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Thidiazuron and silver nitrate enhanced gynogenesis of unfertilized ovule cultures of Cucumis sativusJ. W. Li, S. W. Si, J. Y. Cheng, J. X. Li, J. Q. LiuBiologia plantarum 57:164-168, 2013 | DOI: 10.1007/s10535-012-0269-x Gynogenesis of Chinese long cucumber (Cucumis sativus L.) was obtained from unpollinated ovules cultured on cucumber basal medium (CBM) supplemented with thidiazuron (TDZ) and in some experiments AgNO3. High induction frequencies (7.85-12.14 %) were induced from unpollinated ovules at the time of anthesis at 0.03-0.07 mg dm-3 TDZ. Histological analysis indicated that embryo sacs developed completely at the time of anthesis. Further, the highest plant regeneration rate was achieved at CBM supplemented with 0.05 mg dm-3 a-naphthaleneacetic acid, 0.2 mg dm-3 6-benzyladenine and 5-10 mg dm-3 AgNO3. Flow cytometry analysis showed that 80 % of the regenerated plants were haploid. Histological micrographs and ploidy level analyses clearly revealed initiation, development, and germination of embryos from the unpollinated ovules. |
Early transcription of defence-related genes in Platanus × acerifolia leaves following treatment with cerato-plataninI. Baccelli, A. Scala, L. Pazzagli, R. BernardiBiologia plantarum 57:571-575, 2013 | DOI: 10.1007/s10535-013-0308-2 The protein elicitor cerato-platanin (CP) is known to induce defence-related responses in various plants. Some of these responses occur very quickly. In the present work, transcriptional changes caused by CP in leaves from Platanus × acerifolia (Aiton) Willd. were studied. With a cDNA microarray, 131 differentially regulated transcripts were identified as responsive to CP after 24 h of treatment. Eighty-six of these were cold-or ozone-modulated transcripts, thus revealing a significant overlap between genes responsive to CP and to cold/ozone stress. The transcriptional changes caused by CP were compared with the CP-orthologous protein Pop1 in a time-course analysis performed after 3, 6, 12, and 24 h of treatment by real-time RT-PCR on five defence-related genes. Despite some differences, CP and Pop1 were both able to induce early transcriptional changes (WRKY was overexpressed after only 3 h) confirming that pathogenassociated molecular patterns (PAMPs) act very quickly on gene transcription. |
Cucumber BAX inhibitor-1, a conserved cell death suppressor and a negative programmed cell death regulator under cold stressX. H. Chen, H. Yu, H. J. Deng, J. X. Chen, H. B. Mi, L. C. MaoBiologia plantarum 57:684-690, 2013 | DOI: 10.1007/s10535-013-0347-8 Programmed cell death (PCD) is a genetically controlled and conserved process in eukaryotes during development as well as in response to pathogens and other stresses. BAX inhibitor-1 (BI-1) has been implicated as an anti-PCD factor which is highly conserved in plants. Sequence of putative cucumber BI-1 protein exhibited 77.7 % identity and 91.2 % positive value with the homologue Blast BI-1 protein of Arabidopsis thaliana (AtBI-1). This highly homologous protein to the AtBI-1 protein was named CsBI-1. This protein contains an open reading frame (ORF) of 250 amino acids with a BAX inhibitor domain and five transmembrane regions conserved among members of the BI-1 family. Primers designed by the cDNA of CsBI-1gene were used for further sequencing. Cell death in cold-stored cucumber developed concomitantly with increased expression of the CsBI-1 gene and reached maximum at day 6. However, cell death accelerated significantly after 9 d when sharp decrease of the CsBI-1 expression occurred. After warming to 20 °C, expression of the CsBI-1 gene was the highest at day 3, decreased afterwards, and the lowest expression was detected at day 9 when PCD obviously appeared. The overall results indicate that CsBI-1 is cucumber homologue of Arabidopsis thaliana AtBI-1 gene. CsBI-1 is a conserved cell death suppressor induced by cold stress and a negative regulator of PCD. |
