Workpackage 02 Germination testing Christoph Germeier Zofia Bulinska

Workpackage 02 Germination testing Christoph Germeier, Zofia Bulinska – Radomska, Izabela Borowska, Audrey Didier

Germination methodology Germination and Fusarium counting methodology Germination test results 2008 Germination methodology check test Germination test results 2009 germination test: how many accessions are being tested, when the tests have been started, how long do they all take? AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ), 02 -04 March, 2009, Quedlinburg

Germination methodology General outline of germination tests according to ISTA methodology: Between paper in climate chamber, Continuous light 20°C, 4 replications (recommended 8) a 50 seeds Breaking dormancy with gibberellic acid: GA 3 (1000 mg/l) in phosphate buffer (1. 7799 g Na 2 HPO 4 x 2 H 20 + 1. 3799 g Na. H 2 PO 4 x H 2 O + 1 l aqua dest. ) Count after 5 and 10 days Variants used in the project FRA 2007 / 2008: Round filter paper (diameter: 90 mm) are moistened with gibberellin solution and placed in the petri plate then seeds are laid down onto the filter paper. A second moistened round filter paper is disposed on the top. POL 2007 / 2008: Filter paper envelopes (CHEMPUR, 65 g/m 2, 12, 5 x 18, 5 cm) are moistened with gibberellin solution. Seeds are placed into the envelope, incubated at 200 C with continuous light. AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ), 02 -04 March, 2009, Quedlinburg

Germination methodology: Radzików AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ), 02 -04 March, 2009, Quedlinburg

Germination methodology: Radzików Normal germ Mold AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ), 02 -04 March, 2009, Quedlinburg

Germination methodology: Radzików Fusarium Normal germ AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ), 02 -04 March, 2009, Quedlinburg

DEU 2007 / 2008: Filter paper towels (e. g. Munktell C 160, 15 x 60 cm 2) are pre-folded and moistened with gibberellin solution. Seeds are placed onto the fold, the towel then folded, rolled, stabilized in a plastic bag and incubated at 200 C with continuous light. 2007: climate cabinet side of plastic bag open 2008: climate chamber top of plastic bag open AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ), 02 -04 March, 2009, Quedlinburg

Germination methodology: Quedlinburg Set up of germination tests in climate chamber (2008) AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ), 02 -04 March, 2009, Quedlinburg

Germination and Fusarium counting methodology: Quedlinburg A) Abnormal germ and Molds AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ), 02 -04 March, 2009, Quedlinburg

Germination and Fusarium counting methodology: Quedlinburg B) Fusarium more or less thick mycelium, reddish color, plants affected or not germinated AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ), 02 -04 March, 2009, Quedlinburg

Germination and Fusarium counting methodology: Quedlinburg B) Fusarium more or less mycelium, reddish colour, plants not affected AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ), 02 -04 March, 2009, Quedlinburg

Germination and Fusarium counting methodology: Quedlinburg B) Fusarium mainly reddish color, mycelium, symptoms not detectable AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ), 02 -04 March, 2009, Quedlinburg

Germination methodology check test (wild species) Accession A. barbata ssp. PRT barbata POL DEU side open DEU top open DEU 14 6 AVE 850 75 89, 5 69 80 78 A. clauda BGR 00 1 A 2 E 000 3 52 50, 5 5 62 65 A. damascena POL 00 3 PL 52345 32, 5 97, 5 60, 5 DEU 14 6 AVE 2671 47, 5 79, 5 65, 5 83, 5 82 DEU 14 6 AVE 2804 63 81, 5 36, 5 88 81, 5 DEU 14 6 AVE 3066 77, 5 71, 5 20 A. fatua AUT A. fatua ssp. fatua RO M A. sterilis ssp. ludoviciana ALB 198 4 FRA DEU bag closed 198 6 199 4 AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ), 02 -04 March, 2009, Quedlinburg 94, 5 86

