Uticaj kadmijuma na razviće kalusa u kulturi zigotnog embriona pšenice

Toxicity effects of cadmium on callus growth and callus tissue dry matter content were investigated. Two winter wheat varieties, Evropa-90 and NSR-5 were used as a material in embryo culture. Isolated embryos were cultivated on a modified MS (Murashige and Skoog, 1962) medium to which Cd was added in three different concentrations (10-6, 10-5 and 10-4 M/l Cd). At the highest Cd concentration (10-4 m/l), CV. evropa-90 had 90.3% and CV. nsr-5 52.5% of survived calluses. Fresh callus weight was reduced at the concentration of 10-4 M/l Cd by 51% in cv. Evropa-90 and by 57% in cv. NSR-5 in relation to the control. Cadmium decreased the callus dry matter content in cv. NSR-5 only at the highest concentration (10-4 M/l), while in cv. Evropa-90 Cd had no signifficant effect on callus dry matter content.Ispitivano je toksično dejstvo kadmijuma na porast i sadržaj suve materije u kalusima pšenice. Dve sorte ozime pšenice, Evropa-90 i NSR-5, korišćene su kao materijal u kulturi embriona. Izolovani embrioni su kultivisani na modifikovanu MS podlogu (Murashige and Skoog, 1962) kojoj je Cd dodat u tri različite koncentracije (10-6, 10-5 i 10-4 M/l Cd). Pri najvišoj koncentraciji Cd (10-4 M/l), sorta Evropa-90 je imala 90.3%, a sorta NSR-5 52.5% preživelih kalusa. Pri koncentraciji od 10-4 M/l CdCl2 sveža masa kalusa je bila umanjena za 51% kod sorte Evropa-90, a kod sorte NSR-5 za 57%. Kadmijum je izazvao sniženje sadržaja suve materije u kalusima sorte NSR-5 samo pri najvišoj koncentraciji (10-4 M/l), dok kod sorte Evropa-90 Cd nije imao značajnog uticaja na sadržaj suve materije u kalusima

Visoka produkcija dvostrukih haploida pšenice korišćenjem kulture antera

Androgenous and regeneration abilities of 14 randomly selected F1 hybrids of wheat (Triticum aestivum L.) were analyzed. Anthers were grown in vitro on a modified Potato-2 inductive medium. The hybrid NS111-95/Ana had the highest average values for androgenous capacity (33%) and callus yield (119%), while the hybrid NS 92-250/Tiha had the lowest values for these traits (9 and 21%, respectively). Seven genotypes (50%) had a frequency of green plants relative to the number of isolated anthers of over 10%, with the highest frequency of 21.3% (NS111-95/Sremica). This hybrid produced 12.8 doubled haploid (DH) lines per spike used for isolation. In the other genotypes, the number of produced DH lines per spike ranged from 1 (30­Sc.Smoc.88-89/Hays-2) to 11.2 (NS111-95/Ana). As half of the randomly selected genotypes exhibited high green plant regeneration ability and a high production of DH lines per spike, it can be concluded that in vitro anther culture can be successfully used in breeding programs for rapid production of homozygous wheat lines.Analizirana je androgena i regeneraciona sposobnost 14 slučajno odabranih F1 hibrida pšenice (Triticum aestivum L.). Antere su gajene u in vitro uslovima na modifikovanoj Potatao-2 podlozi. Hibrid NS111-95/Ana imao je najviše prosečne vrednosti za androgeni kapacitet (33%) i prinos kalusa (119%), dok je hibrid NS 92-250/Tiha imao najniže vrednosti za ove osobine (9% i 21%). Sedam genotipova (50%) imalo je frekvenciju zelenih biljaka u odnosu na broj izolovanih antera preko 10%, sa najvišom vrednošću od 21.3% (NS11­95/Sremica). Kod istog hibrida (NS11-95/Sremica) proizvedeno je 12.8 linija dvostrukih haploida (DH) po klasu, korišćenom za izolaciju. Kod drugih genotipova, broj proizvedenih DH linija po klasu varirao je od 1 (30-Sc.Smoc.88­89/Hays-2) do 11.2 (NS111-95/Ana). S obzirom na to da je polovina slučajno odabranih genotipova ispoljila visoku sposobnost za regeneraciju zelenih biljaka i visoku produkciju DH linija po klasu, in vitro kultura antera može se uspešno koristiti u oplemenjivačkim programima za brzu proizvodnju homozigotnih linija pšenice

