A population of 150 F2 plants was derived from a cross between two spring oat individuals, one from cv. Aslak (Boreal Plant Breeding LTd., Finland) and the other from cv. Salo (Svalöf-Weibull AB, Sweden). Cadmium was tested by inductively coupled plasma mass spectrometry (ICP-MS) method

Kalendar, Ruslan

Kiviharju, Elina

Schulman, Alan

Tanhuanpää, Pirjo

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A  A major major gene gene for  for grain grain cadmium cadmium accumulation accumulation in oat in oatPirjo Tanhuanp Pirjo Tanhuanpää ää1 1,  , Ruslan Ruslan Kalendar Kalendar2 2, Alan Schulman , Alan Schulman1,2 1,2, Elina Kiviharju , Elina Kiviharju1 11 1MTT Agrifood Research Finland,  MTT Agrifood Research Finland, Biotechnology Biotechnology and Food  and Food Research Research, FI , FI- -31600 Jokioinen, Finland  ( 31600 Jokioinen, Finland  (email email: first name.last  : first name.last name@mtt.fi name@mtt.fi) )2 2MTT/BI Plant Genomics Laboratory, Institute of Biotechnology, FI MTT/BI Plant Genomics Laboratory, Institute of Biotechnology, FI- -00014 Helsinki, Finland  (email:  00014 Helsinki, Finland  (email: first first name.last name.last name name@ @helsinki.fi helsinki.fi) )INTRODUCTION INTRODUCTIONCadmium is a risk factor in cereal crops due to its high toxicit Cadmium is a risk factor in cereal crops due to its high toxicity and accumulation in the body, particularly to liver and kidney y and accumulation in the body, particularly to liver and kidneys, with  s, with associated osteoporosis and cancer. Crop plants uptake  associated osteoporosis and cancer. Crop plants uptake Cd Cd from soil as a trace of fertilizers. Oat ( from soil as a trace of fertilizers. Oat (Avena Avena  sativa sativa  L.) is widely used in  L.) is widely used in baby foods and also adult consumption is increasing due to reduc baby foods and also adult consumption is increasing due to reduction of the risk of heart disease caused by  tion of the risk of heart disease caused by β β- -glucan glucan. The  . The Cd Cd level  level in plants is directly correlated with that in the soil, but is a in plants is directly correlated with that in the soil, but is also affected by genetic factors. lso affected by genetic factors.MATERIALS AND METHODS MATERIALS AND METHODSA population of 150 F A population of 150 F2 2 plants was derived from a cross  plants was derived from a cross  between two spring oat individuals, one from cv.  between two spring oat individuals, one from cv. Aslak Aslak (Boreal Plant Breeding Ltd, Finland) and the other from cv.  (Boreal Plant Breeding Ltd, Finland) and the other from cv. Salo Salo ( (Sval Svalö öf f- -Weibull Weibull AB, Sweden). The parent  AB, Sweden). The parent Salo Salo was  was known to have the tendency to accumulate  known to have the tendency to accumulate Cd Cd in the field (1).  in the field (1). Cadmium accumulation to the grains of the F2 individuals  Cadmium accumulation to the grains of the F2 individuals was tested by was tested by the developed  the developed Cd Cd feeding pot test in the  feeding pot test in the  greenhouse (Fig 1.). Cadmium was tested by  greenhouse (Fig 1.). Cadmium was tested by inductively  inductively coupled plasma mass spectrometry (ICP coupled plasma mass spectrometry (ICP- -MS) MS) method.  method. Bulked Bulked- -segregant segregant analysis (3) was used to find PCR based  analysis (3) was used to find PCR based markers linked to  markers linked to Cd Cd accumulation.  accumulation. MAPMAKER 3.0 software  MAPMAKER 3.0 software (2) was used for determining linkages between markers. The  (2) was used for determining linkages between markers. The QTL (quantitative trait locus) affecting  QTL (quantitative trait locus) affecting Cd Cd accumulation was  accumulation was localized using MAPMAKER/QTL 1.1. localized using MAPMAKER/QTL 1.1.600 800 1000 1200 1400 1600 1800 2000 2200 2400 2600 2800 3000 3200 3400 3600Number of individuals0510152025Cd concentration µg/kg600 800 1000 1200 1400 1600 1800 2000 2200 2400 2600 2800 3000 3200 3400 3600Number of individuals0510152025Cd concentration µg/kg01000200030004000Roots Straw GrainsRESULTS AND DISCUSSION RESULTS AND DISCUSSIONThe amount of  The amount of Cd Cd accumulation is presented in Figs. 2 (parents), and  accumulation is presented in Figs. 2 (parents), and 3  (F2 progeny). The distribution fits the hypothesis of single  3  (F2 progeny). The distribution fits the hypothesis of single gene  gene inheritance, the allele for low accumulation being dominant.  