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Molecular cloning and expression analysis of a new stress-related AREB gene from Arachis hypogaeaL. Hong, B. Hu, X. Liu, C. Y. He, Y. Yao, X. L. Li, L. LiBiologia plantarum 57:56-62, 2013 | DOI: 10.1007/s10535-012-0236-6 An AREB gene, designated as AhAREB1, was cloned from peanut (Arachis hypogaea L.). The gene contains a 1 338-bp open reading frame that encodes a putative protein of 445 amino acids. The corresponding genomic DNA containing four exons and three introns was isolated and analyzed. An upstream 1 060-bp DNA promoter fragment of the AhAREB1 gene was also amplified from peanut genomic DNA. Multiple sequence alignment of the deduced amino acids of AREB showed that the AhAREB1 protein shares high sequence homology with GmAREB1, S1AREB and ABF2. Quantitative real-time PCR analysis showed that AhAREB1 was induced by polyethylene glycol, NaCl, gibberellic acid, abscisic acid and salicylic acid. The cloning and characterization of the AhAREB1 gene will be useful for further studies establishing the biological role of AhAREB1 in plants. |
Lithium alters elicitor-induced H2O2 production in cultured plant cellsN. Orbán, K. BókaBiologia plantarum 57:332-340, 2013 | DOI: 10.1007/s10535-012-0279-8 Lithium pollution may seriously influence the metabolic and signalling processes of plants. In the present paper, we investigate the effect of lithium chloride on fungal elicitor-triggered H2O2 generation in Rubia tinctorum L. cell cultures. Our results show that Li+ strongly influences elicitor-induced H2O2 formation and time-course in the cells nad culture medium. Neomycin, a phospholipase C inhibitor, and 2-APB, an inositol-1,4,5-triphosphate (IP3) receptormediated Ca2+ release blocker, strongly affected the elicitor-induced H2O2 production and had a similar effect on elicitor-triggered H2O2 formation as Li+. We monitored changes in H2O2 location at subcellular level and our observations confirmed the changes measured by quantitative methods. The obtained results enabled us to deduce that the IP3 pathway might be involved in the early signalling events leading to the moderation of elicitor-induced reactive oxygen species generation. |
Ferulic acid pretreatment enhances dehydration-stress tolerance of cucumber seedlingsD. -M. Li, Y. -X. Nie, J. Zhang, J. -S. Yin, Q. Li, X. -J. Wang, J. -G. BaiBiologia plantarum 57:711-717, 2013 To examine whether ferulic acid (FA) could protect plants from dehydration stress and to investigate a mechanism for the protection, cucumber seedlings were pretreated with 0.5 mM FA for 2 d and then were exposed to dehydration induced by 10 % polyethylene glycol 6000. After pretreatment with FA, the activities of antioxidant enzymes (catalase, superoxide dismutase, and quaiacol peroxidase) in leaves were higher than under dehydration treatment alone which was in accordance with the increased transcript levels of respective genes. Moreover, the combination of FA pretreatment and dehydration reduced the content of superoxide radical, hydrogen peroxide, and malondialdehyde, and increased the relative water content and content of FA, proline, and soluble sugars in comparison with dehydration alone. We propose that pretreatment with FA protects cucumbers against dehydration stress by decrease of lipid peroxidation due to activation of antioxidant enzymes and by increase of proline and soluble sugar content in leaves. |
Lead uptake, toxicity and accumulation in Phaseolus vulgaris plantsA. Piechalak, A. Malecka, D. Bara³kiewicz, B. TomaszewskaBiologia plantarum 52:565-568, 2008 | DOI: 10.1007/s10535-008-0112-6 The effects of lead were investigated in bean plants (Phaseolus vulgaris L. cv. Zlota Saxa) grown hydroponically in nutrient solution and exposed to Pb(NO3)2 (0.1, 0.5, 1 mM) with or without equimolar concentrations of chelator ethylenediaminetetraacetic acid (EDTA). The roots treated only with Pb(NO3)2 accumulated up to 25 g(Pb) kg-1(d.m.), during 4-d exposure. However, in bean plants exposed to 0.5 mM Pb + 0.5 mM EDTA or 1 mM Pb + 1 mM EDTA 2.5 times less Pb was determined. In bean plants treated only with Pb, less than 6 % of total lead accumulated was transported to the aboveground parts, while in the case of plants grown with Pb + EDTA, around 50 % of total Pb was transported to the shoots. |