Genotipska specifičnost F1 hibrida pšenice u pogledu proizvodnje dvostrukih haploida metodom kulture antera

The objective of the study was to investigate the production of homozygous doubled haploid (DH) lines from heterozygous genotypes of wheat (Triticum aestivum L.) using an anther culture. Anthers were isolated from 20 randomly selected F1 wheat combinations and grown in vitro on a modified Potato-2 inductive medium. Plant regeneration was performed on the modified 190-2 medium. The results have shown that all genotypes had the ability to produce pollen calli, as well as, to regenerate green plants. The hybrid Mex.3 x MV 18 had the highest value for androgenous capacity (40.6%), while the combination Kutječanka x Slavija had the lowest androgenous capacity (6.3%). The average androgenous capacity of all genotypes was 19.4%. The regeneration frequency ranged from 1.3 to 21.6 green plants per 100 isolated anthers, with an overall mean of 9.6%. Out of a total of 582 regenerated green plants 279 (47.9%) were spontaneous double haploids. The average production of double haploids of wheat in this study amounted to 4.6 per 100 cultured anthers.Cilj rada bio je da se ispita mogućnost proizvodnje homozigotnih linija dvostrukih haploida (DH) iz heterozigotnih genotipova pšenice (Triticum aestivum L.) korišćenjem kulture antera. Antere su izolovane iz 20 slučajno odabranih F1 kombinacija pšenice i gajene u kulturi in vitro na modifikovanoj potato-2 indukcionoj podlozi. Regeneracija biljaka vršena je na modifikovanoj 190-2 podlozi. Rezultati su pokazali da su svi genotipovi posedovali sposobnost da proizvedu kaluse iz polena, kao i da regenerišu zelene biljke. Hibrid Mex.3 x MV 18 je imao najvišu vrednost za androgeni kapacitet (40,6%), dok je kombinacija Kutječanka x Slavija imala najniži androgeni kapacitet (6,3%). Prosečan androgeni kapacitet za sve genotipove bio je 19,4%. Frekvencija za regeneraciju zelenih biljaka kretala se od 1,3 do 21,6 zelenih biljaka na 100 izolovanih antera, sa prosekom od 9,6%. Od ukupno 582 regenerisane zelene biljke, 279 (47,9%) su bile spontani dvostruki haploidi. U ovom istraživanju prosečno je proizvedeno 4,6 dvostrukih haploida pšenice na 100 izolovanih antera

Korišćenje kalusne kulture za ispitivanje tolerantnosti genotipova pšenice prema suši

