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Comparing results of different genetic diversity estimates can be useful in parental selection for plant breeders. Forty winter wheat cultivars, from three Croatian breeding centres and four foreign countries, were used to utilize and compare agronomic, morphologic and molecular based genetic diversity estimates. Ten morphologic descriptors according to UPOV guidelines and eight agronomic traits were used to establish phenotypic data. Molecular data consisted of 26 SSR and four combinations of AFLP markers, covering all three wheat genomes. Agro-morphologic data showed variability especially regarding plant height (CV=18.44%), yield (CV=22.02%), and ear emergence (range=8). Discriminant analysis confirmed grouping among cultivars was mostly influenced by number of days to heading and yield. The four AFLP primer combinations and 26 SSR markers yielded 108 polymorphic bands. The UPGMA based on phenotypic data, arranged cultivars in four clusters, with one distinctive outlier, cultivar U1. The UPGMA based on molecular data also arranged cultivars in four clusters, with one distinctive outlier, cultivar Antonius. The similarities based on all four genetic diversity estimates reflected, on average, the degree of relatedness of cultivars used. No correlations between phenotypic and molecular data were found implying that both types of data should be used for genetic diversity estimates in order to cover wider variability between tested cultivars.

Mosf commonly used correlation coefficients are Pearson's product moment correlation coefficient and Spearman's rank correlation coefficient. The aim of this paper is to compare a Pearson's and Spearman's coefficient of correlation on the same data set. The winter wheat grain cadmium (Cd) concentration was correlated to grain zinc (Zn) concentration, plant height, plant weight, number of spikelets per spike and 1000 kernel weight. Data were collected from the experiment carried out in semi controlled conditions, where genotypic specificity of winter wheat varieties was tested on the grain Cd and Zn accumulation on uncontaminated and Cd contaminated soil. Results showed that selection of most convenient correlation coefficient mostly depends on the type of variables, presence of outliers normality and linearity of relationship.

Cilj ovoga istraživanja bio je ispitati varijabilnost amiloze i amilopektina kod 24 hrvatske i šest stranih sorata ozime pšenice te otkriti potencijal istih za posebne namjene. Analiza sastava škroba temeljila se na izdvajanju amiloze i amilopektina te određivanju njihove količine i omjera. Analiza količine amiloze i amilopektina utvrdila je statistički visoko opravdane razlike među ispitivanim sortama. Ispitivane sorte većinom su krušne pšenice različite kvalitete te imaju uobičajen sadržaj amiloze i amilopektina. Među njima izdvojile su se pojedine sorte s visokim sadržajem amilopektina i niskim sadržajem amiloze te jedna sorta s visokim sadržajem amiloze, koje imaju potencijal u budućim oplemenjivačkim programima i selekciji za posebne namjene., The aim of this study was to investigate the variability of amylose and amylopectin in 24 Croatian and six foreign winter wheat varieties and to detect the potential of these varieties for special purposes. Starch composition analysis was based on the separation of amylose and amylopectin and the determination of their amounts and ratios. Analysis of the amount of amylose and amylopectin determined statistically highly significant differences between the varieties. The tested varieties are mostly bread wheat of different quality which have the usual content of amylose and amylopectin. Some varieties were identified among them with high amylopectin and low amylose content and one variety with high amylose content. They have the potential in future breeding programs and selection for special purposes.

Razvoj i održivost genetske različitosti germplazme ozime pšenice jedan je od osnovnih uvjeta za uspjeh u budućim oplemenjivačkim programima. Odabir divergentnih i genetski različitih roditelja najvažniji je korak u stvaranju novih superiornih sorata. Opasnost od genetske erozije je pogotovo prisutna u manjim oplemenjivačkim programima koji imaju manji ograničenih proizvodnih površina u kojima se selekcija odvija u sličnim uvjetima uzgoja. Upotreba molekularnih markera u procjeni genetske različitosti sorata ozime pšenice je ograničena u hrvatskim oplemenjivačkim programima. Cilj ovoga istraživanja je bio procijeniti genetsku različitost hrvatske sorata ozime pšenice koristeći SSR i AFLP markere. U istraživanje je bilo uključeni 40 sorata ozime pšenice iz hrvatskih i stranih oplemenjivačkih centara. Korišten je set od 26 mikrosatelitnih početnica s kojima se nastojao pokriti genom pšenice sa 42 kromosoma. Prosječna udaljenost iznosila je 0,66. Najveća udaljenost zabilježena je između sorata Zlatna dolina i Lela (dij=0,98), dok su najsličnije sorte bila Super Žitarka i Barbara s genetskom udaljenosti od dij=0,21. Pomoću četiri kombinacije AFLP početnica proizvedeno je ukupno 108 polimorfnih fragmenata sa prosjekom od 34 fragmenta po kombinaciji. Prosječno je utvrđeno 27 polimorfnih alela po kombinaciji i prosječnom vrijednosti PIC od 0,34. Specifični AFLP fragmenti uspješno su razlučili tri sorte pšenice: Žitarku, Super Žitarku i Barbaru te se stoga mogu koristiti za njihovu identifikaciju. Grupiranje sorata bilo je u skladu s njihovim podrijetlom i podacima o pedigreu., Sustaining and developing of genetic diversity in winter wheat germplasm is one of the main prerequisite for success in future winter wheat breeding programs. Selection of diverse parents is essential for creation of superior new varieties. Danger of genetic erosion specially exists in smaller breeding programs and selection for limited production area with similar growing conditions. Croatian winter wheat breeding has a long tradition but it is relatively small compared to countries with bigger growing areas. Use of molecular markers in evaluating genetic diversity of winter wheat varieties is limited in Croatian breeding programs. Therefore, aim of this study was to evaluate genetic diversity of Croatian winter wheat varieties using SSR and AFLP markers as a powerful tool for assessing genetic diversity. Forty winter wheat varieties, from three Croatian breeding centres and foreign centres were included in the study. A set of 26 microsatellite primers were used, covering three wheat genomes and 42 chromosomes by 0.66 average genetic distance. The largest calculated distance was between the varieties Zlatna Dolina and Lela (dij=0.98), while most similar varieties were Super Žitarka and Barbara with the distance value of dij=0.21. Four AFLP marker combinations generated 108 polymorphic bands with average of 34 specific bands per primer combination. On the average 27 polymorphic bands were generated with average PIC value of 0.34. Specific polymorphic bands were discriminant for the three varieties Žitarka, Super Žitarka and Barbara, which, therefore, can be used for their identification. Grouping of varieties was in accordance with their origin (breeding centre) and pedigree data.

