Biotechnology: reality or dream

The development of molecular biology and molecular genetics, especially of the recombinant DNA technology enabled improvement of experimental methods that provide manipulation within a cell-free system, such as cell and tissue cultures. Such methods resulted in the development of different new technologies with specific properties in relation to the conventional definitions. According to PERSLEY and lantin (2000) the following components are essential for the contemporary biotechnology: (i) genomics - a molecular characterization of all genes and gene products of an organism (ii) bioinformatics - the assembly of data from genomic analysis into accessible forms; (iii) transformation - the introduction of genes controlling a trait of interest into a genome of a desired organism (micro organisms, plants, animal systems). By the application of cotemporary biotechnology new methods in the field of diagnostic are developed such as rapid and more accurate identification of the presence and absence of genes in the genome of the organism of interest (identification of pathogens prenatal diagnostics, molecular markers assisted breeding for plants, etc). The traits of an organism are determined by its genetic material, i.e. by a molecule of deoxyribonucleic acid (DNA). watson and crick (1953) were the first scientists to describe the structure of DNA as a double-stranded helix. Higher organisms contain a set of linear DNA molecules - chromosomes and a full set of chromosomes of an organism is a genome. Each genome is divided into a series of functional units, i.e. genes. The traits of an organism depend on genes, but their expression depends not only on genes but also on many other factors, including whether a gene, controlling the trait, expresses, specific cells in which it expresses and specially the mode by which the gene and its product interact with the environment. A special aspect within the application of biotechnology occurs as an interaction of a foreign gene with a genome of an integrated organism. Also application of biotechnology provides transfer of one or several favorable genes from any evolutionary category into other category of an organism and in such a way it is possible to develop genetically modified organisms (GMO) having expressed desired, target traits. A survey of the application of biotechnology in the world and our country is presented in this paper

Poljoprivedna biotehnologija - stanje i perspektive

The development of the DNA recombinant technology has provided the transfer of a single or several genes within or among species whereby organisms with new traits were developed. Such organisms have been called genetically modified organisms. The first genetically modified varieties of cultivated plants entered the market in 1996 and since then areas sown with such crops has been increasing, amounting to 60 M ha in 2002. During the stated period, genetically modified varieties and hybrids of cultivated plants were developed with a gene introduced for tolerance to herbicides, resistance to insects, prolonged maturity period and improved quality. However, heavy disputes have arisen all over the world relating the possible gain and potential risks from the growth and utilization of modified crops. First of all, there are ethical issues related directly to gene transfer from a species to a species, then effects of the introduced gene on the environment and human health, economical justification of cultivating genetically modified crops, consumers confidence in the legislation, labeling the products encompassing a genetic modification, effects on the global market and ever increasing food requirements. The effect of products derived from genetically modified plants on human health depends on a specific content of a product itself and can potentially be useful if a product contains an increased content of vitamins, with an allergen removed, or potentially harmful, if a new allergen or a toxin were introduced by genetic modifications. Each genetically modified product is subjected to a rigorous testing of its safety prior to its introduction into a food chain. It encompasses molecular, biochemical, toxicological, nutritional and allergenic tests. Many countries apply legislation that stipulates labeling of genetically modified products, whereby the fact that the commodity encompasses products of genetic modification is clearly pointed out.Razvoj tehnologije rekombinantne DNK omogućio je prenos pojedinačnog ili nekoliko gena unutar ili između vrsta pri čemu su dobijeni organizmi sa novim svojstvima, koji se nazivaju genetički modifikovani organizmi. Prve genetički modifikovane sorte gajenih biljaka su se pojavile na tržištu 1996. godine i od tada su površine zasejane ovim usevima povećane na 58.7 miliona hektara u 2002. godini. U navedenom periodu stvorene su genetički modifikovane sorte i hibridi gajenih biljaka sa unetim genom za tolerantnost na herbicide, otpornost na insekte, produženo vreme sazrevanja i poboljšan kvalitet. Ipak, u svetu postoje ogromna neslaganja oko moguće dobiti i potencijalnih rizika koji mogu nastati gajenjem i korišćenjem modifikovanih useva. Pre svega to su etička pitanja vezana za direktno prenošenje gena iz jedne u drugu vrstu, uticaj unetog gena na spoljnu sredinu i zdravlje ljudi, ekonomska opravdanost gajenja genetički modifikovanih useva, poverenje potrošača u zakonsku regulativu, obeležavanje proizvoda koji sadrže genetičku modifikaciju, uticaj na globalno tržište i rastuće potrebe za hranom

