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Abstract
Biolistic gene transfer is one of the preferred methods for both transient gene expression studies and stable genetic transformation for improvement of crops including sugarcane. In preparation for bombardment, DNA is precipitated onto micro-particles in the presence of a cationic polyamine and buffer. Three different DNA coating procedures were compared for biolistic transformation of sugarcane using linear expression cassettes: spermidine (free base), protamine sulfate, and the Seashell DNAdel™ Gold Carrier with proprietary precipitation buffer from Seashell Technology. All three DNA coating ...
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Abstract
The human CD14, a high affinity receptor for lipopolysaccharides (LPS), is involved in the innate immunity system and the inflammatory response. There is increasing interest in using recombinant approaches to produce purified CD14 protein for therapeutic uses. Plants provide ideal expression systems for the production of recombinant proteins, but the levels of expression of recombinant proteins produced in planta are still not high. To improve expression levels of CD14 the 22-kDa alpha -zein signal peptide (ZSP) from maize was fused to ...
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Abstract
Using several explants of Pueraria candollei Grah. ex Benth. var. candollei and two strains of Agrobacterium rhizogenes (ATCC 15834 and 43057), hairy root cultures were established. Including 100 μM acetosyringone in the culture medium enhanced frequency of hairy root induction by up to 58 %. Subsequently, effects of inoculum size (IS) and temperature on growth and production of isoflavonoids in hairy roots were determined. Conditions of 1 % IS and 32 °C promoted the highest accumulation of total isoflavonoid content, up ...
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Abstract
Curcuma attenuata is a highly valued ornamental. This study provides the first report on C. attenuata shoot organogenesis and plant regeneration. Immature anthers derived from 5 to 7 cm long inflorescences were isolated and cultured on different variations of Murashige and Skoog (MS) media to induce callus and then shoot organogenesis. When the 2-mm long anthers in which microspores were at the uninucleate developmental stage were cultured in the dark on MS medium containing 13.6 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 2.3 ...
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Abstract
Somatic embryogenesis was induced from leaf and stem of chrysanthemum ( cv . Yuukou) using various combinations of plant growth regulators. About 93 and 63 % of somatic embryogenesis were respectively induced from leaf and stem cultured in Murashige and Skoog (MS) medium supplemented with 6-benzyladenine (BA) and a-naphthalene acetic acid (NAA). Somatic embryogenesis was more readily induced from leaf, yielding 100 % regeneration rate in MS medium supplement with 0.6 mg/L BA and 1.8 mg/L NAA. Plants were efficiently regenerated ...
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Abstract
Genetic improvement of sunflower ( Helianthus annuus L.) through the use of biotechnological tools requires a reliable in vitro shoot regeneration system. Tissue culture protocols reported to date for sunflower suffer from low efficiency, poor reproducibility, genotype dependence and a tendency for flowering in vitro. The present study describes an efficient protocol system for shoot regeneration via direct adventitious shoot organogenesis from cotyledons of mature seeds of sunflower. About 169 media combinations comprising 12 different growth regulator combinations in various concentrations ...
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Abstract
Shoot organogenesis was successfully achieved in petiole explants excised from 6 to 8-week old in vitro plantlets of yam Dioscorea rotundata P. cv. Kponan fissa, Dioscorea cayenensis L. cv. Krengle IB14 and cv. Krengle IB35, and Dioscorea alata L. cv. Bete bete. Only the basipetal portion of the petiole acquired competence, and plants regenerated within 21 days on MS medium supplemented with 2 % sucrose, 100 mg/l myo-inositol, 10 μM kinetin and 1.5 mM putrescine referred to as yam regeneration medium ...
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Abstract
Bacopa monnieri (L.) Wettst., a traditional Indian medicinal plant with high commercial potential, is used as a potent nervine tonic. A slow growth protocol was developed for medium-term conservation using mineral oil (MO) overlay. Nodal segments of B. monnieri (two genotypes; IC249250, IC468878) were conserved using MO for 24 months. Single node explants were implanted on MS medium supplemented with 0.2 mg l −1 BA and were covered with MO. Subculture duration could be significantly enhanced from 6 to 24 months, ...
