Microsoft Word - 13 Murariu_Gontariu Lucrare.doc Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefan cel MareUniversity - Suceava Volume XI, Issue 1 – 2012 82 COLD TO LER AN CE IN EU RO PEAN O A T GEN ETIC RES OU RC ES Danela MURARIU1, Marius MURARIU2, *Ioan GONTARIU3 1Suceava Genebank , B-dul 1 Decembrie 1918, nr. 17, Suceava, Romania, e-mail dmurariu@suceava.astral.ro 2Agricultural Research and Development Station of Suceava, B-dul 1 Decembrie 1918 nr. 17 Suceava, Romania, e-mail marius_murariu2005@yahoo.com 3Faculty of Food Engineering, Stefan cel Mare University, Street. Universitatii no. 13, 720229, Suceava, Romania, e-mail ioang@fia.usv.ro *Corresponding author Received 19 January 2012, accepted 20 February 2012 Abstract: Oat is a crop with an important European history and tradition. The high value of oat in human nutrition, which is unique among cereals, is widely recognized and confirmed by health claims issued in various countries. It is based on a high content and quality of proteins, considerable content of fat with high proportion of polyunsaturated fatty acids, high contents of dietary fiber, especially the soluble, highly viscous mixed linked (1->3)(1->4)-ß-D-glucans, which hypo-cholesterolemic effects, minerals and antioxidants, especially tocopherols, tocotrienols and avenanthramides. Oat is considered moderately susceptible to Fusarium spp. infection and mycotoxin contamination. Nevertheless T2/HT-2 toxins raise considerable concerns in the European health administration (European Commission, 2006). Among winter cereals, oat is the most frost sensitive and its insufficient level of winterhardiness is the most important factor limiting the sowing of winter oat in Northern and Central Europe (Cattivelli et al., 1998). Resistance to low temperatures, which limited yield potential and sustainable oat growing in many major oat growing regions in Europe, was the main aspect which have been evaluated in this paper. The results of the evaluation of frost tolerance at first leaf stage, accomplished in the laboratory of Experimental Institute for Cereal Research, Fiorenzuola d’Arda, Italy were compared with the data of winter hardiness obtained in the field experiments carried out by Suceava Genebank in environments more frequently subjected to severe frost events, as compared to the Italian site Keywords: cold tolerance, winter hardiness, genotypes 1. Introduction In order to improve the utilisation of the oat germplasm collection in breeding programs, apart from biochemical and molecular characterisation, an extensive evaluation of important agronomic traits is also required. Resistance to abiotic stress is a crucial aspect for cultivar adaptation to agriculture environments. Among winter cereals, oat is the most frost sensitive and its insufficient level of winterhardiness is the most important factor limiting the sowing of winter oat in Northern and Central Europe [1]. Winter hardiness can be defined as the ability to survive throughout the winter. By the virtue of the wide range of stressful conditions that a plant may experience during the cold season, winter hardiness is a complex trait. Freezing temperature is the most relevant stress factor, although other stress situations, such as anoxia due to excess water or to ice encasement and photoinhibition due to the combination of light and low temperature, may also occur. In temperate areas, winter cultivars are preferred, whenever possible, as winter Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefan cel MareUniversity - Suceava Volume XI, Issue 1 – 2012 83 varieties are higher-yielding than spring ones, and the identification of new gene sources for frost tolerance is still an important task for oat improvement [2]. Despite the fact that genetic diversity has also been observed for freezing tolerance of non-acclimated barley plants, the ability of overwintering plants to withstand cold is mainly based on an adaptive response, known as cold acclimation or hardening, activated during growth at low non- freezing temperatures. Frost