Genetički modifikovane biljke - koristi i rizici

Modern biotechnology is applied in agriculture, together with classical technology, in aim to increase production and improve quality of food. Revolutionary event for the plant biotechnology was possibility of creations of genetically modified plants, in fact, possibility of introducing of the genes from any organisms, to the plant genome. A genetic modified plant has the potential to offer very significant improvements in the quantity quality and acceptability of the world's food supply. First genetically modified plant were generated less than 18 years ago and now many GM plants are beginning to widely grown and products of these plants are available in the market place. Since the first commercial introduction of a genetically modified plant growing area has increase rapidly to more than 50 mill ha. The first generation of genetically modified crops with "input "traits as insect resistance, herbicide tolerance, delayed ripening, virus resistance are currently on the market.Moderna biotehnologija se primenjuje u poljoprivredi, zajedno sa klasičnim tehnologijama, u cilju povećanja proizvodnje hrane i poboljšanja njenog kvaliteta. Revolucioni pomak na polju biotehnologije biljaka predstavlja mogućnost stvaranja genetički modifikovanih biljaka tj. mogućnost ugrađivanja gena poreklom iz bilo kog organizma u genom biljke. Genetički modifikovane biljke imaju mogućnost da obezbede više, bolje i bezbednije proizvode koji zajedno sa klasičnom tehnologijom, povećavaju proizvodnju hrane koja će moći da zadovolji potrebe rastuće ljudske populacije. Prva genetički modifikovana biljka je dobijena pre osamnaest godina, a danas se gaje komercijalno mnoge genetički modifikovane biljke i proizvodi od njih su dostupni na tržištu. Prva generacija genetički modifikovanih biljaka za takozvane ²input² osobine obuhvata tolerantnost na totalne herbicide rezistentnost na insekte, rezistentnost na viruse. Od prve komercijalizacije genetički modifikovanih biljaka površine na kojima se gaje su značajno povećane na više od 50 mil ha

Genetička divergentnost samooplodnih linija kukuruza i heterozis

The genetic diversity of six maize inbred lines, as well as, the genetic similarity between parental inbred lines and heterosis for yield in their F1 crosses was studied. Three inbred lines (ZPL 142, ZPL 680 and ZPL 357/3) are of the BSSS origin and another three (ZPL 257/3, ZPL 17/5 and ZPL 173/3) are of the non-BSSS genetic background. Molecular markers provide a direct determination of a number for which two inbred lines are different for a given number of loci. Maize inbred lines were genetically characterized with RAPD markers. Genetic similarity among genotypes was done by a statistical analysis with NTSYSpc v2.0 and by the application of the cluster analysis. Parental inbred lines, 30 F1 crosses with reciprocals, were included in a randomized complete block design with four replications in two densities (44,640 and 64,935 plants ha-1) at the location of Zemun Polje in 2003 and 2004. The differences among genotypes (F1 and inbreds line per se), densities and years were statistically significant. The highest value of mid-parent heterosis (192.2 %), as well as, the best-parent heterosis (178.0 %) was obtained for the cross ZPL 173/3 x ZPL 680 over investigated densities and years. The Spearman's correlation coefficient was used to determine correlations between the values of heterosis obtained in the field experiment and the value of genetic distances based on RAPD markers. The obtained results indicate that RAPD markers can be used to study the genetic diversity of maize inbred lines, although their application in the prediction of heterosis for grain yield is limited.U radu je ispitivana genetička različitost šest samooplodnih linija kukuruza na osnovu RAPD markera i odnos genetičke sličnosti između roditeljskih linija i heterozisa za prinos zrna u njihovim ukrštanjima (F1 hibridima). Roditeljske linije i 30 F1 hibrida, dobijenih ukrštanjem roditeljskih linija u potpunom dijalelu, su testirani po slučajnom blok dizajnu sa četiri ponavljanja u dve gustine useva (G1=44.640 i G2=64.935 biljaka ha-1) u Zemun Polju tokom 2003 i 2004. godine. Razlike u prinosu zrna između ispitivanih genotipova (F1 i samooplodne linije per se), ispitivanih gustina i spoljnih sredina su bile statistički značajne. Najveći heterozis dobijen je kod kombinacije ZPL 173/3 x ZPL 680 i u odnosu na vrednost srednjeg roditelja (192.2 %) i u odnosu na boljeg roditelja (178.0 %) u ispitivanim gustinama i godinama. Za određivanje korelacija između vrednosti za heterozis u kombinacijama dobijenim u poljskom ogledu i vrednosti genetičkih distanci izračunatih na osnovu RAPD markera korišćen je Spearman-ov koeficijent korelacije. Dobijeni rezultati ukazuju da se RAPD markeri mogu koristiti za ispitivanje genetičke divergentnosti samooplodnih linija kukuruza, mada je njihova primena za predviđanje heterozisa za prinos zrna ograničena

