Elektroforetska slika proteina normalnog i tumorskog tkiva boba

Iz lista, stabljike i tumora na stabljici boba ekstrahirani su topivi i ukupni proteini te kvantitativno određeni. Proteinima je najbogatiji list, a u tumorskom tkivu znatno ih je više nego u odgovarajućem tkivu stabljike. Razdvajanje proteina disk-elektroforezom u poliakrilamidnom gelu pokazalo je razlike u proteinskoj slici između normalnog i tumorskog tkiva. Za tumor je karakteristično razdvajanje proteinske frakcije 1 u dvije pruge

Složena biokemija i biotehnološka proizvodnja betalaina

The demand for natural food colourants is increasing because of public awareness of their health benefits. Betalains are nitrogen-containing plant pigments whose colours range from red-violet betacyanins to yellow betaxanthins. They are used for colouring dairy products, meat and frozen desserts. Betalains have attracted additional interest because of their antioxidative, anti-inflammatory and anticarcinogenic properties. The main source of commercially produced betalains is red beet root, but alternative sources are found in plants from the Amaranthaceae and Cactaceae families. Another alternative source is plant cell culture in bioreactors, although optimization of pigment production seems necessary. In this paper we synthesize the results of recent studies on betalain biosynthesis, chemical properties, sources, biotechnology and applications.Sve je veća potražnja za prirodnim bojilima zbog njihovih zdravstvenih pogodnosti. Betalaini su biljni pigmenti koji sadrže dušik, s rasponom boja od crveno-ljubičastih betacijana do žutih betaksantina. Koriste se za bojanje mliječnih proizvoda, mesa i smrznutih deserta. Zanimljivi su i zbog svoga antioksidativog, protuupalnog i antikancerogenog djelovanja. Glavni komercijalni izvor betalaina je korijen cikle, a alternativni izvori pigmenta nalaze se u biljkama iz porodica Amaranthaceae i Cactaceae. Biljne stanice uzgajane u bioreaktorima također su izvor pigmenata iako je potrebno daljnje optimiranje njihove proizvodnje. U ovom su radu sažeti rezultati nedavnih istraživanja biosinteze betalaina, njihovih kemijskih svojstava, izvora, biotehnologije i uporabe

Složena biokemija i biotehnološka proizvodnja betalaina

The demand for natural food colourants is increasing because of public awareness of their health benefits. Betalains are nitrogen-containing plant pigments whose colours range from red-violet betacyanins to yellow betaxanthins. They are used for colouring dairy products, meat and frozen desserts. Betalains have attracted additional interest because of their antioxidative, anti-inflammatory and anticarcinogenic properties. The main source of commercially produced betalains is red beet root, but alternative sources are found in plants from the Amaranthaceae and Cactaceae families. Another alternative source is plant cell culture in bioreactors, although optimization of pigment production seems necessary. In this paper we synthesize the results of recent studies on betalain biosynthesis, chemical properties, sources, biotechnology and applications.Sve je veća potražnja za prirodnim bojilima zbog njihovih zdravstvenih pogodnosti. Betalaini su biljni pigmenti koji sadrže dušik, s rasponom boja od crveno-ljubičastih betacijana do žutih betaksantina. Koriste se za bojanje mliječnih proizvoda, mesa i smrznutih deserta. Zanimljivi su i zbog svoga antioksidativog, protuupalnog i antikancerogenog djelovanja. Glavni komercijalni izvor betalaina je korijen cikle, a alternativni izvori pigmenta nalaze se u biljkama iz porodica Amaranthaceae i Cactaceae. Biljne stanice uzgajane u bioreaktorima također su izvor pigmenata iako je potrebno daljnje optimiranje njihove proizvodnje. U ovom su radu sažeti rezultati nedavnih istraživanja biosinteze betalaina, njihovih kemijskih svojstava, izvora, biotehnologije i uporabe

Electrophoretic protein patterns and peroxidase activity related to morphogenesis in Mammillaria gracillis tissue culture

The cactus Mammillaria gracillis Pfeiff. was propagated in vitro on Murashige and Skoog (MS) medium without any growth regulators. At the bases of some plants abundant callus masses developed. Lower agar and higher MS salt concentrations stimulated callus production. In the culture, morphologically normal and hyperhydric shoots were regenerated. The growth of crown-gall tumour was induced on disc-like explants of in vitro grown plants infected with Agrobacterium tumefaciens, B6S3 strain. Calli appeared on both infected and control explants. The tumorous tissue grew more intensively than the control. The transformed character of the callus was confirmed by PCR amplification of a gene 6a of T-DNA. Gene expression in cactus shoots (grown in vitro or in pot), callus, hiperhydric and morphologically normal regenerated shoots and crown-gall tumour were analysed at the level of the electrophoretic pattern of soluble proteins. Some tumour-specific polypeptides were detected (76, 32-39, and 23 kDa). That of 42 kDa was highly expressed in callus and hyperhydric shoots. A faint 58 kDa band was found in all extracts except in the pot-grown shoot. Relatively high peroxidase activity was detected in callus and shoot regenerants and it was lower in tumour and the lowest in plant shoots

Protein glycosylation in sugar beet cell line can be influenced by DNA hyper- and hypomethylating agents

