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Organogenesis and Agrobacterium tumefaciens-mediated transformation of Eucalyptus saligna with P5CS geneR. Dibax, C. Deschamps, J. C. Bespalhok Filho, L. G. E. Vieira, H. B. C. Molinari, M. K. F. De Campos, M. QuoirinBiologia plantarum 54:6-12, 2010 | DOI: 10.1007/s10535-010-0002-6 The purpose of this research was Eucalyptus saligna in vitro regeneration and transformation with P5CSF129A gene, which encodes Δ1-pyrroline-5-carboxylate synthetase (P5CS), the key enzyme in proline biosynthesis. After selection of the most responsive genotype, shoot organogenesis was induced on leaf explants cultured on a callus induction medium (CI) followed by subculture on a shoot induction medium (SI). Shoots were subsequently cultured on an elongation medium (BE), then transferred to a rooting medium and finally transplanted to pots and acclimatized in a greenhouse. For genetic transformation, a binary vector carrying P5CSF129A and uidA genes, both under control of the 35SCaMV promoter, was used. Leaves were co-cultured with Agrobacterium tumefaciens in the dark on CI medium for 5 d. The explants were transferred to the selective callogenesis inducing medium (SCI) containing kanamycin and cefotaxime. Calli developed shoots that were cultured on an elongation medium for 14 d and finally multiplied. The presence of the transgene in the plant genome was demonstrated by PCR and confirmed by Southern blot analysis. Proline content in the leaves was four times higher in transformed than in untransformed plants while the proline content in the roots was similar in both types of plants. |
Changes in AFLP and SSR DNA polymorphisms induced by short-term space flight of rice seedsJ. Y. Lu, W. L. Zhang, H. Xue, Y. Pan, C. H. Zhang, X. H. He, M. LiuBiologia plantarum 54:112-116, 2010 | DOI: 10.1007/s10535-010-0016-0 Differences of both amplified fragment length polymorphism (AFLP) and simple sequence repeat (SSR) polymorphisms were compared between the 60-d-old rice (Oryza sativa L. cv. DH7) and F3 rice plants (SP3) derived from seed, which endured a 7-d-space flight in March 2002. Total leaf AFLP DNA bands amplified from 22 primer pairs were 537 in DH7, whereas 562 in SP3. From the total 267 SSR DNA bands generated by 267 primer pairs, 39 were polymorphic with 22 larger (56 %) or 17 smaller (44 %) fragment size bands. The greatest numbers of AFLP DNA bands were amplified by primer E1M1 in DH7 (33) and E3M1 in SP3 (35), whilst the least by E4M3 in DH7 (14) and E5M2 in SP3 (16). |
Sensitivity of stem and petiole hydraulic conductance of deciduous trees to xylem sap ion concentrationK. Aasamaa, A. SõberBiologia plantarum 54:299-307, 2010 | DOI: 10.1007/s10535-010-0052-9 Hydraulic conductance of stem and petioles increased in response to an increase in xylem sap ion concentration, and decreased in response to a decrease in the ion concentration in six temperate deciduous tree species. The ion sensitivity of hydraulic conductance of stem and petioles was higher than the ion sensitivity of the stem alone. The ion sensitivity was lowest in the earliest developmental stages of the xylem, and had a seasonal maximum in the second half of summer. The ion sensitivity was highest in slow-growing species and lowest in fast-growing species. The ion sensitivity correlated negatively with mean radius of xylem conduits, hydraulic conductance of stem and petioles, hydraulic conductance of leaf laminae, and stomatal conductance, and positively with response of the hydraulic conductance of leaf laminae to HgCl2, and stomatal response to a decrease in leaf water potential or abscisic acid. It was concluded that the high ion sensitivity of xylem hydraulic conductance is a relevant characteristic of slow growth and a conservative water use strategy. |
Hairy roots formation in recalcitrant-to-transform plant Chenopodium rubrumS. Dmitroviĉ, N. Mitiĉ, S. Zdravkoviĉ-Koraĉ, B. Vinterhalter, S. Ninkoviĉ, L. J. ĈulafiĉBiologia plantarum 54:566-570, 2010 | DOI: 10.1007/s10535-010-0101-4 Susceptibility of C. rubrum to Agrobacterium-mediated transformation was demonstrated by inoculating the petioles of in vitro grown plants with A. rhizogenes strain A4M70GUS. Hairy roots were produced in 8 % of explants. They were isolated and maintained on plant growth regulator-free solid or liquid half-strength Murashige and Skoog medium for two years. Hairy root fresh mass increased 30 - 90 folds when grown in liquid medium, which was superior to solid medium, where most of the hairy roots produced calli. When these calli were grown on medium supplemented with 0.5 mg dm-3 thidiazuron, embryo-like structures were obtained. Transgenic status of long-term callus and hairy root cultures was confirmed by histochemical GUS assay, by PCR specific to the uidA, rolA&B and ags genes and by Southern hybridization. |
