Fulltext search in archive
Results 721 to 750 of 1797:
High efficient transgenic plant regeneration from embryogenic calluses of Citrus sinensisY. X. Duan, W. W. Guo, H. J. Meng, N. G. Tao, D. D. Li, X. X. DengBiologia plantarum 51:212-216, 2007 | DOI: 10.1007/s10535-007-0043-7 Transformation and high efficient regeneration of transgenic plants from embryogenic calluses of Bingtang sweet orange [Citrus sinensis (L.) Osbeck] was reported. Embryogenic calluses were inoculated with Agrobacterium tumefaciens strain EHA105, harboring the binary Ti plasmid pROK II and carrying a neomycin phosphotransferase II (NPTII) gene, an intron β-glucuronidase (GUS) gene and the Arabidopsis APETALA1 (AP1) gene. Transformation treatment was with inoculation time of 30 min, co-culture of 3 d at 23 °C and supplementation of the co-culture medium with 2 mg dm-3 acetosyringone (AS). Kanamycin (50 mg dm-3) was effective to inhibit the growth of non-transformed calluses while it did not affect the transformed ones. The total number of transformed callus lines was 7 with 100 % embryo induction. High efficient regeneration of the transgenic embryos (88 % with 4-5 shoots per embryoid) was realized within 3 months. Integration of the transgene into the citrus genome was confirmed by histochemical GUS staining, polymerase chain reaction (PCR) analysis with AP1-specific primer and Southern blot hybridization with a 712 bp PCR fragment of AP1 as the probe. |
Efficient Regeneration of Tetraploid Isatis indigotica Plants via Adventitious Organogenesis from Hypocotyl ExplantsL. Zhang, G. Kai, T. Xu, Y. Pi, H. Zhang, X. Sun, K. TangBiologia plantarum 48:121-124, 2004 | DOI: 10.1023/B:BIOP.0000024287.23065.e8 An efficient in vitro plant regeneration system via hypocotyl segments of tetraploid Isatis indigotica Fort. was established. Murashige and Skoog's (MS) and Gamborg's (GB5) media were found to be superior to White medium for promoting shoot regeneration. The highest shoot regeneration (92 %) was achieved from hypocotyls cultured on MS medium containing 8.9 μM benzyladenine (BA) and 2.7 μM naphthaleneacetic acid (NAA), with an average of 4.2 shoots developed per explant. Plant regeneration was also improved when the explants were cultured in MS basal medium containing 3 % (m/v) sucrose and grown under a 12-h photoperiod. The developed shoots were well rooted in a half-strength MS medium supplemented with 0.5 μM indole-3-butyric acid (IBA) and were morphologically normal after transfer to soil. |
Activity of secreted cell wall-modifying enzymes and expression of peroxidase-encoding gene following germination of Orobanche ramosaC. Veronesi, E. Bonnin, S. Calvez, P. Thalouarn, P. SimierBiologia plantarum 51:391-394, 2007 | DOI: 10.1007/s10535-007-0084-y Radicle growth of germinated seed of the root parasite O. ramosa is shown to be rapidly accompanied by secretion of proteins including pectinolytic enzymes, polygalacturonase and rhamnogalacturonase. These secretions peaked between 4 to 8 d after induction of germination and remained constant for some further days in the case of polygalacturonases. After 6 d, germinated seeds secreted proteins which exhibit peroxidase activity. The latter may be correlated with expression of OrPOX1, a putative gene encoding for secreted peroxidase. The involvement of these enzymes in host root attack and haustorium formation by the parasite is discussed. |
Multiple Shoot Induction and Plant Regeneration from Embryo Axes of Six Cultivars of Gossypium hirsutumA.K. Banerjee, D.C. Agrawal, S.M. Nalawade, S. Hazra, K.V. KrishnamurthyBiologia plantarum 46:433-436, 2003 | DOI: 10.1023/B:BIOP.0000023889.20631.92 The report describes in vitro plant regeneration from embryo axis explants of six cultivars of cotton. Induction of a maximum number of multiple shoots in all six cultivars could be achieved on Murashige and Skoog's (MS) salts and Gamborg's (B5) vitamins supplemented with 0.4 μM benzyladenine (BA) and 0.1 μM napthaleneacetic acid (NAA). Elongated shoots could be rooted on half strength medium supplemented with 0.5 μM NAA. Rooted shoots survived (92 %) after hardening in the greenhouse and grew to maturity (100 %) after transfer to field. |