Genetički modifikovane biljke - nutritivni i zdravstveni aspekti

Genetically modified plants (GMP) with specific traits, have been developed by transfer of a single or several genes within or among species. Already, genetically modified sorts of cotton, soybean, oilseed rape as well as corn hybrids with gene for herbicide tolerance, insect and virus resistance, improving nutrition characteristics have been created. The use of genetically modified crops as animal feed from nutritional aspects have great potential but otherwise raised concerns about potentional risks in scientific community all over the world. Potential risks include development of allergic reactions, transfer of antibiotic gene to gut bacteria of animal and development of antibiotic resistance and potential adverse health effect in animals. The study of food safety obtained from genetically modified crops in the nutrition of animals has been conducted throughout the whole world. The results obtained have demonstrated no difference in the quality of the milk, meat and eggs of animals that had eaten either transgenic or no-transgenic feed. Of course, long term and thorough research should give more reliable results.Genetički modifikovane biljke (GMB), sa specifičnim karakteristikama, stvorene su zahvaljujući visokom razvoju tehnologije koja je omogućila prenos gena unutar ili između vrsta. Tako su već stvorene genetički modifikovane sorte soje, pamuka, uljane repice kao i hibridi kukuruza sa unetim genom za tolerantnost na herbicide, otpornost na insekte ili sa poboljšanim nutritivnim karakteristikama. Primena GM biljaka u ishrani domaćih životinja sa nutritivnog aspekta pruža velike mogućnosti dok sa druge strane potencijalni rizici njihovog korišćenja izazivaju brojne polemike u naučnim i stručnim krugovima. Kao najveći potencijalni rizik njihove primene navodi se opasnost od pojave rezistentnosti na antibiotike alergijske reakcije ali i razni drugi mogući neželjeni efekti na zdravstveno stanje. U cilju rešavanja ovih dilema sprovedena su brojna istraživanja koja su obuhvatila ispitivanje bezbednosti hrane dobijene od GM biljaka kod ishrane domaćih životinja. Rezultati dosadašnjih istraživanja su pokazala da se životinjski proizvodi dobije-ni od životinja hranjenih genetički modifkovanim biljkama ne razlikuju od onih hranjenih nemodifikovanim biljakama i nemaju uticaj na kvalitet mesa, mleka i jaja. Naravno, pravi potpun odgovor treba da pruže buduća obimnija istraživanja koja bi trebalo da daju relevantne podatke kako bi se ove dileme resile

Innovative solutions in crop utilization biofortification as a function of quality food production

