Plant regeneration of southern Adriatic iris by somatic embryogenesis

A simple protocol has been developed for plant regeneration by somatic embryogenesis of Southern Adriatic iris (Iris pseudopallida Trinajstić), an endemic species of the Balkan Peninsula. Somatic embryogenesis was induced in zygotic embryo culture on media supplemented with 2,4-dichlorophenoxy acetic acid (2-10 mgL-1) as the sole plant growth regulator, where both embryogenic calli and somatic embryos were induced. Subsequent decrease of 2,4-D in the media promoted formation of somatic embryos. Developed somatic embryos germinated on medium without growth regulators. The regenerated plantlets had diploid chromosome number. Planted plantlets acclimatized very well under greenhouse and garden conditions.Jednostavan protokol za regeneraciju biljaka procesom somatske embriogeneze razvijen je za južno-jadranski iris (Iris pseudopallida Trinajstić), endemit Balkanskog poluostrva. Somatska embriogeneza je indukovana u kulturi 'zrelih' zigotskih embriona na hranljivoj podlozi obogaćenoj 2,4-dihlorofenoksi sirćetnom kiselinom (2,4-D), (kao jedinim regulatorom rastenja), gde su embriogeni kalus i somatski embrioni formirali. Smanjenje koncentracije 2,4-D u hranljivoj podlozi dovodi do povećanja broja formiranih somatskih embriona. Klijanje somatskih embriona je postignuto na podlozi bez regulatora rastenja. Regenerisane biljčice su imale diploidan broj hromozoma. Aklimatizacija biljaka je bila dobra u uslovima staklenika i bašte.Projekat ministarstva br. 14263

Superoxide dismutase activity and isoenzyme profiles in bulbs of snake's head fritillary in response to cold treatment

The activities and isoenzyme profiles of superoxide dismutase (SOD) in in vitro Fritillaria meleagris bulbs in response to cold treatment (4°C) were investigated. Differences in SOD activity and isoenzyme profiles in bulbs under standard growth conditions, six weeks chilling, as well as seven days after the completion of cold treatment are presented. SOD activity initially decreased but then rapidly increased seven days after cold treatment. Four isoforms of SOD are active under standard and chilling conditions, while three isoforms are presented 7 days after cold treatment. Native gel electrophoresis indicated the presence of mitochondrial and chloroplast localized SODs.

Superoxide dismutase activity and isoenzyme profiles in bulbs of snake's head fritillary in response to cold treatment

The activities and isoenzyme profiles of superoxide dismutase (SOD) in in vitro Fritillaria meleagris bulbs in response to cold treatment (4°C) were investigated. Differences in SOD activity and isoenzyme profiles in bulbs under standard growth conditions, six weeks chilling, as well as seven days after the completion of cold treatment are presented. SOD activity initially decreased but then rapidly increased seven days after cold treatment. Four isoforms of SOD are active under standard and chilling conditions, while three isoforms are presented 7 days after cold treatment. Native gel electrophoresis indicated the presence of mitochondrial and chloroplast localized SODs.Proučavana je aktivnost i izoenzimski profil superoksid dizmutaze (SOD) u lukovicama Fritillaria meleagris gajenim u uslovima in vitro kao odgovor na tretman hlađenja. Prikazane su razlike u aktivnosti SOD kao i izoenzimskim profilima kod lukovica na standardnim uslovima gajenja, posle šest nedelja hlađenja i sedam dana posle završetka tretmana hlađenja. Aktivnost SOD se smanjuje tokom hlađenja dok se značajno povećava sedam dana posle hlađenja. Četiri SOD izoforme su aktivne tokom gajenja u standardnim uslovima i tokom hlađenja dok su tri izoforme detektovane sedam dana posle hladnog tretmana. Nativna gel elektroforeza je pokazala prisustvo aktivnosti mitohondrijalne i hloroplastne SOD.Projekat ministarstva br. 143026

Efficient genetic transformation of Impatiens hawkerii Bull. (Balsamiaceae) using agrobacterium rhizogenes