Protein glycosylation is a co- and post-translational modification that influences protein function, stability and localization. Changes in glycoprotein pattern during differentiation/dedifferentiation events exist in animal cells and DNA methylation status is closely related to the changes. However, in plant cells this relationship is not yet established. In order to verify whether such a relation exists, hypermethylating drugs 2,4-dichlorophenoxyacetic acid and hydroxyurea, or hypomethylating drug 5-azacytozine were applied to sugar beet (Beta vulgaris L.) cells during 14 days of in vitro subculture, and the glycoprotein patterns of the cells were compared. The applied drugs were not toxic, as observed from cell phenotype and by measuring growth of the control and treated cells. Hyper and hypomethylating treatments influenced the activity of enzymes related to differentiation state of the cells: peroxidases and esterases, and their isoform patterns. Electrophoretic patterns of soluble and membrane proteins were similar between control and treatments, but the treatments modified N- and O-linked glycoprotein patterns as visible from GNAand PNAlectin blots. This suggested that hypermethylation and hypomethylation of genomic DNA in sugar beet cells affect protein glycosylation patterns and cellular metabolism, possibly in a mechanism similar to that existing in animal cells

Crown-Gall Tumors on Bean Leaves Induced by Agrobacterium tumefaciens (I 1-4 i B6)

Agrobacterium tumefaciens, strains B6 and I1-4 and crown-gall tumors, which they induced on bean leaves, were investigated by means of light and electron microscopy. The results show: 1) When the bacteria of the strain I1-4 are reisolated from primary tumors their colonies show the S-forms as a rule. The view, that at low temperatures the R-form changes to the S-form, cannot be confirmed. 2) In the cells of both strains of Agrobacterium tumefaciens it has never been possible to prove any signs of the presence of a bacteriophage. 3) In living tumor cells the only cell organelles, which showed considerable ultrastructural changes, were the plastids. In other cell organelles no ultrastructural changes could be stated with certainty

Crown-gall tumor kao test za određivanje antitumorske aktivnosti proteina imele (Viscum album L.)

The crown gall tumor test system on potato tuber discs was used for the study of antiumor activity of mistletoe protein extracts. By estimating the number and weight of tumors we found that crude mistletoe protein extract inhibits tumor induction and growth. The refined protein fraction was even more efficient. The mistletoe proteins also showed stimulative effects on rhizogenesis.Crown-gall tumori na tkivu gomolja krumpira korišteni su kao test-sistem za ispitivanje antitumorske aktivnosti proteina u ekstraktu imele. Određivanje broja i mase tumora pokazalo je, sa statističkom sigurnošću, da već sirovi ekstrakt inhibira tumorsku indukciju i rast. Inhibitomo djelovanje pročišćenih proteinskih frakcija još je izrazitije. Proteinski ekstrakt stimulirao je rizogenezu u tumorskom tkivu

Analiza nastanka adventivnih pupova u kulturi embrija crnog bora (Pinus nigra Am.)

The development of adventitious buds in Black Pine (Pinus nigra Arn). embryo culture was studied from its beginning to its end, i. e. from the 4th to the 21st day of culturing. The formation of meristemoids brought up in about 12, of primordial buds 16 and of leaf primordia in 19 days. The adventitious buds developed in 21 days and then started to grow. During the culturing of Black Pine embryos the total activity of peroxydases was observed. From the 12th to the 19th day a large increase in peroxidase activity and a characteristic anodic separation pattern of two isoenzymes were stated.Razvitak adventivnih pupova u kulturi embrija crnog bora praćen je histološki i na temelju promjena u aktivnosti i elektroforetskoj slici izoproksidaza. Histološke analize pokazale su da su se adventivni pupovi počeli razvijati oko 4. dana, a njihovo formiranje je bilo okončano oko 21. dana trajanja kulture. Vremenski slijed četiriju sekvencija na koje je razvitak adventivnih pupova bio raščlanjen izgledao je ovako: nastajanje maristemoida bilo je završeno oko 12. dana, primordijalnih pupova oko 16. dana, a primordijalnih listova oko 19. dana nakon čega su rasli sve do kraja kulture. Praćenjem promjena u aktivnosti i elektroforetskoj slici izoperoksidaza utvrđeno je da je ukupna aktivnost peroksidaza u embrijima crnog bora tijekom njihove kulture rasla tako da je između 12. i 19. dana došlo do naglog porasta aktivnosti. Slika izoperoksidaza dobivena anodnim razdvajanjem pokazala je karakteristično razdvajanje na dva izoenzima također između 12. i 19. dana

Tumorska transformacija u tkivu gomolja krumpira

Potato (Solarium tuberosum L. cv. Desirée) tuber tissue was infected with Agrobacterium tumefaciens, strains: B6S3 (wild type), pGV2255 (rooty mutant), pGV2215 (shooty mutant) and with Agrobacterium rhizogenes (8196). Successful tumor induction was achieved by B6S3 (unorganised tumors) and by pGV2255 (rooty tumors). Transformed tuber cells were smaller than normal ones and free from starch. Transformation process caused a significant increase in peroxidase activity which can be taken as a tumorigenetic marker. Electrophoretic protein pattern showed that tumor cells stopped producing a polypeptide of about 40 kd, which was a storage protein dominant in normal tissue.Tkivo gomolja krumpira inficirale smo bakterijom Agrobacterium tumefaciens (sojevi: B6S3, pGV2215, pGV2255) i A. rhizogenes (8196). Uspješnu tumorsku transformaciju postigle smo divljim sojem (B6S3) i mutiranim (pGV2255) koji izaziva tumore s korijenom. Svjetlosnim mikroskopom mogle smo razlikovati normalne stanice od tumorskih. Tumorska transformacija odrazila se u porastu peroksidazne aktivnosti te stoga, ovaj enzim može poslužiti kao pokazatelj transformacijskih procesa. Elektroforetskim razdvajanjem topivih proteina u SDS-poliakrilamidnom gelu utvrdile smo da tumorske stanice prestaju sintetizirati proteinsku prugu na položaju koji odgovara molekularnoj težini od oko 40 kd