44 research outputs found

    Molecular Characterization and Expression of Four Aquaporin Genes in <i>Impatiens walleriana</i> during Drought Stress and Recovery

    No full text
    Aquaporins comprise a large group of transmembrane proteins responsible for water transport, which is crucial for plant survival under stress conditions. Despite the vital role of aquaporins, nothing is known about this protein family in Impatiens walleriana, a commercially important horticultural plant, which is sensitive to drought stress. In the present study, attention is given to the molecular characterization of aquaporins in I. walleriana and their expression during drought stress and recovery. We identified four I. walleriana aquaporins: IwPIP1;4, IwPIP2;2, IwPIP2;7 and IwTIP4;1. All of them had conserved NPA motifs (Asparagine-Proline-Alanine), transmembrane helices (TMh), pore characteristics, stereochemical properties and tetrameric structure of holoprotein. Drought stress and recovery treatment affected the aquaporins expression in I. walleriana leaves, which was up- or downregulated depending on stress intensity. Expression of IwPIP2;7 was the most affected of all analyzed I. walleriana aquaporins. At 15% and 5% soil moisture and recovery from 15% and 5% soil moisture, IwPIP2;7 expression significantly decreased and increased, respectively. Aquaporins IwPIP1;4 and IwTIP4;1 had lower expression in comparison to IwPIP2;7, with moderate expression changes in response to drought and recovery, while IwPIP2;2 expression was of significance only in recovered plants. Insight into the molecular structure of I. walleriana aquaporins expanded knowledge about plant aquaporins, while its expression during drought and recovery contributed to I. walleriana drought tolerance mechanisms and re-acclimation

    Phenotypic performance of transgenic potato (Solanum tuberosum L.) plants with pyramided rice cystatin genes (OCI and OCII)

    No full text
    The evaluation of transgenic plants which is usually carried out under controlled conditions in culture rooms and greenhouses can yield valuable information about the influence of introduced genes on a transgenic plant phenotype. However, an overall assessment of plant performance can only be made by testing transgenic plants in the field environment. Thus, the effects of pyramided rice cystatin genes OCI and OCII on morphological parameters of transgenic potato cv. Desiree, Dragacevka and Jelica lines were compared under in vitro, greenhouse, and field conditions. All analyzed OC co-expressing transgenic lines exhibited normal phenotype, both in vitro and in greenhouse conditions. In the field environment, eight of nine OCI/OCII lines were similar to the wild-type control plants in their general phenotypic appearance. Yield parameters, such as tuber number and tuber weight for these phenotypically normal OCI/OCII lines, were also comparable to the controls. Only transgenic cv. Jelica line 4 plants exhibited slightly reduced growth, atypical leaf morphology and, contrary to the plants of other transgenic lines and untransformed controls, failed to flower. However, despite the phenotypic and developmental changes under field conditions, the OCI/OCII Jelica line 4 did not exhibit a significant decrease in tuber yield. Stacking of OCI and OCII genes preserves important attributes of the parental lines, confirming that this approach could be suitable for improving agronomical traits in potato.Serbian Ministry of Education, Science and Technological Development {[}ON173015

    Effect of phenol on germination capacity and polyphenol oxidase, peroxidase and catalase activities in lettuce

    Get PDF
    In this study we examined the activities of polyphenol oxidase (PPO) and antioxidant enzymes, peroxidase (POX) and catalase (CAT) during lettuce seed germination at different concentrations of phenol. Out of eleven varieties of lettuce, four were chosen according to their germination tolerance to phenol as follows: plants exhibiting high (Ljubljanska ledenka - LJL and Nansen - N) and low toleranace (Little Gem - LG and Majska kraljica - MK). A decrease in germination efficiency after exposure to LD50 of phenol was determined for these four varieties. The effects of phenol treatment on POX, CAT and PPO activities were determined after 4, 5, 6, 7 and 8 days of growth at LD50 concentrations. A trend of increased peroxidase activity was observed in seeds grown on LD50 of phenol compared to control seeds. A significant increase in CAT activity was observed at the beginning of treatment for MK, LG and N in seeds grown on phenol as well as in control seeds. A trend of increased PPO activity was observed in all control seeds. We also investigated the affinity of PPO for two different substrates that were used for the determination of enzyme activity. Our results show that LJL and N are the varieties most tolerant to growth on phenol. Here we report on the activities of their antioxidant enzymes and PPO during seed germination.Serbian Ministry of Education, Science and Technological Development {[}ON173017

    A rapid protocol for DNA extraction and a comparison of tree protocols for RNA extraction from Impatiens walleriana L.