Role of Amino Acids in Plant Responses to StressesV.K. RaiBiologia plantarum 45:481-487, 2002 | DOI: 10.1023/A:1022308229759 Plants subjected to stress show accumulation of proline and other amino acids. The role played by accumulated amino acids in plants varies from acting as osmolyte, regulation of ion transport, modulating stomatal opening, and detoxification of heavy metals. Amino acids also affect synthesis and activity of some enzymes, gene expression, and redox-homeostasis. These roles played by amino acids have been critically examined and reviewed. |
Stability of β-glucuronidase gene expression in transgenic Tricyrtis hirta plants after two years of cultivationS. Mori, E. Oka, H. Umehara, H. Kobayashi, Y. Hoshi, M. Kondo, K. Ogata, M. NakanoBiologia plantarum 52:513-516, 2008 | DOI: 10.1007/s10535-008-0099-z Transgenic plants of Tricyrtis hirta carrying the intron-containing β-glucuronidase (GUS) gene under the control of the CaMV35S promoter have been cultivated for two years. Four independent transgenic plants produced flowers 1-2 years after acclimatization, and all of them contained one copy of the transgene as indicated by inverse polymerase chain reaction (PCR) analysis. All the four transgenic plants showed stable expression of the gus gene in leaves, stems, roots, tepals, stamens and pistils as indicated by histochemical and fluorometric GUS assays, although differences in the GUS activity were observed among different organs of each transgenic plant. No apparent gus gene silencing was observed in transgenic T. hirta plants even after two years of cultivation. |
Resistance of transgenic papaya plants to Papaya ringspot virusS. Kertbundit, N. Pongtanom, P. Ruanjan, D. Chantasingh, A. Tanwanchai, S. Panyim, M. JuříčekBiologia plantarum 51:333-339, 2007 | DOI: 10.1007/s10535-007-0065-1 The coat protein gene isolated from Papaya ringspot virus, Thai isolate, was used to generate transgenic papayas. A binary vector containing the coat protein gene under the control of a 35S promoter, was constructed and transformed into somatic embryos of papaya cultivar Khak Dum by microprojectile bombardment. Eight transgenic lines were identified from 1980 bombarded calli of papaya somatic embryos under kanamycin selection. Integration of the transferred genes into kanamycin resistant papaya calli was verified by PCR amplification of the coat protein gene, GUS assays and Southern blot hybridization. Although the coat protein gene was detected in all transgenic lines, only line G2 was found to be highly resistant to virus. This resistant line showed high degree of rearrangement of the inserted coat protein expression cassette while the coat protein gene itself had a deletion of 166 bp on the 3' end of its sequence. Although the transcription of the coat protein gene was detected in all transgenic lines by RT-PCR, only two transgenic papayas expressed the intact coat protein. Moreover, in the resistant line G2 the amount of the truncated coat protein mRNA was significantly decreased. These results point to an RNA mediated mechanism of coat protein mediated resistance in papaya, probably based on post-transcriptional gene silencing. |
Interactive effects of cadmium and aluminum on growth and antioxidative enzymes in soybeanI. H. Shamsi, K. Wei, G. P. Zhang, G. H. Jilani, M. J. HassanBiologia plantarum 52:165-169, 2008 | DOI: 10.1007/s10535-008-0036-1 The effects of Al, Cd and pH on growth, photosynthesis, malondialdehyde (MDA) content, and some antioxidant enzyme activities of the two soybean cultivars with different Al tolerance were determined using a hydroponic culture. There were six treatments as follows: pH 6.5; pH 4.0; pH 6.5 + 1.0 µM Cd; pH 4.0 + 1.0 µM Cd; pH 4.0 + 150 µM Al; pH 4.0 + 1.0 µM Cd + 150 µM Al. The results showed that the low pH (4.0) and Al treatments caused marked reduction in the growth (root and shoot length and dry mass), chlorophyll content (SPAD value) and net photosynthetic rate. Higher malondialdehyde content, superoxide dismutase (SOD) and peroxidase (POD) activities were detected in the plants exposed to both Al and Cd than in those exposed to Al treatment alone. An expressive enhancement of SOD and POD was observed in the plants exposed to 150 µM Al in the comparison with the control plants, especially in Al-sensitive cv. Zhechun 2 which had also significantly higher Al and Cd content than Al tolerant cv. Liao-1. Cd addition increased Al content in the plants exposed to Al + Cd stress, and cv. Zhechun 2 had relatively lower Al content. The present research indicated that Al and Cd are synergistic in their effects on plant growth and some physiological traits. |