Induction of tetraploidy in Juncus effusus by colchicineL. Xu, U. Najeeb, M. S. Naeem, M. K. Daud, J. S. Cao, H. J. Gong, W. Q. Shen, W. J. ZhouBiologia plantarum 54:659-663, 2010 | DOI: 10.1007/s10535-010-0117-9 Tetraploidy was induced in vitro in mat rush (Juncus effusus L.) cultivar Nonglin-4 by exposure to colchicine (0, 50, 100 and 500 mg dm-3) for 6, 12 and 24 h. Flow cytometric analysis was used to confirm the ploidy level. Anatomical and ultrastructural analyses at cellular and subcellular levels in tetraploid and diploid control plants revealed differences between diploid and tetraploid plants. The leaf epidermis had larger stomata but lower stomatal density in tetraploid plants. In addition, mesophyll cells in tetraploid plants appeared more compact and showed less intercellular spaces along with increased size of vascular bundles. However, a significant reduction of chlorophyll content was observed in tetraploid plants that might be the result of structural modification in the lamellar membranes of chloroplasts. |
Direct plant regeneration from encapsulated nodal segments of Vitex negundoN. Ahmad, M. AnisBiologia plantarum 54:748-752, 2010 | DOI: 10.1007/s10535-010-0134-8 An efficient protocol for encapsulation of nodal segments of Vitex negundo L. has been developed for the production of non-embryogenic synthetic seeds. The encapsulations of nodal segments were significantly affected by the concentrations of sodium alginate and calcium chloride. A 3 % Na2-alginate with 100 mM CaCl2 has been found to be optimum concentration for the production of uniform synthetic seed. For germination, the synseeds were cultured on Murashige and Skoog (MS) basal medium supplemented with kinetin (KIN) and α-naphthalene acetic acid (NAA) either singly or in various combinations. MS medium containing 2.5 μM KIN in combination with 1.0 μM NAA was found to be the optimum for maximum (92.6 ± 3.71 %) plantlet conversion frequency. Well developed regenerated plantlets were hardened, acclimatized and established in field, where they grew well without any detectable variation. |
Development and evaluation of microsatellite markers in Phoenix dactylifera L. and their transferability to other Phoenix speciesA. Akkak, V. Scariot, D. Torello Marinoni, P. Boccacci, C. Beltramo, R. BottaBiologia plantarum 53:164-166, 2009 | DOI: 10.1007/s10535-009-0026-y Forty one simple sequence repeats were isolated from two microsatellite enriched libraries of date palm (Phoenix dactylifera L.). After screening, 17 selected microsatellite loci were characterized and evaluated on a set of 31 cultivars and clones from Algerian and Californian germplasm. All primer pairs produced an amplification product of the expected size and detected high polymorphism among the analysed samples. These nuclear simple sequence repeat (SSR) markers are expected to be a very effective tool for evaluating genetic diversity in date palm germplasm. Acrosstaxa amplification showed the usefulness of most SSR markers in 14 other species across the genus Phoenix. |
Nitric oxide alleviates Fe deficiency-induced stress in Solanum nigrumJ. Xu, H. X. Yin, X. J. Liu, T. Yuan, Q. Mi, L. L. Yang, Z. X. Xie, W. Y. WangBiologia plantarum 53:784-788, 2009 | DOI: 10.1007/s10535-009-0144-6 The possible involvement of nitric oxide in response of Solanum nigrum seedlings to Fe deficiency was investigated. Iron deficiency resulted in decreased shoot height and chlorophyll content and increased proliferation of root hairs and H2O2, K+ and Ca2+ content. NO donor S-nitrosoglutathione (GSNO) was effective in preventing Fe deficiency-induced increase in content of H2O2 and the ion uptake. The protective effects of GSNO were reversed by cPTIO, an NO scavenger, and tungstate, a nitrate reductase (NR) inhibitor. |
