Tropic responses of potato single-node explant cultures

A special in-vitro protocol was elaborated enabling the production of potato single-node explant plantlets that can be used as objects for tropic studies. In light-grown plantlets, achievement of a full (90°) phototropic (PT) curvature required 75 to 120 min of continuous unilateral blue light irradiation or 120-135 min of gravitropic stimulation (GT). Time-lapse photography revealed that the curves describing PT and GT bending have a sigmoid shape. Continuous BL irradiation was necessary for the induction of continuous PT bending. If the BL was turned off after 30-50 min of PT stimulation, the bending gradually decreased and stopped in darkness after 25.0 ± 2.0 min. Within this period, curvature increased by 15.5 ± 1.5°. When the BL was turned off upon completion of PT bending (when the plantlets reached an angle of 90°), the plantlets entered the phase of fast straightening. The 90° PT curvature was significantly exaggerated in darkness by turning the jars from a vertical to horizontal position providing 120.74 ± 2.5° as the final curvature angle after two more hours in darkness

High biomass producing root cultures of Gentiana punctata L. transformed with Agrobacterium tumefaciens C58Cl(pArA4b)

Gentiana punctata L. shoot cultures were transformed with Agrobacterium tumefaciens clone C58C1 harbouring A. rhizogenes A4 Ri plasmid (pRiA4b) to produce hairy roots capable of autonomous growth on growth regulator-free medium. Optimal medium comprised WPM macro-nutrient salts, MS micro-salts and iron, LS vitamins and 2% sucrose. Successful transformation was confirmed by PCR amplification of awe 1 and rol C sequences. Excised, transgenic root cultures characterized by intensive lateral root branching enabled high biomass production. Fast growth required use of explants with small initial weight and frequent subculturing. Cultures were maintained for 10 years (more than 100 subcultures) without changes in morphology or physiological responses.Kulture izdanaka Gentiana punctata L. transformisane su transkonjugovanim sojem Agrobacterium tume-faclens klon C58C1 sa ubačenim A. rhizogenes A4 Ri plazmidom (pArA4b) koji produkuje korenove sposobne za autonoman rast na podlozi bez regulatora rastenja. Uspešna transformacija pokazana je PCR amplikacijom aih 1 i rol C sekvenci u uzorcima transformisanih korenova. Kulture transgenih korenova koje su se karakterisale intenzivnim bočnim grananjem, održavane su tokom 10 godina (90 i više subkultura) bez promena u morfologiji i fiziološkim svojstvima. Optimalni uslovi za rast ovih kultura bili su tečna podloga i 2% saharoza. Brz porast kultura zahtevao je male početne eksplantate i česta pasažiranja na svežu podlogu.nul

Phototropic bending of intact and wounded potato shoots

Intact and wounded shoots of in vitro cultured potato were investigated in situ to determine how their phototropic (PT) bending capacity was affected by water availability and exogenous auxin supplementation. Wounding strongly decreased PT bending but it recovered upon supplementation of water, auxin or both. Intact shoots required neither water nor auxin, while shoot segments required both. Shoot cuttings required only water, while, like shoot segments, decapitated shoots had a strong requirement for auxin. Water supplementation was beneficial in all treatments and PT bending was not affected in cultures that were submerged in water for a short period. Sucrose and inorganic salts present in the medium strongly affected PT bending capacity of cultures, favouring combinations with lower concentrations of both. Sucrose alone strongly promoted PT bending up to a concentration of 5%. Osmotic shock induced by the addition of small volumes of highly concentrated carbohydrate solutions (sucrose, glucose or sorbitol) induced a rapid but transient decline in PT bending capacity. These results indicate that water availability is a major factor that affects PT bending in potato plantlets.This is a post-peer-review, pre-copyedit version of an article published in Plant Cell, Tissue and Organ Culture (PCTOC). The final authenticated version is available online at: [http://dx.doi.org/10.1007/s11240-017-1235-2]Plant Cell, Tissue and Organ Culture (PCTOC) (2017

Chemical analysis and antimicrobial activity of methanol extracts of celandine (Chelidonium majus L.) plants growing in nature and cultured in vitro