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Abstract
The capacity of regeneration of adventitious shoots from leaf explants was studied in sour cherry ‘Čačanski Rubin’ ( Prunus cerasus L.) and cherry rootstock Gisela 5 ( P. cerasus × P. canescens ). Regeneration assay included thirty different combinations of plant growth regulators. 6-benzyladenine (BA) and thidiazuron (TDZ) were applied either individually or each combined with different concentrations of indole-3-butyric acid, α-naphthaleneacetic acid (NAA) and 2,4-dichlorophenoxyacetic acid (2,4-D). ‘Čačanski Rubin’ showed higher regeneration capacity in comparison with Gisela 5 regarding the ...
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Abstract
Ruta graveolens shoot cultures were maintained on static medium supplemented with 0, 1, 2 and 3 % mannitol. The cultures were grown in vessels that ensured a ventilation rate of 7.44, 10.82 or 62.83 air exchanges per day (V1, V2 or V3, respectively). The growth index and fresh weight were significantly increased at 1 % mannitol and decreased with increasing mannitol concentrations, whereas the dry weight (DW) and DW % increased at higher concentrations of mannitol. Improving the culture ventilation significantly ...
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Abstract
Aloe species are highly-prized for their ornamental value and have been utilized for centuries in traditional medicine. Due to their habitat loss and exploitation for medicinal and ornamental plant trade, many species in this genus have become threatened. One of the most important globally rated medicinal species in Aloe genus is A. arborescens . The current study evaluated the roles of different aromatic cytokinin types and concentrations on direct organogenesis, in vitro bioactive secondary metabolite production and antioxidant activity of regenerated ...
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Abstract
Protoplast fusion between sweet orange and mandarin/mandarin hybrids scion cultivars was performed following the model “diploid embryogenic callus protoplast + diploid mesophyll-derived protoplast”. Protoplasts were isolated from embryogenic calli of ‘Pera’ and ‘Westin’ sweet orange cultivars ( Citrus sinensis ) and from young leaves of ‘Fremont’, Nules’, and ‘Thomas’ mandarins ( C. reticulata ), and ‘Nova’ tangelo [ C. reticulata × ( C. paradisi × C. reticulata )]. The regenerated plants were characterized based on their leaf morphology (thickness), ploidy level, ...
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Abstract
Acquired thermotolerance in plants refers to the ability to cope with lethal high temperatures following acclimatization at sublethal high temperatures. Acquired thermotolerance reflects an actual thermotolerance mechanism naturally occurring in plants and has been extensively used in thermotolerant line identification. In recent years, great progress has been achieved in the elucidation of biochemical, physiological, and molecular mechanisms of thermotolerance acquisition by using genomic approaches, including microarray analysis and mutation, knockout, and overexpression of related genes. Heat shock proteins (HSPs), such as ...
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Abstract
The somatic embryogenesis (SE) capacity of megagametophytes belonging to Continental and Mediterranean Spanish provenances of maritime pine ( Pinus pinaster Aiton) was studied, noting factors (megagametophyte developmental stage and culture medium) that enhanced the induction and establishment of SE lines. In both provenances, initiation and establishment of embryogenic calli was higher on megagametophytes in which the dominant zygotic embryo had begun to develop. In the Mediterranean provenance, however, SE lines were also established from megagametophytes enclosing zygotic embryos with well-developed cotyledons. ...
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Abstract
Inositol polyphosphate kinase (IPK; EC 2.7.1.151) participates in inositol metabolism, calcium signaling, stress response, gene transcription and other physiological and biochemical processes. In the present work, ThIPK2 (an inositol polyphosphate kinase from Thellungiella halophila ) was transferred into soybean through an Agrobacterium tumefaciens -mediated transformation. Transgenic plants with ThIPK2 gene displayed water deficit-, salt- and oxidative-tolerance compared to non-transformed controls. Furthermore, the expression of ThIPK2 altered the ratio of fatty acid components in soybean seeds, resulting in an increase of oleic ...
