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

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

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

Dimethyl sulfoxide improves sensitivity and specificity of RT-PCR and qRT-PCR amplification of low-expressed transgenes

The expression of transgenes in a host plant may be low for a number of reasons. Both low expression and poor specificity of amplification were encountered during analysis of the expression of the Arabidopsis cytokinin oxidase/dehydrogenase (AtCKX1) gene in transgenic Centaurium erythraea. The optimization of the PCR protocol involved a gradient of annealing temperatures, as well as the application of seven PCR enhancers: formamide, dimethyl sulfoxide (DMSO), glycerol, ethylene glycol, trehalose, BSA and Tween-20. The best results for AtCKX1 amplification were obtained at 55.1ºC, with the addition of 5% DMSO. Glycerol and trehalose also improved the sensitivity of amplification, while formamide, ethylene glycol and BSA enhanced only the amplification of control purified targets, but not the transcripts. Tween-20 inhibited PCR. DMSO enhanced AtCKX1 PCR amplification and improved the specificity of qPCR amplification, as well as the assay reproducibility. This work emphasizes the usefulness of additives, which are rarely used for PCR optimization in real-time experiments.Projekat ministarstva br. ON173015 i br. ON17302

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

Production of neutron-rich nuclei in fragmentation reactions of 132Sn projectiles at relativistic energies

The fragmentation of neutron-rich 132Sn nuclei produced in the fission of 238U projectiles at 950 MeV/u has been investigated at the FRagment Separator (FRS) at GSI. This work represents the first investigation of fragmentation of medium-mass radioactive projectiles with a large neutron excess. The measured production cross sections of the residual nuclei are relevant for the possible use of a two-stage reaction scheme (fission+fragmentation) for the production of extremely neutron-rich medium-mass nuclei in future rare-ion-beam facilities. Moreover, the new data will provide a better understanding of the "memory" effect in fragmentation reactions.Comment: 5 pages, 3 figure

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

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

Solar neutrino detection sensitivity in DARWIN via electron scattering

We detail the sensitivity of the proposed liquid xenon DARWIN observatory to solar neutrinos via elastic electron scattering. We find that DARWIN will have the potential to measure the fluxes of five solar neutrino components: pp, $$^7$$ 7 Be, $$^{13}$$ 13 N, $$^{15}$$ 15 O and pep. The precision of the $$^{13}$$ 13 N, $$^{15}$$ 15 O and pep components is hindered by the double-beta decay of $$^{136}$$ 136 Xe and, thus, would benefit from a depleted target. A high-statistics observation of pp neutrinos would allow us to infer the values of the electroweak mixing angle, $$\sin ^2\theta _w$$ sin 2 θ w , and the electron-type neutrino survival probability, $$P_{ee}$$ P ee , in the electron recoil energy region from a few keV up to 200 keV for the first time, with relative precision of 5% and 4%, respectively, with 10 live years of data and a 30 tonne fiducial volume. An observation of pp and $$^7$$ 7 Be neutrinos would constrain the neutrino-inferred solar luminosity down to 0.2%. A combination of all flux measurements would distinguish between the high- (GS98) and low-metallicity (AGS09) solar models with 2.1–2.5 $$\sigma$$ σ significance, independent of external measurements from other experiments or a measurement of $$^8$$ 8 B neutrinos through coherent elastic neutrino-nucleus scattering in DARWIN. Finally, we demonstrate that with a depleted target DARWIN may be sensitive to the neutrino capture process of $$^{131}$$ 131 Xe.DARWIN Collaboration (ukupan broj autora: 166