Ovaj rad istražuje učinkovitost primjene različitih razina digestiranog ostatka, dobivenog nakon anaerobne digestije (AD) mesno-koštanog brašna (MKB), na morfološka i mineralna svojstva kukuruza. Istraživanje je provedeno u trajanju od dvije godine, a sastojalo se od osam tretmana: kontrola (bez gnojiva); samo mineralna gnojidba; tretmani s najmanjom, srednjom i maksimalnom dozom digestiranog MKB-a; gnojidba minimalnom, srednjom i maksimalnom dozom digestiranog ostatka u kombinaciji s mineralnim gnojivom. Kombinacija tretmana s mineralnim gnojivom i različitom dozom ostatka anaerobne digestije, povećala je prinos mase biljke, bile su veće i imale više lišća. Sadržaj mineralnih tvari bio je prosječne vrijednosti, kako u biljnom materijalu tako i u jezgri. Varijacije vrijednosti uvjetovane su ukupnom masom i ionskim interakcijama. MKB nakon tretmana AD moguće je primijeniti kao organsko gnojivo. Ima veće hranjive vrijednosti i, kao takvo, prikladno je za tretiranje poljoprivrednih biljaka, posebno u ratarstvu., The present paper investigates the effectiveness of different levels of applied digested residue obtained after anaerobic digestion (AD) of meat-bone meal (MBM) on the morphologic and mineral characteristics of corn. The experiment was conducted in duration of two years and consisted of eight treatments: control (without any fertilizer); mineral fertilization only; treatments with minimum, medium and maximum dosage of digested MBM; fertilization of minimum, medium and maximum dosages of AD residue combined with mineral fertilizer. By combination of treatments with mineral fertilizer and different rates of AD residue, plants increased mass yield, they were larger and had more leaves. Content of mineral substances was within average values, both in plant material and in kernel, and variations in values were conditioned by total mass and ion interactions. MBM after AD treatment is possible to be applied as organic fertilizer. It has higher nutritive value and, as such, is suitable for treatment of agricultural plants, especially in crop production.

Spelt (Triticum spelta L.), also known as dinkel wheat or hulled wheat, is one of the oldest known grains and it originates from Asia. It was known even to the Egyptians, the oldest findings of this type of wheat were found in the Nile valley and they date to the fourth millennium BC. This edible, chaff-like, type of wheat was used for food by the ancient Romans who grew it throughout the empire. This cereal was very important from the Bronze Age to the Middle Ages in Europe. However, in recent times it has largely been replaced by other types of wheat (Triticum aestivum and Triticum durum). This grain has again started attracting attention in the past two decades because of the possibility of organic breeding, since it requires lesser use of fertilizers and pesticides compared to the common wheat. Due to the increasing awareness of consumers who pay more attention every day to the variety and quality of food, spelt is increasingly gaining importance with the tendency of multiplying sown areas so the goal of this study was to determine the nutritional characteristics of spelt shelled beans of studied varieties Bc Vigor and Ostro (water content, ash, starch, protein and crude fat) for nutritional use.

Većina postojećih sorti rajčice nastala je klasičnim oplemenjivanjem. Međutim, mnogobrojni problemi koji se javljaju pri procesu oplemenjivanja doveli su do razvoja i primjene novih tehnologija u oplemenjivanju bilja. Jedna od važnijih tehnologija je tehnologija molekularnih markera, koja bi mogla doprinijeti povećanju učinkovitosti procesa oplemenjivanja. Zbog svoje jednostavne genetske strukture, rajčica je jedna od najviše istraživanih vrsta porodice Solanaceae te jedna od prvih kultura za koju su razvijeni molekularni markeri i mape. Rajčica danas ima jednu od najdetaljnijih genetskih mapa te je gotovo nemoguće odrediti točan broj gena/QTL-a koji su mapirani na kromosomima rajčice. Svakako najraširenija primjena molekularnih markera u oplemenjivanju rajčice odnosi se na genetsku kontrolu patogena., Although most of the existing tomato cuitivars have been developed by conventional breeding, numerous problems that occur during the breeding process have led to the development and application of new technologies in plant breeding. Certainly one of the most important is molecular markers technology, which might have an important role in increasing the efficiency of breeding process. Because of its simple genetic structure, tomato is one of the most studied species of the family Solanaceae, and one of the first culture for which molecular markers and maps have been developed. Tomato has one of the most detailed genetic maps and it is almost impossible to determine the exact number of genes/QTLs that have been mapped on tomato chromosomes. Certainly the most widespread application of molecular markers in tomato breeding is the genetic control of pathogens.

Main aim of plant breeding is creation of new genotypes with superior agronomic traits. Genetic difference is base for breeding success and it can be find in plant genetic resources (from wild relatives to breeding material). Plant genetic resources are important for agriculture, nutrition, environment protection, sustainable production, preservation of rural areas, lowering level of poverty in rural areas, market stability. Genetic erosion is loss of genetic difference or variability in plant/agricultural species and it has high impact on future improvement of cultivars and food production. For future food production it is essential to determine the level and sources of genetic erosion in plant species and to create plans for preservation and development of new genetic difference.

The aim of this paper is to present Installation research project “Creating wheat for the future–quest for the new genes in the old gene pool” financed by Croatian Science Foundation. Project goals are: assessment of phenotypic and molecular genetic diversity of wheat breeding germplasm ; identification of existing functional markers for agronomical important traits ; identification of marker- trait associations ; outlining the most suitable parents for future crossing and breeding programs. The knowledge gained through this project will provide the scientific training in biotechnology for young scientist from the Institute for Seed and Seedlings, as well as education and mentorship of his doctoral thesis at the Faculty of Agriculture in Osijek.

Aim of the study was to examine and determine distribution of glutenin (Glu-A1, Glu-B1 and Glu-D1) and gliadin (Gli-B1 and Gli-D1) loci in twenty Croatian winter wheat varieties using SDS-PAGE. Highest frequency at Glu-A1 loci was recorded for subunit 2*(50%). At Glu-B1 loci subunit 7+9 was dominant with frequency of 45% while the subunit 7+8 was at second place with 40%. At these loci lowest frequency (5%) had subunit 14+15. Subunit 5+10 prevailed at Glu-D1 with frequency of 70%. At Gli-B1 loci we determined prevalence of subunits 63+67 combination with a frequency of 50%, while the lowest prevalence had subunit 61 with a frequency of 5%. Subunits 60, 66 and null allele (N) were also present. At Gli-D1 locus, the most common subunit was 55 with a frequency of 90%, combination of subunits 55+56+59 and the subunit 59 were also present with frequency of 5%.

