Sugar beet micropropagation

Vegetative in vitro multiplication is one of the most efficient methods for sugar beet (Beta vulgaris L.) propagation. The usual steps in this procedure are sterilization of explant, multiplication, rhizogenesis and acclimatization. In the paper is presented development of regeneration and multiplication techniques from different explants. It gives a detailed description of further micropropagation steps and presents necessary conditions for their realization. The paper also discuss different ways of micropropagation application especially in sugar beet breeding, but in other plant sciences as well

Korelacije korenskih svojstava monogermnih genotipova šećerne repe iz slobodne oplodnje i njihova varijabilnost

The aim of this study was to analyze six open-pollinated monogerm sugar beet genotypes for the important root traits, to examine their hybrids with one (CMS) tester and to estimate the correlation between the traits. Root weight, dry matter content, dry matter yield per root, circumference of root and height of the root head were analyzed. All of the tested hybrids showed better results for root weight and dry matter yield per root than their parents. It was not possible to predict superior hybrid combination based solely on the characteristics of parents. Parents with the lowest root weight per se produced the test hybrid with the highest root weight. Parents showed a positive correlation between root weight and dry matter content and this could help sugar beet breeders to choose the most suitable selection criteria.Cilj ovog rada je bilo ispitivanje šest novih jednokličnih genotipova šećerne repe poreklom iz slobodne oplodnje, kao i njihovi hibridi za najvažnija korenska svojstva. Određeni su koeficijenti korelacije između svojstava. Nakon završene vegetacije analizirani su: masa korena, sadržaj suve materije, prinos suve materije po korenu, obim korena i visina glave korena. Utvrđeno je povećanje mase korena i prinosa suve materije po korenu kod svih test hibrida u poređenju sa roditeljima. Za razliku od hibrida, kod roditelja je ustanovljena pozitivna korelacija između mase korena i sadržaja suve materije. Na osnovu karakteristika roditelja per se nije moguće predvideti superiorno hibridno potomstvo, jer su roditelji sa najmanjom masom korena dali hibride sa najvećom masom korena

Effect of inoculation on azotobacter population size in sugarbeet rhizosphere depending on fertilization

The objective of this study was to assess the effect of inoculation on Azotobacter population size in dependence of fertilizer dose and fertilization method. Differences were registered in Azotobacter population size which depended on both, nitrogen dose and fertilization method. On average, the highest percentage of increase in Azotobacter population size in relation to the non-inoculated variant, was registered in the variant with nitrogen, liquid manure and harvest residues. The largest increase in Azotobacter population size was obtained in the inoculation variants

Indukcija i potencijal za mikropropagaciju haploida šećerne repe

The aim of research was obtaining sugar beet haploids via gyno-genesis and their micropropagation. Haploids were obtained by ovule culture from fourteen diploid, monogerm, fertile genotypes. On the tested nutrient media genotypes exhibited different gynogenic potential. Eight haploid plant were chosen for further investigation and after development of first leaves put on micropropagation medium. The presence of cyto-kinin in medium stimulated development of axillary buds, while in some genotypes adventitious buds developed as well. Multiplication rate was not consistent, although number of developed plants grew after each sub-cultivation. Differences in plant multiplication started to differ after four subcultures. By testing of differences between correlation coefficients, i.e. multiplication rate during six subcultivations, it was determined that they significantly differ between tested genotypes.Cilj istraživanja je bio dobijanje haploida šećerne repe putem ginogeneze i njihova mikropropagacije. Haploidi su dobijeni kulturom semenog zametka iz Četrnaest diploidnih, monogermnih i muški fertilnih genotipova šećerne repe. Ispitivani genotipovi su na podlogama sa različitom koncentracijom stimulatora rasta ispoljili različit potencijal za ginogenezu. Za dalja ispitivanja je odabrano osam haploida koji su nakon razvića prvog para listova postavljeni na podlogu za mikropropagaciju. Prisustvo citokinina u hranljivoj podlozi je stimulisalo pojavu i razviće bočnih pupoljaka u pazuhu listova, dok je kod nekih genotipova takođe došlo do pojave adventivnih pupoljaka. Porast broja biljaka tokom mikropropagacije nije bio kontinuiran iako je broj novostvorenih biljaka rastao posle svake sub-kultivacije. Razlike u intenzitetu multiplikacije biljaka počele su da se značajno ispoljavaju nakon Četvrte subkultivacije. Testiranjem značajnosti razlika korelacionih koeflcijenata, odnosno intenziteta mikropropagacije ispitivanih genotipova tokom šest subkultivacija, utvrđeno je da se one međusobno statistički značajno razlikuju