Pepper morphological traits related with resistance to Phytophthora capsiciC. Egea-Gilabert, G. Bilotti, M. E. Requena, M. Ezziyyani, J. M. Vivo-Molina, M. E. CandelaBiologia plantarum 52:105-109, 2008 | DOI: 10.1007/s10535-008-0019-2 Inheritance of 10 morphological and quantitative traits related to plant and fruit development and resistance to the pathogen Phytophthora capsici was studied in an intraspecific cross between a non-pungent, susceptible Capsicum annuum parent (cv. Americano) and a wild, pungent and resistant line (Serrano Criollo de Morelos-334). Data were obtained from the segregation of 166 F2 plants and 50 F3 plants in four years. Three of the traits analyzed (necrosis length, leaf width and leaf length) exhibited a transgressive segregation. A multiple linear regression analysis was applied in order to establish a relationship between necrosis length and some of the morphological traits measured such as length and width of leaf, length, diameter and mass of fruit, capsaicin content in fruits, and presence of hair on leaves and stems. The results identified a linear dependence between necrosis length (as an inverse measurement of resistance) and leaf width, fruit diameter and hair presence in the stem. Pungency was not related with resistance. |
Exogenous salicylic acid alleviates NaCl toxicity and increases antioxidative enzyme activity in Lycopersicon esculentumY. He, Z. J. ZhuBiologia plantarum 52:792-795, 2008 | DOI: 10.1007/s10535-008-0155-8 Effects of exogenous salicylic acid (SA) on plant growth, contents of Na, K, Ca and Mg, activities of superoxide dismutase (SOD), guaiacol peroxidase (GPX), ascorbate peroxidase (APX), dehydroascorbate reductase (DHAR), glutathione reductase (GR) and catalase (CAT), and contents of ascorbate and glutathione were investigated in tomato (Lycopersicon esculentum L.) plants treated with 100 mM NaCl. NaCl treatment significantly increased H2O2 content and lipid peroxidation indicated by accumulation of thiobarbituric acid reactive substances (TBARS). A foliar spray of 1 mM SA significantly decreased lipid peroxidation caused by NaCl and improved the plant growth. This alleviation of NaCl toxicity by SA was related to decreases in Na contents, increases in K and Mg contents in shoots and roots, and increases in the activities of SOD, CAT, GPX and DHAR and the contents of ascorbate and glutathione. |
Cadmium mitigates ultraviolet-B stress in Anabaena doliolum: Enzymatic and non-enzymatic antioxidantsP. Bhargava, N. Atri, A. K. Srivastava, L. C. RaiBiologia plantarum 51:546-550, 2007 | DOI: 10.1007/s10535-007-0118-5 Impact of ultraviolet-B (UV-B) and Cd, applied individually and in combination, measured in terms of oxygen-evolution, chlorophyll (Chl) and protein contents, lipid peroxidation, and enzymatic and non-enzymatic antioxidants of Anabaena doliolum, revealed a greater oxidative damage induced by UV-B than by Cd. While superoxide dismutase (SOD) showed a greater stimulation by UV-B than Cd, the activities of catalase (CAT) and glutathione reductase (GR) declined after UV-B treatment. Cd treatment, however, enhanced the activities of ascorbate peroxidase (APX) and GR. CAT activity increased at low but decreased at high dose of Cd. Increase in carotenoid (Car) content in UV-B treated cells suggested a shielding effect of Car against UV-B stress. A 15-and 10-fold rise in α-tocopherol (α-TOC) content at high dose of Cd and/or UV-B offered testimony to the antioxidant role of α-TOC. |