Agrobacterium-Mediated Transformation and Plant Regeneration of Triticum aestivum L.N. Mitiæ, R. Nikoliæ, S. Ninkoviæ, J. Milju¹-Djukiæ, M. Ne¹koviæBiologia plantarum 48:179-184, 2004 | DOI: 10.1023/B:BIOP.0000033442.15611.7d The use of two Agrobacterium tumefaciens strains for transformation of Triticum aestivum L. cv. Vesna was studied. Immature embryos, isolated 15 d after pollination, were co-cultivated with the super-binary LBA4404/pTOK233 and the binary AGL1/pDM805 vectors. While the transient GUS-intron expression was high (69.9 and 80.0 %), the number of plants regenerated on selective media containing hygromycin or phosphinotricin did not exceed 0.4 and 0.13 %, respectively. Nevertheless, the regenerated plants were fertile and produced seeds. The T0 plants, as well as the T1 seedlings, displayed the activity in the β-glucuronidase histochemical assay and a positive signal in PCR analysis for the presence of uidA gene sequences in their genomes. The data suggest that the transformation of wheat cv. Vesna with both Agrobacterium strains is feasible. |
Metabolism and aluminum accumulation in Plantago almogravensis and P. algarbiensis in response to low pH and aluminum stressN. Martins, S. Gonçalves, A. RomanoBiologia plantarum 57:325-331, 2013 | DOI: 10.1007/s10535-012-0271-3 We investigated the impact of low pH and aluminum on the metabolism and capacity for Al accumulation in shoots of the plantain species Plantago algarbiensis and P. almogravensis. We found that increasing the concentration of Al in the medium increased accumulation of it in the shoots of both plants (although more in P. almogravensis than in P. algarbiensis). The presence of Al in the medium induced proline and saccharide synthesis in P. almogravensis without affecting lipid peroxidation, but increased proline synthesis and lipid peroxidation in P. algarbiensis without affecting the saccharide content. Lipid peroxidation in P. algarbiensis was also enhanced at pH 4.0. The activity of antioxidant enzymes was increased as a response to low pH and Al in both species. Our data indicate that both species can accumulate high levels of Al but they have different sensitivities to low pH and/or the presence of Al in the growth medium. |
Effect of Pb ions on superoxide dismutase and catalase activities in leaves of pea plants grown in high and low irradianceE. Romanowska, B. Wróblewska, A. Dro¿ak, M. Zienkiewicz, M. SiedleckaBiologia plantarum 52:80-86, 2008 | DOI: 10.1007/s10535-008-0012-9 The role of irradiance on the activity of antioxidant enzymes: superoxide dismutase (SOD) and catalase (CAT) was examined in the leaves of Pisum sativum L. plants grown under low (LL) or high (HL) irradiance (PPFD 50 or 600 µmol m-2 s-1) and exposed after detachment to 5 mM Pb (NO3)2 for 24 h. The activities of both enzymes increased in response to LL compared with HL and no effect of Pb ions was observed. Photosystem (PS) 1 and PS 2 activities were also investigated in chloroplasts isolated from these leaves. LL lowered PS 1 electron transport rate and changes in photochemical activity of PS 1 induced by Pb2+ were visible only in the chloroplasts isolated from leaves of LL grown plants. PS 2 activity was influenced similarly by Pb ions at both PPFD. This study demonstrates that leaves of HL grown plants were less sensitive to lead toxicity than those from LL grown plants. Changes in electron transport rates were the main factors responsible for the generation of reactive oxygen species in the chloroplasts and as a consequence, in induction of antioxidant enzymes. |
The plant activator BTH promotes Ornithogalum dubium and O. thyrsoides differentiation and regeneration in vitroO. M. Tun, A. Lipsky, T. Luzzatto Knaan, Z. Kerem, I. YedidiaBiologia plantarum 57:41-48, 2013 | DOI: 10.1007/s10535-012-0254-4 Benzothiadiazole (BTH) is a structural analogue of salicylic acid (SA) which is widely recognized for its role in elicitation of systemic acquired resistance in a broad range of plant species. Here, BTH was applied to cell cultures of the bulbous ornamental plants Ornithogalum dubium and O. thyrsoides, showing a strong effect on rates of differentiation and morphogenesis. Morphogenic cell clusters in liquid Murashige and Skoog (MS) medium containing 1-naphthaleneacetic acid (NAA) and 6-benzylaminopurine (BAP) were used for all treatments. The calluses were washed thoroughly and activated with increasing concentrations of BTH. Following the induction, calli were grown on a solid MS medium without growth regulators (MS) or on a comparable media with NAA and BAP (M-206). The calli treated with BTH displayed a dose dependent increase in formation of meristematic centres followed by enhanced shoot formation compared to controls. Microscopic analyses revealed increased differentiation to cell organelles and a strengthening of the cell wall. A stronger response to BTH was observed in MS than in M-206 medium. A similar effect on calli differentiation was obtained by three weeks darkness followed by light exposure. The dark/light positive effect on differentiation was further augmented by BTH in a synergistic fashion. It is suggested that BTH enhances the rates of morphogenesis in Ornithogalum cultures by triggering a plant regulator-like activity. |