Characterization of the antioxidant system during the vegetative development of pea plantsP. Díaz-Vivancos, G. Barba-Espín, M. J. Clemente-Moreno, J. A. HernándezBiologia plantarum 54:76-82, 2010 | DOI: 10.1007/s10535-010-0011-5 The antioxidative system was studied during the development of pea plants. The reduced glutathione (GSH) content was higher in shoots than in roots, but a greater redox state of glutathione existed in roots compared with shoots, at least after 7 d of growth. The 3-d-old seedlings showed the highest content of oxidised ascorbate (DHA), which correlated with the ascorbate oxidase (AAO) activity. Also, the roots exhibited higher DHA content than shoots, correlated with their higher AAO activity. The activities of antioxidant enzymes were much higher in shoots than in roots. Ascorbate peroxidase (APX) activity decreased during the progression of growth in both shoots and roots, whereas peroxidase (POX) activity strongly increased in roots, reflecting a correlation between POX activity and the enhancement of growth. Catalase activity from shoots reached values nearly 3 or 4-fold higher than in roots. The monodehydroascorbate reductase (MDHAR) activity was higher in young seedlings than in more mature tissues, and in roots a decrease in MDHAR was noticed at the 11th day. No dehydroascorbate reductase (DHAR) was detected in roots from the pea plants and DHAR values detected in seedlings and in shoots were much lower than those of MDHAR. In shoots, GR decreased with the progression of growth, whereas in roots an increase was seen on the 9th and 11th days. Finally, superoxide dismutase (SOD) activity increased in shoots during the progression of growth, but specific SOD activity was higher in roots than in shoots. |
Characteristics of chlorophyll fluorescence and antioxidative system in super-hybrid rice and its parental cultivars under chilling stressY. H. Zhang, L. J. Chen, J. L. He, L. S. Qian, L. Q. Wu, R. F. WangBiologia plantarum 54:164-168, 2010 | DOI: 10.1007/s10535-010-0027-x Characteristics of chlorophyll fluorescence and antioxidative system were investigated in rice (Oryza sativa L.) super-hybrid Liangyoupeijiu (LYPJ), maternal cultivar Peiai64s, and paternal cultivar indica rice 9311 under chilling stress. During 6-d chilling treatment, chlorophyll content of all three genotypes was gradually declined. However, the decrease in photosystem 2 (PS 2) maximum photochemical efficiency (Fv/Fm) and quantum yield of PS 2 (ΦPS2) was less expressive in LYPJ than in parental cultivars The electrolyte leakage and malondialdehyde content in all cultivars increased after chilling treatment, but LYPJ exhibited the least increasing tendency. Activities of superoxide dismutase (SOD), ascorbate peroxidase (APX) and glutathione reductase (GR) were higher in LYPJ than in parental cultivars. The results demonstrated that tolerance to chilling stress in LYPJ might be adopted mostly from its maternal cultivar. |
Assessment of genetic diversity and relationships among Coix lacryma-jobi accessions using microsatellite markersK. -H. Ma, K. -H. Kim, A. Dixit, I. -M. Chung, J. -G. Gwag, T. -S. Kim, Y. -J. ParkBiologia plantarum 54:272-278, 2010 | DOI: 10.1007/s10535-010-0047-6 The present study describes the assessment of genetic diversity and relationships among 79 Job's tears (Coix lacrymajobi L.) accessions collected from China and Korea using 17 microsatellite markers. A total of 57 alleles were detected with an average of 3.4 alleles per locus. A high frequency of rare alleles (36.3 %) was observed within the collection. Values for observed (HO), expected heterozygosity (HE) and Shannon's information index (I) within the analysis ranged from 0.00 (GBssrJT183) to 0.81 (GBssrJT130), from 0.01 (GBssrJT170) to 0.65 (GBssrJT130) and from 0.034 (GBssrJt170) to 1.13 (GBssrJT130), respectively. The locus GBJT130 was the most informative marker with the highest values for observed and effective alleles as well as for HO, HE and I. Based on the UPGMA algorithm, the majority of the Chinese accessions grouped in one cluster, whereas all the Korean accessions grouped together in a separate cluster, indicating that Chinese accessions are genetically quite distinct from Korean accessions. No relation between genetic relatedness among Job's tears accessions and their place of collection was observed. Chinese accessions exhibited greater within population polymorphism (P = 95 %, HE = 0.30, I = 0.52) than the accessions from Korea (P = 68 %, HE = 0.13, I =0.24), indicating their potentiality as a reservoir of novel alleles for crop improvement. However, in general the low diversity within each population indicates a narrow genetic base within our collection. |