Ecophysiological Characterization of Dormancy States in Turions of the Aquatic Carnivorous Plant Aldrovanda vesiculosaL. AdamecBiologia plantarum 46:395-402, 2003 | DOI: 10.1023/B:BIOP.0000023883.62127.5e Two main dormancy states, innate and imposed dormancy, were characterized in turions (winter buds) of the aquatic carnivorous plant Aldrovanda vesiculosa L. (Droseraceae) kept at 3 ± 1 °C in a refrigerator over the winter. As a result of the breaking of imposed dormancy by a temperature increase (at 15 - 20 °C), some of the turions rose to the water surface within 1 - 3 d and germinated. Turion leaves contained large lacunae with a slimy reticulum and were filled by water over winter. As a result of breaking imposed dormancy, the proportion of gas volume in inner turion leaves rose from 10 - 20 % to 100 % of leaf lacunae volume. The aerobic dark respiration rate of the turions [0.74 - 1.5 μmol O2) kg-1(FM) s-1] slightly increased during innate dormancy after 1 - 2 d at 20 °C, while it was almost constant during the breaking of imposed dormancy. The anaerobic fermentation rate of the turions was only 1.5 - 7 % of the oxygen respiration rate and also was constant during the breaking of imposed dormancy. In turions, the content of glucose, fructose, and sucrose was the same for the two states of dormancy, but starch content was greatly reduced for the imposed dormancy (10 - 11 vs. 32 % DM). It may be suggested that a temperature increase causes an increase of fermentation or respiration which is responsible for the evolution of gas in turion lacunae and, thus, for turion rising. |
Genetic diversity in in vitro-conserved germplasm of Curcuma L. as revealed by RAPD markersZ. Hussain, R. K. Tyagi, R. Sharma, A. AgrawalBiologia plantarum 52:627-633, 2008 | DOI: 10.1007/s10535-008-0123-3 A set of 30 accessions of five Curcuma species-C. latifolia, C. malabarica, C. manga and C. raktakanta and 13 morphotypes (identified on the basis of morphological markers) of C. longa conserved in the In Vitro Genebank at National Bureau of Plant Genetic Resources, New Delhi, were subjected to RAPD analysis. Of the 200 RAPD primers screened, 21 polymorphic primers were selected for further study. Mean genetic similarities based on Jaccard's similarity coefficient ranged from 0.18 to 0.86 in accessions of cultivated species, i.e., C. longa and from 0.25 to 0.86 in wild species. The dendrogram derived from the RAPD data corroborated the morphological classification of the morphotypes. The efficiency of individual RAPD primers was also compared; primers OPC-20, OPO-06, OPC-01 and OPL-03 were adjudged highly informative in discriminating the germplasm of Curcuma. |
Effect of drought on biomass, protein content, lipid peroxidation and antioxidant enzymes in two sesame cultivarsF. Fazeli, M. Ghorbanli, V. NiknamBiologia plantarum 51:98-103, 2007 | DOI: 10.1007/s10535-007-0020-1 The effects of drought on growth, protein content, lipid peroxidation, superoxide dismutase (SOD), peroxidase (POX), catalase (CAT) and polyphenol oxidase (PPO) were studied in leaves and roots of Sesamum indicum L. cvs. Darab 14 and Yekta. Four weeks after sowing, plants were grown under soil moisture corresponding to 100, 75, 50 and 25 % field capacity for next four weeks. Fresh and dry masses, and total protein content in leaves and roots decreased obviously under drought. However, several new proteins appeared and content of some proteins was affected. Measurement of malondialdehyde content in leaves and roots showed that lipid peroxidation was lower in Yekta than in Darab 14. Severe stress increased SOD, POX, CAT and PPO activities in leaves and roots, especially in Yekta. According to the present study Yekta is more resistant to drought than Darab 14. |
Optimum storage conditions for product of transiently expressed epitopes of Human papillomavirus using Potato virus X-based vectorN. Čeřovská, H. Hoffmeisterová, T. Moravec, H. Plchová, J. Folwarczna, R. HadámkováBiologia plantarum 52:184-186, 2008 | DOI: 10.1007/s10535-008-0041-4 We describe the optimized storage conditions of recombinant Potato virus A coat protein (ACP) carrying two different epitopes from Human papillomavirus type 16 (HPV-16). Epitope derived from minor capsid protein L2 was expressed as N-terminal fusion with ACP while an epitope derived from E7 oncoprotein was fused to its C-termini. The construct was cloned into Potato X potexvirus (PVX) based vector and transiently expressed in plants using Agrobacterium tumefaciens mediated inoculation. The effect of storage conditions on the serological activity of L2ACPE7 was studied by ELISA using IgG anti PVX, PVA and L2. Purified L2ACPE7 stored freeze-dried (at -20 °C), frozen at various temperatures (-20 °C, -70 °C) and at +4 °C were tested. Purified L2ACPE7 was most stable as lyophilized material stored at -20 °C. Our study demonstrates suitable way for the storage of plant material containing foreign viral epitopes for the purposes of edible vaccination. |