inheritance, the allele for low accumulation being dominant. The parents,  The parents, Aslak Aslak and  and Salo Salo, were analyzed with 218 RAPD, 31 SRAP , were analyzed with 218 RAPD, 31 SRAP,  , and 337 REMAP primers or primer combinations, of which 78 (36%), and 337 REMAP primers or primer combinations, of which 78 (36%), 26 (84%), and 73 (22%) produced polymorphisms, respectively. Fou 26 (84%), and 73 (22%) produced polymorphisms, respectively. Four  r markers (two  markers (two RAPDs RAPDs, one REMAP, and one SRAP) were analyzed in  , one REMAP, and one SRAP) were analyzed in the F the F2 2 population population, and  , and they were found to be statistically highly  they were found to be statistically highly significantly associated with grain  significantly associated with grain Cd Cd concentration. concentration. The markers  The markers were converted into SCAR ( were converted into SCAR (sequence characterized amplified region)  sequence characterized amplified region) markers.  markers.  SCAR AF20 (Fig.4) and SCAR AF15 were the best  SCAR AF20 (Fig.4) and SCAR AF15 were the best markers owing to their capability to identify  markers owing to their capability to identify homozygotes homozygotes for the  for the low ( low (Aslak Aslak) allele (Fig.3).  ) allele (Fig.3). T The four markers were assigned to one linkage group which  he four markers were assigned to one linkage group which exhibited a QTL for grain  exhibited a QTL for grain Cd Cd accumulation (LOD score > 16, the  accumulation (LOD score > 16, the variance variance- -explained  ~50 %). explained  ~50 %).FURTHER PLANS FURTHER PLANSIn future, we intend to identify candidate genes for  In future, we intend to identify candidate genes for Cd Cd accumulation by using bioinformatics, genetics, and  accumulation by using bioinformatics, genetics, and  genomics tools. Our overall goal to understand the  genomics tools. Our overall goal to understand the  mechanisms and key genes responsible for  mechanisms and key genes responsible for Cd Cd accumulation in oat. Practical goal is to produce  accumulation in oat. Practical goal is to produce  molecular markers tightly linked to the gene to enable  molecular markers tightly linked to the gene to enable the marker assisted the marker assisted selection of low selection of low- -accumulating  accumulating cultivars in breeding programmes for healthy oat  cultivars in breeding programmes for healthy oat  cultivars. cultivars.Fig. 2. Cd concentration of roots, straw and grains in Aslak (□) and Salo (■) after Cd feeding pot test. Means with standard error bars, except in roots where no replicates existed.Fig. 3. Grain Cd concentration in the F2 population based on markers SCAR AF20 (a) and SCAR AF15 (b). Marker genotypes are presented as fills in the columns:  □ = homozygotes for Aslak allele, hatched = homozygotes for Salo allele, ■ = heterozygotes (SCAR AF20) or homo- and heterozygotes (SCAR AF15) for Salo allele.Fig. 4. Codominant SCAR marker AF20. A=Aslak, S=SaloREFERENCES1. Eurola M, Hietaniemi V, Kontturi M, Tuuri H, Pihlava J-M, Saastamoinen M, Rantanen O, Kangas A, Niskanen M (2003) Cadmium contents of oats (Avena  sativa  L.) in official variety, organic cultivation, and nitrogen fertilization trials during 1997-1999. J Agric Food Chem 51:2608-26142. Lander ES, Green P, Abrahamson J, Barlow A, Daly MJ, Lincoln SE, Newburg L (1987) MAPMAKER: An interactive computer package for constructing primary genetic linkage maps of experimental and natural populations. Genomics 1:174-1813. Michelmore RW, Paran I, Kesseli RV (1991) Identification of markers linked to disease-resistance genes by bulked segregant analysis: a rapid method to detect markers in specific genomic regions by using segregating populations. Proc Natl Acad Sci USA 88:9828-9832a. b.A S12001500bpF2  individualsFig. 1. A preliminary experiment of cadmium accumulation into the oat individuals in the Cd-feeding pot test in the greenhousewww.mtt.fi

A major gene for grain cadmium accumulation in oat

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