In this paper, we studied the drought tolerance under in vitro conditions of two winter wheat (T. aestivum L.) cultivars, Košuta and Renesansa. The tolerance was tested on a modified MS (Murashige and Skoog, 1962) nutrient medium to which polyethylene glycol (PEG) was added at three different concentrations: 10%, 20% and 30%. Calluses from the control group were grown on a medium without PEG. After four months of growing on these mediums, fresh weight and dry matter content in the callus tissue were determined. We found significant differences in genotype response to different PEG concentrations The highest concentration (30%) was lethal to the isolated embryos, so at this concentration no calluses formed in any of the genotypes. At 10 and 20% PEG, there was a significant decrease in the fresh callus weight in both genotypes. Thus, at the lowest concentration ( 10% PEG), the fresh weight decreased by 90% in Košuta and by 93% in Renesansa relative to the control. Due to callus dehydration in the presence of PEG, the dry matter content in the calluses of both genotypes increased with increasing PEG concentrations in the medium. In Košuta, the dry matter content increased from 8.7% (control) to 24.9% (20% PEG), while in Renesansa it increased from 8.6% (control) to 39.7% (20% PEG).Ispitivana je tolerantnost prema suši u in vitro uslovima, kod dve sorte ozime pšenice (T. aestivum L.), Košuta i Renesansa.Tolerantnost je testirana na modifikovanoj MS(Murashige and Skoog, 1962) hranljivoj podlozi, kojoj je dodat polietilen glikol (PEG) u tri različite koncentracije: 10%, 20% i 30%. Kontrolna grupa kalusa je gajena na podlozi bez PEG-a. Nakon četiri meseca gajenja na ovim podlogama utvrđena je sveza masa i sadržaj suve materije u kalusnom tkivu. Utvrđene su značajne razlike u pogledu reakcije genotipova na različite koncentracije PEG-a. Najviša koncentracija (30% PEG-a) je bila letalna za izolovane embrione, lako da pri ovoj koncentraciji, ni kod jednog genotipa nije došlo do formiranja kalusa. Pri koncentracijama od 10 i 20% PEG-a došlo je do naglog snižavanja vrednosti sveže mase kalusa kod oba genotipa. Tako je pri najnižoj koncentraciji (10% PEG) kod sorte Košuta sveza masa smanjena za 90% u odnosu na kontrolu, a kod sorte Renesansa za 93%. Usled dehidratacije kalusa u prisustvu PEG-a, sa povećanjem njegove koncentracije u podlozi, došlo je i do povećanja sadržaja suve materije u kalusima kod oba genotipa. Kod sorte Košuta sadržaj suve materije je povećan sa 8.7% (kontrola) na 24.9% (20% PEG), dok je kod sorte Renesansa povećan sa 8.6% (kontrola) na 39.7% (20% PEG)

Efikasnost tehnike kulture antera u proizvodnji dvostrukih haploida pšenice

The objective of the study was to investigate efficiency of anther culture in the production of spontaneous double haploids from randomly selected heterozygous genotypes of wheat (Triticum aestivum L.). Anthers of 20 F1 wheat combinations were grown in vitro on a modified Potato-2 medium. All of the examined genotypes have shown the ability to produce pollen calluses as well as to regenerate green plants. On average for the whole experiment material, 47.2 calluses were produced per 100 cultured anthers. The green plant regeneration ranged from 0.8 to 13.4 green plants per spike, with an overall mean of 5.8. From the total of 582 regenerated green plants, 47.9% (279) were spontaneous double haploids. The final average yield from the study was 2.8 double haploids per spike.Cilj rada bio je da se ispita efikasnost kulture antera u proizvodnji spontanih dvostrukih haploida iz slučajno odabranih heterozigotnih genotipova pšenice (Triticum aestivum L.). Antere iz 20 F1 kombinacija pšenice gajene su in vitro na modifikovanoj Potato-2 podlozi. Svi ispitivani genotipovi pokazali su sposobnost da proizvedu kaluse, kao i da regenerišu zelene biljke. U proseku za ceo eksperimentalni materijal, proizvedena su 47.2 kalusa na 100 izolovanih antera. Regeneracija zelenih biljaka kretala se od 0.8 do 13.4 zelene biljke po klasu, sa ukupnim prosekom od 5.8. Od ukupno 582 regenerisane zelene biljke, 47.9% (279) bile su spontani dvostruki haploidi. U proseku, u ovom istraživanju, proizvedeno je 2.8 DH biljaka po klasu

