Apple seeds treated with variable intensity doses of irradiation produce  alterations of apple heredity, which are generally characterized by the  reduction of tree vigour, change of fructification type, crop capacity,  fruit shape, colour and biochemical fruit content. Morphological and  productive features of trees derived through mutagenesis and fruit  quality were studied on twelve apple genotypes, using Jonathan and  Golden Delicious cultivars as control. The analysis of apple genotypes  growing potential obtained through mutagenesis showed that there were  significant differences in the tree height, trunk diameter, crown  diameter and fructification type among the apple trees. Several  genotypes (G-1/7, G-1/8, G-3/123, G-4/131, G-9/11) were registered with a  high content of total nitrogen and protein in the fruits and G-9/11  genotype had the highest content of phosphorus (0.408%). Genotypes  G-4/101 and G-4/131 accumulated a high quantity of nitrates, over the  maximum admitted levels of 60 ppm NO3- for fresh fruits. Cu and Zn  contents were low, under the maximum admitted limits

Gabriela NEATA

Gheorghe CAMPEANU

Gina DARJANSCHI

Rodica STAN

Notulae Botanicae Horti Agrobotanici Cluj-Napoca

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Available online at www.notulaebotanicae.roNot. Bot. Hort. Agrobot. Cluj 38 (1) 2010, 248-251Print ISSN 0255-965X; Electronic 1842-4309Notulae Botanicae Horti AgrobotaniciCluj-NapocaTree and Fruit Characteristics of Various Apple Genotypes Obtained through MutagenesisGheorghe Campeanu1), Gabriela neaţa1), Gina DaRJanSCHI1), Rodica STan2)1)University of Agronomic Sciences and Veterinary Medicine, Blvd. Mărăşti 59, sect. 1, 011464, Bucharest, Romania; neatag@yahoo.com2)University of Agricultural Sciences and Veterinary Medicine, Cluj-Napoca, 3-5 Manastur St., 400372, Romania; ro7si@yahoo.com AbstractApple seeds treated with variable intensity doses of irradiation produce alterations of apple heredity, which are generally characterized by the reduction of tree vigour, change of fructification type, crop capacity, fruit shape, colour and biochemical fruit content. Morphological and productive features of trees derived through mutagenesis and fruit quality were studied on twelve apple genotypes, using ‘Jonathan’ and ‘Golden Delicious’ cultivars as control. The analysis of apple genotypes growing potential obtained through mutagenesis showed that there were significant differences in the tree height, trunk diameter, crown diameter and fructification type among the apple trees. Several genotypes (‘G-1/7’, ‘G-1/8’, ‘G-3/123’, ‘G-4/131’, ‘G-9/11’) were registered with a high content of total nitrogen and protein in the fruits and ‘G-9/11’ genotype had the highest content of phosphorus (0.408%). Genotypes ‘G-4/101’ and ‘G-4/131’ accumulated a high quantity of nitrates, over the maximum admitted levels of 60 ppm NO3- for fresh fruits. Cu and Zn contents were low, under the maximum admitted limits.Keywords: fruit features, mutation, chemical contentIntroductionIn apple breeding, obtaining new genotypes by mu-tagenesis induction represents an important and useful method for creating new cultivars (Shidakov, 1991; Jan-ick et al., 1996; Predieri, 2001). Unlike natural mutations, which occur with a very low rate of frequency, artificial mutations significantly increase the frequency of mutant plants appearance ( Jain, 2010), but the same characteris-tics are affected. Induced mutations, as natural ones, can occur in a single gene that can cause a distinct trait, or act altering the degree of ploidy.Mutations that produce single gene differences can be most easily induced peculiarities affecting fruit colour and quality, type of growth, plant size (reduction), blooming time, fructification of trees (Broertjes, 1977; Janick et al., 1996), despite the low number of cultivars that became established commercial varieties (Predieri, 2002). “Spur” forms (e.g. ‘Golden spur’, ‘Yellow spur’, ‘Wellspur’ etc.), various fruit colours, quality and maturation