Germination methodology check test (marginally cultivated species) Accession A. abyssinica 193 9 FRA POL DEU bag closed DEU side open BGR 00 BGR 1 8133 25 54, 5 25 35 44 78 78 60, 5 80 82 A. abyssinica cult. Finca-san-jose ESP DEU 14 AVE 6 4711 A. strigosa var. nigra BEL 199 8 RUS 00 2001146 1 78 25, 5 87, 5 61, 5 A. strigosa var. typica Local NO R RUS 00 2001125 1 62 73, 5 80 17 A. strigosa ssp. brevis var. nudibrevis ESP 198 9 POL 00 3 PL 51277 60, 5 88 64, 5 A. strigosa ssp. brevis var. nudibrevis cult. Saia DE U DEU 14 AVE 436 6 13, 5 78 24 197 8 195 9 AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ), 02 -04 March, 2009, Quedlinburg DEU top open 74 79 77 84 67, 5 80, 5

Germination methodology check test (cultivated oat) Accession FRA POL DEU bag closed A. byzantina ssp. HR 199 byzantina cult. Zutna V 6 Germination methodology kisa DEU 14 6 AVE 3095 29, 5 76 11 A. byzantina ssp. byzantina var. aurea AUT DEU 14 6 AVE 2741 45 73 49, 5 A. sativa var. nigra cult. Stormogul Ii SW E 191 8 SWE 00 2 NGB 486 6 82 95, 5 69 92, 5 A. sativa ssp. nudisativa var. chinensis cult. Platek POL 197 4 POL 00 3 PL 50731 42, 5 86, 5 28, 5 88, 5 DEU 14 6 AVE 674 31 70 35 65, 5 77, 5 99, 5 85 98 98 check test A. sativa ssp. DE nudisativa var. U chinensis Cult. Weißer Nackter Fahnen DEU side open DEU top open 77, 5 76, 5 81 Standard cultivars from breeder A. sativa Cult. Aintree FRA 199 2 FRA 26 1 AINTRE E 65, 5 GENRES 061: Avena genetic resources for quality in human consumption (AVEQ), 02 -04 March, 2009, Quedlinburg A. AGRI sativa Cult. SW GB SWE 00 SW 78 94, 5 79 94, 5

Germination tests total summary Fusarium Infestation Germinability Avg Accessions per test (day) Total Accessions Tests/ Days Lab Year 2008 DEU 16 385 24, 1 54, 9 57, 2 3, 1 FRA 10 374 37, 4 50, 2 0, 1 1, 9 POL 8 211 26, 4 81, 1 13, 8 6, 7 2009 DEU 10 449 44, 9 68, 8 45 3, 1 FRA 7 300 42, 9 58, 5 0 0 POL 16 320 20 80, 8 14, 5 6, 3 AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ), 02 -04 March, 2009, Quedlinburg

Germination results species wise – wild species DEU FRA POL 79 90 1 2008 3 69 82 96 2 43 50 58 0 A. brevis 2008 2 74 74 74 2 84 84 84 0 A. canariensis 2007 1 46 0 2008 1 80 1 A. clauda 2007 1 65 0 A. damascena 2007 0 A. fatua 2007 8 49 74 88 9 9 56 76 0 2008 11 55 71 88 7 21 49 73 0 2007 1 2008 3 A. hybrida 2007 1 A. sterilis 2007 4 A. hirtula 57 70 83 92 74 42 56 74 86 10 0 0 46 2 0 0 10 0 0 0 1 59 Max 68 Avg Max 3 N Min Avg 2007 N Max Min A. barbata N Avg Year Min Species 83 0 0 AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ), 02 -04 March, 2009, Quedlinburg

Germination results species wise DEU FRA POL Min Avg Max 59 82 3 30 49 82 0 A. abyssinic a 2008 5 50 61 74 3 32 39 52 2 78 83 88 A. strigosa 2007 21 21 51 80 21 20 62 88 5 50 80 92 A. strigosa 2008 39 49 69 90 32 34 62 90 29 0 77 97 Min Max 40 N Avg 3 N Max 2007 Year Avg A. abyssinica Species N Marginally cultivated species Cultivated oats A. byzantina 2007 16 37 60 81 16 14 48 82 6 62 75 84 A. byzantina 2008 25 50 63 82 22 16 55 82 24 71 85 96 A. sativa 2007 261 for quality 10 in 57 90 257 2 March, 49 2009, 97 17 AGRI GENRES 061: Avena genetic resources human consumption (AVEQ), 02 -04 Quedlinburg 6 80 10