Indirect regeneration of Withania somnifera and comparative analysis of withanolides in in vitro and greenhouse grown plantsY. H. Dewir, D. Chakrabarty, S. -H. Lee, E. -J. Hahn, K. -Y. PaekBiologia plantarum 54:357-360, 2010 | DOI: 10.1007/s10535-010-0063-6 The present study reports an efficient protocol for indirect shoot organogenesis and plantlets regeneration of Withania somnifera (L.) Dunal. Leaf explants were cultured on Murashige and Skoog (MS) medium supplemented with different concentrations and combinations of 6-benzylaminopurine (BAP) and indole-3-acetic acid (IAA). The highest callus induction rate (89.5 %) and shoot regeneration rate (92 %) were obtained when 2 mg dm-3 BAP was combined with 0.5 mg dm-3 IAA. Three major withanolides (withaferine A, 12-deoxywithastramonolide and withanolide A) were investigated in different plant organs from in vitro and greenhouse grown plants. Leaves contained higher contents of withanolides and phenolics than roots or stems, whereas roots contained the highest contents of flavonoids and polysacharides. In vitro grown plants contained greater contents of phenolics, flavonoids and polysaccharides while lower contents of withanolides than greenhouse grown plants. |
PCR-based molecular markers for assessment of somaclonal variation in Pinus pinea clones micropropagated in vitroC. Cuesta, R. J. Ordás, A. Rodríguez, B. FernándezBiologia plantarum 54:435-442, 2010 | DOI: 10.1007/s10535-010-0079-y Four different markers [random amplified polymorphic DNA (RAPD), inter simple sequence repeat (ISSR), amplified fragment length polymorphism (AFLP), and selective amplified microsatellite polymorphism length (SAMPL)] were applied for evaluating somaclonal variation of micropropagated genotypes of stone pine (Pinus pinea L.). The total number of primers tested was 130, with 223 combinations assayed. A high number of them amplified successfully (178), representing 79.82 % of the total, and the average number of amplified fragments ranged from 2.47 (ISSR) to 65.76 (SAMPL). Based on internal controls, no problem of reproducibility was detected. Almost no somaclonal variation was detected within the clones. Of the tested markers, ISSR, AFLP, and SAMPL showed monomorphic amplification profiles, with only RAPD markers showing some interclonal variation. |
Using morphological diagnosis and molecular markers to assess the clonal fidelity of micropropagated Echinacea purpurea regenerantsS. J. Chuang, C. L. Chen, J. J. Chen, J. M. SungBiologia plantarum 54:539-542, 2010 | DOI: 10.1007/s10535-010-0095-y Both morphological characteristics and amplified fragment length polymorphism (AFLP) markers were used to validate the genetic fidelity of 1 080 field-grown Echinacea purpurea plants regenerated from leaf explants of donor T5-9. Morphological diagnosis revealed that 1 067 out of 1 080 regenerants were normal, while 13 regenerants were aberrant. AFLP analysis was further performed to assess DNA variations among donor, 43 sampled normal regenerants and all 13 aberrant regenerants. Seven primer combinations generated 471 fragments among donor and normal regenerants, of which 9 fragments were polymorphic. The same primer pairs generated 484 fragments for aberrant regenerants, of which 417 fragments were polymorphic. UPGMA clustering indicated that 42 normal regenerants and donor fell into same cluster at similarity scale of > 0.99, while all 13 aberrant regenerants and one morphologically normal regenerant comprised the other clusters. AFLP analysis indicated that these 14 regenerants are off-types. |