Uticaj hladnog pretretmana na androgenu sposobnost antera pšenice

In this paper the effect of cold pretreatment on androgenous ability of wheat anthers was investigated. Anthers were isolated from 12 different genotypes of wheat (Triticum aestivum L.). Before isolation, a half of anthers were exposed to a cold pretreatment on 4±1°C for 5-10 days, while the other half of anthers were inoculated onto the Potato-2 induction medium without the cold pretreatment. The results have shown that absence of cold pretreatment had significant effect on androgenous capacity in 5 cultivars, while on callus yield had the effect in 4 from the total of 12 examined wheat genotypes. It can be concluded that the effect of cold pretreatment on androgenous ability of wheat is genotype specific.U radu je ispitan uticaj hladnog pretretmana na androgenu sposobnost antera pšenice u kulturi in vitro. Korišćeno je 12 genotipova ozime pšenice. Pre izolocije, polovina antera bila je izložena hladnom pretretmanu na 4±1°C 5-10 dana, a druga polovina je bez pretretmana inokulisana na Potato-2 indukcionu podlogu. Rezultati su pokazali da je odsustvo hladnog pretretmana imalo uticaja na adrogeni kapacitet kod 5, dok je na prinos kalusa imalo uticaja kod 4 od 12 ispitivanih genotipova. Može se zaključiti da je uticaj pretretmana na androgenu sposobnost genotipski specifičan

Aspekti oplemenjivanja pšenice na niske temperature

Wheat resistance to low temperatures is complex in nature and dependant on variety as well as on interaction with the environment. In unfavorable conditions for hardening, even varieties resistant to cold may exhibit higher susceptibility to low temperatures. Over a two-year period (2003-04), tests were conducted in cold chamber at-15˚C lasting 24, 36, 48 and 60 hours. The 23 wheat varieties analyzed were divided into four groups according to year of release. In the first group, which was mostly made up of foreign introductions, the average plant survival rate across treatments was 87%. The different origins of the varieties from this group were a significant factor behind the differences in their resistance levels. The second group, comprised of the first domestic varieties, had an average plant survival rate of 89%, while the third, containing the dominant varieties from the 1980s, had a plant survival rate of 88.4% on average. In the fourth group, consisting mostly of varieties that are currently commercially grown in the country, the average plant survival rate was 86.6%. Thanks to high temperatures present during the process of hardening, there was greater plant damage and percent winterkill in the second year than in the first. The resistance of the varieties to low temperatures has been maintained at a level enabling successful and stable production under the cold temperature stress conditions of southeastern Europe.Otpornost pšenice prema niskim temperaturama je kompleksnog karaktera, pri čemu pored sorte, do izražaja dolazi i interakcija sa spoljnom sredinom. Pri nepovoljnim uslovima za kaljenje biljaka, i otporne sorte mogu ispoljiti veću osetljivost prema niskim temperaturama. U dvogodišnjem periodu (2004-2005), izvršeno je testiranje u hladnim komorama na temperaturi -15˚C u trajanju od 24, 36, 48 i 60 časova. Analizirano je 23 sorte pšenice koje su podeljene u četiri grupe, u zavisnosti od godine priznavanja. U prvoj grupi, u kojoj se uglavnom nalaze introdukovane sorte, procenat preživelih biljaka na svim tretmanima iznosio je u proseku 87%. Različito poreklo sorti koje se nalaze u ovoj grupi, značajno je uticalo na međusobne razlike u otpornosti prema niskim temperaturama. U drugoj grupi, koju čine prve domaće sorte, procenat preživelih biljaka iznosio je 89%, dok su u treću grupu uvrštene sorte koje su bile dominantne 80-ih godina, sa 88,4% preživelih biljaka. Većina sorti koje čine četvrtu grupu, trenutno je zastupljena na proizvodnim površinama u našoj zemlji. Kod ovih sorti, procenat preživelih biljaka iznosio je 86,6%. Visoke temperature u periodu kaljenja biljaka, uslovile su veća oštećenja, odnosno veći procenat izmrzlih biljaka u drugoj godini ispitivanja. Otpornost sorti pšenice prema niskim temperaturama, zadržana je nivou koji obezbeđuje uspešnu i stabilnu proizvodnju u uslovima hladnog temperaturnog stresa jugoistočne Evrope