Development and application of qRT-PCR for sugar beet gene expression analysis in response to in vitro induced water deficit

Sugar beet is a significant industrial crop, often grown in the areas where summer drought can severely limit root yield and sugar content. In order to improve development of sugar beet cultivars with increased drought tolerance it is necessary to understand plant response to water stress at the genomic level. Since recent research efforts have focused on the molecular response of the plant in order to identify water deficit inducible genes, the aim of this investigation was to develop qRT-PCR methodology for the quantification of gene expression in sugar beet under conditions of water deficiency in vitro. Sugar beet genotypes, selected for different response to water deficit, were grown and multiplied in vitro. Axilary shoots were placed on micropropagation media with 0%, 3% and 5% PEG, for 28 days. To determine reaction of sugar beet genotypes to in vitro induced water deficit changes in number of axillary shoots, shoot fresh weight and dry matter content were measured. Total RNA was extracted from leaves and reverse transcribed into cDNA, which served as matrix in real-time PCR reaction using TaqMan technology. The housekeeping gene for glutamine synthetase was used as endogenous control, while the genes for alpha amylase and osmotin-like protein were target genes. The relative quantification values for each target gene were calculated by the 2(-Delta Delta Ct) method. Selected candidate genes differed in relative gene expression among genotypes and applied PEG treatments. The obtained results indicated that qRT-PCR protocol was efficient and accurate, showing the potential to be used in further expression analysis of candidate genes involved in sugar beet reaction to water stress

Uticaj rokova vađenja na kvantitativna svojstva šećerne repe

Results of the investigation regarding influence of harvest dates on quantitative traits of sugar beet are presented in this paper. The three-year trial was conducted on five varieties developed by Institute of Field and Vegetable Crops, Novi Sad. Analysis of variance showed significant differences between harvest dates in 2008 and 2010 for all analyzed traits, but in 2009, due to severe drought differences were not found. With the delay of harvest date, root and raw sugar yield increased, while sugar content was dependent on climatic conditions (precipitation) between two harvest dates. There were no differences among the analyzed genotypes for measured traits in 2010. Interaction between genotype and harvest date has not been confirmed. Variety Drena had the highest root and raw sugar yields in 2008 and 2010.U radu su predstavljeni rezultati trogodišnjeg ispitivanja uticaja tri roka vađenja šećerne repe na kvantitativna svojstva korena pet komercijalno gajenih sorata Instituta za ratarstvo i povrtarstvo u Novom Sadu. Analizom varijanse utvrđene su značajne razlike između rokova vađenja u 2008. i 2010. za sva ispitivana svojstva. Zbog suše u 2009. nisu utvrđene razlike između različitih datuma vađenja. Odlaganjem roka vađenja prinos korena i prinos polarizacionog šećera se linearno povećavao u 2008. i 2010. godini. Sadržaj šećera je varirao u zavisnosti od vremenskih uslova (padavine) između dva različita roka vađenja. U pogledu ispitivanih svojstava između genotipova nije uočena razlika samo u 2010. godini. Nije potvrđeno postojanje interakcije između genotipa i roka vađenja za ispitivana svojstva. Od ispitivanih sorata u 2008. i 2010. najbolja je bila sorta Drena, koja je u tim godinama ostvarila najveće prinose korena i polarizacionog šećera