CDPK gene expression in salt tolerant rolB and rolC transformed cell cultures of Panax ginsengK. V. Kiselev, O. V. Grishchenko, Y. N. ZhuravlevBiologia plantarum 54:621-630, 2010 | DOI: 10.1007/s10535-010-0112-1 CDPKs (calcium-depended protein kinases) are of great importance for the activation of defense reactions in plants. In this study, we aimed to find a connection between CDPK expression and increased salt tolerance in Panax ginseng. Treatment of P. ginseng cell cultures with W7 (CDPK protein inhibitor) showed that CDPK proteins were necessary for salt tolerance. Expression of PgCDPK1c, PgCDPK2c and PgCDPK4a was significantly increased in the cells treated with 60 mM NaCl compared to control cells, whereas expression of PgCDPK1b and PgCDPK3a was decreased. In the NaCl-treated cells, new CDPK transcripts also appeared (PgCDPK3c, PgCDPK4as). We also used rolC and rolB transformed cultures and the effects of the rol genes on CDPK expression were similar to the effects of salt stress: they caused a significant increase in the expression of PgCDPK1c, PgCDPK2c, and PgCDPK4a and decreased expression of PgCDPK3a, in addition to the appearance of the "short" CDPK transcripts. |
Chromosomal location of 45S rDNA and dfr gene in Citrus sinensisX. Lu, W. Zhou, F. GaoBiologia plantarum 54:798-800, 2010 | DOI: 10.1007/s10535-010-0146-4 Dihydroflavonol 4-reductase (DFR) is a key enzyme in the anthocyanin biosynthesis. In this study, the localization of 45S rDNA and dfr gene (named as CsDFR-bo) of the blood orange (Citrus sinensis Osbeck cv. Ruby) on chromosomes was investigated by fluorescence in situ hybridization (FISH). A karyotype of C. sinensis was reconstructed based on the length of mitotic metaphase chromosomes. The 45S rDNAs were localized on chromosomes 2p and 7q. The detection ratios of 45S rDNA in the two chromosomes were 69.5 and 77.3 %, respectively. The CsDFR-bo was proved to be a single-copy gene and localized on chromosome 3p. The detection ratio of CsDFR-bo was 8.2 %. |
Photosynthetic and anatomic responses of peanut leaves to zinc stressG. R. Shi, Q. S. CaiBiologia plantarum 53:391-394, 2009 | DOI: 10.1007/s10535-009-0074-3 In this study, photosynthetic performance, pigment content, chlorophyll a fluorescence, and leaf anatomy in peanut (Arachis hypogaea) subjected to zinc (Zn) stress were investigated. Zn stress resulted in reduction of photosynthetic and transpiration rates, pigment contents and root biomass. Zn-induced xerophyte structure in peanut leaves (i.e. thick lamina, upper epidermis, and palisade mesophyll, as well as abundant and small stomata) also contributed to decreased transpiration rate and stomatal conductance. This in turn, partially contributed to the limitation of photosynthesis. |
Transformation of potato via Agrobacterium coated microparticle bombardmentT. T. Nguyen, P. J. Dix, G. D. NugentBiologia plantarum 54:141-144, 2010 | DOI: 10.1007/s10535-010-0022-2 The transformation of potato (Solanum tuberosum L. cv. Désirée) was extended by the Agrobacterium-mediated biolistic method. Using this approach transgenic shoots could be obtained at a similar frequency to that achieved through conventional biolistics. Leaves from shoot cultures were bombarded with gold particles coated in Agrobacterium tumefaciens cells harboring a binary plasmid encoding three genes of interest in the T-DNA. Nine shoots were obtained from 20 shots, with selection of transgenic shoots on a series of media containing progressively increasing concentrations of hygromycin from 5 to 20 mg dm-3. |
High efficiency organogenesis and analysis of genetic stability of the regenerants in Solanum melongenaY. Xing, Y. Yu, X. Luo, J. -N. Zhang, B. Zhao, Y. -D. GuoBiologia plantarum 54:231-236, 2010 | DOI: 10.1007/s10535-010-0041-z A novel protocol for plant regeneration from cotyledon explants of eggplant (Solanum melongena) reducing concentration of sucrose was established. The most efficient bud induction medium consisted of Murashige and Skoog (MS) medium supplemented with 2.0 mg dm-3 zeatin, 0.1 mg dm-3 indoleacetic acid and 10 g dm-3 sucrose. After 15 d, the shoot buds were fragmented and transferred to the shoot elongation MS supplemented with 1.0-2.0 mg dm-3 gibberellic acid and 4.0-8.0 mg dm-3 AgNO3, which promoted shoots elongation. The genetic stability of the regenerated plants was analyzed by flow cytometry, RAPD and SSR molecular markers. The results indicated that almost no somaclonal variation was detected among the regenerants. |