were ob-tained in apple by induced mutagenesis (Sestras, 2004).Mutations can be induced through physical or chemi-cal treatments on seeds, buds, stems, pollen. Physical mu-tagenic agents like X-rays can be used, gamma rays or ther-mal neutrons respectively; treatments can also be applied in dormant branches (3-5 Kr), or active buds (2-4 Kr). Thermal neutron treatments are carried out with doses of 3.9-15.6 x 1012 neutroni/cm2 ( Janick et al., 1996).Although scientific interest in mutation breeding has drastically decreased during the last years, mutation in-duction continues to be an attractive method for creating genetic variability (Schum, 2003) and to produce basic material for intravarietal selection on apple (Zwintzscher, 1977). New apple cultivars, as ‘Redix’, ‘Irisem’, ‘Iris’, ‘Real’ were obtained and promoted in Romania, using mutagen-esis (Petre, 2008). Using gamma rays from 60Co source, mutants of apple rootstocks were obtained and have been registered in Poland (Przybyla et al., 2009).Materials And MethodsThe main characteristics of tree and fruit quality re-garding useful chemical components of twelve apple gen-otypes obtained through mutagenesis were studied at the Fruit Research Station Voinesti, Romania. The biological material was obtained through post-maturated seeds, ir-radiated with 5000-80000 R doses, from open pollinated ‘Prima’ cultivar, with genetically resistance to apple scab (Vf), which has a good response in Romanian ecological conditions (Sestras et al., 2005).From the irradiated biological material, existing in the selection field, the most valuable elites were selected, grafted on MM 106 rootstock and studied in a compara-tive trial with 1250 trees/ha (Petre, 2008). Measurements of trees growth were height of trees, trunk diameter and crown diameter. Biochemical determination were water content, total dry matter, total sugar, acidity, vitamin C and mineral elements - total nitrogen, protein, phospho-rus, potassium, calcium, iron,  and heavy metals Cu, Zn, Pb, Cd.Campeanu, G. et al. / Not. Bot. Hort. Agrobot. Cluj 38 (1) 2010, 248-251249differences in height, trunk diameter, crown diameter and fructification type of apple trees.According to the trunk diameter there are small vig-orous genotypes like ‘G-6/42’, middle ‘G-1/8’, large in ‘G-4/101’ and super large in ‘G-3/123’. The highest trees have upright trunks, with ‘G-1/123’ genotype and the ones with small trunks are from ‘G-6/42’ genotype. Large crowns were presented ‘G-3/123’ and ‘G-9/11’ genotypes and tight crowns ‘G-6/42’ and ‘G-5/56’ genotypes.It is likely that tree force differences recorded among varieties be due to gene recombinations that arise from crosses among various genotypes, as the force of the apple tree is a feature with a polygenic determinism ( Janick et al., 1996; Sestras et al., 2009).Biochemical features were also studied. Medium values of biochemical characteristics of apple genotypes and were In order to obtain water content and dry matter the drying closet method at 105°C, sugar content Abbe meth-od, total acidity through volumetric method with NaOH 0.1n, vitamin C iodometric method, nitrogen content with Kjeldahl, phosphorus with colorimetric method and potassium with flam photometric method were used. Heavy metal contents were determined with wet mineral-ization and spectrophotometer method.The genotypes considered mutants, were studied along with ‘Jonathan’ and ‘Golden Delicious’ cultivars, untreat-ed, two of the most cultivated and well-known apple culti-vars in Romania (Mitre et al., 2009).Results and discussionsThe growing potential analyses of some apple genotypes obtained through mutagenesis induction show significant Tab. 1. The vigour of apple trees and productivity (fruit yield)No Genotypes Trunk diameter(cm)Tree height(cm)Crown diameter(cm) Tree vigourFruit yield(kg/tree)1 ‘Jonathan’ 34.2b 485ab 350b Medium 18.7c2 ‘Golden Delicious’ 35.8ab 486ab 310bc Medium 23.4b3 ‘G-1/7’ 34.1b 456b 275c Medium 28.3ab4 ‘G-1/8’ 34.3b 420c 230d Medium 25.6b5 ‘G-1/26’ 33.1c 