The 26S Proteasome of the Resurrection Plant Tortula ruralis: Cloning and Characterization of the TrRPT2 SubunitX. Chen, A.J. WoodBiologia plantarum 46:363-368, 2003 | DOI: 10.1023/A:1024369917715 Tortula ruralis is an important experimental system for the study of plant desiccation tolerance. EST gene discovery efforts utilizing desiccated gametophytes have identified a cDNA TrRpt2 encoding a predicted polypeptide with significant similarity to the 26S proteasome regulatory subunit IV. TrRPT2, the 446 amino acid deduced polypeptide, has a predicted molecular mass of 49.6 kDa, and a predicted pI of 8.15. Phylogenetic analysis demonstrated that previously characterized RPT2 polypeptide sequences could be reproducibly grouped into 3 major clades and that TrRPT2 forms a discrete evolutionary group. RNA blot hybridizations were used to analyze TrRpt2 expression in response to: 1) desiccation and rehydration, 2) abscisic acid-treatment, 3) increased NaCl concentration, and 4) NaCl-shock. TrRpt2 steady-state mRNA transcript levels are unchanged in response to all treatments and the gene is constitutively expressed. |
ISSR primer screening and preliminary evaluation of genetic diversity in wild populations of Gycyrrhiza uralensisH. Yao, Y. Zhao, D. F. Chen, J. K. Chen, T. S. ZhouBiologia plantarum 52:117-120, 2008 | DOI: 10.1007/s10535-008-0022-7 Fourteen efficient inter-simple sequence repeat (ISSR) primers were screened and optimized for detecting the genetic diversity in wild populations of Glycyrrhiza uralensis Fisch. By using these primers, 249 polymorphic bands out of a total of 270 (92.2 %) were generated from 70 individuals of 4 wild G. uralensis populations sampled from Inner Mongolia Province of China. Nei's gene diversity (h) and Shannon index (I) calculated from the data matrix of the ISSR phenotypes revealed a high level of genetic diversity with h = 0.268 and I = 0.415 within this plant. Analysis of molecular variation (AMOVA) showed that most of the genetic variation (81 %) occurred within the populations, whereas the variance among populations was only 19 %. The UPGMA tree based on Nei's unbiased genetic diversity illustrated that populations from Bulage and Bayanwusu were genetically close related, while the population from Shanghaimiao was found to be the most diverse from the other three. The high genetic diversity implies that the wild resources of this species could be restored soon if an appropriate and efficient protection strategy was employed. Our results also provided an optimized method for evaluating genetic diversity of G. uralensis using ISSR markers which was useful for further investigation. |
Responses of the cherry rootstock to salinity in vitroU. Erturk, N. Sivritepe, C. Yerlikaya, M. Bor, F. Ozdemir, I. TurkanBiologia plantarum 51:597-600, 2007 | DOI: 10.1007/s10535-007-0132-7 The in vitro response of sweet cherry rootstock Gisela 5 (Prunus cerasus × Prunus canescens) to increasing concentrations of NaCl (0, 50, 100 and 150 mM) in the Murashige and Skoog culture medium was studied. Induced salinity reduced growth and chlorophyll content in shoots but had no effect on water content. The increase in malondialdehyde content indicated that salinity induced oxidative stress which was accompanied with the visible symptoms of salt injury in the shoots. Antioxidant enzymes, such as superoxide dismutase, ascorbate peroxidase, peroxidase, catalase, and glutathione reductase were also significantly elevated. Although no change was observed in the Cl concentration, Na concentration of shoots significantly rose and NaCl treatments impaired K, Ca and Mg nutrition and induced imbalance in K:Na and Na:Ca ratios. |