The translocation between the short arm of chromosome 1R of rye and the long arm of chromosome 1A, 1B or 1D of wheat represents practically the most common introduced foreign genes into the genome of hexaploid wheat. 1RS chromosome arm represents a source of different useful genes, associated with increased disease resistance, improved adaptability, stress tolerance, and increased stability and level of yield. On the other hand, translocation is associated with poor technological quality of wheat as a result of the presence of secalin and reduced number of gluten loci. The aim of this study was to determine the prevalence of 1RS translocation among some Croatian winter wheat varieties using molecular markers. The study included 40 varieties of which 23 Croatian. Four pairs of primers: RIS, SCM9, RYE-NOR and PAW161 were used for determination of translocation. The presence of translocation was determined in 12 of 40 (30%) varieties, while among the Croatian wheat varieties translocation had 8 of 23 (34.78%) varieties (Zlatna Dolina, Barbara, Nova Žitarka, Marija, Prima, Kuna, Koleda and Dea)., Translokacija između kratkoga kraka kromosoma 1R raži i dugoga kraka kromosoma 1A, 1B ili 1D pšenice predstavlja praktično najzastupljenije unesene strane gene u genom heksaploidne pšenice. 1RS krak kromosoma predstavlja izvor različitih korisnih gena, koji se povezuju s povećanom otpornošću na bolesti, povećanom adaptabilnosti, tolerantnosti na stres te povećanom stabilnošću i visinom prinosa. S druge strane, translokaciju se povezuje i s lošom tehnološkom kvalitetom pšenice, kao posljedicom prisutnosti sekalina i smanjenja broja glutenskih lokusa. Cilj ovoga istraživanja bio je utvrditi zastupljenost 1RS translokacije u hrvatskom sortimentu ozime pšenice korištenjem molekularnih markera. U istraživanje je bilo uključeno 40 kultivara pšenice, od čega 23 hrvatska kultivara. Za identifikaciju translokacije korištena su četiri para početnica: RIS, SCM9, RYE-NOR i PAW161. Istraživanjem je utvrđena prisutnost translokacije u 12 od 40 (30%) ispitivanih kultivara, dok je među hrvatskim kultivarima pšenice translokaciju imalo 8 od 23 (34,78%) kultivara (Zlatna Dolina, Barbara, Nova Žitarka, Marija, Prima, Kuna, Koleda i Dea).

The translocation between the short arm of chromosome 1R of rye and the long arm of chromosome 1A, 1B or 1D of wheat represents practically the most common introduced foreign genes into the genome of hexaploid wheat. 1RS chromosome arm represents a source of different useful genes, associated with increased disease resistance, improved adaptability, stress tolerance, and increased stability and level of yield. On the other hand, translocation is associated with poor technological quality of wheat as a result of the presence of secalin and reduced number of gluten loci. The aim of this study was to determine the prevalence of 1RS translocation among some Croatian winter wheat varieties using molecular markers. The study included 40 varieties of which 23 Croatian. Four pairs of primers: RIS, SCM9, RYE-NOR and PAW161 were used for determination of translocation. The presence of translocation was determined in 12 of 40 (30%) varieties, while among the Croatian wheat varieties translocation had 8 of 23 (34.78%) varieties (Zlatna Dolina, Barbara, Nova Žitarka, Marija, Prima, Kuna, Koleda and Dea).

Mosf commonly used correlation coefficients are Pearson's product moment correlation coefficient and Spearman's rank correlation coefficient. The aim of this paper is to compare a Pearson's and Spearman's coefficient of correlation on the same data set. The winter wheat grain cadmium (Cd) concentration was correlated to grain zinc (Zn) concentration, plant height, plant weight, number of spikelets per spike and 1000 kernel weight. Data were collected from the experiment carried out in semi controlled conditions, where genotypic specificity of winter wheat varieties was tested on the grain Cd and Zn accumulation on uncontaminated and Cd contaminated soil. Results showed that selection of most convenient correlation coefficient mostly depends on the type of variables, presence of outliers normality and linearity of relationship., Za opisivanje veze između dvaju svojstava najčešće se upotrebljavaju Pearsonov i Spearmanov koeficijent korelacije. Cilj je ovoga rada usporediti vrijednosti Pearsonovog i Spearmanovog koeficijenta korelacije na istom setu podataka. Podatci su prikupljeni iz pokusa provedenog u polukontroliranim uvjetima, u kojemu je ispitivana sortna specifičnost 51 genotipa ozime pšenice s obzirom na akumulaciju Cd i Zn u zrno na nekontaminiranome i Cd kontaminiranome tlu. Ispitivana je veza između koncentracije Cd u zrnu te koncentracije Zn u zrnu, visine biljke, mase biljke, broja klasića po klasu i mase 1000 zrna na nekontaminiranome i Cd kontaminiranome tlu. Na temelju rezultata, možemo zaključiti da izbor odgovarajućega koeficijenta korelacije najviše ovisi o vrsti podataka, prisustvu netipičnih vrijednosti i ispunjavanju parametrijskih pretpostavki.

Estimates of genetic diversity can be based on different types of data. The aim of this research were to study genetic diversity among Croatian wheat cultivars by random amplified polymorphic DNA (RAPD) markers, morphological traits and pedigree records; to analyse differences between wheat cultivars from two breeding centres; and to evaluate usability of RAPD markers for estimation of genetic diversity among wheat cultivars in comparison with morphological traits and pedigree record data. Studies were conducted on 14 wheat cultivars and breeding lines from two breeding centres in Croatia. For the RAPD analysis 36 primers were screened and the 14 most polymorphic ones yielded 341 polymorphic bands. Twelve morphological traits were used for morphological analysis. Pedigrees were composed of seven generations of ancestors. RAPD markers showed a high level of polymorphism among the cultivars examined and the breeding lines. No significant correlations were observed among the methods tested.