380de 240d Medium 26.5ab6 ‘G-3/123’ 36.7a 460b 460a Very large 32.5a7 ‘G-4/50’ 32.1c 510a 204de Large 18.0c8 ‘G-4/56’ 34.8b 495a 215de Medium-Large 16.6cd9 ‘G-4/101’ 32.6c 489ab 240d Large 15.5d10 ‘G-4/131’ 33.1c 401cd 310bc Medium 28.1ab11 ‘G-5/56’ 35.2b 495a 210de Large 14.5d12 ‘G-5/79’ 34.3b 480ab 350b Medium-Large 17.4cd13 ‘G-6/42’ 30.6d 365e 190e Low 25.6b14 ‘G-9/11’ 32.5c 385de 330b Medium 18.3cTab. 2. The biochemical features of apple treesNo Genotypes Water(%)Dry matter(%)Total sugar(%)Acidity(%)Vitamin C(mg/100g fruit)Mineral substances(%)1 ‘Jonathan’ 86.30 13.70 10.61 0.210 9.21 1.662 ‘Golden Delicious’ 85.90 14.10 11.36 0.156 12.63 1.933 ‘G-1/7’ 85.60 14.40 12.10 0.188 8.31 1.864 ‘G-1/8’ 85.10 14.90 11.80 0.195 15.20 1.255 ‘G-1/26’ 86.90 13.10 12.03 0.244 17.31 1.766 ‘G-3/123’ 87.34 12.66 9.84 0.345 7.66 1.587 ‘G-4/50’ 88.46 11.54 9.53 0.173 7.89 1.738 ‘G-4/56’ 86.20 13.80 9.75 0.214 8.75 2.109 ‘G-4/101’ 86.30 13.70 10.96 0.310 19.21 2.3010 ‘G-4/131’ 85.90 14.10 9.81 0.293 9.35 2.3011 ‘G-5/56’ 87.62 12.38 10.00 0.195 7.56 1.9812 ‘G-5/79’ 85.18 14.82 9.53 0.157 17.40 1.2613 ‘G-6/42’ 89.32 10.68 10.94 0.232 7.67 1.3514 ‘G-9/11’ 90.15 9.85 9.53 0.127 7.19 1.36Campeanu, G. et al. / Not. Bot. Hort. Agrobot. Cluj 38 (1) 2010, 248-251250influenced by the mutagenesis process. They are presented in Tab. 2.The highest values of the water content of fruits were registered in genotypes ‘G-9/11’, ‘G-6/42’ and ‘G-4/50’; values were between 88.46-90.15%. The highest content of dry matter were in genotypes ‘G-5/79’, ‘G-5/56’ and ‘G-6/42’ and the smallest contents were in genotypes ‘G-9/11’, ‘G-6/42’.Regarding the sugar content, there was normal accu-mulation in all genotypes comparatively with values ob-tained in ordinary orchards (Sestras et al., 2005) or even as biological crop (Campeanu et al., 2009). High contents of sugar were registered in ‘G-1/7’ and ‘G-1/26’.Vitamin C contents had superior values in all vari-ants comparatively with ‘Jonathan’. In genotypes ‘G-1/8’, ‘G-1/26’, and ‘G-5/79’ vitamin C contents were double and triple comparatively with the content of ‘Jonathan’ and ‘Golden Delicious’.The values of acidity contents were near the control values with the exception of genotypes ‘G-1/8’, ‘G-5/56’ and ‘G-6/42’ in which acidity had higher values.Mineral elements contents were higher in value in fruits from genotypes ‘G-4/56’ , ‘G 4/131, and ‘G-4/101’.The mineral elements analyses (Tab. 3) showed a high content of total nitrogen and protein in the apples of genotypes ‘G-1/7’, ‘G-1/8’, ‘G-3/123’, ‘G-4/131’, ‘G-9/11’ and a small content in variants ‘G-6/42’, ‘G-4/56’, ‘G-4/50’ and ‘G-5/79’. With the exception of genotype ‘G-9/11’ which had the higher content of phosphorus, with the value 0.408%, apples of the other genotypes presented the same values.The content of other mineral elements analysed were in normal variability for apple species but there were not high differences between genotypes.Apples from genotypes ‘G-4/101’ and ‘G-4/131’ were predicted to accumulate high quantities of nitrates and ‘G-6/42’ and ‘G-1/26’ accumulated small quantities of nitrates. All nitrates quantities accumulated in apple geno-types were over the maximum admitted levels of 60 ppm NO3- mentioned in the legislation regarding security con-sumption conditions for fresh fruits.The accumulation levels of Cu and Zn were low (Tab. 4), under the maximum admitted limits for those elements. The levels of lead and cadmium were in trace values, which showed the good fruit quality.ConclusionsThe analyses for the growing potential of some apple genotypes obtained through mutagenesis induction show that there are significant differences in height, trunk di-ameter, crown diameter, and yield among apple variants. The mutagenesis process influenced medium values of biochemical features of apple genotypes. The values of dry matter, acidity, sugar and vitamin C had large variability among variants. Apples from genotypes ‘G-4/101’ and ‘G-4/131’ were mentioned to accumulate high quantities of nitrates. All nitrates