Specific activation of PR-10 pathogenesis-related genes in apple by an incompatible race of Venturia inaequalisM. Chevalier, L. Parisi, B. Gueye, C. Campion, P. Simoneau, P. PoupardBiologia plantarum 52:718-722, 2008 | DOI: 10.1007/s10535-008-0138-9 Apple (Malus × domestica Borkh. cv. Golden Delicious) trees were inoculated with fungal pathogens, corresponding either to an incompatible strain of Venturia inaequalis or to non-host pathogens (Venturia pyrina, Alternaria brassicicola) in order to characterize the regulation of PR-10 genes in these different situations in relationship to symptom development. Macro-and microscopic observations of the plant-fungus interactions revealed typical symptoms of resistance with V. inaequalis and symptoms of hypersensitivity (HR) on around 5 % of leaves with V. pyrina. No HR was microscopically observed with A. brassicicola. In the non-host situations, the PR-10 expression in leaves at the transcriptional level was not (A. brassicicola) or very slightly (V. pyrina) activated. This strongly suggests that PR-10 is not a component of the non-host resistance in the interactions studied here. By contrast specific up-regulation of PR-10 was evident after inoculation with V. inaequalis. |
Photosynthetic rate and yield formation in different maize hybridsL. Ding, K. J. Wang, G. M. Jiang, M. Z. Liu, L. M. GaoBiologia plantarum 51:165-168, 2007 | DOI: 10.1007/s10535-007-0032-x The relationship between photosynthetic rate and yield formation processes of the newer and older maize hybrids were investigated. Leaf area at flowering (source) and kernel number (sink) of the newer hybrids were greater than the older ones although their light-saturated photosynthetic rate (Psat) were not greater than the older ones before flowering. After flowering, Psat and chlorophyll content of the newer hybrids declined more slowly than the older ones. They not only distributed almost all photosynthates produced after flowering to grain but also reallocated some reserved photosynthates produced before flowering to grain. The newer hybrids exhibited greater grain mass than the older ones mostly because they could optimally regulate the photosynthetic rate and yield formation processes to maximize grain mass. |
In vivo activity of recombinant human lewis fucosyltransferase III in leaves of Nicotiana tabacum L.S. Lhernould, P. Labrousse, D. Lejolly, R. Léonard, M. Carlué, G. CostaBiologia plantarum 52:267-274, 2008 | DOI: 10.1007/s10535-008-0057-9 Fucosylation in plants occurs in glycoproteins and polysaccharides but the function of fucosylation is largely unknown. We aimed to analyze the effects of over-expression of human fucosyltransferase III (hFucT III) on in vivo N-glycan accumulation in tobacco plant leaves and focused on comparing the amount of Lewis a (Lea)-epitope accumulation in transgenic and in wild-type plants. Fucosyltransferase assays, Western blot and mass spectrometry were used to identify, quantify and analyse Lea N-glycans. We found that constitutive overexpression of hFucT III activity had no effect on Lea complex type N-glycans accumulation. Our results suggest that tobacco recombinant hFucT III acts more as a hydrolase than as a transferase. |
Effects of Capsaicin on Plant GrowthH. Kato-Noguchi, Y. TanakaBiologia plantarum 46:157-159, 2003 | DOI: 10.1023/A:1027317906839 Capsaicin, a possible allelochemical, caused growth inhibition of roots and shoots of alfalfa (Medicago sativa), cress (Lepidium sativum), lettuce (Lactuca sativa), crabgrass (Digitaria sanguinalis), timothy (Phleum pratense) and ryegrass (Lolium multiflorum), and suppressed their germination. Increasing the dose of capsaicin increased the inhibition. The concentrations for 50 % inhibition of the root growth were 2.7, 0.32, 2.1, 0.27, 0.29 and 0.57 mM for alfalfa, cress, lettuce, crabgrass, timothy and ryegrass, respectively, and the concentrations for 50 % inhibition of the shoot growth were 17, 0.87, 6.7, 2.3, 1.4 and 6.2 mM for alfalfa, cress, lettuce, crabgrass, timothy and ryegrass, respectively. Germination percentage was inhibited 50 % at the concentrations 82, 88, 68, 4.8, 22 and 11 mM for alfalfa, cress, lettuce, crabgrass, timothy and ryegrass, respectively. Thus, effectiveness of capsaicin on the plant growth differed with species and targets, and suggests that capsaicin may act as an allelochemical to other plants. |