Model glavnih efekata i multiplikativne interakcije (AMMI) pokazao se kao vrlo učinkovit u oplemenjivanju bilja, jer obuhvaća veliki dio sume kvadrata interakcije genotip x okolina (GEI). Kod suncokreta, analiza GEI ne bi se trebala usredotočiti samo na prinos zrna već i na prinos ulja koja su dva oplemenjivački najznačajnija svojstva. Istraživanja su provedena s ciljem objašnjenja GEI pomošu AMMI modela za prinos ulja i prinos zrna za deset hibrida suncokreta istraživanih na šest okolina u istočnoj Hrvatskoj tijekom 2004., 2005. i 2006. godine. Analiza varijance pokazala je visoku značajnost za glavne efekte (genotip, okolina), dok je GEI za prinos zrna bila značajna na razini P=0.05, a za prinos ulja na razini P=0.10 što ukazuje da su glavni efekti genotip i okolina važniji nego GEI za oba svojstva. Općenito, GEI ima manji značaj za prinos ulja nego za prinos zrna. Iako vrlo liberalan, Gollobov F test je detektirao samo po dvije signifikantne multiplikativne komponente u AMMI analizi za oba svojstva, objašnjavajući 90.3% sume kvadrata GEI za prinos zrna i 89.5% za prinos ulja. Biplot dvije glavne komponente pokazao je vrlo jasno grupiranje okolina i pojedinih godina za oba svojstva ; gdje su niskoprinosne okoline 2005. očigledno odvojene od drugih istraživanih okolina. Za prinos zrna nijedan genotip nije povezan s niskoprinosnim okolinama dok su za prinos ulja dva genotipa grupirana unutar visokoprinosnih okolina.

Tretiranje sjemena hibrida kukuruza je neizostavan dio procesa dorade sjemena. Sjeme kukuruza najčešće je tretirano nekim od fungicida, a u poslijednjih par godina zbog problema sa žičnjacima i kukuruznom zlaticom u ponudi je sjeme tretirano i insekticidom. Cilj ovih istraživanja bio je utvrditi promjenu energije klijanja i standardne klijavosti kukuruza tijekom tri godine skladištenja na dva genetski različita hibrida (OSSK-596 i OSSK-602) kreiranih na Poljoprivrednom institutu Osijek. U istraživanjima oba hibrida testirano je netretitano sjeme te sjeme tretirano u tri varijante: 1.fungicid A ; 2.fungicid B ; i 3.fungicid + insekticid. Rezultati istraživanja pokazuju da tijekom tri godine skladištenja dolazi do pada energije klijanja i standardne klijavosti. Najveći gubitak kvalitete sjemena tijekom trogodišnjeg skladištenja zabilježen je s varijantama sjemena oba hibrida tretiranih suspenzijom fungicida i insekticida. Značajan pad energije klijavosti iskazan je već nakon 12 mjeseci skladištenja kod oba istraživana hibrida, dok je značajni pad standardne klijavosti uočljiv nakon 24 mjeseca skladištenja. Na temelju dobivenih rezultata kvalitete sjemena kukuruza ispitivanih tretmana, sjeme tretirano suspenzijom fungicida i insekticida nije pogodno za skladištenje duže od 12 mjeseci u skladištima koja nemaju mogućnost kontroliranja temperature i vlage zraka, te bi se takovo sjeme trebalo zasijati u godini tretiranja.

During the process of plant breeding and selection of new winter wheat cultivars a lot of attention is paid to the improvement of the main components of grain yield-number of ears/m2, number of grains per ear and 1000 seed weight. Production of cultivars that achieve a large number of fertile ears per unit area, a large number of grains per ear and a heavy 1000 seed weight, even with a lower density, would mean that a high grain yield is achieved with lower costs. It is of great importance for wheat production in Croatia. Along with the research on sowing rate influence (400, 500, 600 and 700 germinable seeds/m2) on winter wheat grain yield, other trials were done on the grain yield components (number of ears per unit area, number of grains per ear and 1000 seed weight) during periods 1996–1997 and 1997–1998 both on three AG cultivars of winter wheat Lara, Kruna and Lenta and on the check cultivar Ẑitarka. The trial was done according to the split-plot method in 4 replications. Statistically speaking, various sowing rates had a very significant influence on the number of ears per unit area. The largest number was achieved with the sowing rate of 700 germinable seeds/m2 (783,38 ears/m2). This number of ears is statistically more significant (P

Morfološka svojstva klasa korištena su za utvrđivanje genetske različitosti 50 sorti heksaploidne ozime pšenice porijeklom iz hrvatskih oplemenjivačkih programa. Poljski pokusi postavljeni su na dvije lokacije tijekom dvije vegetacijske godine (2008./09. i 2009./10.). Ocjene u poljskim pokusima i laboratoriju provedene su na 13 svojstava klasa koja se koriste u DUS ispitivanju. Temeljem provedenog istraživanja genetske različitosti hrvatske germplazme pšenice na osnovi morfoloških svojstava klasa, utvrđena je visoka razina različitosti (0,625) između ispitivanih sorata, te su se izdvojile sorte Super Žitarka i AFZG Karla, između kojih je utvrđen najveći koeficijent različitosti (0,94). Primjenom UPGMA metode, sve su sorte u različitim skupinama pokazale prisustvo značajne genetske različitosti. Na osnovi analiziranih podataka, utvrđene su najudaljenije sorte najboljih morfoloških svojstava klasa, što će pomoći pri uspješnom izboru novih roditeljskih kombinacija u budućim oplemenjivačkim programima., Morphological characteristics of ear were used for estimation of genetic diversity in 50 varieties of hexaploid winter wheat originated from Croatian breeding programs. Field trials were set at two locations in two vegetation years (2008/09 and 2009/10). Observations in field trials and laboratory were done on 13 ear characteristics used in DUS testing. Genetic diversity research of Croatian wheat germplasm according to ear morphological characteristics, showed a high level of dissimilarity (0.625) among the tested varieties. Varieties Super Žitarka and AFZG Karla are pointed out with highest coefficient of dissimilarity (0.94). Application of UPGMA method showed that all varieties in different groups had significant genetic diversity. On the basis of data analysis the most distant varieties with the best morphological characteristics of ear were determined and it will be help in the selection of new parent combinations in future breeding programs.