Allelic differentiations and effects of the Rf3 and Rf4 genes on fertility restoration in rice with wild abortive cytoplasmic male sterilityJ. Cai, Q. P. Liao, Z. J. Dai, H. T. Zhu, R. Z. Zeng, Z. M. Zhang, G. -Q. ZhangBiologia plantarum 57:274-280, 2013 | DOI: 10.1007/s10535-012-0294-9 To reveal the allelic differentiations at the two genes for fertility restoration (Rf) on chromosomes 1 (Rf3) and 10 (Rf4), 15 chromosome single segment substitution lines (SSSLs) with the Rf3 locus and 18 SSSLs with the Rf4 locus were crossed with Bobai A (BbA), a cytoplasmic male sterility line with wild abortive type of cytoplasm (WA-CMS), respectively. Based on the pollen and seed fertility of the F1 hybrids, the Rf3 and Rf4 genes were each classified into four alleles, namely Rf3-1, Rf3-2, Rf3-3, and Rf3-4 for Rf3, and Rf4-1, Rf4-2, Rf4-3, and Rf4-4 for Rf4. Out of the 33 SSSLs, an SSSL W23-19-06-06-11 carrying the genotype Rf3-4Rf3-4/Rf4-4Rf4-4 possessed the strongest restoring ability for BbA. To determine the genetic effects of Rf3 and Rf4 for WA-CMS, one BC3F2 population possessing the genetic background of W23-19-06-06-11 was generated from the cross between W23-19-06-06-11 and BbA by backcrossing and marker-assisted selection. In the BC3F2 population, the plants carrying the Rf3Rf3/Rf4Rf4, Rf3Rf3/rf4rf4, and rf3rf3/Rf4Rf4 genotypes were selected and their phenotyping for pollen and spikelet fertility were evaluated. The result showed that under the genetic background of SSSL W23-19-06-06-11, the effect of Rf4 appeared to be slightly larger than that of Rf3 and their effects were additive for WA-CMS system. These studies will lead to the transfer of Rf genes into adapted cultivars through marker-assisted selection in active hybrid rice breeding programs. |
Metabolism of glutathione and ascorbate in lingonberry cultivars during in vitro and ex vitro propagationP. Vyas, S. C. Debnath, A. U. IgamberdievBiologia plantarum 57:603-612, 2013 | DOI: 10.1007/s10535-013-0339-8 Lingonberry (Vaccinium vitis-idaea L. ssp. vitis-idaea Britton) cultivars Regal, Splendor, and Erntedank were obtained by conventional softwood cuttings (taken as a control), by in vitro shoot proliferation of node explants, and by adventitious shoot regeneration from excised leaves of micropropagated shoots. In the plants propagated in vitro, the total ascorbate content increased and its pool was more oxidized, the total glutathione content also increased but its pool became more reduced. The leaves of plants obtained from the in vitro culture showed significantly higher antioxidant enzyme activities except for dehydroascorbate reductase which was at a similar level in all plants. Total soluble phenolics, tannins, and flavonoids were enhanced in fruits of in vitro-propagated plants whereas in leaves, the levels of these metabolites (except flavonoids) were higher in ex vitro derived plants. The total radical scavenging capacity was enhanced in berries of the in vitro propagated plants. It is suggested that the active morphogenetic process, characterized by intensive formation and scavenging reactive oxygen species is reflected in the activities of antioxidant enzymes and metabolites. The reduction potential of glutathione is the most important parameter which determines patterns of growth and differentiation in the investigated plants. |
Direct shoot regeneration from Arabidopsis thaliana shoot apical meristemsW. Xin, Z. Liu, Y. Song, T. Hou, F. XiangBiologia plantarum 56:601-606, 2012 | DOI: 10.1007/s10535-012-0127-x In Arabidopsis thaliana in vitro culture, shoots were induced from the shoot apical meristem (SAM) of germinating seeds in the presence of 2,4-dichlorophenoxyacetic acid. Primary shoot primordia developed leaf-like structures, from which secondary shoot primordia were produced. Regenerated shoots were recovered when the material was transferred to a medium lacking auxin. Adventitious roots formed from a callusing basal region of the secondary shoots. The CUC1 transcription factor was expressed at the apex of the primary shoot primordium and at the boundary between the regenerated SAM and the developing leaf primordia. The DR5::GUS transgene was used to localize sites of maximum auxin occurrence. Auxin was firstly detected in the dividing cells beneath the SAM epidermis, which coincided with sites where primary shoot primordia were initiated. In the regenerated shoots, auxin response was not detected in the basal region of the stem, suggesting that the regenerating structures were shoots rather than somatic embryos. Direct shoot regeneration from the A. thaliana SAM requires a localized accumulation of auxin. |