Preparation of HMW DNA from Plant Nuclei and Chromosomes Isolated from Root TipsH. ©imková, J. Číhalíková, J. Vrána, M.A Lysák, J. DoleľelBiologia plantarum 46:369-373, 2003 | DOI: 10.1023/A:1024322001786 Simple, fast and cost-effective method for preparation of DNA with high molecular weight (HMW DNA) from plant nuclei and mitotic chromosomes has been developed. The technique involves mechanical homogenization of formaldehyde-fixed root tips, purification of nuclei and/or chromosomes on sucrose gradient, embedding in low-melting-point agarose, and DNA isolation in agarose plugs. Alternatively, nuclei and chromosomes may be purified using flow cytometry. Majority of DNA obtained is megabase-sized and well digestible by restriction endonucleases. The method is highly efficient as microgram amounts of DNA can be obtained from only several milligrams of plant tissue. Handling negligible amounts of plant material reduces the consumption of chemicals. Furthermore, the use of root tips makes it possible to obtain high-quality DNA even from plant species with leaves that are rigid or rich in secondary metabolites such as polyphenols. It is expected that preparation of HMW DNA from root tip nuclei will facilitate long-range mapping and construction of large-insert DNA libraries also in these species. Successful isolation of HMW DNA from flow-sorted chromosomes opens a way for construction of chromosome-specific large-insert libraries in plants. |
Improvement of in vitro androgenesis in niger using amino acids and polyaminesB. P. Hema, H. N. MurthyBiologia plantarum 52:121-125, 2008 | DOI: 10.1007/s10535-008-0024-5 The effects of amino acids (arginine, aspargine, cysteine, glutamine, glycine and proline) and polyamines (putrescine and spermidine) on embryogenesis and plant regeneration from cultured anthers of Guizotia abyssinica (L. f.) Cass. cv. Ootacamund was studied. Supplementation of amino acids (0.5-5.0 mM) to the induction medium individually and in combination, improved embryo yield. B5 medium supplemented with 2 mM proline, 10 µM 2,4-dichlorophenoxyacetic acid, 2 µM kinetin and 0.2 M sucrose induced highest number of embryos (63 per 60 anthers cultured). Addition of polyamines (5-200 µM) to the same medium also enhanced the rate of embryogenesis. |
Assessment of genetic stability of in vitro grown Dictyospermum ovalifoliumM. Chandrika, Thoyajaksha, V. Ravishankar Rai, K. Ramachandra KiniBiologia plantarum 52:735-739, 2008 | DOI: 10.1007/s10535-008-0142-0 In the present study, a polymerase chain reaction (PCR)-based method namely inter simple sequence repeat (ISSR) was employed to assess genetic stability in tissue culture-derived Dictyospermum ovalifolium plantlets. To study genomic stability of micropropagated plants, 14 individuals were randomly tagged among a population of 2500 regenerants and were compared with single donor mother plant. A total of 51 clear and reproducible bands ranging from 200 bp to 2.1 kb were scored corresponding to an average of 3.64 bands per primer. Two of the 51 bands were polymorphic (3.92 %) among 14 individuals, thus indicating the occurrence of low level genomic variation in the micropropagated plants. Cluster analysis indicates that genetic similarity values were 0.978 which allows classification of the plants to distinct groups. Further an attempt was made to reintroduce the micropropagated plants into their natural habitat. Over one thousand six hundred fifty plants were successfully established. |
Effect of NaCl and CaCl2 on growth and contents of minerals, chlorophyll, proline and sugars in the apple rootstock M 4 cultured in vitroT. E. SotiropoulosBiologia plantarum 51:177-180, 2007 | DOI: 10.1007/s10535-007-0035-7 The apple (Malus domestica Borkh) rootstock M 4 shoots were grown in vitro for 4 weeks on Murashige and Skoog (MS) medium containing three NaCl concentrations (35, 100 and 200 mM) in combination with two CaCl2 concentrations (5 and 10 mM). Inclusion of 10 mM CaCl2 in the medium, in the presence of 35 mM NaCl, significantly increased the number of shoots and the fresh mass compared to 5 mM CaCl2. The number of shoots, length of shoots, and the fresh mass of cultures were very low in the presence of 100 and 200 mM NaCl, independently of CaCl2 concentration of the medium. By increasing NaCl and CaCl2 concentrations in the culture medium, contents of N, Na, Cl, proline and soluble sugars in plantlets increased, whereas K, Mg, B, Zn and chlorophyll content decreased in comparison to the control. |