Investigations of the influence of hermetic cereal seed storage on germination were carried out in the laboratory of the Faculty of Agriculture in Osijek (Croatia)over 5 years (1992-96). Seeds of four species (winter wheat, winter barley, spring oat, and maize) were stored in hermetic glass containers at an air temperature of 20 degrees C and a relative humidity of 65%. The moisture level in stored seeds was 13%. After 5 years, statistically highly significant associations (P

Kod 50 sorti heksaploidne ozime pšenice (Triticum aestivum L.) porijeklom iz Republike Hrvatske utvrđena je i analizirana kompozicija ω – glijadina. Poliakrilamid gel elektroforeza (PAGE) u prisutnosti natrijeva dodecil sulfata (SDS – sodium dodecyl sulphate) korištena je za identifikaciju podjedinica na Gli-B1 i Gli-D1 genlokusima. Utvrđena je najčešća kombinacija podjedinica 63+67 s frekvencijom od 64% na genlokusu Gli-B1, zatim podjedinica 66 sa frekvencijom od 16%, te nulti alel (N) s frekvencijom od 14%. Na lokusu Gli-D1 najzastupljenija podjedinica je bila 55 s frekvencijom od 94%. Broj alela po lokusu kretao se od 3 (Gli-D1) do 7 (Gli-B1), veća genetska različitost (He=0,557) kao i veći PIC (0,529) utvrđeni su na lokusu Gli-B1, dok je znatno manja genetska različitost utvrđena na lokusu Gli-D1 (He=0,114) kao i PIC (0,110)., The composition of ω – gliadins was identified and analyzed in 50 varieties of hexaploid winter wheat (Triticum aestivum ssp. vulgare L.) originated from the Republic of Croatia. Polyacrylamide gel electrophoresis (PAGE) in presence of sodium dodecyl sulfate (SDS) was used for determination of subunits at Gli-B1 i Gli-D1 genloci. The most common combination of subunits at genloci Gli-B1 was 63+67 with frequency of 64%, then subunit 66 with frequency of 16% and null allele (N) with frequency of 14%. At Gli-D1 loci the most frequent subunit was 55 with presence at 94% in tested varieties. Number of allels per loci was from 3 (Gli-D1) to 7 (Gli-B1), high genetic diversity (He = 0,557) and high PIC (0,529) were estimated at loci Gli-B1, while significantly lower genetic diversity was estimated at loci Gli-D1 (He = 0,114) and PIC (0,110).

Cilj istraživanja bio je proučiti utjecaj genotipa i vodnog stresa pri različitim koncentracijama dušika na najznačajnija svojstva biljke i korijena u ranijim fazama razvoja pšenice. Za tu svrhu postavljen je staklenički mikropokus s pet genotipova pšenice, dvije razine opskrbe vodom i tri razine koncentracije dušika. Utvrđeno je da su se broj zelenih listova, ukupna masa biljke, masa zelenih listova, masa stabljike i površina zelenog lista povećavali dodavanjem dušika i vode. Broj i masa otpalih listova povećavali su se dodavanjem dušika, ali su se smanjivali dodavanjem vode. Statistički značajne interakcije zabilježene su najčešće između dušika i vode te između genotipa i dušika.

Opsežne spoznaje o genetskoj raznolikosti genetskog materijala pšenice vrlo su važne za daljnje uspješno i održivo genetsko unapređivanje kultivara kako unutar pojedinih programa oplemenjivanja, tako i unutar pojedinih država i šire. Analize genetske raznolikosti omogućavaju i bolje razumijevanje utjecaja modernog oplemenjivanja bilja na genetsku raznolikost pojedine biljne kulture kao i učinkovitije korištenje, upravljanje i očuvanje biljnih genetskih resursa. Zahvaljujući visokoj razini polimorfizma i informativnosti kao i relativnoj jednostavnosti primjene, mikrosatelitni biljezi ili jednostavne ponavljajuće sekvence (simple sequence repeats ; SSR) predstavljaju važno oruđe za analize genetske raznolikosti. Mikrosatelitni biljezi su u posljednjih dvadeset godina uspješno primjenjeni u brojnim genetskim studijama na različitim biljnim vrstama. Slijedom navedenog, cilj ovog istraživanja bio je procijeniti: (1) genetsku raznolikost 98 hrvatskih i 24 inozemna kultivara pšenice priznatih i uvedenih u proizvodnju od 1905. do 2008. godine i (2) procijeniti mogućnost primjene mikrosatelitnih biljega u sjemenarstvu i oplemenjivanju pšenice. Ukupno 122 kultivara pšenice analizirano je pomoću 45 mikrosatelita. Unutar skupine od 122 kultivara umnoženo je ukupno 413 polimorfnih alela. Unutar skupine hrvatskih kultivara (n=98) umnožena su ukupno 354 alela, dok su unutar skupine kultivara kreiranih na Poljoprivrednom institute Osijek (n=74) utvrđena 303 alela. Broj alela po lokusu (Na) kretao se od 3 do 20, a prosječno je iznosio 9.18, 7.87 i 6.73 alela unutar skupina s n=122, n=98 i n=74 kultivara. Prosječna vrijednost genske raznolikosti (Gene diversity ; HE) i Informacijskog sadržaja polimorfizma (Polimorfic Information Content ; PIC) iznosila je od HE=0.69 i PIC=0.65 (n=122) do HE=0.63 i PIC=0.58 (n=74). Alelno bogatstvo (Allelic richness ; Nar), kao mjera broja alela po lokusu neovisna o veličini uzorka kretala se od Nar=8.78 unutar skupine od n=122 do Nar=6.73 unutar skupine od n=74 kultivara. Ukupna utvrđena genska raznolikost i Informacijski sadržaj polimorfizma po biljegu varirao je od HE=0.34 i PIC=0.32 (biljeg gwm493) do HE=0.86 i PIC=0.85 (biljeg wmc634). Matrica genetske udaljenosti između parova kultivara utvrđena je temeljem udjela zajedničkih alela (Proportion of Shared Alleles Distance ; DPSAM) iznosile su od 0.02 do 0.96 pri čemu je veća prosječna vrijednost DPSAM utvrđena između svih (0.69), a zatim između hrvatskih (0.66) i kultivara Poljoprivrednog instituta Osijek (0.63). Na osnovi matrice udaljenosti između svih parova kultivara, u svrhu razvrstavnog i grafičkog prikaza genetske raznolikosti između kultivara, provedena je izrada stabla pomoću Neponderirane metode za sparivanje skupina na temelju prosječnih vrijednosti (Unweighted Pair-Group Method Using Arithmetic Averages ; UPGMA). Na izrađenom UPGMA stablu bilo je moguće uspješno razlikovati sve kultivare u analizi. Nije utvrđeno svrstavanje kultivara u skupine prema oplemenjivačkom program ili vremenskom razdoblju priznavanja kultivara. Općenito se može ustvrditi da je na tvorbu skupina i podskupina na stablu UPGMA najviše utjecala genetska osnova kultivara, najčešće na način da većina članova skupine posjeduje u određenoj mjeri zajedničke genetske poveznice. Potvrđen je važan potencijal primjene mikrosatelitnih biljega u provjeri genetske uniformnosti i čistoće unutar i između blisko srodnih kultivara pšenice, koji implementiran zajedno s klasičnim metodama umnažanja sjemena može značajno doprinijeti unapređenju sjemenarstva kao i zaštiti oplemenjivačkih prava. Ukupno, rezultati ovog istraživanja omogućavaju bolji uvid u genetsku raznolikost hrvatskih kultivara pšenice na DNA razini. Procijenjene genetske udaljenosti (DPSAM) između kultivara mogu pomoći u preciznijem odabiru roditelja za križanja i stvaranje novih oplemenjivačkih populacija i s obzirom na očuvanje genetske raznolikosti pšenice. Ovo istraživanje je jedna od pretpostavki uvođenju molekularnim biljezima potpomognutog oplemenjivanja (Molecular Marker Assisted Breeding ; MMAB) u program oplemenjivanja pšenice na Poljoprivrednom institute Osijek kao nadopune klasičnim oplemenjivačkim metodama.