An assessment of genetic fidelity of micropropagated plants of Chlorophytum borivilianum using RAPD markersS. Samantaray, S. MaitiBiologia plantarum 54:334-338, 2010 | DOI: 10.1007/s10535-010-0058-3 Rapid micropropagation was achieved in Chlorophytum borivilianum Santapau and Fernandes using shoot base as explants. Multiple shoots were induced on Murashige and Skoog's (MS) medium supplemented with 3.0 mg dm-3 6-benzylaminopurine, 0.1 mg dm-3 1-naphthaleneacetic acid, 150 mg dm-3 adenine sulphates and 3 % saccharose. Rooting was readily achieved upon transferring the shoots onto half strength MS medium supplemented with 0.1 mg dm-3 indolebutyric acid and 2 % saccharose. Micropropagated plantlets were hardened in the greenhouse and successfully established in soil. Random amplified polymorphic DNA (RAPD) markers were used to evaluate the genetic stability of the micropropagated plants. Thirty one arbitrary decamers were used to amplify genomic DNA from in vitro and in vivo plant material to assess the genetic stability. All RAPD profile analysis from micropropagated plants was genetically similar to mother plants. |
Peter Apel Member of Editorial Board 1936-2009Martin Peisker, Armin MeisterBiologia plantarum 54:401-402, 2010 | DOI: 10.1007/s10535-010-0074-3 |
Morpho-histological study of direct somatic embryogenesis in endangered species Frittilaria meleagrisA. Subotiĉ, M. Trifunoviĉ, S. Jevremoviĉ, M. PetriĉBiologia plantarum 54:592-596, 2010 | DOI: 10.1007/s10535-010-0107-y Direct somatic embryogenesis of Frittilaria meleagris L. was induced using leaf base explants excised from in vitro grown shoots. Somatic embryos occurred at the basal part of leaf explants 4 weeks after culture on a Murashige and Skoog (MS) medium supplemented with various concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) or kinetin (KIN). The highest number of somatic embryos (SEs) were formed (9.74) from leaf explant on MS medium supplemented with 0.1 mg dm-3 2,4-D after 4 weeks of culture initiation. An initial exposure to a low concentration of KIN in the medium also enhanced SEs induction. Our observations by light and scanning electron microscopy revealed that SEs originate directly from the epidermal and subepidermal layers of leaf explant. The developmental stages of somatic embryogenesis from the first unequal cell division through the meristematic clusters, multi-cellular globular somatic embryos to the fully formed cotyledonary embryos were determined. After 4 weeks on MS medium without plant growth regulators, SEs developed into bulblets. |
Microwave treatment induced mutations and altered gene expression in Vigna aconitifoliaR. K. Jangid, R. Sharma, Y. Sudarsan, S. Eapen, G. Singh, A. K. PurohitBiologia plantarum 54:703-706, 2010 | DOI: 10.1007/s10535-010-0124-x Primary leaf explants of aseptically grown seedlings of moth bean [Vigna aconitifolia (Jacq.) Marechal] immersed in water or not were treated in microwave oven (2450 MHz, 800 W cm-2) for 1, 3, 5 and 7 s before culturing. Callusing and shoot emergence from these explants were enhanced up to microwave exposure lasted 5 s while longer treatment of water-immersed explants delayed callusing. One polypeptide (26.6 kD) was up regulated in the callus derived from microwave treatment in water-immersed explants. RAPD analysis detected alteration in DNA sequences due to microwave treatment in water-immersed explants for 7 s. The frequency of mutation was 1.6 % (4 bands out of 248) over all the cultures analyzed and the same was 13 % (4 bands out of 31), if amplicons generated at 7 s treatment alone were considered. |
Organization of a dispersed repeated DNA element in the Zamia genomeD. Cafasso, S. Cozzolino, N. J. Vereecken, P. De Luca, G. ChinaliBiologia plantarum 53:28-36, 2009 | DOI: 10.1007/s10535-009-0005-3 Occurrence and genomic organization of dispersed elements containing ZpS1 satellite repeats have been investigated in a wide representation of species of the old plant genus Zamia (Zamiaceae, Cycadales). In Z. paucijuga, the ZpS1 repeat is organized as long satellite DNA arrays and as short arrays inserted into AT-rich dispersed elements. A comparative study by Southern analysis shows that these unusual dispersed elements containing the ZpS1 repeat are present with different organizations in all investigated Zamia species. In some species these elements are present with a low copy number, while in other species secondary amplification events, involving specific sequence clusters, appear to have generated characteristic dispersed elements in a high copy number. Among Zamia species, several groups share similar restriction patterns, as the Zamia loddigesii complex and the Caribbean species suggesting a general correlation between organization and genomic representation of the dispersed repeated sequence and the pattern of phyletic relationships in the genus. However, the finding of different patterns also among closely related species suggests a complex history of amplifications and losses of these dispersed repetitive elements that cannot be always easily traced through the phylogenetic reconstruction of this ancient plant group. |