resistance can be assessed through field evaluation methods, a strategy depending of the occurrence of natural conditions which satisfactorily differentiated genotypes, or through artificial freezing tests such as percentage of post-stress survival, LT50 (temperature at which 50% of the population is killed), integrity of cell membranes after freezing [3], evaluation of the Fv/Fm ratio the maximum quantum yield of the PSII photochemistry after freezing [4]. A number of physiological traits such as proline accumulation, ABA and crown fructan content have been associated with the development of frost tolerance. Generally, the amount of these metabolites increases during acclimation, being higher in frost-tolerant genotypes versus frost- susceptible ones [5]. A comparison of a number of frost resistance evaluation methods in different crop species was carried out by [6]. The value of the laboratory tests appears to vary with the plant species, although for cereals frost resistance evaluation methods based on freezing assays provide the highest correlation with field survival. 2. Materials and methods Winter hardiness was assessed in a field experiment at Suceava for 104 oat accessions (including 9 standards - Mures, Ivory, Jaak, Argentina, Mina, Belinda, Saul, Genziana, Auteuil). The seeds were sown manually into one row of 1m length (50 seeds per row) in two replications with a distance between accessions of 25 cm. Time of sowing was October 1st 2009. The following data were recorded:  Number of emerged seeds.  Number of survived plants after winter  The field injury have been visually estimated on a 0-9 scale according to Rizza et al. (1994), where: 0: no damage; 1: slightly yellowed leaf tips; 2: half yellowed basal leaves; 3: fully yellowed basal leaves; 4: whole plants slightly yellowed; 5: whole plants yellowed and some plants withered; 6: whole plants yellowed and 10% plant mortality; 7: whole plants yellowed and 20% plant mortality; 8: whole plants yellowed and 50% plant mortality; 9: all plants killed.  The environmental conditions were registered in the target period (October 2009 – March 2010). All obtained results in the field were compared with obtained results in the laboratory of the Experimental Institute for Cereal Research, Fiorenzuola d’Arda, Italy The used method for frost tolerance was the chlorophyll fluorescence evaluation (Fv/Fm) on a set of 104 oat cultivars, at the temperature on -10°C and -12°C. 3. Results and discussion A minimum air temperature of -24.7°C (-31°C soil temperature) was measured in January and severe freezing conditions persisted from December 2009 to March 2010 (Table1). Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefan cel MareUniversity - Suceava Volume XI, Issue 1 – 2012 84 Table 1 The decadal temperatures registered at Suceava Meterological Centre, Romania, during period October 2009- March 2010 High damage occurred for most accessions. For 91 out of 104 accessions all plants were killed showing a visual score of 9 (Table 2): For 8 accessions a score of 8 (whole plants yellowed and 50% plant mortality) was recorded. Only five accessions were identified as superior for winter hardiness capacity showing a value October, 2009 Aer temperatures Soil temepratures P D ec ad es Average Max Min 7 h. a.m avera ge Max Min RH (%) Sun light (h) mm Decade 1 12.4 19.3 7.3 8.7 12.2 23.4 5.9 79 39.0 12.4 Decade 2 6.0 10.3 2.5 4.4 5.9 12.3 2.1 83 27.7 35.2 Decade 3 7.8 11.8 5.0 5.1 7.6 14.6 4.0 86 40.4 15.0 Monthly average 8.7 13.7 4.9 6.0 8.5 16.7 4.0 83 107.1 62.6 November 2009 Decade 1 3.3 5.5 1.4 2.7 3.6 6.4 1.8 93 4.4 7.9 Decade 2 6.8 10.5 3.7 4.2 6.0 10.8 3.0 87 22.3 4.4 Decade 3 6.5 12.8 2.2 1.0 4.3 12.3 0.1 80 50.2 6.3 Monthly average 5.5 9.6 2.4 2.6 4.7 9.8 1.6 87 76.9 18.6 December 2009 Decade 1 2.4 4.7 0.6 1.4 2.5 5.2 0.9 97 9.2 0.2 Decade 2 -8.0 -6.1 -9.8 -1.2 -0.9 -0.2 -1.3 93 6.0 20.9 Decade 3 -1.3 2.4 -5.3 -4.1 -2.8 1.4 -5.9 87 22.7 16.0 Monthly average -2.3 0.4 -4.8 -1.4 -0.5 2.1 -2.2 92 37.9 37.1 January 2010 Decade 1 -2.4 0.0 -4.6 -3.8 -2.8 -0.6 -5.2 96 3.8 8.7 Decade 2 -5.1 -3.7 -5.8 -5.0 -4.4 -2.3 -6.0 97 0.0 19.0 Decade 3 -13.1 -9.2 -16.9 -17.5 -13.5 -6.0 -20.3 86 30.3 3.7 Monthly average -7.0 -4.4 -9.4 -9.1 -7.1 -3.1 -10.8 93 34.1 31.4 February 2010 Decade 1 -7.9 -4.3 -11.4 -12.3 -8.9 -2.6 -14.6 87 36.5 1.4 Decade 2 -2.1 0.8 -5.3 -4.6 -2.6 1.6 -6.6 98 15.4 30.0 Decade 3 0.3 4.4 -2.0 -2.3 -0.6 3.4 -3.4 93 18.6 3.8 Monthly average -3.5 0.0 -6.5 -6.7 -4.3 0.6 -8.5 92 70.5 35.2 March 2010 Decade 1 -2.9 1.3 -5.8 -5.9 -3.2 2.7 -8.4 86 27.4 Decade 2 0.6 5.8 -4.2 -3.9 -0.9 6.0 -7.0 73 49.6 2.2 Decade 3 9.2 15.5 4.3 5.6 8.7 18.1 2.6 69 81.9 16.8 Monthly average 2.6 7.8 -1.7 -1.2 1.8 9.2 -4.0 76 158.9 28.6 Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefan cel MareUniversity - Suceava Volume XI, Issue 1 – 2012 85 of 6 (plants yellowed and up to 10% plant mortality) for 83/200-CR (coming from Bulgaria) and Local 7 (coming from Russia) and value of 7 (whole plants yellowed and between 10 to 20% plant mortality) for Millennium (coming from Great Britain), Ava and Donata (coming from Italy). Table 2 The winter hardiness capacity of oat analysed genotypes St at us o f s am pl e G en ot yp e N o. o f e m er ge d se ed s N o. o f s ur vi va l pl an ts a ft er w in te r Sc or e ac co rd in g R iz za , 1 99 4, im m ed ia te ly a ft er w in te r St at us o f s am pl e G en ot yp e N o. o f e m er ge d se ed s N o. o f s ur vi va l pl an ts a ft er w in te r Sc or e ac co rd in g R iz za , 1 99 4, im m ed ia te ly a ft er w in te r modern Cavallo 14.5 0 9 modern Raven 19.5 0 9 modern Monarch 14.5 0 9 modern Flaming 30 0 9 modern Efesos 18.5 0 9 modern Flamingsg 21 0 9 modern Effektiv 21.5 0 9 modern Nelson 27 0 9 modern Ehostar 21 0 9 Standard Ivory 34 0.5 9 Standard Mina 19.5 0 9 modern Tyfhon 20.5 0 9 cultivated 83/200-C 11.5 8.5 6 modern Pergamon 18 0 9 cultivated BGR 250 17.5 0 9 cultivated Konradin 14.5 0 9 cultivated Sofia 121 14 0 9 cultivated Nuernberg 25.5 0 9 cultivated BGR 7982 22 3.5 8 cultivated Mayer A 22.5 0 9 modern Veli 11 0 9 cultivated Lueneburg 31.5 0 9 cultivated Brnensky 20.5 1 8 modern Sandokan 32.5 0 9 modern Abel 21 0 9 modern Kaplan 25.5 0 9 modern Auron 22 0 9 modern Leo 22 0 9 modern Neklan 16.5 0 9 cultivated Alo 29.5 0 9 modern Istra 14.5 0.5 9 modern Nelson 27 0 9 modern Izak 21.5 0 9 cultivated Platek 18.5 0 9 cultivated Dalimil 19.5 0 9 Modern Breton 15 0 9 Standard Saul 17 0 9 Modern Cwal 9.5 0 9 cultivated Konradin 14.5 0 9 Modern Hetman 3 0 9 cultivated Nuernberg 25.5 0 9 Modern Sam 11.5 0 9 cultivated Mayer A 22.5 0 9 cultivated Acmariu 3 29.5 0 9 cultivated Lueneburg 31.5 0 9 Standard MURES 30.5 0 9 Modern Sandokan 32.5 0 9 cultivated Sacel 25 0 9 Modern Kaplan 25.5 0 9 cultivated Lunca Il 9.5 0 9 Modern Leo 22 0 9 cultivated Baisoara 1 24.5 0 9 cultivated Alo 29.5 0 9 cultivated Local 1 21 0 9 Standard JAAK 37 0 9 cultivated Winterhaf 17 0 9 cultivated Jogeva 33 0 9 cultivated Local 2 16.5 0 9 Modern Miku 26 0 9 cultivated Local 3 10.5 0 9 Modern Villu 24.5 0 9 cultivated Local 4 19.5 0.5 9 cultivated Noire 31 0.5 9 cultivated Mulyat 22.5 0 9 cultivated Noire Ri. 15 0 9 cultivated Jari Oves 28.5 0 9 cultivated Neu Gro 14 2 8 cultivated Beltckii 1 23.5 0 9 cultivated Grise 31.5 0 9 cultivated Kinelskjj 29 1.5 8 cultivated Jaune 16 0 9 cultivated Local 5 20 0.5 9 Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefan cel MareUniversity - Suceava Volume XI, Issue 1 – 2012 86 The results of the evaluation of frost tolerance at first leaf stage, accomplished in the laboratory of Experimental Institute for Cereal Research, Fiorenzuola d’Arda, Italy were compared with the data of winter hardiness obtained in the field experiments carried out by Suceava Genebank in environments more frequently subjected to severe frost events, as compared to the Italian site. Correlation coefficients (r) are reported in Table 3 between the frost tolerance data obtained in two independent experiments under controlled conditions at stress temperature of -10 and -12°C and the winter hardiness evaluated during 2009- 2010 in