    No full text
    In order to detect and identify viruses in the plant material or to confirm the expression of a gene after genetic transformation it is necessary to have a fast, simple and reliable method for the extraction of DNA and RNA. In this study, we tested a protocol for DNA and three protocols for RNA extraction from plants of I. walleriana L. RNA obtained by different extraction methods significantly differed in its yield and purity. The optimum method proved to be the one applicable to plant species that contain large amounts of secondary metabolites (polyphenols and polysaccharides). Nucleic acid extracted by the application of these methods showed satisfactory results in PCR and RT-PCR analysis.Za detekciju i identifikaciju virusa u biljnom materijalu ili potvrdu ekspresije nekog gena nakon genetičkih transformacija neophodno je imati brzu, jednostavnu i pouzdanu metodu za ekstrakciju DNK i RNK. U ovom radu smo testirali jedan protokol za izolaciju DNK i tri za ekstrakciju RNK iz biljaka Impatiens walleriana L. RNK dobijena primenom različitih metoda za ekstrakciju znatno se razlikovala po prinosu i čistoći. Kao optimalna metoda se pokazala ona koja se primenjuje za biljne vrste koje sadrže velike količine sekundarnih metabolita (polifenola i polisaharida). Nukleinske kiseline ekstrahovane primenom ovih metoda pokazale su zadovoljavajuće rezultate u PCR i RT-PCR analizama.Projekat ministarstva br. TR 3101

    Establishing a model system for transformation of I. hawkerii Bull. plants

    No full text
    One of the leading areas of modern biotechnology is to develop plants resistant to pathogens using genetic engineering methods. The genetic transformation of I. hawkerii Bull. with Agrobacterium tumefaciens C58C1pac1 was observe in this study, and the aim was to derive plants carrying the gene for pac1 resistance to tomato spotted wilt virus and other plant RNA viruses. I. hawkerii Bull. transformants on a selective medium rapidly failed, and not a single clone survived the fifth passage. This decline was due to great susceptibility of this type to kanamycin or failure of the transformation.Jedna od vodećih oblasti savremene biotehnologije je dobijanje biljaka otpornih na patogene pomoću metoda genetičkog inženjeringa. U ovom radu je proučavana genetička transformacija Impatiens hawkerii Bull. pomoću Agrobacterium tumefaciens C58C1pac1, a cilj je dobijanje biljaka koje nose pac1 gen za rezistentnost na virus bronzavosti paradajza i druge biljne RNK viruse. Transformanati I. hawkerii Bull. na selektivnoj podlozi su brzo propadali, a ni jedan klon nije preživeo peti pasaž. Ovo propadanje je posledica velike osetljivosti ove vrste na kanamicin ili neuspešnosti transfomacije.Projekat ministarstva br. TR 3101

    Cyclisation oxydative d’hexadiène-1,5.

    No full text
    Cytokinin oxidase/dehydrogenase (CKX) is the only known enzyme involved in cytokinin catabolism. Genes coding for two Arabidopsis CKX isoforms, AtCKX1 and AtCKX2, were introduced separately into a binary cloning vector, immobilized into Agrobacterium tumefaciens strain GV3101, and introduced into root explants of centaury (Centaurium erythraea Rafn.). The integration of each transgene was confirmed by genomic PCR. Of the total transformed explants, 30 and 28.2 % of the transformants carried AtCKX1 and AtCKX2 transgenes, respectively. Of these transformants, 50 % exhibited expression of the AtCKX1 transgene, while 64 % of transformants exhibited expression of the AtCKX2 transgene. For all analysed AtCKX transgenic centaury lines, as well as for untransformed control plants, CKX activity was higher in roots than in shoots. Expression of AtCKX in most transgenic lines contributed to enhanced levels of CKX activity in root tissues; whereas, only a few lines demonstrated increased CKX activity in shoot tissues compared to those of control plants. Moreover, overexpression of AtCKX resulted in reduced morphogenetic potential in transgenic plants, but did not significantly affect biomass production in comparison to untransformed control plants.Ministry of Education, Science and Technological Development of the Republic of Serbia [ON173015]; Czech Science Foundation [P506/11/0774

    Agrobacterium-mediated transformation of ornamental species: A review

    No full text
    Integration of desirable traits into commercial ornamentals using genetic engineering techniques is a powerful tool in contemporary biotechnology. However, these techniques have had a limited impact in the domain of ornamental horticulture, particularly floriculture. Modifications of the color, architecture or fragrance of the flowers as well as an improvement of the plant tolerance/resistance against abiotic and biotic stresses using plant transformation techniques, is still in its infancy. This review focuses on the application of Agrobacterium-mediated transformation, a major plant genetic engineering approach to ornamental plant breeding and the impact it has had to date.Serbian Ministry of Education, Science and Technological Research {[}TR 31019

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

    Get PDF
    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

    Development of the model system for ex vitro transformation of Impatiens Walleriana

    No full text
    We investigated possibility of ex vitro genetic transformation of Impatiens walleriana using Agrobacterium rhizogenes A4M70GUS and Agrobacterium tumefaciens C58C1pac1 bacterial suspension mixture with aim to obtain composite plants carrying GUS and pac1 genes. Compared to the control, treated plants exhibited increase of 27% in number of roots, while a difference in shoot length and number of leaves was not statistically significant. The PCR analysis did not confirmed integration of transgenes in the plant genome. The reason for this can be inadequate sampling or low transformation efficiency.U ovom radu je proučavana mogućnost ex vitro genetičke transformacije Impatiens walleriana pomoću smeše bakterijskih suspenzija Agrobacterium rhizogenes A4M70GUS i Agrobacterium tumefaciens C58C1 pac1, a cilj je dobijanje kompozitnih biljaka koje nose GUS i pac1 gene. Tretirane biljke su imale 27% veći broj korenova, a razlike u dužini izdanaka i broju listova u odnosu na kontrolu nisu bile statistički značajne. PCR analizom nije potvrđena inkorporacija navedenih gena u biljni genom, što može biti posledica lošeg izbora uzoraka ili malog procenta transformacije.Projekat ministarstva br. TR 3101
    corecore