Somatic embryogenesis from immature zygotic embryos and monitoring the genetic fidelity of regenerated plants in grapevineX. M. Yang, L. Z. An, Y. C. Xiong, J. P. Zhang, Y. Li, S. J. XuBiologia plantarum 52:209-214, 2008 | DOI: 10.1007/s10535-008-0047-y Somatic embryogenesis and plant regeneration were successfully established on Nitsch and Nitsch (NN) medium from immature zygotic embryos of six genotypes of grapevine (Vitis vinifera). The optimum hormone combinations were 1.0 mg dm-3 2,4-dichlorophenoxyacetic acid (2,4-D) for callus induction and 1.0 mg dm-3 α-naphthalene acetic acid (NAA) + 0.5 mg dm-3 6-benzyladenine (BA) for embryos production and 0.03 mg dm-3 NAA + 0.5 mg dm-3 BA for embryos conversion and plant regeneration. The frequency of somatic embryogenesis varied from 10.5 to 37.5 % among six genotypes and 15.5-42.1 % of somatic embryos converted into normal plantlets. The analysis of DNA content determined by flow cytometry and chromosome counting of the regenerated plantlets clearly indicated that no ploidy changes were induced during somatic embryogenesis and plant regeneration, the nuclear DNA content and ploidy levels of the regenerated plants were stable and homogeneous to those of the donor plants. RAPD markers were also used to evaluate the genetic fidelity of plants regenerated from somatic embryos. All RAPD profiles from regenerated plants were monomorphic and similar to those of the field grown donor plants. We conclude that somaclonal variation is almost absent in our grapevine plant regeneration system. |
Accumulation of WCS120 and DHN5 proteins in differently frost-tolerant wheat and barley cultivars grown under a broad temperature scaleK. Kosová, P. Vítámvás, P. Prá¹ilová, I. T. Prá¹ilBiologia plantarum 57:105-112, 2013 | DOI: 10.1007/s10535-012-0237-5 Proteins WCS120 and DHN5 are known as the major cold-inducible dehydrins in wheat and barley plants, respectively. WCS120 and DHN5 relative accumulation increased exponentially along with a growth temperature decline in the range from optimum to cold temperatures. Even at optimum growth temperatures, the most frost-tolerant wheat and barley cultivars can be distinguished from the remaining ones according to dehydrin relative accumulation. The highly tolerant wheat and barley cultivars started accumulating dehydrins at higher growth temperatures and reached higher dehydrin amounts than the less tolerant ones. Statistically significant correlations between lethal temperature for 50 % of the samples (LT50) and dehydrin relative accumulation have been found at all growth temperatures (5, 10, 15 and 20 °C) for WCS120 in wheats and at 5 and 10 °C for DHN5 in barleys. Analogous relationships between dehydrin relative accumulation at different growth temperatures and plant acquired frost tolerance have been proved for wheat WCS120 and barley DHN5. |
Lipid peroxidation, H2O2 content, and antioxidants during acclimatization of Abrus precatorius to ex vitro conditionsS. Perveen, M. Anis, I. M. ArefBiologia plantarum 57:417-424, 2013 | DOI: 10.1007/s10535-013-0328-y An efficient, rapid, and reproducible plant regeneration protocol was successfully developed for Abrus precatorius L. using mature nodal explants excised from a 5-year-old field grown plant. The highest shoot regeneration frequency (87 %) with maximum number of multiple shoots (15.0) and shoot length (4.8 cm) were recorded on Murashige and Skoog (MS) medium amended with 2.5 μM thidiazuron, 120 mg dm-3 polyvinylpyrrolidone, and 0.5 μM α-naphthalene acetic acid. The best treatment for maximum root (4.0) induction was half strength MS medium supplemented with 1.5 μM indole-3-butyric acid. The in vitro plantlets with well-developed shoots and roots were successfully transferred into plastic cups with Soilrite and acclimatized in a culture room under photon flux density (PFD) of 150 μmol m-2 s-1, thereafter transferred to a greenhouse with PFD of 300 μmol m-2 s-1, and finally to a field with 70 % survival rate. During the acclimatization period (0-49 d), leaf chlorophyll and carotenoid content increased whereas malondialdehyde and H2O2 content decreased probably due to increasing activities of antioxidant enzymes (catalase, superoxide dismutase, glutathione reductase, and ascorbate peroxidase). Our work suggests that micropropagated plants developed an antioxidant enzymatic protective system to avoid oxidative stress during establishment under ex vitro environment. |