Globalni trend industrijalizacije poljoprivrede, naročito od pojave „zelene revolucije“, karakteriše povećanje prinosa biomase, zrna i plodova useva. Ovakav trend ima za posledicu paralelno smanjenje hranljive vrednosti poljoprivrednih proizvoda, koje se ogleda u smanjenoj koncentraciji minerala i vitamina, posebno kada su u pitanju gvožđe, magnezijum, cink i selen. Kada se uzme u obzir visok stepen erozije, kao i narušavanje zemljišnog ekosistema, uz intenzivnu upotrebu mineralnih đubriva, baziranih uglavnom na azotu, fosforu i kalijumu, a bez unosa organskih đubriva, zamljišta se „ispošćuju“ i rapidno se gubi plodnost. Važno je istaći da je skoro polovina svetskih zemljišta deficitarna mineralima. Ovo se dalje, preko ciklusa ishrane, vrlo nepovoljno odražava na zdravstveno stanje ljudi i životinja, dovodeći do sistemske neishranjenosti i pojave brojnih hroničnih bolesti. Da bi se stalo na put navedenim trendovima, potrebno je proizvesti poljoprivredne proizvode bogate hranivima. Akumulacija minerala u biljkama, posebno u zrnu, kao i sinteza vitamina je kontrolisan brojnim biohemijskim procesima. Biofortifikacija ima za cilj povećanje koncentracije minerala i vitamina u jestivim delovima biljaka. Mere gajenja koje utiču na povećanje apsorpcije minerala i sintezu vitamina, kao i metode oplemenjivanja i genetičkog inženjeringa koje imaju za cilj stvaranje genotipova poželjnih osobina, mogu povoljno uticati na povećanje koncentracije minerala i vitamina u jestivim delovima gajenih biljaka. Paralelno sa povećanjem koncentracije esencijalnih minerala i vitamina, potrebno je razviti i mere kojima se utiče na smanjenje anti-nutritiva, koji sprečavaju resorpciju minerala i vitamina iz organa za varenje, odnosno njihovo iskorišćenje od strane animalnih organizama. Povećanje koncentracije esencijalnih minerala i vitamina, kao i njihova iskoristljivost od strane ljudskih i životinjskih organizama je vrlo kompleksna problematika, koja se bazira na primeni brojnih inovativnih rešenja i koja bi trebalo da predstavlja integralni deo i sponu između agronomskih i medicinskih nauka, sa ciljem poboljšanja kvaliteta agroekositema, gajenih biljaka i života, uopšte.The globally present trend of agriculture industrialization, particularly from arising of "green revolution" was characterised with increase of biomass, grain and fruit yields. In parallel, this trend had as a consequence decrease of nutritional quality of agricultural products, reflected through reduced concentration of minerals and vitamins, especially of iron, magnesium and selenium. When high intensity of erosion and depletion of soil ecosystem was taken into account, together with intensive usage of mineral fertilizers, based on nitrogen, phosphorus and potassium, without incorporation of organic fertilizers, soils became exhausted, rapidly losing their fertility. It is important to underline that almost half of soils worldwide is deficient in minerals. In further, through the nutrition cycle, this situation is transmitting to health depletion of humans and animals, towards systemic malnutrition and arising of numerous chronically diseases. To combat present trends, it is necessary to produce agricultural commodities rich in essential nutrients. Various biochemical processes control accumulation of mineral nutrients in plant tissues, particularly in grains, as well as vitamins synthesis. The goal of bio-fortification is increase of concentration of essential minerals and vitamins in edible parts of plants. Thus, growing measures that enhance absorption of minerals and vitamins synthesis, as well as methods of breeding and genetic engineering, having as a target creation of genotypes with desirable traits, could be positively reflect to increase in concentration of minerals and vitamins in edible parts of agricultural plants. Correspondingly to the increase in concentration of essential minerals and vitamins, it is necessary to develop measures that will reduce concentration of anti-nutrients, which diminish absorption of minerals and vitamins from digestive organs, thus obstruct their utilization by animals and humans. The increase of concentration of essential minerals and vitamins, as well as their bioavailability and utilization by animal and human organisms is very complex issue, based on the application of various innovative solutions and it should present integral part, i.e. connection between agricultural and medical sciences, aimed to increase quality of agro-ecosystem, agricultural plants, and life, in general