Suceava, Romania, (by visual score and number of plants that survived) The correlations between the different evaluations of winter hardiness are also reported to compare the results obtained in the different fields and with different methods. Table 3 Correlations of frost tolerance results on the oat accessions analyzed in the laboratory conditions (Experimental Institute for Cereal Research, Fiorenzuola d’Arda, Italy) and in field conditions (Suceava Genebank) (2009 – 2010) WINTERHARDINESS Suceava Genebank field 2009-10 Visual score (0-9 Rizza et al.) WINTERHARDINESS Suceava Genebank field 2009- 10) number survived plant FROST TOLERANCE Fv/Fm means two experiments (-10 and -12°C) -0.447*** 0.432*** WINTERHARDINESS Suceava Genebank field 2009-10 Visual score (0-9 Rizza et al.) -0.951*** *,**,*** significant at the 0.05, 0.01 and 0.001 probability level, respectively; ns non significant ( 0.05 level) cultivated Joanette 26.5 0 9 cultivated Local 6 33.5 0 9 Modern Chantilly 23.5 0 9 cultivated Local 7 19 12.5 6 Standard Auteuil 32 0 9 cultivated Signal 26.5 0 9 Modern Aintree 0 0 9 cultivated Anchar 10 0 9 Modern Lennon 11 0 9 cultivated Vendelin 22.5 0 9 Modern Millenium 10.5 5.5 7 cultivated Gagybator 21 3 8 Modern Ava 10.5 4 7 modern Detvan 8.5 0 9 Modern Donata 12.5 4.5 7 cultivated Zvolen 14.5 0 9 Modern Primula 9.5 0 9 cultivated SVKPOL 16.5 0.5 9 Modern Bionda 9.5 2 8 cultivated UKRKAR 25.5 0 9 Modern Teo BD40 16.5 0 9 cultivated Sisko 26 0 9 Standard Gentiana 13 2.5 8 cultivated Fyris 16.5 0 9 Modern Novella 6.5 0 9 cultivated Seger li 13 0 9 Standard ARG 14.5 2 8 cultivated Purhavre 12 0 9 cultivated 1404-11 15 0 9 cultivated Nidar li 24.5 0 9 Modern Jaugila 17.5 0 9 modern Cilla 27 0 9 cultivated Stendes 21.5 0 9 Standard Belinda 26 0 9 cultivated Pulawski 28 0 9 modern SW Betw 12 0 9 cultivated Gorski 25.5 0 9 modern Kentucky 22 0 9 cultivated Persidskij 13.5 0 9 modern SW Iborg 20.5 0 9 cultivated Platek 18.5 0 9 cultivated OMSKIJ 27.5 0 9 Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefan cel MareUniversity - Suceava Volume XI, Issue 1 – 2012 87 4. Conclusion 1. The high severity of the conditions recorded in the site of Suceava Genebank did not allow to record high variability among the accessions (visual score 9 for 87.5% of the accessions) but it was very useful in showing few genotypes with plant survival (6, 7) that were identified as superior for frost tolerance also in the laboratory tests recorded in the Experimental Institute for Cereal Research, Fiorenzuola d’Arda, Italy. 2. Results obtained from laboratory tests on frost tolerance of 104 accessions were highly correlated with data for winter hardiness obtained in Romania for the same group of accessions. 5. References [1]. Cattivelli L., Crosatti, C., Rizza F., 1998, - Oats. In: Italian Contribution to Plant Genetics and Breeding. (G.T. Scarascia-Mugnozza and M.A. Pagnotta, eds.). Tipografia Quatrini A. & F. snc - Viterbo, Italy, Publisher. pp. 257-308. [2]. Stanca A M, Romagosa I, Takeda K, Lundborg T, Terzi V, Cattivelli L, 2003, - Diversity in abiotic stresses. In: von Bothmer R, Knüpffer H, van Hintum T, Sato K (eds.) Diversity in barley (Hordeum vulgare L.). Elsevier, pp. 179-199. [3]. Rizza F., Crosatti C., Stanca A. M., Cattivelli L., 1994 - Studies for assessing the influence of hardening on cold tolerance of barley genotypes. Euphytica, 75: 131-138. [4]. Rizza F., Pagani D., Stanca A. M., Cattivelli L., 2001 - Use of chlorophyll fluorescence to evaluate the cold acclimation and freezing tolerance of winter and spring oats. Plant Breeding, 120: 389- 396. [5]. Murelli C., Rizza F., Marinone F., Albini A., Dulio V., Terzi V., and Cattivelli L, 1995, - Metabolic changes associated with cold- acclimation in contrasting cultivars of barley. Physiol. Plant. 94: 87-93. [6]. Pulli S., Hjortsholm K., Larsen A., Gudleifsson B., Larsson S., Kristiansson B., Hömmö L., Tronsmo A.M., Ruuth P., and Kristensson C., 1996, - Development and evaluation of laboratory testing methods for winterhardiness breeding. Nordic Gene Bank.