Tolerantnost genotipova pšenice prema olovu u kulturi in vitro

We studied the effects of five lead concentrations (10-7, 10-6, 10-5, 10-4 and 10-3 M) on callus growth and dry matter content in the callus tissue. Two winter wheat (Triticum aestivum L.) cultivars, Balkan and Proteinka, were used to isolate mature embryos. The embryos were grown on a modified MS (Murashige and Skoog, 1962) nutrient medium to which lead in the form of Pb(NO3)2 was added. Calluses from the control group were grown on a lead-free medium. During cultivation, the growth of callus tissue was observed until, 30 days alter isolation, fresh callus weight and dry matter content were measured The results showed that there were significant differences between the genotypes with regard to their response to certain lead concentrations. The highest concentration (10-3 M) significantly decreased fresh callus weight relative to the control- by 43% in Balkan and 22% in Proteinka. At the same lead concentration, the callus tissue dry matter content of Balkan increased significantly (23% relative to the control), while the increase of the same parameter in Proteinka was not significant (8.6% relative to the control). The lower lead concentrations had a lees pronounced effect, although the 10-6 M dose had a stimulatory effect on callus tissue growth in Balkan, while the 10-7 M one had the same effect in Proteinka.Ispitivan je uticaj pet koncentracija olova (10-7, 10-6, 10-5, 10-4 i 10-3 M) na porast kalusa i sadržaj suve materije u kalusnom tkivu. Za izolaciju zrelih embriona korišćene su dve sorte ozime pšenice (Triticum aestivum L.), Balkan i Proteinka. Embrioni su gajeni na modifikovanoj MS (Murasbige and Skoog, 1962) branljivoj podlozi, kojoj je dodalo olovo u obliku. Pb(NO3)2. Kontrolna grupa kalusa je gajena na podlozi, koja nije sadržala olovo. Za vreme kultivacije, praćenje porast kalusnog tkiva, da bi se 30 dana nakon izolacije pristupilo merenju sveže mase kalusa, kao i sadržaju suve materije. Rezultati su pokazali da su postojale značajne razlike između genotipova, u pogledu njihove reakcije na pojedine koncentracije olova. Najviša koncentracija (10-3 M) olova je imala značajan uticaj na smanjenje sveže mase kalusa. Ovo smanjenje je kod sorte Balkan iznosilo 43%, a kod sorte Proteinka 22%, u odnosu na kontrolu. Najviša koncentracija olova, takođe je imala značajan uticaj na sadržaj suve materije u kalusnom tkivu. Tako je kod sorte Balkan došlo do značajnog povećanja sadržaja suve materije (23% u odnosu na kontrolu) , dok kod sorte Proteinka povećanje (8,6% u odnosu na kontrolu) nije bilo značajno. Efekat nižih koncentracija olova, koje sit ispitivane u ovom eksperimentu, bio je slabije izražen, mada su doze 10-6M kod sorte Balkan i 10-7M kod sorte Proteinka, imale stimulativno dejstvo na porast kalusnog tkiva

Povezanost disaharidnih profila odredenih GC/EI-qMS sistemom sa genomima sorti pšenice, raži i tritikalea