Transformation of Impatiens hawkerii Bull. mediated by Agrobacterium rhizogenes strain A4M70GUS was studied. Hairy roots developed 10 days after inoculation were excised from the shoot explants and transferred onto Murashige and Skoog's (MS) basal medium lacking plant growth regulators. More than 20 hairy root clones were established and eight of them were further analyzed. Each clone differed significantly from the others in growth capacity and lateral branching. Clone C2 showed the highest biomass (20.6 g L-1) as well as the highest number of lateral roots (37 ± 2.2). The transgenic nature of the established hairy root clones was confirmed by GUS assay and PCR analysis. In conclusion, hairy roots were developed for the first time in I. hawkerii Bull., and transgenic hairy root clones showed a distinct morphological nature and growth patterns.Proučavana je genetička transformacija Impatiens hawkerii Bull. posredstvom Agrobacterium rhizogenes soja A4M70GUS. Deset dana posle inokulacije formirali su se transgeni korenovi na eksplantatima izdanaka, a zatim gajeni na Murashige and Skoog's (MS) osnovnoj hranljivoj podlozi bez biljnih regulatora rastenja. Uspostavljene su kulture više od 20 klonova, a 8 je dalje analizirano. Klonovi su se međusobno značajno razlikovali u odnosu na kapacitet rastenja i bočnog grananja. Klon C2 je imao najveću biomasu (20.6 g L-1), kao i najveći broj bočnih korenova (37 ± 2.2). Prisustvo stranih gena u klonovima transgenih korenova je potvrđeno GUS eseja i PCR analize. Transgeni korenovi su dobijeni prvi put kod Impatiens hawkerii Bull. i pokazuju značajne razlike u morfologiji i parametrima rastenja.Projekat ministarstva br. TR-2301

Clonal fidelity of chrysanthemum regenerated from long term cultures

Morphological characteristics of flowers of long term regenerated chrysanthemum, cv. "White Spider", after ten years of micropropagation are investigated. Shoot cultures are established and maintained more than ten years by stem segment culture on MS medium supplemented with BAP and NAA (1.0, 0.1 mgL-1, respectively). Rooting of shoots (100 %) has done on MS medium without hormones and it was very successful after ten years, as well as, after two or eight years of micropropagation. Acclimation of rooted chrysanthemum plantlets at greenhouse conditions was excellent and after appropriate photoperiod "in vitro" plants flowered 90.3 % and have the same flower color, shape and size as mother plants. Flower color changes of "in vitro" plants are observed during another flowering cycle one year after acclimatization. Observed variations of chrysanthemum flowers could be attributed to epigenetic factors.Praćene su morfološke karakteristike cvetova regeneranata cv. "White Spider" posle deset godina gajenja u kulturi in vitro. Izdanci su umnožavani u kulturi segmenata stabla preko aksilarnih pupoljaka na MS podlozi sa α-naftilsirćetnom kiselinom i benzilaminopurinom (0,5 odnosno 1,0 mgL-1), . Praćeno je umnožavanje izdanaka u periodu od devetnaest pasaža (tri godine) kada je došlo do sukcesivnog smanjenja indeksa umnožavanja. Oživljavanje izdanaka je praćeno na MS podlozi bez hormona i bilo je podjednako uspešno posle dve, kao i posle osam i deset godina mikropropagacije. Oživljene biljke hrizantema su uspešno aklimatizovane na uslove staklenika (97 %). Nakon odgovarajućeg fotoperioda, 90,3 % biljaka je cvetalo i imale su identičnu boju cveta matičnim biljkama hrizantema. Promene u boji cveta su uočene kod "in vitro" biljaka u sledećem ciklusu cvetanja, tj. godinu dana posle aklimatizacije. Ove promene bi se mogle pripisati epigenetskim faktorima.nul

Effect of low temperature on rooting rate and carbohydrate content of Fritillaria meleagris bulbs formed in culture in vitro

Projekat ministarstva br. 143026

Effect of nitrogen salts on the growth of Ceratonia siliqua L. Shoot cultures

Effects of reduced nitrogen salt nutriton on the growth, lenticel hypertrophy and anthocyanin accumulation of carob (Ceratonia siliqua L.) shoot cultures were investigated in conditions of light and darkness. Growth of shoot cultures was not significantly affected until nitrogen salts were reduced to less than ¼ of full-strength MS (Murashige and Skoog, 1962) values. Cultures in darkness were less affected and their main shoots even increased in length. Appearance of hypertrophied lenticels in light decreased, while in darkness they were absent in all treatments. Reduced nitrogen salt nutrition strongly affected anthocyanin accumulation of shoots and leaves, which greatly increased in both light and darkness. .Kod kultura izdanaka rogača ispitivani su efekti smanjenja koncentracije azotnih soli u podlozi MS mineralnog rastvora na multiplikaciju i izduživanje izdanaka, hipertrofiju lenticela i sintezu i akumulaciju antocijanina na svetlosti i u mraku. Smanjenje koncentracije N u podlozi bitno menja parametre rastenja izdanaka (multiplikaciju i izduživanje) tek na ¼ (na svetlosti) tj. 1/10 (u mraku). Hipertrofirane lenticele se ne razvijaju na etioliranim izdancima gajenim u mraku dok su kod izdanaka gajenim na svetlosti brojne. Smanjenjem koncentracije N u podlozi broj HL se smanjuje kao i njihovo akropetalno prostiranje. Sinteza i akumulacija antocijanina u izdancima i listovima rogača bila je direktno zavisna od koncentracije N u podlozi i na svetlosti i u mraku. .nul

