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

Comparative analysis of microspore size variability in the genus Aesculus (Hippocastanaceae)

Pollen size varies extensively among angiosperm species and partially reflects evolutionary adaptation of each species to the pollination and fertilization environment. Size of uninuclear microspores in Aesculus parviflora was analyzed and compared with the size of microspores in Aesculus hippocastanum, Aesculus carnea, and Aesculus flava. The microspores came from closed flower buds of different size (3, 4, and 5 mm) isolated from lower (female flowers), middle (bisexual flowers), and upper (male flowers) segments of inflorescences. Aesculus parviflora had smaller microspores than Aesculus carnea and Aesculus flava, but larger microspores than Aesculus hippocastanum. All analyzed microspores showed bimodal distribution in all investigated species of the genus Aesculus.Projekat ministarstva br. 14302

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

Secondary embryogenesis in androgenic embryo cultures of Aesculus hippocastanum L.

Secondary somatic embryos appeared on the cotyledons and radiculi of embryos derived from suspension and anther cultures of Aesculus hippocastanum L. The highest number of secondary somatic embryos formed on a hormone-free medium.nul

Genetic modification of horse chestnut as a tool for the ornamental traits improvement

Reduced plant size, modified habitus and changes in the flowering time may be interesting and economically important in the breeding of ornamentals. These traits could be achieved by genetic transformation with the rol genes of Agrobacterium rhizogenes. Transformed plants, more or less, exhibit traits typical for the hairy root syndrome, including the shortening of internodes abolished apical dominance, increased axilary branching, more compact habitus, changes in leaf and flower morphology, and flowering time shift. Horse chestnut was transformed by using the agropine A. rhizogenes strain A4GUS, with the uidA reporter-gene inserted in the TL-DNA. Plantlets were regenerated from the acquired hairy root lines via the process of somatic embryogenesis, and, to a lesser extent, organogenesis. Transformed plantlets displayed the characteristic hairy root syndrome at varying degrees. Plants of all lines were shorter than control ones, although branching did not increase. The line 36 had almost normal leaf morphology, while lines 43 and 31 had narrow leaves. Lines 39 and 47 displayed the most severe phenotype stunted growth, very narrow dark-green leaves and short petioles. It seems that at least line 36 may be interesting, for example, for the bonsai plants production.Redukovana veličina biljke, modifikovani habitus i promene u vremenu cvetanja mogu biti zanimljive i ekonomski značajne u oplemenjivanju ornamentalnih vrsta. Ove osobine se mogu postići genetičkom transformacijom ml genima Agrobacterium tiiizogenes. Transfoimisane biljke, u manjoj ili većoj meri pokazuju osobine tipične za "hairy root" sindrom: skraćene internodije ukinutu apikalnu dominaciju, izraženo bočno grananje, kompaktniji habitus izmenjenu morfologiju listova i cvetova i pomereno vreme cvetanja. Divlji kesten je transfonnisan pomoću agropinskog soja A. rhizogenes A4GUS, sa uidA reporter genom insertovanim u TL-DNK. Iz dobijenih linija transformisanih korenova su regenerisane biljčice procesom somatske embriogeneze i, u manjoj meri procesom organogeneze. Transformisane biljčice su u različitom stepenu pokazivale karakterističan "hairy root" sindrom. Biljčice svih transfonnisanih linija su bile kraće od netransformisanih, mada grananje nije bilo povećano. Linija 36 je imala skoro normalnu morfologiju lista, dok su linije 43 i 31 imale uzane listove. Kod linija 39 i 47 sindrom je bio najizraženiji; biljčice su bile zakržljale, sa vrlo uzanim tamno-zelenim listovima i kratkim petiolama. Čini se da bi bar linija 36 mogla biti zanimljiva, npr. za produkciju bonsai-biljaka.Projekat ministarstva br. 157

Genetic modification of horse chestnut as a tool for the ornamental traits improvement