Cultivars of small grain species: wheat (Triticum aestivum), rye (Secale cereale) and triticale (Triticosecale) were selected because of their specific biochemical composition of carbohydrates for gas chromatographic analysis. Samples were grounded to flour and extracted with 96% ethanol solution. Extracted mono- and disaccharides were first derivatized into corresponding trimethylsilyl-oximes (TMSO) and analyzed on a GC/EI-qMS device. Four eluting peaks at specific retention times (18.57 min, 18.80 min, 20.85 min, and 21.39 min), were identified as trimethylsilyl-oximes of turanose, melibiose and two peaks of sucrose using Wiley 275 and NIST 14 mass spectra libraries, and were further selected as discrimination factors. Their presence/absence in certain small grain cultivar was labeled with 1/0 thus creating binary numerical matrices, introduced into multivariate analysis. Exploratory data analysis techniques were applied, thus providing a successful background for wheat, rye and triticale species differentiation, regardless of the specific cultivar they belong to, avoiding quantitative determinations of defined discrimination markers. Higher similarities between wheat and triticale cultivars, in comparison with rye species, may be due to the fact that most of the triticale genome structure originates from wheat species.Sorte strnih žita: pšenice (Triticum aestivum), raži (Secale cereale) i tritikalea (Triticosecale), analizirane su primenom sistema gasne hromatografije sa maseno-spektrometrijskom detekcijom zbog specifičnog biohemijskog sastava ugljenih hidrata. Uzorci su prvo samleveni u brašno, a potom ekstrahovani 96%-tnim rastvorom etanola. Ekstrahovani disaharidi su zatim derivatizovani u odgovarajuće trimetilsilil-oksime i analizirani pomoću GC/EI-qMS uređaja. Bibliotečkim pretraživanjem Wiley 275 i NIST 14 baza masenih spektara četiri eluirajuća pika, sa specifičnim vremenima retencije (18,57 min, 18,80 min, 20,85 min, i 21,39 min), identifikovana su kao trimetilsilil-oksimi turanoze, melibioze i saharoze, i odabrana kao diskriminirajući faktori. Njihovo prisustvo, odnosno, odsustvo u određenim sortama strnih žita označeno je sa '1', odnosno, '0', kreirajući pritom binarne numeričke matrice, koje su podvrgnute multivarijantnoj analizi. Primenjene istraživačke tehnike analize podataka pružile su dobru osnovu za diferencijaciju pšenice, raži i tritikalea, bez obzira na specifičnu sortu kojoj pripadaju, izbegavajući pritom kvantitativnu analizu definisanih markera diskriminacije. Pretpostavlja se da uočena izraženija sličnost između sorti pšenice i tritikalea, u odnosu na raž, može biti rezultat činjenice da najveći deo strukture genoma sorti tritikalea vodi poreklo upravo od sorti pšenice

In vitro evaluacija tolerantnosti genotipova pšenice (Triticum aestivum L.) prema boru

Wheat tolerance to high boron concentrations was determined using mature embryo culture. The testing was performed on a modified MS nutrient medium to which boric acid was added in two concentrations: 15 mM and 30 mM. The control medium contained no excess boric acid. The experiment involved 14 Serbian varieties. After one month of cultivation callus fresh weight (CFW) was measured and reductions of fresh callus weight (RFCW) at boron (B) concentration of 15 mM in relation to the control were calculated. The genotypes differed significantly according to their reaction to different B concentrations. Six genotypes had RFCWs below 50.0 %, while eight genotypes had RFCWs above 50.1 %. Cultivars Nevesinjka and Pesma were considered the most tolerant genotypes, while cultivars Balada and Vila were the most sensitive. The results confirmed that differences in genotype reactions to excess B are visible at the cellular level and that they may serve as a selection criterion.Tolerantnost pšenice prema visokim koncentracijama bora ispitana je primenom kulture zrelog embriona. Testiranje je vršeno na modifikovanoj MS hranljivoj podlozi, kojoj je borna kiselina dodata u dve koncentracije: 15 mM i 30 mM. Kontrolna podloga nije sadržala bornu kiselinu u suvišku. Eksperiment je izveden na 14 sorti pšenice proizvedenih u Srbiji. Posle mesec dana gajenja na podlogama izmerena je sveža masa kalusa (SMK) i izračunata redukcija sveže mase kalusa (RSMK) na podlozi sa 15 mM borne kiseline, u odnosu na kontrolu. Genotipovi su se značajno razlikovali u pogledu njihove reakcije na različite koncentracije bora. Šest genotipova imalo je RSMK ispod 50,0 %, dok je osam genotipova imalo RSMK iznad 50,1 %. Sorte Nevesinjka i Pesma ocenjene su kao najtolerantnije, dok su sorte Balada i Vila bile najosetljivije na prisustvo bora u podlozi. Rezultati su potvrdili da su reakcije genotipova pšenice na suvišak bora vidljive i na ćelijskom nivou, te da se ova reakcija može koristiti kao selekcioni kriterijum za procenu tolerantnosti na bor