Brassinosteroid regulates secondary metabolism in tomato towards enhanced tolerance to phenanthreneG. J. Ahammed, Y. H. Zhou, X. J. Xia, W. H. Mao, K. Shi, J. Q. YuBiologia plantarum 57:154-158, 2013 | DOI: 10.1007/s10535-012-0128-9 We investigated the role of 24-epibrassinolide (EBR) in the amelioration of phenanthrene (PHE) stress in tomato (Solanum lycopersicum L.). Exposure to PHE (300 μM) significantly decreased shoot and root length (19 and 16 %, respectively), fresh mass (35 and 43 %, respectively), contents of chlorophyll a (26 %), chlorophyll b (27 %) and carotenoids (18 %) in tomato plants. In addition, PHE increased the malondialdehyde (MDA) content (57 %) and activity of secondary metabolism related enzymes glutathione-S-transferase (GST), glucose-6-phosphate dehydrogenase (G6PDH), shikimate dehydrogenase (SKDH), phenylalanine ammonia-lyase (PAL) and cinnamyl alcohol dehydrogenase (CAD). The expression levels of GST1, PPO, SKDH, PAL and CAD genes were also induced by PHE. Importantly, EBR (0.1 μM) alone and in combination with PHE increased the growth, biomass and activity of those enzymes significantly over control and PHE alone, respectively. Consistent with enzymes activities transcript levels of GST1, PPO, SKDH, PAL and CAD were further increased in PHE+EBR over PHE alone. However, MDA content was remarkably decreased in PHE+EBR than PHE alone. Meanwhile, content of phenols, flavonoids and antioxidant activity were increased by PHE and PHE+EBR further increased all those parameters. These observations suggest that EBR regulates secondary metabolism in tomato which might enhance tolerance to PHE. |
Isolation and characterization of cold inducible genes in carrot by suppression subtractive hybridizationS. R. Kumar, S. Anandhan, S. Dhivya, A. Zakwan, R. SathishkumarBiologia plantarum 57:97-104, 2013 | DOI: 10.1007/s10535-012-0250-8 Daucus carota is cultivated widely but grows best in cool climates. Suppression subtractive hybridization (SSH) is a PCR based method used to selectively amplify differentially expressed cDNAs and simultaneously suppress non-target cDNA. A subtraction forward library was constructed using RNA isolated from the leaves of unstressed and cold stressed carrot plants to determine the genes upregulated during cold stress. Out of the hundreds of clones obtained, sequences of 41 promising clones were submitted to the NCBI EST database. Sequence analyses revealed that these genes have significant roles in signal transduction, osmolyte synthesis and transport, regulation of transcription, translation and protein folding. Semiquantitative real-time polymerase chain reaction analysis (sqRT-PCR) of Dc cyclin, Dc WD and Dc profilin shows that the first two genes were upregulated while Dc profilin was constitutively expressed, but the analyses of the same with SSH, a much more sensitive technique showed an upregulation of all three genes. |
Sex-related differences of two ecologically divergent Salix species in the responses of enzyme activities to atmospheric CO2 enrichmentT. Ruuhola, L. Nybakken, R. Julkunen-TiittoBiologia plantarum 57:732-738, 2013 | DOI: 10.1007/s10535-013-0338-9 Sex-related differences in the responses of plants to CO2 enrichment are still rarely studied. In this study, we examined the effects of elevated atmospheric CO2 (720 μmol mol-1) on the activities of polyphenoloxidases (PPOs) and guaiacol peroxidases (PODs) in male and female plants of two ecologically divergent willow species Salix repens and S. phylicifolia. We detected that females invested more in PPO-based defence than did males, whose PPO activity decreased as a result of CO2 enrichment. Moreover, we found that the inherently slow-growing S. repens had markedly higher POD activity than did the more rapid-growing S. phylicifolia. The PODs of these two species also differed in their biochemical properties. |
Expression of the BBM gene during somatic embryogenesis of Arabidopsis thalianaK. Kulinska-Lukaszek, M. Tobojka, A. Adamiok, E. U. KurczynskaBiologia plantarum 56:389-394, 2012 | DOI: 10.1007/s10535-012-0105-3 The relationship between somatic embryogenesis (SE) and the expression of the BABY BOOM (BBM) gene was studied in cultured immature zygotic embryos (IZEs) using a transgenic line of Arabidopsis thaliana containing a BBMPro::GUS construct. Results showed spatio-temporal differences in BBM expression in explants during culture. BBM promoter activity was observed in freshly isolated IZEs except distal parts of cotyledons. At the beginning of culture, considerable increase of GUS staining intensity was observed in all parts of explants, which maintained at high level over next few days and coincide with cell divisions. Gradual decrease of GUS distribution in explants was observed at about the 5th day of culture. BBM promoter activity became largely restricted to dividing cells, then to developing somatic embryos, shoot-like structures and callus. In parts of explants not involved in morphogenesis BBM promoter activity was absent or hardly seen. Thus the in vitro expression of BBM coincides with cell proliferation and morphogenesis. |