Agrobacterium-mediated transformation in Citrullus lanatusM.-A. Cho, C.-Y. Moon, J.-R. Liu, P.-S. ChoiBiologia plantarum 52:365-369, 2008 | DOI: 10.1007/s10535-008-0076-6 Agrobacterium tumefaciens-mediated transformation was used to produce transgenic watermelon. Cotyledonary explants of Citrullus lanatus Thumb (cv. Daesan) were co-cultivated with Agrobacterium strains (LBA4404, GV3101, EHA101) containing pPTN289 carrying with bar gene and pPTN290 carrying with nptII gene, respectively. There was a significant difference in the transformation frequency between bacteria strains and selective markers. The EHA101/pPTN289 showed higher transformation frequency (1.16 %) than GV3101/pPTN289 (0.33 %) and LBA4404/pPTN289 or /pPTN290 (0 %). The shoots obtained (633 and 57 lines) showed some resistance to glufosinate and paromomycin, respectively. Of them, the β-glucuronidase positive response and PCR products amplified by bar and nptII specific primers showed at least 21 plants resistant to glufosinate and at least 6 plants to paromomycin. Southern blot analysis revealed that the bar gene integrated into genome of transgenic watermelon. Acclimated transgenic watermelons were successfully transplanted in the greenhouse and showed no phenotypic variation. |
Improved Performance of Transgenic Glycinebetaine-Accumulating Rice Plants under Drought StressW. SawahelBiologia plantarum 46:39-44, 2003 | DOI: 10.1023/A:1027372629612 Plasmid DNA (pChlCOD), containing the selectable hygromycin phosphotransferase hpt gene for hygromycin B resistance and the Arthrobacter globiformis codA gene for choline oxidase which catalyzes the direct conversion of choline to glycinebetaine, was delivered into rice plants using Agrobacterium-mediated gene transfer via scutellum-derived calli. Southern, Northern and Western blot analyses demonstrated that the foreign gene had been transferred, integrated into rice chromosomal DNA and expressed. Drought test indicated that glycinebetaine acts as an osmoprotectant and its production in transgenic rice plant helped the cells to maintain osmotic potential and increased root growth, and thus enhanced the ability of the plants to tolerate water deficit |
In vitro clonal propagation of Nyctanthes arbor-tristisG. R. Rout, A. Mahato, S. K. SenapatiBiologia plantarum 52:521-524, 2008 | DOI: 10.1007/s10535-008-0101-9 Rapid shoot multiplication of Nyctanthes arbor-tristis L. was achieved from axillary meristems on Murashige and Skoog (MS) basal medium supplemented with 1.0-1.5 mg dm-3 6-benzylaminopurine (BA), 50 mg dm-3 adenine sulfate (Ads) and 3 % (m/v) sucrose. Inclusion of indole-3-acetic acid (IAA) in the culture medium along with BA + Ads promoted a higher rate of shoot multiplication. Maximum mean number of microshoots per explant (6.65) was achieved on the MS medium supplemented with 1.5 mg dm-3 BA, 50 mg dm-3 Ads and 0.1 mg dm-3 IAA after 4 weeks of culture. The elongated shoots rooted within 13 to 14 d on half-strength MS medium supplemented with either indole-3-butyric acid (IBA), IAA or 1-naphthaleneacetic acid (NAA) with 2 % sucrose. Maximum percentage of rooting was obtained on medium having 0.25 mg dm-3 IBA and 0.1 mg dm-3 IAA. About 70 % of the rooted plantlets survived in the greenhouse. The in vitro raised plants were grown normally in the field. |
Plant regeneration from mesophyll protoplasts of Agrobacterium rhizogenes-transformed Astragalus melilotoidesG. N. Zhang, J. F. Jia, J. G. Hao, X. R. Wang, T. HeBiologia plantarum 52:373-376, 2008 | DOI: 10.1007/s10535-008-0078-4 Plant regeneration from mesophyll protoplasts of Agrobacterium rhizogenes-transformed Astragalus melilotoides Pall. was here developed. The protoplasts were isolated directly from the leaves of the hairy root-induced plants. The highest yield of protoplasts was obtained from fully expanded leaves of young plants. Their viability was up to 72 ± 2.3 %. The highest division frequency (32.4 ± 0.13 %) and sustained divisions were obtained in Durand, Potrykus and Donn (DPD) medium supplemented with 2.0 mg dm-3 2,4-dichlorophenoxyacetic acid, 0.2 mg dm-3 6-benzylaminopurine, 0.3 M mannitol, 2 % sucrose and 500 mg dm-3 casein hydrolysate at the plating density of 3.0 × 105 cm-3. The frequency of shoot differentiation from protocalli reached to 91.75 ± 3.1 %. Opine synthesis and polymerase chain reaction analysis confirmed that T-DNA still existed in the protoplast regenerated plants. |