Construction of plant transformation vectors carrying beet necrotic yellow vein virus coat protein gene (i) - transformation vectors

Coat protein gene of beet necrotic yellow vein virus (BNYVV) was isolated from inoculated sugar beet roots and leaves of Chenopodium quinoa and Tetragonia expansa, by RT-PCR and imuno capture RT-PCR. Specific primers were made to complement coat protein gene and untranslated leader sequence, so that two fragments were obtained: long (731 bp), which contained coat protein gene and leader sequence, and short (587 bp), with coat protein gene. Fragments were cloned in two plant transformation vectors: pCAMBIA 3301M and pCAMBIA 1304M, which were modified by removing multicloning site and NcoI restriction site at the 5' end of the reporter genes. Vector pC3301M had bar gene which confers resistance against the herbicide gluphosinate ammonium as selectable marker, and pC1304M had gene for resistance to antibiotic hygromycin. Three constructs were made from each vector: CPL, containing coat protein gene with leader sequence; CPS with gene for coat protein, and CPSas with coat protein gene in antisense orientation. All constructs were transfered to Agrobacterium tumefaciens strain LBA4404

Uticaj đubrenja i distribucije nitratnog azota u profilu zemljišta na prinos i kvalitet korena šećerne repe

Researches, which have lasted for two years, were carried out on long-term trial field at Rimski Šančevi, Novi Sad, Serbia. In this trial, the eight fertilization variants of N, P2O5 and K2O increased amounts were studied. Sugar beet root and tops yields were determined, as well as the elements of technological sugar beet root quality. Based on these results, percentage of sugar utilization and refined sugar yield was defined. In the spring, before applying of N fertilizer, amount of nitrate nitrogen in the soil and its influence on yield and quality was determined. The highest root yield in 2002 was produced at the variant N100 P150 K150, and in 2003 at the variant N150 P150 K150. However, in both years, referring to the variant N100 P100 K100, the differences were not statistically significant. Increasing of nitrogen amounts had negative effects on refined sugar yield. Amounts of NO3-N in the soil in spring, before sugar beet sowing, in 2002 had significant influence on root yield and refined sugar yield. In the year 2003, which was highly dry, high correlation ratio were gained between amounts of NO3-N in the soil and root quality parameters, but it wasn't significant between nitrogen amounts and root and refined sugar yield.Dvogodišnja istraživanja uticaja rastućih količina NPK hraniva na prinos i kvalitet korena šećerne repe izvedena su na stacionarnom poljskom ogledu na Rimskim Šančevima. U proleće, pre primene N đubriva, praćena je količina nitratnog azota po slojevima zemljišta, te njegov uticaj na navedena svojstva. U obe godine, razlika u prinosu korena postignutog pri najvećim količinama NPK hraniva nije bila statistički značajna u odnosu na varijantu N100 P100 K100. Povećanje količine azota delovalo je negativno na prinos rafinisanog šećera. Količina NO3-N u zemljištu u proleće pre setve u 2002. godini imala je značajan pozitivan uticaj na prinos korena i rafinisanog šećera. U izrazito sušnoj 2003. godini visoki koeficijenti korelacije dobijeni su između količine NO3-N u zemljištu i parametara kvaliteta korena, ali nisu bili značajni između količine azota i prinosa korena i rafinisanog šećera

Razvoj transgene otpornosti šećerne repe na virus nekrotičnog žutila nerava repe (BNYVV)