A mutation affecting symbiosis in the pea line Risnod27 changes the ion selectivity filter of the DMI1 homologK. Novák, J. Felsberg, E. Biedermannová, J. VondrysBiologia plantarum 53:451-460, 2009 | DOI: 10.1007/s10535-009-0085-0 After identifying regions of cDNA conserved between the symbiotic gene DMI1 of the model species Medicago truncatula and the homologous genomic region of Arabidopsis thaliana, universal primers were designed from 8 of 12 exons to allow the routine amplification of plant homologs. As an example, the complete homologous sequence from the pea (Pisum sativum L.) was amplified and sequenced, although the poorly conserved 5'-end and 5'-flanking region of the gene had to be amplified using a modified TAIL-PCR strategy. The identity of this amplified homolog with the SYM8 gene was independently confirmed by the presence of a single nucleotide change in the coding sequence of the mutant line Risnod27 (sym8) that cosegregated with the asymbiotic phenotype. Five insertions in pea introns responsible for increasing the total length of SYM8 by 1443 bp, compared to the M. truncatula homolog DMI1, belong to known transposon and retrotransposon families of pea and legumes in general. In view of the predicted function of SYM8 as an ion channel, the Risnod27 mutation (His309Tyr) appears to be localized in the selectivity filter domain. This finding confirms the essential role of histidine 309 in the symbiotic function of SYM8 and provides a guide to its ionic specificity. In view of the Risnod27 symbiotic phenotype, we hypothesize that SYM8 does not have identical functions in the transduction of rhizobial and mycorrhizal signals. The variability of the N-proximal region of the known legume homologs of DMI1 suggests an interaction with a variable ligand. |
Efficient in vitro plant regeneration from shoot apices and gene transfer by particle bombardment in Jatropha curcasJ. Purkayastha, T. Sugla, A. Paul, S. K. Solleti, P. Mazumdar, A. Basu, A. Mohommad, Z. Ahmed, L. SahooBiologia plantarum 54:13-20, 2010 | DOI: 10.1007/s10535-010-0003-5 An efficient and reproducible in vitro plant regeneration system from shoot apices was developed in Jatropha curcas. Benzylaminopurine (BAP; 2.5 μM) was most effective in inducing an average of 6.2 shoots per shoot apex. Incorporation of gibberellic acid (GA3; 0.5 μM) to basal medium was found essential for elongation of shoots. The BAP-habituated mother explants continuously produced shoots during successive subculture without any loss of morphogenic potential. The shoots rooted efficiently on half-strength MS medium. The rooted plantlets were acclimatized with more than 98 % success and the plants transferred to soil:compost in nursery showed no sign of variation compared to the seed-grown plants. The whole process of culture initiation to plant establishment was accomplished within 5-6 weeks. A genetic transformation system in J. curcas was established for the first time, using bombardment of particles coated with plasmid pBI426 with a GUS-NPT II fusion protein under the control of a double 35S cauliflower mosaic virus (CaMV) promoter. The β-glucuronidase (GUS) activity in J. curcas shoot apices was significantly affected by the gold particle size, bombardment pressure, target distance, macrocarrier travel distance, number of bombardments, and type and duration of osmotic pre-treatment. The proliferating bombarded shoot apices were screened on medium supplemented with 25 mg dm-3 kanamycin and surviving shoots were rooted on medium devoid of kanamycin. The integration of the transgene into genomic DNA of transgenic plants was confirmed by PCR and Southern blot hybridization. The transgenic plants showed insertion of single to multiple copies of the transgene. |