Adventitious bud induction in pinus heldreichii christ: Seedling explant culture

By using tissue culture methods multiple shoot regeneration was achieved from seedling expiants of Pinus heldreichii in the presence of 1 mg/1 benzyladenine. The age of seedlings was found to be important for bud induction and shoot elongation. Histological events associated with bud primordium formation were also examined.Regeneracija izdanaka u kulturi klijanaca Pinus hrldreichii postignta je u prisustvu l mg/1 benziladenina, korišćenjem metode kulture tkiva. Nađeno je da starost eksplantata utiče na proces indukcije i razvića adventivnih pupoljaka. Razviće adventivnih pupoljaka je histološki analizirano.nul

Plant regeneration of southern Adriatic iris by somatic embryogenesis

A simple protocol has been developed for plant regeneration by somatic embryogenesis of Southern Adriatic iris (Iris pseudopallida Trinajstić), an endemic species of the Balkan Peninsula. Somatic embryogenesis was induced in zygotic embryo culture on media supplemented with 2,4-dichlorophenoxy acetic acid (2-10 mgL-1) as the sole plant growth regulator, where both embryogenic calli and somatic embryos were induced. Subsequent decrease of 2,4-D in the media promoted formation of somatic embryos. Developed somatic embryos germinated on medium without growth regulators. The regenerated plantlets had diploid chromosome number. Planted plantlets acclimatized very well under greenhouse and garden conditions.Jednostavan protokol za regeneraciju biljaka procesom somatske embriogeneze razvijen je za južno-jadranski iris (Iris pseudopallida Trinajstić), endemit Balkanskog poluostrva. Somatska embriogeneza je indukovana u kulturi 'zrelih' zigotskih embriona na hranljivoj podlozi obogaćenoj 2,4-dihlorofenoksi sirćetnom kiselinom (2,4-D), (kao jedinim regulatorom rastenja), gde su embriogeni kalus i somatski embrioni formirali. Smanjenje koncentracije 2,4-D u hranljivoj podlozi dovodi do povećanja broja formiranih somatskih embriona. Klijanje somatskih embriona je postignuto na podlozi bez regulatora rastenja. Regenerisane biljčice su imale diploidan broj hromozoma. Aklimatizacija biljaka je bila dobra u uslovima staklenika i bašte.Projekat ministarstva br. 14263

Virus elimination from ornamental plants using in vitro culture techniques

Viruses are responsible for numerous epidemics in different crops in all parts of the world. As a consequence of their presence great economic losses are being incurred. In addition to the development of sensitive techniques for detection, identification and characterization of viruses, substantial attention has also been paid to biotechnological methods for their elimination from plants. In this review article, the following biotechnological in vitro culture techniques for virus elimination from ornamental plants are presented: meristem culture, thermotherapy, chemotherapy, cryotherapy or a combination of these methods. The plant species, as well as the type of virus determine the choice of a most suitable method. The state of the art in investigation of virus elimination from Impatiens sp. in Serbia is summarized.Virusi su odgovorni za brojne epidemije na različitim usevima u svim delovima sveta. Posledica njihovog prisustva su velike ekonomske štete, pa osim razvoju osetljivih tehnika za detekciju, identifikaciju i karakterizaciju virusa, velika pažnja se poklanja i biotehnološkim metodama za njihovu eliminaciju. U ovom preglednom radu predstavljene su tehnike in vitro kulture za eliminaciju virusa iz biljnog materijala: kultura meristema, termoterapija, hemoterapija, krioterapija ili kombinacija ovih metoda. Koja će metoda biti primenjena zavisi od biljne vrste, kao i od vrste virusa. U radu je dat pregled istraživanja na eliminaciji virusa iz Impatiens sp. u Srbiji.Projekat ministarstva br. TR-31019 and III4300