Fragments of viral cDNA containing the coat protein gene of beet necrotic yellow vein virus were cloned in plant transformation vector pCAMBIA3301M with the bar gene as selectable marker. Vector pC3301MCPL carrying coat protein gene with leader sequence, and pC3301MCPS with coat protein gene, were used in Agrobacterium - mediated transformation of sugar beet. The transformation method used was based on the fact that sugar beet develops axillary shoots in in vitro conditions, when placed on media with citokinins. Since this ability is not genotype or ploidy dependant it is widely used for sugar beet vegetative multiplication. Sterile seedlings, with removed cotyledons and lower half of hypocotyl, were used as starting material. After transformation ex-plants were put on micropropagation medium with cephotaxime and phosphinotricyn (ppt), where axillary shoots started to develop. Since concentration of ppt was not selective enough, after two subcultivations it was increased twofold. Only one sample, transformed with pC3301MCPS preserved morphogenetic potential for micropropagatio, and it was tested for presence of COS fragment and bar gene bz PCR with soecific primers.Fragmenti virusne cDNK sa genom za protein omotača virusa nekrotičnog žutila nerava repe su klonirani u vektor za transformaciju biljaka pCAM-BIA3301M koji je sadržao bar gen kao selektivni marker. Vektori pC3301MCPL, sa genom za protein omotača virusa i njegovom lider sekvencom, i pC3301MCPS, sa genom za protein omotača, su korišćeni u tramsformaciji repe pomoću Agrobacterium-a. Metod transformacije se zasniva na sposobnosti repe da u uslovima in vitro razvije aksilarne pupoljke na podlozi sa citokininima. Pošto ova sposobnost ne zavisi od genotipa ili od nivoa plodnosti, postala je standardni metod za vegetativno umnožavanje repe. Kao početni materijal su korišćeni sterilni ponići kojima su odstranjeni kotiledoni i donja polovina hipokotila. Nakon transformacije eskplantati su postavljeni na selektivnu podlogu za mikropropagaciju sa cefotaksimom i fosfinotricinom (ppt) gde je došlo do razvoja bočnih pupoljaka. Po.sto koncentracija fosfinotricina nije bila dovoljno selektivna, ona je nakon dve subkultivacije dvostruko povećana. Samo je jedan uzorak, transformisan vektorom pC3301MCPS, nakon dve subkultivacije sačuvao mofrogenetski potencijal za mikropropagaciju, i bio testiran na prisustvo CPS fragmenta i bar gena PCR reakcijom sa specifičnim prajmerima

Doprinos nauke u promeni genetičke konstitucije sorata šećerne repe tokom četrdeset godina seminara agronoma

In the forty years of seminars of agronomists, the genetic composition of sugar beet hybrids and varieties has been completely changed, mainly due to development of different scientific methods. Every change lead to increase of genetic yield potential for important quantitative and qualitative traits, or improvement of efficiency and profitability of production. Sugar beet breeding in Institute of field and vegetative crops always managed to follow European and world trends, in order to create hybrid and varieties whose performance was equal to hybrids made by international companies. Increase in yield potential of 2% per year in the last few decades, is mostly due to genotype improvement, which presents huge success of scientists involved in sugar beet genetics and breeding.U proteklih četrdeset godina u vremenu trajanja seminara agronoma zahvaljujući rezultatima fundamentalne nauke mnogo češće nego kod drugih gajenih biljaka kod šećerne repe je nekoliko puta dolazilo do kompletne promene genetičke kompozicije gajenih sorti šećerne repe. Svaka promena dovodila je do povećanja genetičkog potnecijala rodnosti za najvažnije kvantitativne karakteristike ili do povećanja ekonomičnosti i profitabilnosti proizvodnje. Oplemenjivanje šećerne repe u Naučnom institutu za ratarstvo i povrtarstvo je uglavnom vrlo efikasno pratilo evropske i svetske trendove u smislu stvaranja sorti koje po svojim karakteristikama nisu zaostajale za sortama multinacionalnih kompanija. U zadnjih nekoliko dekada povećanje potencijala rodnosti koje se duguje isključivo poboljšanom genotipu iznosi 2% godišnje što predstavlja nesumnjivo veliki uspeh genetičara i oplemenjivača šećerne repe