ABA or cadmium induced phytochelatin synthesis in potato tubersA. Stroiñski, T. Chadzinikolau, K. Giżewska, M. ZieleziñskaBiologia plantarum 54:117-120, 2010 | DOI: 10.1007/s10535-010-0017-z Short-term treatment of potato (Solanum tuberosum L.) tuber discs with CdCl2 solution elevated both the StPCS1 transcript level, phytochelatin synthase (PCS) activity and contents of phytochelatin (PC) and abscisic acid (ABA). Similar effects but less marked were noticed after treatment of tuber tissues with ABA solution. Cd-treatment increased also cysteine (CYS) content but did not change glutathione content. If ABA treatment preceded Cd-treatment, the elevation of CYS and PC contents were limited. The data suggest the participation of ABA in the regulation of PCS. |
In vitro responses of grape rootstocks to NaClM. Alizadeh, S. K. Singh, V. B. Patel, R. C. Bhattacharya, B. P. YadavBiologia plantarum 54:381-385, 2010 | DOI: 10.1007/s10535-010-0069-0 An investigation was undertaken to adjudge the in vitro salt tolerance and biochemical changes due to NaCl on four grape rootstocks (Dogridge, SO4, H-144 and 3309C). The in vitro two-node micro-cuttings from the established cultures were sub-cultured on rooting medium comprising Murashige and Skoog (MS) basal medium supplemented with 0.5 μM indolebutyric acid (IBA), 200 mg dm-3 activated charcoal and different NaCl concentrations ranging from 0 to 125 mM. The surviving and proliferated cultures were further sub-cultured four times on respective media. Dogridge and H-144 tolerated 125 and 100 mM NaCl, respectively, while SO4 and 3309C survived only up to 75 mM NaCl. Contents of proteins, proline, K+ and Na+ in tissue increased in all genotypes due to NaCl supplementation, while contents of chlorophyll and total soluble sugars declined. Higher K+/Na+ ratio was registered in Dogridge and H-144 than in SO4 and 3309C. The relative NaCl tolerance for different grape rootstocks under study could be ranked as Dogridge > H-144 > SO4 and 3309C. |
Contents of polyamines during vernalization in wheat and the effect of zearalenoneM. Filek, J. Biesaga-Koĥcielniak, I. Marciñska, M. Cvikrová, I. Macháèková, J. KrekuleBiologia plantarum 54:483-487, 2010 | DOI: 10.1007/s10535-010-0085-0 The contents of endogenous free and conjugated polyamines, putrescine (Put) and spermidine (Spd), were determined during 9 week of vernalization (at 5 °C) in winter wheat seedlings cultivated on Murashige and Skoog media without (MS0) and with 2 mg dm-3 zearalenone (MSZEN). At the 4th week of chilling treatment, which is sufficient to induce generative development in 30 % of plants, the marked increase in free and conjugated forms of Put and free Spd were observed. The presence of ZEN in medium significantly accelerated the vernalization. About 20 % of plants treated with ZEN flowered already after 2 weeks and 40 % after 3 weeks of chilling. Significantly higher content of free Put was determined in roots grown on MSZEN compared with MS0 during the first 5 weeks of vernalization with maximum at the 4th week. After germination, a marked decrease in free Spd content was observed both in plants grown on MS0 and MSZEN. Application of ZEN significantly slowed down the Spd decline in leaves and roots during the first and second week of vernalization. The content of Spd and its conjugates decreased in vernalized plants after 1 week of cultivation at 20 °C. |
UV-B induced stress responses in three rice cultivarsI. Fedina, J. Hidema, M. Velitchkova, K. Georgieva, D. NedevaBiologia plantarum 54:571-574, 2010 | DOI: 10.1007/s10535-010-0102-3 UV-B responses of three rice (Oryza sativa L.) cultivars (Sasanishiki, Norin 1 and Surjamkhi) with different photolyase activity were investigated. Carbon dioxide assimilation data support that Sasanishiki was less sensitive to UV-B than Norin 1 and Surjamkhi. UV-B radiation sharply decreased the content of Rubisco protein in Surjamkhi and has no effect in Sasanishiki. The photochemical activities of photosystem (PS) 1 and PS 2 was slightly affected by UV-B treatment. The content of H2O2 and the activities of antioxidant enzymes, catalase (CAT), peroxides (POX) and superoxide dismutase (SOD) were enhanced after UV-B treatment. The activities of CAT and POX isoenzymes in Sasanishiki were more enhanced by UV-B radiation than those in Norin 1 and Surjamkhi. |