Efficient Genetic Transformation of Lotus corniculatus L. and Growth of Transformed Plants in FieldR. Nikoliæ, N. Mitiæ, S. Ninkoviæ, J. Milju¹-Djukiæ, M. Ne¹koviæBiologia plantarum 46:137-140, 2003 | DOI: 10.1023/A:1027357620951 An efficient protocol for shoot regeneration and genetic transformation was applied to root segments of a new Lotus corniculatus L. cultivar Bokor. The shoots, that regenerated on root segments, were inoculated with Agrobacterium rhizogenes A4M70GUS, and produced hairy roots, which on media with 0.2 mg dm-3 benzylaminopurine, regenerated shoots. After rooting and acclimation, the transformed plants were planted in the experimental field. Their morphological traits were compared to controls. No signs of the rol genes phenotype were present. The transformants were significantly taller than controls, while there were no significant differences in the leaf area. The glucuronidase activity and the presence of uidA gene was demonstrated in transformed plants of T0 and in seedlings of T1 generations. It is concluded that A. rhizogenes could be a vector of choice for the transfer of desirable genes into the bird's foot trefoil genome. |
In vitro regeneration of medicinal plant Centella asiaticaH. Mohapatra, D. P. Barik, S. P. RathBiologia plantarum 52:339-342, 2008 | DOI: 10.1007/s10535-008-0069-5 This paper describes multiple shoot regeneration from leaf and nodal segments of a medicinally important herb Centella asiatica L. on Murashige and Skoog's (MS) medium supplemented with a range of growth regulators. The highest number of multiple shoots was observed on MS augmented with 3.0 mg dm-3 N6-benzylaminopurine (BAP) and 0.05 mg dm-3 α-naphthaleneacetic acid (NAA). Leaf explant showed maximum percentage of cultures regenerating shoots (81.6 %), with the highest shoot number (8.3 shoots per explant) and the shoot length (2.1 cm) whereas, nodal explant showed less number of shoots with callus formation at the base cut end. Successive shoot cultures were established by repeatedly sub-culturing the original explant on a fresh medium. Rooting of in vitro raised shoots was best induced on half strength MS supplemented with 0.5 mg dm-3 indole-3-butyric acid (IBA) with highest percentage of shoot regenerating roots (76.8 %) with 3-4 roots per shoot. Plantlets were acclimated in Vermi-compost and eventually established in soil. Contents of chlorophyll, total sugars, reducing sugars and proteins were estimated in leaf tissue from both in vivo and in vitro raised plants. Chlorophyll content was higher in in vivo plants, whereas other three components were higher in in vitro plants. |
Analysis of genetic diversity among Swertia chirayita genotypesP. Joshi, V. DhawanBiologia plantarum 51:764-768, 2007 | DOI: 10.1007/s10535-007-0156-z Inter simple sequence repeat (ISSR) markers were used to analyse genetic diversity of Swertia chirayita genotypes collected from the temperate Himalayas of India. Allied species of Swertia chirayita were used in the study as outliers. Nineteen UBC primers generated a total of 315 ISSR bands, revealing 98.7 % polymorphism among the genotypes assayed. This was reduced to 42.5 % when the outliers were excluded. The results revealed a high genetic diversity within the genotypes. |
Transgenic Tobacco Plants Constitutively Overexpressing a Rice Thaumatin-like Protein (PR-5) Show Enhanced Resistance to Alternaria alternataR. Velazhahan, S. MuthukrishnanBiologia plantarum 46:347-354, 2003 | DOI: 10.1023/B:BIOP.0000023876.55053.5e Overexpression of antifungal pathogenesis-related (PR) proteins in crop plants has the potential for enhancing resistance against fungal pathogens. Thaumatin-like proteins (TLPs) are one group (PR-5, permatins) of antifungal PR-proteins isolated from various plants. In the present study, a plasmid containing a cDNA of rice tlp (D34) under the control of the CaMV-35S promoter was introduced into tobacco plants through Agrobacterium-mediated transformation system. A considerable overproduction of TLP was observed in transformed tobacco plants by Western blot analysis. There was a large accumulation of tlp mRNA in transgenic plants as revealed by Northern blot analysis. Southern blot analysis of the DNA from transgenic tobacco plants confirmed the presence of the rice tlp gene in the genomic DNA of transgenic tobacco plants. Immunoblot analysis of intracellular and extracellular proteins of transgenic tobacco leaves using a Pinto bean TLP antibody demonstrated that the 23-kDa TLP was secreted into the extracellular matrix. T2 progeny of regenerated plants transformed with TLP gene were tested for their disease reaction to Alternaria alternata, the brown spot pathogen. Transgenic tobacco plants expressing TLP at high levels showed enhanced tolerance to necrotization caused by the pathogen. |