Projekat ministarstva br. 143026B i 143041

High biomass producing root cultures of Gentiana punctata L. transformed with Agrobacterium tumefaciens C58Cl(pArA4b)

Gentiana punctata L. shoot cultures were transformed with Agrobacterium tumefaciens clone C58C1 harbouring A. rhizogenes A4 Ri plasmid (pRiA4b) to produce hairy roots capable of autonomous growth on growth regulator-free medium. Optimal medium comprised WPM macro-nutrient salts, MS micro-salts and iron, LS vitamins and 2% sucrose. Successful transformation was confirmed by PCR amplification of awe 1 and rol C sequences. Excised, transgenic root cultures characterized by intensive lateral root branching enabled high biomass production. Fast growth required use of explants with small initial weight and frequent subculturing. Cultures were maintained for 10 years (more than 100 subcultures) without changes in morphology or physiological responses.Kulture izdanaka Gentiana punctata L. transformisane su transkonjugovanim sojem Agrobacterium tume-faclens klon C58C1 sa ubačenim A. rhizogenes A4 Ri plazmidom (pArA4b) koji produkuje korenove sposobne za autonoman rast na podlozi bez regulatora rastenja. Uspešna transformacija pokazana je PCR amplikacijom aih 1 i rol C sekvenci u uzorcima transformisanih korenova. Kulture transgenih korenova koje su se karakterisale intenzivnim bočnim grananjem, održavane su tokom 10 godina (90 i više subkultura) bez promena u morfologiji i fiziološkim svojstvima. Optimalni uslovi za rast ovih kultura bili su tečna podloga i 2% saharoza. Brz porast kultura zahtevao je male početne eksplantate i česta pasažiranja na svežu podlogu.nul

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

In vitro shoot regeneration from seedling explants in Brassica vegetables: Red cabbage, broccoli, Savoy cabbage and cauliflower

Brassica oleracea varieties (red cabbage, broccoli, Savoy cabbage and cauliflower) were tested for their ability to regenerate shoots in vitro. Cotyledon, hypocotyl and root explants of 7 day-old seedlings were incubated on Murashige and Skoog's (MS) medium supplemented with 1 mg l-1 6-benzyladenine (BA) or 6-furfurylaminopurine (KIN) in combination with 0, 0.1, and 0.2 mg l-1 indole-3-butyric acid (IBA). Hypocotyls showed the best explants in almost all varieties tested with a minimum regeneration potential of 75% and producing 3.5-7.4 shoots per explant. The BA-supplemented media were optimal for both shoot regeneration and multiplication. Shoots rooted maximally (100%) on plant growth regulator-free MS medium containing 2% or 4% sucrose. Increased sucrose content improved plant acclimation in the greenhouse.Ispitivana je sposobnost četiri varijeteta Brassica oleracea (crveni kupus, brokoli, kelj i karfiol) da regenerišu pupoljke in vitro. Eksplantati kotiledona, hipokotila i korenova, uzetih sa 7 dana starih klijanaca, su gajeni na Murashige i Skoog (MS) hranljivoj podlozi sa dodatkom 1 mg l-1 6-benziladenina (BA) ili 6- furfurilaminopurina (KIN) u kombinaciji sa 0, 0.1, i 0.2 mg l-1 indol-3-butirične kiseline (IBA). Eksplantati hipokotila su se pokazali kao najbolji za regeneraciju kod skoro svih testiranih varijeteta sa minimalnim regenerativnim potencijalom od 75% i sa produkcijom 3.5- 7.4 pupoljaka po eksplantatu. Podloge koje su sadržale BA su bile optimalne, kako za regeneraiju pupoljaka, tako i za njihovu kasniju multiplikaciju. Maksimalan procenat oživljavanja izdanaka (100%) je postignut na MS medijumu bez dodatih regulatora rastenja, a koji je sadržao 2% ili 4% saharozu. Povećan sadržaj saharoze u medijumu za ožiljavanje uticao je na poboljšanu aklimatizaciju biljaka u stakleniku.Projekat ministarstva br. 143026 i TR-20072

Chemical analysis and antimicrobial activity of methanol extracts of celandine (Chelidonium majus L.) plants growing in nature and cultured in vitro

Projekat ministarstva br. 143026B i 143041

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