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Abstract
Genes belonging to the phosphoethanolamine binding protein (PEBP) superfamily play important roles in controlling the switch between vegetative and reproductive growth in higher plants. Here we reported the isolation of two genes from the PEBP superfamily in Dendrobium nobile Lindl homologous to FLOWERING LOCUS T ( FT ) and MOTHER OF FT ( MFT ). These two genes, designated as DnFT and DnMFT , were predominantly expressed in the auxiliary buds and leaves of the plant. In auxiliary buds, DnFT was ...
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Abstract
In this study, an efficient procedure was established for successful induction of tetraploid Arachis paraguariensis by treating diploid explants with colchicine. Quartered-seed, callus and shoot-tips were treated with colchicine at concentrations of 0.05, 0.1, 0.2 and 0.5 % (w/v) for 4, 8, 16, 20 and 24 h before they were transferred unto modified Murashige and Skoog medium for either callus induction or shoot regeneration. Results showed that quartered-seed displayed the highest frequency of in vitro plantlet regeneration and tetraploid induction, as ...
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posted by
1 person
vostroo
Abstract
The effects of five topolins ( meta -Topolin = m T; meta -Topolin riboside = m TR; meta -Methoxy topolin = Me m T; meta -Methoxy topolin riboside = Me m TR and meta -Methoxy topolin 9-tetrahydropyran-2-yl = Me m TTHP) on shoot and root regeneration of micropropagated ‘Williams’ bananas were compared to benzyladenine (BA). Sterile shoot-tip explants were cultured on modified Murashige and Skoog (MS) media containing 10, 20 or 30 μM of the above cytokinins (CKs) for 42 days. ...
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Abstract
Boesenbergia rotunda belongs to the Zingiberaceae family. It is widely found throughout Southeast Asia and is commonly used as a food ingredient and in folk medicine. Extracts from this plant contain a number of important bioactive compounds such as boesenbergin, cardamonin, pinostrobin, pinocembrin, panduratin A and 4-hydroxypanduratin A. These compounds have been shown to exhibit anti-HIV protease, anti-dengue NS2B/ NS3 protease, antibacterial, antifungal, anti-inflammatory, anticancer, and antioxidant activity. Here we report the use of proteomic approaches to identify proteins that may ...
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Abstract
Two methods of in vitro pollination of aspen were tested and compared, first with moist pollen by transferring swollen pollen grains from a wet agar surface to the stigma and, second, using dry pollen grains. In vitro pollination with dry pollen grains appeared to be more efficient and this method was applied in three subsequent years. Additionally, pollination was performed with selected pollen grains in order to obtain triploid plants. Diploid pollen was induced by heat-treatment of pollen mother cells, but ...
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Abstract
Vinca minor is the sole source of vincamine, an alkaloid known to be used in a variety of cerebral disorders. Three stable variant shoot lines (V10, V20 and V30) with tolerance thresholds of 10, 20 and 30 mg/l 5-methyltryptophan (5-MT; analogue of tryptophan), respectively, were selected. These lines showed twofold to threefold increase in tryptophan content and 1.5- to 2-fold increment in the total alkaloids in comparison to the wild line shoots. A maximum of 16-fold enhancement in vincamine production was ...
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Abstract
To evaluate genetic homogeneity of 1-year-old guava ( Psidium guajava L.) plants developed from in vitro somatic embryogenesis, DNA from leaf tissues of seven randomly selected plants along with the mother plant, was isolated and subjected to molecular analysis. A total of six Simple Sequence Repeat (SSR) primer pairs, producing reproducible and clear bands ranging from 100 to 300 bp in size, resulted in amplification of single band (allele), corresponding homozygous individuals. Moreover, of 10 different inter-simple sequence repeat (ISSR) primers ...
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Abstract
A protocol for in vitro propagation of cineraria ( Senecio cruentus ) was developed. The highest frequency of shoot proliferation was obtained from nodal explants cultured on Murashige and Skoog (MS) medium supplemented with 2.0 mg L −1 6-benzyladenine (BA) and 0.5 mg L −1 α-naphthalene acetic acid (NAA), with a mean number of 14 shoots per explant. A high concentration of BA (4.0 mg L −1 ) and repeated subcultures resulted in hyperhydric shoots. Decreasing the BA concentration to 1.0 ...