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. |
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. |
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. |
Analysis of tolerance to copper and zinc in Aechmea blanchetiana grown in vitroP. Giampaoli, F. Tresmondi, G. P. P. Lima, S. Kanashiro, E. S. Alves, M. Domingos, A. R. TavaresBiologia plantarum 56:83-88, 2012 | DOI: 10.1007/s10535-012-0020-7 The aim of this study was to evaluate the growth and development of Aechmea blanchetiana Baker L.B. Sm. in vitro on medium with 0.0, 0.145, 1.45 and 14.5 μM Cu and 0.0, 2.75, 27.5 and 275 μM Zn. Significant accumulation of Cu and Zn occurred at 14.5 μM Cu and 27.5 and 275 μM Zn, respectively, and there were no significant changes in contents of the other macro- and micronutrients. Superoxide dismutase (SOD) activity significantly changed in the presence of both metals. Spermine content increased as Zn concentration increased and decreased with increasing concentrations of Cu. There was an accumulation of H2O2 in the leaf tissue of plants grown in 1.45 and 14.5 μM Cu and 27.5 and 275 μM Zn. A. blanchetiana was found tolerant to the Cu and Zn in concentrations used in this study and displays the capacity to accumulate these metals. |
Salinity affects indirectly nitrate acquisition associated with glutamine accumulation in cowpea rootsJ. A. G. Silveira, A. R. B. Melo, M. O. Martins, S. L. Ferreira-Silva, R. M. Aragão, E. N. Silva, R. A. ViégasBiologia plantarum 56:575-580, 2012 | DOI: 10.1007/s10535-012-0065-7 The aim of this study was to test the hypothesis that salinity can affect indirectly the nitrate acquisition by a negative modulation triggered by glutamine accumulation. Cowpea plants were exposed to a mild NaCl concentration (50 mM) in order to restrict growth and N-demand. After 21 d, pretreated plants and control plants were supplied with 0, 5 and 10 mM of Ca(NO3)2 for 3 d in absence of NaCl. Salt pretreated plants showed a great limitation in acquisition of NO3 -, indicated by decline in the nitrate uptake rate, NO3 - accumulation, nitrate reductase activity and protein content. The restriction of NO3 - utilization was positively associated with increased glutamine synthetase activity and glutamine accumulation, especially in roots. |
In vitro regeneration of Anethum graveolens, antioxidative enzymes during organogenesis and RAPD analysis for clonal fidelityS. Jana, G. S. ShekhawatBiologia plantarum 56:9-14, 2012 | DOI: 10.1007/s10535-012-0009-2 An efficient in vitro regeneration protocol was developed for medicinally important aromatic plant Anethum graveolens. Nodal segments were cultured onto Murashige and Skoog (MS) basal medium supplemented with different auxins and cytokinins singly as well as in combinations. The optimum callus induction (93.33 %) was obtained on medium fortified with 2.2 μM N6-benzyladenine (BA) and 0.21 μM α-naphthaleneacetic acid. The best shoot regeneration (85.7 %) with 12.86 shoots per explant was achieved in two weeks when callus was subcultured on MS medium amended with 2.2 μM BA and 1.85 μM kinetin. The average length of regenerated shoots varied from 3.15 to 4.8 cm. The rooting of regenerated shoots was nearly 100 % on 1/4 MS augmented with 4.9 μM indolebutyric acid with a maximum root length of 5.1 cm. Plantlets were successfully acclimatized with 60 % survival rate. During organogenesis, catalase and ascorbate peroxidase activity increased while superoxid dismutase activity decreased. Clonal fidelity of in vitro raised plants has been checked by random amplified polymorphic DNA using 10 selected decamer primers. It has been found that regenerated plants are true to type plants. |