Leaf gas exchange, water relations, nutrient content and growth in citrus and olive seedlings under salinityJ. C. Melgar, J. P. Syvertsen, V. Martínez, F. García-SánchezBiologia plantarum 52:385-390, 2008 | DOI: 10.1007/s10535-008-0081-9 The effects of salinity on growth, leaf nutrient content, water relations, gas exchange parameters and chlorophyll fluorescence were studied in six-month-old seedlings of citrus (Citrus limonia Osbeck) and rooted cuttings of olive (Olea europaea L. cv. Arbequina). Citrus and olive were grown in a greenhouse and watered with half strength Hoagland's solution plus 0 or 50 mM NaCl for citrus, or plus 0 or 100 mM NaCl for olive. Salinity increased Cl- and Na+ content in leaves and roots in both species and reduced total plant dry mass, net photosynthetic rate and stomatal conductance. Decreased growth and gas exchange was apparently due to a toxic effect of Cl- and/or Na+ and not due to osmotic stress since both species were able to osmotically adjust to maintain pressure potential higher than in non-salinized leaves. Internal CO2 concentration in the mesophyll was not reduced in either species. Salinity decreased leaf chlorophyll a content only in citrus. |
Effects of freezing on plasma membrane H+-ATPase of the callus from Chorispora bungeanaJ. M. Wu, Z. G. Zhao, H. Xing, H. P. Guo, W. X. Li, L. Z. An, S. J. Xu, T. ChenBiologia plantarum 51:229-234, 2007 | DOI: 10.1007/s10535-007-0046-4 The influence of freezing treatment on plasma membrane (PM) H+-ATPase was investigated using plasma membrane vesicles isolated from calluses from Chorispora bungeana Fisch. & C.A. Mey. by the discontinuous sucrose gradient centrifugation. Freezing treatment (-4 °C) for 5 d resulted in significant increases in the ATPase activity and the activity of p-nitrophenyl phosphate (PNPP) hydrolysis, decreases in the Km for ATP hydrolysis and PNPP hydrolysis, and the shift of optimal pH from 6.5 to 7.0. Also, the activity PNPP hydrolysis was less sensitive to vanadate after freezing treatment compared to control, while the inhibition of ATP hydrolysis by hydroxylamine was more sensitive. In addition, freezing treatment also decreased the activation effects of trypsin on PNPP hydrolysis, but increased the activation effects of lysophosphatidylcholine on ATP hydrolysis. Taken together, these results suggested that PM H+-ATPase might play an important role during adaptation to freezing and enhancing the frost hardness in C. bungeana. |
Plant Micropropagation and Callus Induction of Some Annual Salsola SpeciesB. Stefaniak, A. WoĽny, V. LiBiologia plantarum 46:305-308, 2003 | DOI: 10.1023/A:1022879400747 Micropropagated plants of two annual haloxerophytic Asiatic Salsola species (S. pestifer and S. lanata) were obtained from zygotic embryos cultured on Murashige and Skoog (MS) agar medium supplemented with 0.5 µM benzylamino-purine (BAP) and 0.3 µM indole-3-acetic acid (IAA) or with 0.5 µM 6 γ, γ-dimethylallylaminopurine and 0.3 μM IAA. The callus induction from shoot and leaf explants derived from plants propagated in vitro were obtained on MS agar medium with various concentration of auxins and cytokinins. The best medium for growth and proliferation of calluses of both studied species was MS medium containing 9.0 µM 2,4-dichlorophenoxyacetic acid. It was also determined that beginning of plant regeneration from callus of S. lanata was induced by 8.8 µM BAP. |
Headspace-SPME of in vitro shoot-cultures and micropropagated plants of Lavandula viridisS. Gonçalves, H. Serra, J. M. F. Nogueira, R. Almeida, L. Custódio, A. RomanoBiologia plantarum 52:133-136, 2008 | DOI: 10.1007/s10535-008-0027-2 In this work the volatiles emitted from in vitro shoot-cultures and micropropagated plants of Lavandula viridis L'Hér. were characterized and compared with those obtained from the field-grown mother-plant, using headspace solid phase micro-extraction following by capillary gas chromatography coupled to mass spectrometry (HS-SPME-GC/MS). The headspace composition consisted mainly in oxygenated monoterpenes (66.7-79.2 %), where the major constituents emitted by the mature field-grown mother-plant, in vitro shoot-cultures and micropropagated plants were 1,8-cineole (74.0, 51.9 and 57.8 %) and camphor (2.9, 15.3 and 8.7 %), respectively. The headspace of in vitro shoot-cultures and micropropagated plants showed greater amount of α-pinene, camphene, β-pinene, β-selinene and selina-3,7(11)-diene, when compared with the field-grown mother-plant. |