The plant height of wheat is an important quantitative trait, controlled by several genes with strong effect. However, in worldwide wheat breeding, only several of those genes have been used. Rht8 (Reduced Height Gene) is especially important in agro-climatic conditions of South-East Europe. Because of its close linkage with dwarfing gene Rht8, microsatellite marker gwm261 has been accepted as the diagnostic molecular marker for gene Rht8. In this study, allelic variability at the locus Xgwm261 for 122 Croatian and foreign wheat cultivars by means of microsatellite marker gwm261 was determined. A 192 base pairs allele at the locus Xgwm261 was found for 84 Croatian cultivars. The genetic heritage of Croatian cultivars at the locus Xgwm261 is the consequence of new parental components usage, carriers of short plant and early maturity attributes and the consequent selection of progeny with these traits during breeding process. The results of this research will be helpful in characterization of domestic wheat cultivars, as well as in more accurate selection of parents for hybridization purposes., Visina biljke pšenice važno je kvantitativno svojstvo kontrolirano s više gena jakog učinka. Međutim, u oplemenjivanju pšenice u svijetu, najčešće se koristi samo nekoliko takvih gena. U agroklimatskim uvjetima jugoistoka Europe posebno je važan gen Rht8 (Reduced Height Gene). Mikrosatelit gwm261 je, zbog svoje bliske vezanosti s lokusom za gen Rht8, prihvaćen kao dijagnostički molekularni biljeg gena Rht8. U ovome istraživanju je pomoću mikrosatelita gwm261 utvrđena alelna varijabilnost na lokusu Xgwm261 za 122 hrvatska i strana kultivara pšenice. Alel sa 192 para baza na lokusu Xgwm261 utvrđen je za 84 hrvatska kultivara. Genetsko naslijeđe hrvatskih kultivara na lokusu Xgwm261 posljedica je korištenja novih roditelja, nositelja svojstava niže stabljike i ranozrelosti te odabira potomstva s tim svojstvima tijekom oplemenjivačkoga procesa. Rezultati istraživanja pomoći će u karakterizaciji domaćih kultivara pšenice, kao i u preciznijem odabiru roditelja u svrhu križanja.

Cilj rada je procijeniti i utvrditi razlike u prinosu između sorata soje različitih skupina dozrijevanja u različitim okolinama na području Republike Hrvatske. Zbog sve većih klimatskih promjena koje utječu na smanjenje prinosa, potrebno je izvršiti klasifikaciju pojedinih okolina prema stabilnostima prinosa. Tijekom četiri vegetacijske godine (2004. – 2007.) na tri lokacije Osijek (N 45°33', E 18°40'), Kutjevo (N 45°25', E 17° 52') i Nova Gradiška (N 45°15', E 17°22') obavljena su ispitivanja o utjecaju genotipa, okoline i interakcije genotip okolina na visinu prinosa. Dobiveni rezultati pokazali su statistički opravdane razlike između ispitivanih sorata. Godina 2004. je bila najbolja za ostvarenje visokih prinosa (3,94 t/ha), dok je najzapadnija lokacija (N. Gradiška) u svim godinama imala prosječno najveće prinose od 3,80 t/ha, što je bilo znatno više u odnosu na ostale dvije lokacije (Osijek 3,43 t/ha i Kutjevo 3,45 t/ha). Rezultati ukazuju na mogućnost povećanja prinosa pravilnim odabirom sorata i najbolje skupine dozrijevanja za pojedine okolina., The main goal was to define and estimate yield between standards soybean cultivars of different maturity groups on different locations in Republic of Croatia. Because of growing climate changes which impact on yields, it is necessary to classify certain environments to yield stability. During four vegetation years (2004 - 2007) on 3 locations Osijek (N 45°33', E 18°40'), Kutjevo (N 45°25', E 17° 52') i Nova Gradiška (N 45°15', E 17°22') research was conducted on influence of genotype, environment and interaction genotype x environment on soybean yield. The results have shown statistically high difference between soybean cultivars. 2004 was the best for achieving high yield (3.94 t / ha), while western location (N. Gradiška) in all years had averaged highest yields of 3.80 t/ha, which was significantly higher compared to other two locations (Osijek 3.43 t/ha and Kutjevo 3.45 t/ha). The results indicate possibility of increasing yield by proper selection of varieties and the best group of maturation for certain environments.