Korišćenje proteinskih markera za karakterizaciju i utvrđivanje genetičke čistoće semena kukuruza

Purity control and genetically characterization of maize inbred lines and hybrids have great importance in every phase of experimental or commercial hybrid seed production and inbred maintenance. Embryo proteins as genetic markers for maize genotypic identification as well as genetic purity determination were used. By electrophoresis embryo proteins have been separated into numerous components and showed great heterogeneity. All analyzed genotypes have specific protein pattern. Some of protein fraction appeared for each genotype at the same position and could be use as reference bands. The differences of protein complex among various maize variet­ies could be use to detect relationship between hybrids and their parental lines. The method is rapid, cheap and date can be use to form genetically identification card of inbred lines and hybrids.Kontrola genetičke čistoće i identifikacija genotipa je od velikog značaja u svim fazama proizvodnje eksperimentalnih i komercijalnih hibrida i održavanju linija kukuruza. U radu su korišćeni proteini klice za genetičku karakterizaciju genotipova kukuruza kao i za utvrđivanje genetičke čistoće semena. Elektroforezom proteini klice su razdvojeni na veliki broj frakcija koje pokazuju visoku heterogenost. Svi ispitani genotipovi kukuruza imali su jedinstvenu proteinsku sliku pri čemu se određen broj proteinskih frakcija nalazi na približno istoj poziciji u svim ispitanim uzorcima tako da se koriste kao referentne. Razlike u proteinskom sastavu linija mogu da se koriste i za utvrđivanje veze između hibrida i roditeljskih linija. Metod je brz, jeftin i podaci mogu da se koriste za formiranje genetičke identifikacione karte linija i hibrida kukuruza

The conventional and contemporary technologies in maize (Zea mays L) breeding at Maize Research Institut Zemun Polje

Broad genetic variability of starting biological material is crucial prerequisite for the successful breeding program. Maize Research Institute, owning Gene bank with more than 6,000 accessions consisted of the local and introduced collection, has the opportunity for the investigation both fundamental genetic processes, pre-breeding and breeding for commercial purposes. To search for new sources of maize drought tolerance Gene bank accessions were scored visually on stay-green phenotype and total appearance. More than 50 genotypes have been identified as a potential source for drought tolerance. Besides breeding on improved storage protein quality decreasing of phytate content in kernel is also under the consideration, because the most phosphorous is bound in phytate. In breeding program it is desirable to have genotypes with higher content of available phosphorous (P) not bound in phytate. Among 60 analyzed populations from Gene bank only one has been determined to have very low phytate content and will be used in maize breeding program on low phytate in kernel. Process of maize breeding, to get high yielding hybrids, begins by the genetic variability determination of starting breeding material, either populations or selected inbred lines. Genetic divergence of parental inbred lines is main step to get high heterotic effect in yield after crossing. Use of different molecular markers allowed cluster analysis by use of UPGMA methods and select high number of genotypes to be included in crossing process

