diversity in modern cultivars with trend to better yield and hight TGW Diversity of agro-morphologic descriptors in the collection Modern cultivars Old cultivars

AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ) March Quedlinburg Allelic pattern Ave1: b Ave2: a Ave3: b 12 % Ave1: o Ave2: o Ave3: c 19 % Ave1: b Ave2: o Ave3: c 13 % Comparison between agro-morphologic and avenins diversity (91 accessions) No relation between the both