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Abstract
A complete protocol of in vitro selection and greenhouse screening for glyphosate-tolerant variants in manilagrass ( Zoysia matrella [L.] Merr) was established in this study. Newly subcultured calli of more than 5 years’ old were transferred to selection medium containing 2 mM glyphosate. After two rounds of selection, 220 calli survived out of 840 and were transferred to regeneration medium without glyphosate. Regenerated plantlets were then transferred to regeneration medium containing 0.5 mM glyphosate to select tolerant plantlets. After 1-month growth, ...
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Abstract
Production of hybrid seeds and pursuing heterosis breeding of many crops have been accomplished using male sterile lines. However, not all crops have valuable male sterile lines due to instability of male sterility and absence of a restorer system. In this study, male sterile lines have been induced using a two-component system. The extracellular ribonuclease Barnase was cleaves into two inactive yet complementary fragments, designated as ‘Bn-5’ and ‘Bn-3’. Both components were controlled by a TA29 promoter. They were transferred into ...
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Abstract
An efficient protocol for secondary somatic embryogenesis in mountain ash is reported. Secondary somatic embryos (SSEs), initially obtained from primary embryos, were proliferated and maintained for more than 2 years via cyclic secondary somatic embryogenesis. SSEs were produced on the surfaces of cotyledons and radicles of maternal somatic embryos. Histological observations of the various stages of SSE development revealed four typical stages: globular, heart-shaped, torpedo, and cotyledon. Addition of a low concentration of naphthaleneacetic acid (NAA) to Murashige and Skoog (MS) ...
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Abstract
An efficient protocol was developed for short-term storage and conservation of a woody medicinal climber, Decalepis hamiltonii , using encapsulated nodal segments. The encapsulation of nodal segments was significantly affected by the concentrations of sodium alginate (Na-alginate) and calcium chloride (CaCl 2 ·2H 2 O). A gelling matrix of 4 % Na-alginate and 100 mM CaCl 2 ·2H 2 O was found most suitable for the production of ideal Ca-alginate beads. Maximum shoot re-growth (77.00 ± 2.09 %) was recorded on ...
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Abstract
Juvenile trees of Malus xiaojinensis produce leaves with three to five lobes; however, adult trees do not. Thus, M. xiaojinensis serves as a model plant for studies on rejuvenation of woody perennials. To better understand effects of in vitro culture, number of passages, and plant growth regulators on observed somaclonal variations in leaf morphology, total levels of DNA methylation and expression of associated genes, including DRM2 and MET1 , of apical shoot meristems of M. xiaojinensis , were evaluated. Following 17 ...
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Abstract
Malus baccata is widely used as a rootstock in cold regions of the world because of its cold hardiness. In this study, a highly efficient Agrobacterium rhizogenes strain 8196 transformation system was developed using in vitro-derived stem segments of M. baccata . Approximately 37 % agro-infected explants produced hairy roots when they were incubated on Murashige and Skoog (MS) medium without plant growth regulators. A total of 95 % of hairy roots exhibited glucuronidase activity. Calli were induced from putatively-transformed hairy ...
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Abstract
An efficient method has been developed for somatic embryogenesis, plant regeneration and transformation of the important banana cultivar ‘Dwarf Cavendish’ ( Musa AAA). A high embryogenic response was obtained in 1.36 % of immature male flower explants. Once embryogenic structures were transferred to liquid medium, embryogenic cell suspensions (ECSs) with high regeneration capacity were obtained. ECSs were incubated under different conditions with Agrobacterium tumefaciens strain EHA101 harboring vector pFAJ3000 that contains pNos-nptII-tOcs and p35S-uidAintron-t35S expression cassettes. The effect of spermidine and ...