Primena molekularnih markera i bioinformatike u oplemenjivanju biljaka

In the past decade development of molecular genetics brought new dimension of a plant breeding. Molecular markers as universally methods for all biological system virtually effect on a success of directly examination of structure and function of genome and well as determination of genetic polymorphism of plant genomes. The potential applications of molecular markers in plant breeding are: analysis of molecular basis of evolution; germ plasma identification classification and management; assessing genetic diversity; identification of genes underlying agronomy important traits as yield, resistance to stress and disease as well as heterosis. High density genetic linkage maps for a number of plant species as a basis for marker assisted selection of agronomically useful traits and isolation of these genes have been established. Important factor in the successes of the genetic improvement of crops was the development of faster and more reliable methods, which allowed easier analysis of date as well as rapid information exchange. The application of information technology and development of statistical techniques to analyze genomic information is know as 'bioinformatics'. By integrating genetics with informatics investigations of whole genomes aims to elucidate the structure function and evolution of plant genomes are faster. Together these technologies as integral part of classical breeding programs contributing significantly to shorting of plant breeding process and cycles of selection.Tokom poslednje decenije razvoj molekularne biologije uneo je novu dimenziju u oplemenjivanje biljaka. Molekularni markeri kao univerzalna metoda za sve sisteme je značajno uticala na uspeh direktnog ispitivanja strukture i funkcije genoma kao i ispitivanje genetičkog polimorfizma. Potencijalne primene molekularnih markera u oplemenjivanju biljaka su: ispitivanje molekularne osnove evolucije; identifikacija, klasifikacija i održavanje germplazme; ispitivanje genetičke raznovrsnosti; identifikacija gena koji kontrolišu prinos, heterosis, stres ili bolesti. Formirane su visoko zgusnute genetičke mape za brojne biljne vrste kao osnova selekcije zasnovane na markerima za agronomski važne osobine kao i izolovanje gena. Važan faktor u uspehu genetičkog poboljšanja biljaka je razvoj brzih i pouzdanih metoda koji omogućavaju lakšu analizu podataka kao i brzu razmenu informacija. Primena informatike i razvoj statističkih metoda za analizu genetičkih informacija poznata je kao 'bioinformatika'. Integracijom genetike sa informatikom ispitivanja na nivou genoma sa ciljem da se rasvetli struktura, funkcija i evolucija biljnog genoma su ubrzana. Zajedno ove tehnologije kao sastavni deo konvencionalnih programa oplemenjivanja značajno doprinose skraćenju procesa selekcije

Antagonističko dejstvo saprofitne bakterije serratia liquefaciens prema fitopatogenim gljivama kukuruza i potencijalni biohemijski i molekularni mehanizmi njenog dejstva

The objective of our study was to investigate effect of saprophytic bacterium Serratia liquefaciens derived from sunflower on maize phytopathogenic fungi in vitro and in vivo conditions, as well as, potential biochemical and molecular mechanisms. Obtained results indicate that production of an antibiotic is not a biochemical background of the antagonistic effect. Concurrence advantage of the saprophytic bacterial isolate S. liquefaciens isolated from sunflower, over the parasitic fungus Ph. ambiguum when limited quantities of carbon compounds were used in the joint culture, caused antagonistic effect between these two groups of microorganisms. The de novo synthesis new polypeptides of different molecular weight occurred in interactions with phytopathogenic fungi. The successful activities of isolates of the saprophytic bacterium under in vivo conditions were observed and manifested as lack of disease symptoms (root and stalk rots), and stimulatory effects on the rowth of both, roots and above-ground plant parts. The bacterial isolate S. liquefaciens showed the greatest inhibition rate of colonies growth of fungus Ph. ambiguum under in vivo conditions.Ispitivan je antagonistički efekat saprofitne bakterije Serratia liquefaciens, poreklom iz rizosfere suncokreta na fitopatogene gljive kukuruza u in vitro i in vivo uslovima, kao i potencijalni mehanizmi dejstva na biohemijskom i molekularnom nivou. Uočeno je da bakterijska vrsta S. liquefaciens ne poseduje sposobnost sinteze antibiotika. Antagonistički efekat koji je ova vrsta ispoljila prema Ph.ambiguum pri gajenju na hranljivoj podlozi sa različitim koncentracijama glukoze rezultat je postojanja kompeticijskog odnosa. U interakciji sa fitopatogenim gljivama došlo je do de novo sinteze proteinskih frakcija različitih molekulskih težina. Uspešno delovanje ovog bakterijskog izolata na razvoj fitopatogene gljive Ph. ambiguum ustanovljeno je i u in vivo uslovima, koje se ispoljavalo izostankom simptoma bolesti (truleži korena i stabla) i stimulativnom dejstvu na porast korena kao i nadzemnih delova biljke