quantities accumulated in apple genotypes were over the maximum admitted levels of 60ppm NO3- mentioned in the legislation regarding fresh fruits security consumption conditions. The accumulation levels of Cu and Zn were low and were under the maxi-mum admitted limits for those elements.ReferencesBroertjes, C. (1977). Artificially Induced Genetic Variation in Fruit Trees. Acta Hort. 75:19-26.Campeanu, G., G. Neata and G. Darjanschi (2009). Chemical Composition of the Fruits of Several Apple Cultivars Growth as Biological Crop. Notulae Botanicae Horti Agrobotanici Cluj-Napoca 37(2):161-164.Jain, S. M. (2010). Mutagenesis in Crop Improvement under the Climate Change. Romanian Biotechnological Letters. Tab. 3. Mineral elements apples contentsNo Genotypes Nt%Protein%P2O5%K2O%CaO%FeO%1 ‘Jonathan’ 1.7514 10.60 0.295 0.65 0.451 2.3012 ‘Golden Delicious’ 1.8513 10.75 0.346 0.68 0.255 2.3413 ‘G-1/7’ 2.0131 11.21 0.321 0.75 0.265 2.3074 ‘G-1/8’ 1.9615 10.95 0.314 0.65 0.265 2.2055 ‘G-1/26’ 1.8048 11.24 0.363 0.65 0.148 2.2046 ‘G-3/123’ 1.8514 11.75 0.310 0.55 0.246 2.3057 ‘G-4/50’ 1.5792 9.83 0.3285 0.55 0.276 2.3418 ‘G-4/56’ 1.7776 11.61 0.276 0.45 0.275 2.2059 ‘G-4/101’ 1.9766 11.45 0.2176 0.30 0.252 2.20410 ‘G-4/131’ 2.1714 13.52 0.2765 0.50 0.2450 2.35211 ‘G-5/56’ 1.7672 11.00 0.3285 0.55 0.260 2.21212 ‘G-5/79’ 1.7390 10.83 0.3460 0.65 0.158 1.89613 ‘G-6/42’ 1.2690 7.90 0.3805 0.55 0.210 2.48814 ‘G-9/11’ 2.0304 12.64 0.4150 0.45 0.269 2.526Tab. 4. Components and potential toxic elementsNo Genotypes NO3-(ppm)Cu(ppm)Zn(ppm)Pb(ppm)Cd(ppm)1 ‘Jonathan’ 210.1 3.5 2.3 Trace Trace2 ‘Golden Delicious’ 296.9 2.1 4.2 Trace Trace3 ‘G-1/7’ 189.3 3.2 3.1 Trace Trace4 ‘G-1/8’ 175.4 3.3 3.2 Trace Trace5 ‘G-1/26’ 132.3 3.2 3.8 0.21 Trace6 ‘G-3/123’ 272.1 3.8 3.6 Trace Trace7 ‘G-4/50’ 170.1 4.1 2.2 Trace Trace8 ‘G-4/56’ 189.2 3.5 3.8 0.25 Trace9 ‘G-4/101’ 151.2 2.8 4.2 Trace Trace10 ‘G-4/131’ 173.2 3.1 3.6 Trace Trace11 ‘G-5/56’ 189.0 3.5 3.8 0.25 Trace12 ‘G-5/79’ 170.1 4.3 4.8 0.15 Trace13 ‘G-6/42’ 151.2 3.7 4.9 Trace Trace14 ‘G-9/11’ 170.1 3.6 3.7 Trace TraceCampeanu, G. et al. / Not. Bot. Hort. Agrobot. Cluj 38 (1) 2010, 248-251251Sp. Iss. SI Suppl. S 15(2):88-106.Mitre, I., V. Mitre, M. Ardelean, R. Sestras and A. Sestras (2009). Evaluation of Old Apple Cultivars Grown in Central Transylvania, Romania. Notulae Botanicae Horti Agrobotanici Cluj-Napoca 37(1):235-237.Petre, V. (2008). Apple Tree Breeds and Elites Obtained by Induced Mutagenesis, Suitable for Ecological Cultures. Analele Universitatii din Craiova: Biology, Horticulture, Food Produce Processing Technology, Environmental Engineering XIII (XLIX):193-196.Predieri, S. (2001). Mutation induction and tissue culture in improving fruits. Plant Cell Tissue and Organ Culture 64 (2-3):185-210.Predieri, S. (2002). The Importance of Induced Mutations in Pear Improvement. Acta Hort. 596:161-166.Przybyla, A. A., P. Bielicki and A. Czynczyk (2009). New Polish Vegetative Mutant Rootstocks for Apple. Acta Hort. 814:315-318.Schum, A. (2003). Mutation Breeding in Ornamentals: an Efficient Breeding Method? Acta Hort. 612:47-60.Sestras, R. (2004). Horticultural plant breeding (in Romanian). Ed. AcademicPres, Cluj-Napoca pp.334.Sestras, R., A. Barbos, V. Ghidra, C. Botez, G. Ropan, I. Chis and B. Patrascu (2005). The response of several apple varieties with genetic resistance to apple scab in the centre of Transylvania environment. Bulletin of the University of Agricultural Science and Veterinary Medicine. Horticulture 62:95-99.Sestras, R., D. Pamfil, A. Sestras, L. Jäntschi, S. Bolboaca and C. Dan (2009). Inheritance of Vigour Tree in F1 Apple Interspecific Hybrids. Notulae Botanicae Horti Agrobotanici Cluj-Napoca 37(1):70-73.Shidakov, R. S. (1991). Hybridization Efficiency in Combination with Chemical Mutagenesis in Selection of Apple Trees. Genetika 27(3):556-560.Zwintzscher, M. (1977). Examples for Induced Variation by X- and -Rays in Cox Orange Pippin and other Apple Varieties. Acta Hort. 75:35-42.

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