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Abstract
To pursue genetic improvement of lily, efficiency of both regeneration and transformation from callus cultures induced from different explants were evaluated in multiple cultivars. Thirty-five callus lines induced from filaments or styles and one control callus line derived from bulb scales of in total twenty lily cultivars representing Lilium longiflorum , Oriental × Trumpet and Longiflorum × Asiatic hybrids were maintained on a medium with 8.3 μM picloram (PIC). In this study, they were tested for their regeneration potential by transferring ...
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Abstract
The comparison of direct and indirect somatic embryogenesis (DSE and ISE) for biolistic transformation of sugarcane by minimal expression cassettes indicated the highest transformation efficiency with ISE (2.2 independent transgenic plants per shot) and the most rapid production of transgenic plants with DSE (12 weeks from explant to plants in soil). Microprojectiles of 0.3 μm diameter produced 5 times more transgenic lines than 1 μm microprojectiles when used at the same weight basis per shot. A significant reduction of the number ...
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posted by
1 person
rafaelcarvalho
Abstract
To assess the potential of different genotypes of Brazilian oil palm ( Elaeis guineensis Jacq.) to somatic embryogenesis and somatic embryo proliferation, mature zygotic embryos of nine commercial genotypes of E. guineensis (BRSC2001, BRSC2328, BRSC2301, BRSC3701, BRSCM1115, BRSC7201, BRSC2528, BRSC2501, and BRSCN1637) were used. Explants were incubated on Murashige and Skoog (MS) supplemented with 450 μM picloram, 3.0 % sucrose, 500 mg l −1 glutamine, and 2.5 g l −1 activated charcoal, and gelled with 2.5 g l −1 Phytagel. After ...
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Abstract
This study aimed to characterize the anatomical events and ultrastructural aspects of direct and indirect in vitro organogenesis in Passiflora edulis . Root explants were cultured on induction medium, supplemented with 4.44 μM 6-benzyladenine. Roots at different stages of development were collected and processed for observation by light microscopy and scanning and transmission electron microscopy. Patterns of direct and indirect regeneration were observed in the explants. During direct organogenesis, the organogenic buds and nodules, formed from meristemoids, originated from the pericycle ...
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Abstract
DNA becomes methylated in vivo through the action of a specific group of enzymes known as methyltransferases or methylases. Plants are known to possess the methyltransferases (Met), chromo methyltransferases (CMT), and domainrearranged methyltransferases (DRM) methylase families, which affect cytosine methylation within different contexts. DNA methylation has been proposed to play a role in secondary plant metabolism, but there is a lack of valid data connecting these two processes. In this study, we treated control and transformed with rolB gene from Agrobacterium ...
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Abstract
In Vitis spp. where somatic embryogenesis-based regeneration predominates, an efficient, reproducible and robust method of direct shoot organogenesis from leaf explant material has been established in the dwarf wine grape ‘Pixie’ ( Vitis vinifera ). This regeneration system was achieved by testing the response of leaf material in two stages of development, and pre-conditioning the explant material in dark conditions and/or in liquid media prior to excising from the plant and placing it on solidified media. The pre-excision treatments included (1) ...
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Abstract
Plant growth and development depend on a precise coordination between cell division and cell expansion. In this study, a rice cell cycle switch 52 B ( OsCCS52B ) was functionally characterized using two approaches: overexpression of the gene product in fission yeast and characterization of an insertion mutant line 1B-10423. In wild-type plants, OsCCS52B is highly expressed in generative organs such as flowers and kernels. Overexpression of OsCCS52B induces cell elongation and slower cell proliferation in fission yeast. Characterization of the ...
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posted by
1 person
rs
Abstract
Alstroemeria is multiplied in vitro by forced outgrowth of lateral rhizomes from rhizome explants. The multiplication rate is very low because of strong apical dominance and poor rhizome growth. We report here that moderate abiotic stresses stimulate both rhizome growth and outgrowth of lateral rhizomes, and accordingly increase multiplication. Rhizome explants were exposed to heat by a hot-water treatment (HWT) or by a hot-air treatment. Both increased rhizome growth when applied for 1 or 2 h in the range of 30–40 ...
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