Overexpression of AtHsp90.3 in Arabidopsis thaliana impairs plant tolerance to heavy metal stressH. M. Song, H. Z. Wang, X. B. XuBiologia plantarum 56:197-199, 2012 | DOI: 10.1007/s10535-012-0042-1 The functions of cytosolic heat shock protein AtHsp90.3 in response to heavy metal stress were characterized by using expression of AtHsp90.3 gene in yeast and Arabidopsis thaliana. AtHsp90.3 supported the Saccharomyces cerevisiae Hsp90 knockout strain R0005 growth and maintaining cells membrane integrity under cadmium and arsenic stresses, which was compatible with the components of ScHsc82 machinery. However, constitutive overexpression of AtHsp90.3 in Arabidopsis impaired plant tolerance to Cd stress with lower germination rate and shorter root length, decreased contents of phytochelatins (PCs) and glutathione (GSH), inhibited activities of superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD), and increased content of malondialdehyde (MDA). These results suggested that proper homeostasis of Hsp90 was critical for cellular response and/or tolerance to heavy metal stress in plants. |
Sodium nitroprusside modulates gene expression involved in glutathione synthesis in Zea mays leavesC. Souza Mello, V. S. Hermes, M. P. Guerra, A. C. M. ArisiBiologia plantarum 56:383-388, 2012 | DOI: 10.1007/s10535-012-0104-4 To investigate a possible involvement of nitric oxide in gene regulation of glutathione and flavonoid synthesis pathways, maize seedlings were treated with sodium nitroprusside (SNP), a NO donor, and apocynin (APO), an inducer of NO production. After 12-h treatment, the transcripts of γ-glutamylcysteine synthetase (γ-ecs), glutathione synthetase (gsh-s), chalcone synthase (chs), phenylalanine ammonia lyase (pal.1), myb-related protein P (P1) and actin 1 (act) genes were quantified in maize leaves by real time PCR, using α-tubulin as standard transcript. The level of γ-ecs and gsh-s transcripts in maize leaves were increased 9-fold and 12-fold, respectively, following SNP treatment, while after APO treatment, those transcripts were not significantly different from control plants. SNP-treated maize leaves did not show significant changes in pal.1 and chs expression. NO content in maize leaves was increased in SNP and APO treated plants in comparison to control plants. In conclusion, our experiments suggested that genes involved in glutathione synthesis could be modulated by SNP in maize leaves. On the other hand, APO had no effect on γ-ecs and gshs gene expression. |
Pretreatment with 5-aminolevulinic acid mitigates heat stress of cucumber leavesJ. Zhang, D. -M. Li, Y. Gao, B. Yu, C. -X. Xia, J. -G. BaiBiologia plantarum 56:780-784, 2012 | DOI: 10.1007/s10535-012-0136-9 Cucumber seedlings were pretreated with 3 μM 5-aminolevulinic acid (ALA) followed by cultivation at normal (25/18 °C) or high (42/38 °C) day/night temperature to investigate the protective effects of ALA on heat stress in plants. Heat elevated the contents of malondiadehyde (MDA), superoxide radical (O2 .-) and hydrogen peroxide (H2O2) in leaves of all plants but less in ALA-pretreated plants. Heat treatment resulted in higher antioxidant enzyme activities and proline and soluble sugar contents and weaker growth inhibition in ALA-pretreated plants than in those treated with heat alone. These results indicate that ALA pretreatment increased the tolerance of seedlings to heat stress. |
Dehydrin and proline content in Brassica napus and B. carinata under cold stress at two irradiancesM. Klíma, P. Vítámvás, S. Zelenková, M. Vyvadilová, I. T. Prá¹ilBiologia plantarum 56:157-161, 2012 | DOI: 10.1007/s10535-012-0034-1 The accumulation of cold-induced dehydrin and proline was related to the frost tolerance (FT) in several Brassica species or cultivars. A dehydrin of molecular mass 47 kDa was detected in the leaves of an Ethiopian mustard (B. carinata) and a pair of dehydrins of similar molecular mass in the three (two winter, one spring) oilseed rape (B. napus) cultivars, when plants were maintained at 4 °C for one-month under two different irradiances. More dehydrin was accumulated in oilseed rape than in Ethiopian mustard under the high irradiance. A significant correlation was observed between leaf dehydrin content and FT, and no relationship between proline content and FT or between the proline and dehydrin contents. Protoplast-derived callus cells behaved differently from leaves sampled from intact plants, as they did not accumulate dehydrin and proline in response to cold stress. |