Reduced plant size, modified habitus and changes in the flowering time may be interesting and economically important in the breeding of ornamentals. These traits could be achieved by genetic transformation with the rol genes of Agrobacterium rhizogenes. Transformed plants, more or less, exhibit traits typical for the hairy root syndrome, including the shortening of internodes abolished apical dominance, increased axilary branching, more compact habitus, changes in leaf and flower morphology, and flowering time shift. Horse chestnut was transformed by using the agropine A. rhizogenes strain A4GUS, with the uidA reporter-gene inserted in the TL-DNA. Plantlets were regenerated from the acquired hairy root lines via the process of somatic embryogenesis, and, to a lesser extent, organogenesis. Transformed plantlets displayed the characteristic hairy root syndrome at varying degrees. Plants of all lines were shorter than control ones, although branching did not increase. The line 36 had almost normal leaf morphology, while lines 43 and 31 had narrow leaves. Lines 39 and 47 displayed the most severe phenotype stunted growth, very narrow dark-green leaves and short petioles. It seems that at least line 36 may be interesting, for example, for the bonsai plants production.Redukovana veličina biljke, modifikovani habitus i promene u vremenu cvetanja mogu biti zanimljive i ekonomski značajne u oplemenjivanju ornamentalnih vrsta. Ove osobine se mogu postići genetičkom transformacijom ml genima Agrobacterium tiiizogenes. Transfoimisane biljke, u manjoj ili većoj meri pokazuju osobine tipične za "hairy root" sindrom: skraćene internodije ukinutu apikalnu dominaciju, izraženo bočno grananje, kompaktniji habitus izmenjenu morfologiju listova i cvetova i pomereno vreme cvetanja. Divlji kesten je transfonnisan pomoću agropinskog soja A. rhizogenes A4GUS, sa uidA reporter genom insertovanim u TL-DNK. Iz dobijenih linija transformisanih korenova su regenerisane biljčice procesom somatske embriogeneze i, u manjoj meri procesom organogeneze. Transformisane biljčice su u različitom stepenu pokazivale karakterističan "hairy root" sindrom. Biljčice svih transfonnisanih linija su bile kraće od netransformisanih, mada grananje nije bilo povećano. Linija 36 je imala skoro normalnu morfologiju lista, dok su linije 43 i 31 imale uzane listove. Kod linija 39 i 47 sindrom je bio najizraženiji; biljčice su bile zakržljale, sa vrlo uzanim tamno-zelenim listovima i kratkim petiolama. Čini se da bi bar linija 36 mogla biti zanimljiva, npr. za produkciju bonsai-biljaka.Projekat ministarstva br. 157

Secondary embryogenesis in androgenic embryo cultures of Aesculus hippocastanum L.

Secondary somatic embryos appeared on the cotyledons and radiculi of embryos derived from suspension and anther cultures of Aesculus hippocastanum L. The highest number of secondary somatic embryos formed on a hormone-free medium.nul

The horse chestnut lines harboring the rol genes

An Agrobacterium rhizogenes-mediated transformation system for Aesculus hippocastanum L. has been developed. Wounded androgenic embryos of A. hippocastanum were inoculated with bacteria containing the pRiA4 plasmid, with the uid A sequence as a reporter gene. The hairy roots emerging from the wounded sites of androgenic embryos were isolated and maintained in Murashige and Skoog's (MS) liquid hormone-free medium. Five hairy root lines have been maintained in vitro for 4 years with unchanged growth rate and might be a suitable source for secondary metabolite production. The transformation events have been confirmed by a polymerase chain reaction specific to the rol A, B, C and D genes. The absence of residual contaminating bacteria has been shown by a polymerase chain reaction specific to the vir D1 sequence.nul

Impact of genotype, age of tree and environmental temperature on androgenesis induction of Aesculus hippocastanum L.

Influence of ten different genotypes, age of trees and environmental temperature on induction of androgenesis and apperance albino horse chestnut embryos were studied. Efficiency of in vitro androgenesis via anther and microspore culture had been investigated. Microspores and anthers were used from the same closed flower bud. Androgenic response of different genotypes was measured and compared. Anther induction rates were from 5 to 37.6%, depending on genotype. The number of embryos per isolated anther varied between 0.5 to 5.0 embryos in anther culture, while in microspore culture varied between 3.0 to 27 embryos, depending on genotype. A microspore culture was 5 - 6 times efficient than anther culture for same genotype. Age of the trees had no influence on androgenesis induction. Temperature of about 4 - 5 degrees C was optimal for androgenic embryo induction. Albino horse chestnut embryos phenomenon depended on genotype. The number of albino appearing in anther was higher than in microspore culture. The same correlation of appearing albino was observed between short and long day. Flow cytogenetic analysis of androgenic embryos originating from anther and microspore culture was done after a first generation of regenerants. All androgenic embryos, the first generation from microspore culture were haploid, while 50% of the regenerants originating from anther culture were haploid, and the other half diploid.Ministry of Science and Environmental Protection of Serbia [143026, EE18031

Impact of genotype, age of tree and environmental temperature on androgenesis induction of Aesculus hippocastanum L.

Influence of ten different genotypes, age of trees and environmental temperature on induction of androgenesis and apperance albino horse chestnut embryos were studied. Efficiency of in vitro androgenesis via anther and microspore culture had been investigated. Microspores and anthers were used from the same closed flower bud. Androgenic response of different genotypes was measured and compared. Anther induction rates were from 5 to 37.6%, depending on genotype. The number of embryos per isolated anther varied between 0.5 to 5.0 embryos in anther culture, while in microspore culture varied between 3.0 to 27 embryos, depending on genotype. A microspore culture was 5 - 6 times efficient than anther culture for same genotype. Age of the trees had no influence on androgenesis induction. Temperature of about 4 - 5 degrees C was optimal for androgenic embryo induction. Albino horse chestnut embryos phenomenon depended on genotype. The number of albino appearing in anther was higher than in microspore culture. The same correlation of appearing albino was observed between short and long day. Flow cytogenetic analysis of androgenic embryos originating from anther and microspore culture was done after a first generation of regenerants. All androgenic embryos, the first generation from microspore culture were haploid, while 50% of the regenerants originating from anther culture were haploid, and the other half diploid.Ministry of Science and Environmental Protection of Serbia [143026, EE18031