Effect of Darkness on Growth and Flowering of Chenopodium rubrum and C. murale Plants in vitroA. Mitroviæ, B. ®ivanoviæ, Lj. ÆulafiæBiologia plantarum 46:471-474, 2003 | DOI: 10.1023/A:1024367311781 Chenopodium rubrum, a short-day plant, and C. murale, a long-day plant, were grown in vitro in continuous darkness. Control C. rubrum plants exposed to continuous darkness for 15 d at cotyledonary phase, did not flower, while 80 % of plants flowered on the medium with 5 % glucose and 10 mg dm-3 GA3. Control C. murale plants exposed to continuous darkness for 10 d at the age of 4th pair of leaves, did not flower, while GA3 (1 - 5 mg dm-3) stimulated flowering up to 65 %. |
Cyclic somatic embryogenesis and efficient plant regeneration from callus of safflowerJ. Vijaya Kumar, B. D. Ranjitha Kumari, E. CastañoBiologia plantarum 52:429-436, 2008 | DOI: 10.1007/s10535-008-0087-3 Efficient plant regeneration through somatic embryogenesis was established for safflower (Carthamus tinctorius L.) cv. NARI-6. Embryogenic calli were induced from 10 to 17-d-old cotyledon and leaf explants from in vitro seedlings. High frequency (94.3 %) embryogenic callus was obtained from cotyledon explants cultured on Murashige and Skoog's germination (MSG) basal medium supplemented with thidiazuron, 2-isopentenyladenine and indole-3-butyric acid. Primary, secondary and cyclic somatic embryos were formed from embryogenic calli in a different media free of plant growth regulators, however, 100 % cyclic somatic embryogenesis was obtained from cotyledon derived embryogenic calli cultured on MSG. Somatic embryos matured and germinated in quarter-strength MSG medium supplemented with gibberellic acid. Cotyledons with root poles or non root poles were converted to normal plantlets and produced adventitious roots in rooting medium. Rooted plants were acclimatized and successfully transferred to the field. |
Transgenic Rice Plants Expressing Bacillus subtilis Protoporphyrinogen Oxidase Gene Show Low Herbicide Oxyfluorfen ResistanceS.B. Ha, S.B. Lee, D.E Lee, J.O. Guh, K. BackBiologia plantarum 46:277-280, 2003 | DOI: 10.1023/B:BIOP.0000022265.66891.73 Transgenic rice plants harbouring Bacillus subtilis protoporphyrinogen oxidase (Protox) gene, which is targeted into plastid, were generated by Agrobacterium-mediated transformation using a rice (Oryza sativa L. cv. Dongjin) and their gene integration at T1 generation by Southern and mRNA expression in T2 generation by Northern blotting were analyzed. Their herbicide-resistant trait was further confirmed by in vitro leaf segment assay and in planta bioassays such as seed germination assay and measurement of growth inhibition. The herbicide oxyfluorfen resistance in transgenic rice plants was not very high. The results showed that the Protox from B. subtilis can not be applicable as a gene source to generate a high level oxyfluorfen tolerance in plants. |
Special origin of stem sequence influence the resistance of hairpin expressing plants against PVYF. Jiang, B. Wu, C. Zhang, Y. Song, H. An, C. Zhu, F. WenBiologia plantarum 55:528-535, 2011 | DOI: 10.1007/s10535-011-0119-2 In this study, 16 hairpin RNA (hpRNA) vectors were constructed, each harboring 50 bp viral RNA sequence as the stem. They all targeted the coat protein (CP) gene of Potato virus Y (PVY). Virus resistance assay revealed that hairpin constructs targeting the anterior 200 bp regions of the CP gene were unable to induce virus resistance, while the 12 hpRNA constructs targeting posterior 600 bp regions induced high virus resistance up to 77.78 %. Northern blot analysis revealed that 50 bp-length hpRNA constructs could be transcribed efficiently and processed into siRNAs; however, no correlation between siRNA accumulation and degree of antiviral defense was observed. Results presented here indicated that the middle and 3' end of the CP cDNA was important for hpRNA-mediated PVY resistance, improving the design of pathogen-derived hpRNA expression cassettes for transgenic plant against viruses. |