Knowledge about genetic diversity pattern in existing wheat germplasm is essentially important for further sustainable genetic improvement and successfulness of breeding programs. Genetic diversity analysis can provide better understanding of plant breeding influence on crop genetic diversity and more efficient utilization, management and conservation of germplasm resources. Owing to their high level of polymorphisms, microsatellite markers (simple sequence repeats ; SSR) representing important tool for genetic diversity analysis and have been successfully applied for numerous assessments of genetic relationships in different plant species. Consequently, the objective of this study was to estimate genetic diversity of 98 Croatian wheat cultivars released since 1905 to 2008 year and 24 foreign wheat cultivars. Foreign cultivars were included in this research regarding to their pre-ancestral and ancestral role in Croatian cultivars, as well as important widely grown cultivars and as a control. A total of 122 cultivars were screened by 45 wheat microsatellite markers, 1 to 3 from each wheat chromosome. Within group of 122 cultivars a total of 413 polymorphic alleles were amplified. Within group of Croatian cultivars (n=98) were amplified 354 alleles, while within group of cultivars created on Agricultural Institute Osijek (n=74) were found 303 alleles. Allele number per locus (Na) was varied from 3 to 20 with an average of 9.18, 7.87 and 6.73 alleles within group of n=122, n=98 and n=74 cultivars, respectively. Average value of gene diversity (HE) and polymorphic information content (PIC) were ranged from HE=0.69 and PIC=0.65 (n=122) to HE=0.63 and PIC=0.58 (n=74). Allelic richness (Nar) as the measure of the number of alleles per locus independent of sample size showed decreasing trend from 8.78 within group of n=122 to 6.73 within group of n=74 cultivars. Overall observed gene diversity and polymorphic information content per marker were varied from HE=0.34 and PIC=0.32 (marker gwm493) to HE=0.86 and PIC=0.85 (marker wmc634). Genetic distances among pairs of cultivars based on proportion of shared alleles distance matrix (DPSAM) were ranged from 0.02 to 0.96 with higher average value among all (0.69), then among Croatian (0.66) and Agricultural Institute Osijek cultivars (0.63). The cultivars were grouped by cluster analysis using the unweighted pair- group method with arithmetic averages (UPGMA). At the constructed UPGMA tree was possible to successfully differentiate all investigated cultivars. However, no clear clustering of cultivars by breeding program or year of release was observed. Cultivars that belong to the same group generally share the concerned proportion of common pre-ancestral or ancestral genetic basis. Results of this research are providing better insight in genetic diversity of Croatian cultivars at DNA level. Estimated genetic distances (DPSAM) among cultivars can be helpful for more accurate parent assortment for hybridization and creation of new wheat breeding populations with respect to genetic diversity preservation. This research is one of the prerequisites for adoption of marker assisted selection (MAS) and molecular breeding in Agricultural Institute Osijek wheat breeding program as complement to the classical breeding methods.

Mikotoksini (mykes - grč. gljiva, toxikon - grč. otrov) su sekundarni metaboliti kemijski vrlo različite strukture s različitim biološkim učincima. Ovisno o Fusarium vrsti, na zrnima se mogu očekivati mikotoksini: zearalenol, deoksinivalenol (F. graminearum), T-2 toksin, neosolaniol, scirpenol (F. avenaceum), T-2 toksin, HT-2 toksin, tetraol (F. poae). Cilj je ovoga rada prikazati varijabilnost genotipova pšenice kod procijenjenih količina DON-a, a na temelju simptoma bolesti ocijenjenih na 22. dan nakon umjetnih infekcija. Najmanje procijenjene količine DON-a izračunate su kod genotipova Renan (0,8 mg/kg), Soissons (1,0 mg/kg), Sirban Prolifik (1,3 mg/kg) i Libellula (1,5 mg/kg), a najveće kod genotipova Osk.553/98 (7,5 mg/ kg), Osk.554/97 (7,1 mg/kg), Žitarka (6,4 mg/kg), i Zlatna Dolina (5,8 mg/kg)., Mycotoxins are secondary metabolites of very different chemical structure with different biological effects. Depending on the Fusarium species in grains we can expect mycotoxins: zearalenol, deoxynivalenol (F. graminearum), T-2 toxin, neosolaniol, scirpenol (F. avenaceum), T-2 toxin, HT-2 toxin, tetraol (F. poae) . The aim of this paper is to present the variability of wheat genotypes with an estimated amount of DON based on symptoms of disease assessed at 22nd day after artificial infections. The lowest estimated amounts of DON in genotypes were calculated for Renan (0.8 mg/kg), Soissons (1.0 mg/kg), Sirban Prolific (1.3 mg/kg) and Libellula (1.5 mg/kg), and the highest in Osk.553/98 (7.5 mg/kg), Osk.554/97 (7.1 mg/kg), Žitarka (6.4 mg/kg) and Zlatna Dolina (5.8 mg/kg).

Hemoglobin (Hb) was found to be modified in patients with prolonged hypoglycemia caused by hyperinsulinism. This modification was of a nonpolar nature, did not affect the overall charge of the Hb and caused its aggregation in aqueous solution. After normal concentrations of glucose and insulin were reestablished, this modification slowly disappeared. Hemoglobin from normal erythrocytes incubated in vitro with high concentrations of insulin showed similar behavior.

The objective of the investigation was testing of sunflower hybrids in different environmental conditions. Trials were set up at three locations (Osijek – Eastern Slavonia county, Karanac – Baranja county, and Feričanci – Western Slavonia county) during the 2002 and 2003 growing season. Fourteen sunflower hybrids were tested. Stability and adaptability analysis was performed using two parameters – regression coefficient (bi) and deviations mean square (s2 di). Determined were hybrids characterized by high and stable grain yield and wide general adaptability and hybrids for low and high yielding environments. The investigation showed no significant differences among single cross and three-way cross hybrids regarding grain yield stability.

Stability of cultivars across different environments is one of the main goals in winter wheat breeding programs. Much attention has been devoted to analysing cultivar by environment (C x E) interactions to improve crop breeding efforts (Thomason and Philips, 2006 ; Sio-Se Mardeh et al., 2006 ; Hoffman et al., 2005 ; Pepo and Gyi ri, 2005). Ideal genotype would have, in all environments same high yield. Research work was conducted on four test locations in Eastern Croatia - Zupanja, Nova Gradiska, Kutjevo and Osijek - during four vegetation years, 2002/2003 – 2005/2006. Three winter wheat cultivars - Zitarka, Sana and Divana were chosen for the research due to their high yielding performance and good quality. Cultivars were planted in the randomised block design with four replications. Basic plot was 10 m2 with 25 rows and spaced 40 cm apart from other plot.Genotype-environment interaction proved to be very important in winter wheat breeding. For the selection of high yielding and stable genotypes several locations should be included in trials. Selection of locations should be made according to several stability parameters. In this research work all three stability parameters showed low stability of location Kutjevo. Also, there was high correlation between locations Kutjevo and Nova Gradiska. Therefore, location Kutjevo can be excluded from further winter wheat trials in Eastern Croatia.