Fusarium [%] results species wise – wild species DEU FRA POL 0, 5 3, 5 6, 5 1 2, 0 4 2, 04 0 2008 3 1, 5 3 5, 5 2 0 0 0 A. brevis 2008 2 0, 5 0, 75 1 2 0 0 0 A. canariensis 2007 1 2 2 2 0 2008 1 2 2 2 1 A. clauda 2007 1 3 3 3 0 A. damascena 2007 0 A. fatua 2007 8 0, 5 2008 1 1 0, 5 2, 41 2007 1 0, 5 2008 3 0 Min 0 0 0 1 3, 5 0, 5 4 0, 54 6 9 0, 0 6 0, 5 0 4 7 0 0, 5 0 0 0, 83 2 Max 3 N Avg 2007 N Min A. barbata A. hirtula Max Year N Avg Species 2 3, 5 AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ), 02 -04 March, 2009, Quedlinburg 4 4, 5

Fusarium results [%] species wise – cultivated species DEU POL 0 0 0 2 Max 6, 5 Avg 2, 33 Min Max N Avg N Min Max Year N Avg Min Species FRA 1 4, 25 7, 5 Marginally cultivated species A. 2007 3 3, 5 4, 17 2008 5 0, 5 5 3 6, 5 3 abyssini ca A. 2, 7 1 abyssini ca A. strigosa byzantina 2007 21 0, 5 2, 36 Cultivated 6 21 oats 0, 5 0, 62 2007 16 0, 5 1 2, 32 3, 47 5, 5 6, 5 32 16 03 1, 78 2008 39 0 A. byzantina 2008 25 1 3, 88 A. sativa 2007 261 0 3, 39 11 257 8 22 0, 5 0 1 2, 49 4, 53 5 6 6 0 29 0 0, 5 03 7, 8 17, 5 9, 1 2, 1 15, 5 8 7 6, 9 18 24 0, 5 20, 5 173 9 1 7, 5 28 GENRES 061: 2008 Avena genetic resources A. AGRI sativa 295 0 for quality 3, 7 in human 13 consumption 263 (AVEQ), 0 02 -04 0 March, 2009, 0 Quedlinburg 263 0 6, 8 26

Germination check results compared with germination test 2007/2008 Species Accession barbata DEU 14 6 clauda BGR 00 A 2 E 0003 1 AVE 850 FRA Germinabili Chec ty k (2007/2008) POL Chec k DEU Check Side open DEU Check Top open 74, 5 (DEU) 75 89, 5 80 78 19 (DEU) 52 50, 5 62 65 damascen POL 00 a 3 PL 52345 100 (POL) 32, 5 97, 5 fatua DEU 14 6 AVE 2671 80 (DEU) 47, 5 79, 5 83, 5 82 fatua DEU 14 6 AVE 2804 52 (DEU) 63 81, 5 88 81, 5 sterilis DEU 14 6 AVE 3066 69, 5 (DEU) 77, 5 71, 5 AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ), 02 -04 March, 2009, Quedlinburg 94, 5 86

Germination check results compared with germination test 2007/2008 Species FRA Germinabili Chec ty k (2007/2008) Accession POL Chec k DEU Check Side open DEU Check Top open Marginally cultivated species abyssinica BGR 00 BGR 1 8133 48 (DEU) 25 54, 5 35 44 abyssinica DEU 14 6 AVE 4711 57 (DEU) 78 78 80 82 strigosa RUS 00 1 20011256 2 13 (DEU) 73, 5 80 79 77 strigosa POL 00 3 PL 51277 84 (POL) 60, 5 88 strigosa DEU 14 6 AVE 436 24 (DEU) 13, 5 78 29, 5 76 84 67, 5 80, 5 Cultivated oats byzantina DEU 14 6 AVE 3095 84 (POL) AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ), 02 -04 March, 2009, Quedlinburg 77, 5

Conclusion Main problem reducing germinability: Excess of water! AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ), 02 -04 March, 2009, Quedlinburg

Thanks for your attention AGRI GENRES 061: Avena genetic resources for quality in human consumption (AVEQ), 02 -04 March, 2009, Quedlinburg