Constitutive expression of Arabidopsis DREB1B in transgenic potato enhances drought and freezing toleranceS. Movahedi, B. E. Sayed Tabatabaei, H. Alizade, C. Ghobadi, A. Yamchi, G. KhaksarBiologia plantarum 56:37-42, 2012 | DOI: 10.1007/s10535-012-0013-6 DNA cassette consisting of an Arabidopsis dehydration-responsive element binding factor 1 (DREB1B) cDNA, driven by a cauliflower mosaic virus 35S promoter, was introduced into potato plants (Solanum tuberosum L.) through Agrobacterium tumefaciens-mediated gene transfer. The presence and expression of the gene in transgenic plants were confirmed by the PCR and RT-PCR techniques, respectively. Northern hybridization using a DREB1B cDNA probe revealed high levels of DREB1B expression among the most transgenic lines. Overexpression of DREB1B imparted a significant freezing and drought tolerance gain in the transgenic potato lines. In comparison with the wild-type plants, the transgenic potatoes contained higher proline content under drought and freezing conditions, and maintained their relative water content higher under water stress. The enhancement of tolerance in transgenic potato highlights the presence of genes responding to the transcription factor DREB1B in this plant. |
Improved drought resistance in a wheat stay-green mutant tasg1 under field conditionsF. X. Tian, J. F. Gong, G. P. Wang, G. K. Wang, Z. Y. Fan, W. WangBiologia plantarum 56:509-515, 2012 | DOI: 10.1007/s10535-012-0049-7 We investigated the drought resistance of a wheat (Triticum aestivum L.) stay-green mutant tasg1 and its wild-type (WT) in field experiments conducted for two years. Drought stress was imposed by controlling irrigation and sheltering the plants from rain. Compared with the WT, tasg1 exhibited a distinct delayed senescence under both normal and drought stress conditions, as indicated by slower degradation of chlorophyll and decrease in net photosynthetic rate than in WT. At the same time, tasg1 mutants maintained more integrated chloroplasts and thylakoid ultrastructure than did WT plants under drought stress. Lower malondialdehyde content and higher antioxidative enzyme activities in tasg1, compared to WT, may be involved in the stay-green phenotype and drought resistance of tasg1. |
Amelioration of salt-induced oxidative stress in eggplant by application of 24-epibrassinolideH. -D. Ding, X. -H. Zhu, Z. -W. Zhu, S. -J. Yang, D. -S. Zha, X. -X. WuBiologia plantarum 56:767-770, 2012 | DOI: 10.1007/s10535-012-0108-0 The effects of exogenous 24-epibrassinolide (EBR) on the growth, oxidative damage, antioxidant system and ion contents in eggplant (Solanum melongena L.) seedlings under salt stress were investigated. Eggplant seedlings were exposed to 90 mM NaCl with 0, 0.025, 0.05, 0.10 and 0.20 mg dm-3 EBR for 10 d. EBR, especially at concentration 0.05 mg dm-3, alleviated growth suppression caused by NaCl stress, decreased electrolyte leakage, superoxide production and content of malondialdehyde and H2O2 in NaCl-treated plants. EBR also increased activities of superoxide dismutase, guaiacol peroxidase, catalase and ascorbate peroxidase and the contents of ascorbic acid and reduced glutathione. Furthermore, we also found that Na+, Cl- contents were decreased, K+, Ca2+ contents and K+/Na+, Ca2+/Na+ ratios were increased in the presence of EBR under salt stress. |
Transgenic tobacco plants containing Bt and GNA genesH. Y. Zhang, X. Z. Liu, L. Wei, L. Y. Zhou, Y. M. YangBiologia plantarum 51:746-748, 2007 | DOI: 10.1007/s10535-007-0152-3 A new plant expression vector (pBSbtCry1Ac-GNA) containing two insect resistant genes, a synthetic chimeric gene SbtCry1Ac encoding the insecticidal protein CrylAc and a gene GNA encoding snowdrop lectin (Galanthus nivalis agglutinin) was constructed. Transgenic tobacco plants containing these two genes were obtained through Agrobacterium-mediated transformation of tobacco leaf discs. Results from PCR detection and genomic DNA Southern blot analysis indicated that both SbtCrylAc gene and GNA gene were integrated into the genome of these plants. Results of Western blot analysis indicated that these two proteins were expressed in the analyzed plants. Bioassays of Myzus persicae and Helicoverpa assulta on detached leaves of transformed tobacco plants were carried out. The average aphid inhibition rate of these plants tested at 12 d post-infestation was 71.9 %. The average H. assulta mortality of these plants tested at 6 d post-infestation was up to 89.8 %. The kanamycin resistance of the T1 progeny of these transgenic plants was analyzed and a typical 3:1 segregation was observed. |