Root adaptations to cadmium-induced oxidative stress contribute to Cd tolerance in the hyperaccumulator Sedum alfrediiS. K. Tian, L. L. Lu, X. E. Yang, H. G. Huang, K. Wang, P. H. BrownBiologia plantarum 56:344-350, 2012 | DOI: 10.1007/s10535-012-0096-0 Short-term responses of Sedum alfredii roots to Cd exposure was compared in Cd hyperaccumulator (HE) and nonhyperaccumulating ecotype (NHE). Cadmium exposure significantly inhibited root elongation and induced loss of plasma membrane integrity and lipid peroxidation of roots tips in the NHE, whereas these effects were much less pronounced in the HE plants. A strong accumulation of reactive oxygen species with increasing Cd concentration was noted in the NHE root tips, but not in HE. After Cd exposure, a dose-dependent decrease in oxidized glutathione and marked increase in reduced glutathione and non-protein thiols were observed in root tips of HE, but were not seen in the NHE plants. These results suggest that the HE tolerates high Cd in the environment through the differential adaptations against Cd-induced oxidative stress. |
The influence of the N- and C- terminal modifications of Potato virus X coat protein on virus propertiesH. Hoffmeisterova, T. Moravec, H. Plchova, J. Folwarczna, N. CerovskaBiologia plantarum 56:775-779, 2012 | DOI: 10.1007/s10535-012-0130-2 The Potato virus X (PVX)-based vector was used for the construction of N- and C-terminally modified PVX coat protein (XCP) chimeras. N-terminal XCP modifications do not influence the viral life cycle, whereas the simple XCP C-terminal fusion impedes the viral replication. We designed several C-terminally modified XCP chimeras and tested their viabilities in various Nicotiana benthamiana genotypes. Our results showed the negative impact of 3'-terminal modification of XCP on the chimera's life cycle. To ensure chimeric constructs stability, the second copy of the last 60 nucleotides of XCP followed by the 3'-untranslated region (UTR) was added downstream of the recombinant sequence. Simultaneously, the first copy of the last 60 nucleotides of XCP was mutated in order to prevent recombination between the two identical sequences. The movement protein of Tobacco mosaic virus expressed in transgenic N. benthamiana plants positively affected the cell-to-cell spread of C-terminally modified XCP chimeras. |
The antioxidative role of anthocyanins in Arabidopsis under high-irradianceQ. Zhang, L. -J. Su, J. -W. Chen, X. -Q. Zeng, B. -Y. Sun, C. -L. PengBiologia plantarum 56:97-104, 2012 | DOI: 10.1007/s10535-012-0022-5 To uncover the potential antioxidative role of anthocyanins in vivo in protecting photosynthetic tissues from photoinhibition, the effects of high irradiance [HI, 1300 μmol(photon) m-2 s-1] were studied using detached leaves derived from Arabidopsis wild-type (WT) and the mutant deficient in anthocyanin biosynthesis (tt3tt4). HI stress caused decreased chlorophyll content and photochemical efficiency, but increased cell-membrane leakage and contents of hydrogen peroxide and superoxide radical in the leaves of both Arabidopsis phenotypes, but the WT plants showed better HI tolerance than tt3tt4 mutant. HI caused a significant increase in the 1,1-diphenyl-2-picrylhydrazyl scavenging capacity in WT but not in the tt3tt4 mutant. The anthocyanins could not contribute substantially to light-shielding during the periods of HI stress, because the anthocyanin content in WT was very low and the colour of leaves was the same as in the tt3tt4 mutant. Thus, it was assumed that the better HI tolerance in WT was mostly related to the potential antioxidative role of anthocyanins. |
Effects of 5-aminolevulinic acid on the H2O2-content and antioxidative enzyme gene expression in NaCl-treated cucumber seedlingsA. Zhen, Z. L. Bie, Y. Huang, Z. X. Liu, M. L. FanBiologia plantarum 56:566-570, 2012 | DOI: 10.1007/s10535-012-0118-y The potential of 5-aminolevulenic acid (ALA) to enhance the salt tolerance of cucumber (Cucumis sativus L.) seedlings was investigated. ALA was applied at various concentrations (0, 1, 10, 25, 50, and 100 mg dm-3) as foliar spray or root watering. Then the seedlings were exposed to 0 or 75 mM NaCl for 5 d. NaCl stress reduced the root and leaf dry masses, leaf area, and the leaf net CO2 assimilation rate. These reductions were counteracted by exogenous ALA, and the most efficient was 50 mg dm-3 concentration via foliar spray. ALA decreased the H2O2 contents and increased the activities of ascorbate peroxidase (APX) and glutathione reductase (GR) in NaCl-treated cucumber roots and leaves and the activity of catalase (CAT) in leaves. The ALA application also up-regulated the expressions of CAT and cAPX genes in roots and leaves and the expression of GR gene in roots of the NaCl treated cucumber plants. |