Tijekom dvogodišnjeg razdoblja (2004. i 2005. godine) obavljeno je istraživanje o visini prinosa i ekonomskoj isplativosti proizvodnje tri rane sorte soje (Dubravka 0, Marija 00 i Danica 000) različitih grupa dozrijevanja. Pokusi su postavljeni po split-plot shemi na poljima Zavoda za sjemenarstvo i rasadničarstvo iz Osijeka, Kutjevo d.d. i PPK Nova Gradiška uz primjenu standardne agrotehnike za soju. Jaz koji nastaje porastom broja stanovnika i istovremenim smanjenjem poljoprivrednih površina iz godine u godinu sve je veći, a soja zbog velikog postotka bjelančevina (od 35-50%) i ulja (od 18-24%) preuzima vodeće mjesto u proizvodnji i ishrani čovječanstva. Soja zbog naknadne i postrne sjetve postaje zanimljiva poljoprivrednim proizvođačima diljem svijeta. Uzimajući u obzir agrotehničke mjere koje se koriste, uz pomoć tablica i grafikona dana je analiza čimbenika i usporedba isplativosti proizvodnje pojedine sorte soje u Hrvatskoj na površini preko tri hektara, kako bi se odredila najpogodnija sorta za domaću poljoprivrednu proizvodnju., During the two vegetation years (2004 and 2005) an examination was conducted on influence of three early breeds of soybean (Dubravka 0, Marija 00 and Danica 000) belonging to different groups of ripening in terms of level of crop yield and cost effectiveness. The tests were undertaken by split-plot scheme on the crop fields originated from Institution for seed and seedlings Osijek, Kutjevo d.d. and PPK Nova Gradiška using standardized agro measures valid for soybean in The Republic of Croatia. The difference between increase of the number of inhabitants and decrease in the crop fields area has risen significantly. Soybean, because of its high percentage of proteins (35-50%) and oil (18-24%), slightly for sure overtakes the first position in agricultural production and nutrition of mankind. Soybean is becoming more and more interesting to producers worldwide because of its rich seeding capability. Taking into account the agricultural measures used the analysis and comparison of production effectiveness between each breed of soybean in Croatian on the area more than three ha are presented in tables and graphs. In this way the best breed can be ultimately determined for domestic farm production.

Istraživanja su provedena na 14 hibrida stvorenih u okviru oplemenjivačkog programa Poljoprivrednog instituta Osijek. Pokusi su postavljeni po slučajnom bloknom rasporedu u četiri ponavljanja tijekom tri godine (2002-2004) na lokalitetu Osijek. Analizirana su svojstva: visina biljke, masa 1000 zrna, hektolitarska masa, prinos zrna, sadržaj ulja i prinos ulja. Procjenjena je heritabilnost, genetska dobit i provedena path analiza povezanosti istraživanih svojstava i prinosa ulja. Istraživani hibridi su se značajno razlikovali u svim ana-liziranim svojstvima. Procjenjene su visoke vrijednosti heritabilnosti za visinu biljke, masu 1000 zrna, hektolitarsku masu i sadržaj ulja. Heritabilnost za svojstva prinos zrna i prinos ulja je pokazala niže vrijednosti. Najveću izravnu genetsku dobit imala su svojstva visina biljke i sadržaj ulja, a najmanju prinos zrna i prinos ulja. Najveće direktne utjecaje na prinos ulja ostvarili su prinos zrna i sadržaj ulja., Investigations were carried out with 14 hybrid create in frame of the Agricultural Institute Osijek breeding program. Trials were established as randomize block design in four replication during three years (2002-2004) in region of Osijek. Following traits were analyzed: plant height, 1000 kernel weight, hectolitre mass, grain and oil yield and oil content. Heritability, genetic gain and path analysis correlation between investigated traits and oil yield were evaluated. Investigation hybrids showed significant difference for all analyzed traits. High value heritability for plant height, 1000 kernel weight, hectolitre mass and oil content were evaluated. Heritability for grain and oil yield showed lower value. The biggest genetic gain showed plant height and oil content, and the lowest grain and oil yield. The strongest direct effects to oil yield were assessed for grain yield and oil content.

Cilj je istraživanja bio procijeniti visinu, stabilnost i adaptabilnost prinosa ulja nekoliko domaćih hibrida suncokreta. Pokusi su provedeni na tri lokaliteta (Osijek, Karanac i Feričanci) tijekom 2002. i 2003. godine, a obuhvaćali su 9 eksperimentalnih i pet priznatih hibrida suncokreta Poljoprivrednog instituta Osijek. U analizi stabilnosti i adaptabilnosti korištena su četiri parametra: ekovalenca, koeficijent regresije, odstupanje od regresije te model prema Francisu i Kannenbergu. Dobiveni rezultati ukazali su da postoje razlike u visini prinosa ulja te stabilnosti i adaptabilnosti između pojedinih hibrida. Visokim prinosima, stabilnošću i širokom adaptabilnošću izdvojili su se hibridi: Apolon, Olio, (59A x 5B) x O3 i (253A x 5B) x O3. Nisu utvrđene značajne korelacije između prinosa zrna i ulja i parametara stabilnosti, što upućuje da je moguće stvoriti visokog i stabilnog prinosa, široke opće adaptabilnosti. Jaka visoko značajna korelacija između ekovalence i varijance odstupanja od regresije upućuje na to da se procjena stabilnosti može izvršiti samo s jednim parametrom., The aim of this research work was to assess height, stability and adaptability of oil yield in several Croatian sun- flower hybrids. Trials were conducted on three locations (Osijek, Karanac and Feri anci) during vegetation years 2002 and 2003. In the research work nine experimental and five recognized sunflower hybrids of Agricultural Institute Osijek were included. Four parameters were used for analysis of stability and adaptability: ecovalence, regression coefficient, deviation from regression, Francis and Kannenberg model. The results showed differences between hybrids in oil yield, stability and adaptability. High oil yield, stability and wide adaptability have hybrids: Apolon, Olio, (59A x 5B) x O3 and (253A x 5B) x O3. There were no significant correlations between kernel and oil yield and stability parameters. It indicates possibility for creation of hybrids with high and stabile yield and wide general adaptability. Highly significant correlation between ecovalence and deviation of regression showed that it is possible to assess stability with just one parameter.