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Öğe Clonal propagation and synthetic seed production from nodal segments of Cape Gooseberry (Physalis Peruviana L.), a tropical fruit plant(2015) Yücesan, Bahtiyar Buhara; Mohammed, Aliyu; Arslan, Merve; Gürel, EkremPhysalis peruviana L. contains polyphenols and carotenoids with antiinflammatory and antioxidant activities used against diabetes. To establish an efficient regeneration system using nodal segments excised from 4-week-old germinated seedlings, direct plant regeneration, without additional rooting stage, was achieved on LS medium containing 0.5 mg/L 6-benzylaminopurine (BAP), kinetin (KIN), thidiazuron (TDZ), or gibberellic acid (GA3), alone or in combination with 0.25 mg/L indole-3-acetic acid (IAA) or indole-3- butyric acid (IBA), after 2 weeks of incubation. The highest mean numbers of shoots and well-developed roots were obtained on LS medium containing solely 0.5 mg/L TDZ, producing 5.3 shoots and 3.3 roots per explant after 2 weeks of incubation. Direct shoot and root formation were also recorded on LS medium containing no plant growth regulators. Due to the high regeneration capacity of nodal segments, synthetic seed production was also investigated using the sodium alginate (NaAlg) encapsulation technique. Four different matrix compositions, including NaAlg with or without LS medium containing 3% (w/v) sucrose alone or in combination with 0.5 mg/L abscisic acid (ABA) as a growth retardant were tested for the regrowth performance of synthetic seeds after storage at 4 °C up to 70 days. The highest regrowth (100%) was observed at 28 days of storage for all matrix compositions. All plantlets were acclimatized to the soil and then progressively transferred to the field. The fruits were harvested after 5 months. This study might provide a new insight through protocol development for micropropagation and synthetic seed production of many solanaceous species with economical relevance.Öğe Digitalis ferruginea L. türünün kardenolit sentez metabolizmasının moleküler yöntemlerle araştırılması ve kardenolit içeriğinin karşılaştırmalı analizi(2015) Gürel, Ekrem; Yücesan, Bahtiyar Buhara; Eker, Ismail; Ordu, Öznur Demir; Sammeullah, Muhammad; Mohammed, Aliyu; Ilal, Ayşe Tuğçe-Öğe Effects of plant growth regulators, different culture media and strength MS on production of volatile fraction composition in shoot cultures of Ocimwn basilicum(Elsevier Science Bv, 2018) Monfort, Lucila Elizabeth Fragoso; Bertolucci, Suzan Kelly Vilela; Lima, Andreisa Fabri; de Carvalho, Alexandre Alves; Mohammed, AliyuThe influence of different culture media, strength MS media and plant growth regulators (PGR) was assessed through the growth and production of the volatile fraction composition in Ocimwn basilicum (basil). The treatments were: the use of different culture media (MS, B5 and WPM), strength MS media (2MS, MS, 1/2MS and 1/4MS) and different PGR combinations or only IAA, NAA, IBA, BAP and TDZ. The leaf number, nodal segment, shoot number, lenght, shoot dry weight, root number, root size, root dry weight, as well as the volatile fraction analysis through Head-Space - GC/MS were assessed at the 40th culture day. The present results suggest that media type, MS concentration and the growth regulator types have qualitative and quantitatively influenced the growth and volatile composition. A higher amount of methyleugenol in the media 2MS and 1/4MS, and higher content of linalool and 1,8-cineole in MS and WPM were observed. The PGR improved biomass production and induced quantitative modifications in the production of major volatile constituents in Ocimwn basilicum plant lets, although the same major compounds were produced in all treatments. There was a tendency to estragole synthesis in the presence of auxin combined with cytokinin. There was a tendency to linalool synthesis in the presence of auxin isolates.Öğe An efficient regeneration system and steviol glycoside analysis of Stevia rebaudiana Bertoni, a source of natural high-intensity sweetener(Springer, 2016) Yücesan, Buhara; Büyükgöçmen, Refik; Mohammed, Aliyu; Sameeullah, Muhammad; Altuğ, Cevher; Gürel, Songül; Gürel, EkremStevia is a natural, zero-calorie, intensively sweet extracted from the leaves of Stevia rebaudiana. Its sweet taste derives from a group of compounds known as steviol glycosides. In this study, an efficient micropropagation protocol for S. rebaudiana was developed for possible commercial implementation using Murashige and Skoog (MS) basal medium without plant growth regulators for most of the process. Direct shoot formation was achieved after cultivation of nodal segments on MS medium with or without plant growth regulators (benzyl amino purine [BAP], kinetin [KIN]) at various concentrations (0.1, 0.5, 1.0, or 2.0 mg L-1). Although all treatments produced two shoots per explants after 3 wk of culture, high concentrations of KIN and BAP (1.0 or 2.0 mg L-1) induced more callus formation. Rooting was achieved on MS medium containing 0.25 mg L-1 indole-3-acetic acid (IAA), which produced 8.1 roots per shoot after 3 wk of cultivation. After acclimatization of the regenerants in a portable greenhouse for 3 wk, all regenerants and seed-derived seedlings were transferred to field conditions for 16 wk. Steviol glycoside contents (% leaf dry weight) did not differ between leaves collected from regenerants and seed-derived plants. Rebaudioside A content ranged from 4.7 to 5.0% (w/w), while stevioside ranged from 6.4 to 6.9% (w/w). There was no significant difference between the two sampling periods (late vegetative and flowering stages) for the plants grown in the field. In this study, a cost-effective in vitro regeneration protocol was established that enables efficient, large-scale in vitro production of S. rebaudiana for field cultivation.Öğe In vitro and ex vitro propagation of Stevia rebaudiana Bertoni with high Rebaudioside-A content-A commercial scale application(Elsevier, 2016) Yücesan, Buhara; Mohammed, Aliyu; Büyükgöçmen, Refik; Altuğ, Cevher; Kavas, Özge; Gürel, EkremStevia rebaudiana Bertoni is a self-incompatible and the pollination is an entomophilous short-day plant. Rebaudioside A, as an individual steviol glycoside, is of particular interest in global sweetener market due to its most desirable flavour profile as compared to stevioside having aftertaste bitterness. In the present investigation, regenerant formation was performed for 6 weeks, in which randomly selected nodes excised from in vitro germinated seedlings were cultured on solid Murashige and Skoog medium (MS) with or without growth regulators (6-benzylaminopurine- BAP or kinetin- KIN) at various concentrations (ranging from 0.1 to 2.0 mg/L). Irrespective of the medium composition, the results showed that all treatments were effective for shoot induction, producing an average of 2 shoots per explant after three weeks of culture. Following subsequent sub-culturing on MS medium at 3-week intervals, all the shoots regenerated from the nodes were transferred to MS medium with or without indole-3-acetic acid (IAA), indole-3-butyric acid (IBA) or naphthalene acetic acid (NAA) at two different concentrations (0.25 or 0.50 mg/L) for root formation for 3 weeks. IAA was more effective for root formation, producing 7.6 roots per shoot with 100% rooting frequency. All the regenerants (similar to 13.5 cm in length) were potted and successfully acclimatized in a greenhouse for 2 weeks, and then transferred to the field for 14 weeks with a high survival rate (>99%). Similarly, seedlings (8 weeks-old) derived from seed germination in pots were also transferred to the same field in a different plot. There was no significant difference in terms of morphological, yield performance and steviol glycoside compositions between regenerants and seedlings sampled from two developmental periods (late vegetative and flowering period). It was clearly shown that clonal propagation using nodal explants was effective for superior stock plant production with high Reb-A content (11.7% w/w). (C) 2016 Elsevier B.V. All rights reserved.Öğe In vitro propagation and cardenolide profiling of Digitalis ferruginea subsp schischkinii, a medicinally important foxglove species with limited distribution in Northern Turkey(Springer, 2016) Yücesan, Buhara; Mohammed, Aliyu; Eker, İsmail; Sameeullah, Muhammad; Demir-Ordu, Öznur; Cihangir, Cansu; Şahbaz, Nevin; Kaya, Özge; Gürel, EkremCardenolides isolated from genus Digitalis are natural compounds used in treating cardiac insufficiencies. Recent investigations have also suggested the use of cardenolides in oncology. Due to the medicinal importance of the genus Digitalis, in vitro regeneration techniques would be useful for propagating the best genotypes selected from natural sources. To achieve this, Digitalis ferruginea subsp. schischkinii (Ivanina) K. Werner was collected from different locations along the Black Sea region of Turkey. In the cardenolide analysis, samples from Giresun and Trabzon showed a greater diversity than those from other locations, and thus, the Trabzon population was selected for plant tissue culture studies. In vitro regeneration was performed using cotyledonary leaves and hypocotyl segments on Murashige and Skoog (MS) medium containing various concentrations (0.5, 1.0, or 2.0 mg L-1) of 6-benzyladenine (BA), kinetin (KIN), or thidiazuron (TDZ) alone or in combination with indole-3-acetic acid (IAA) at 0.5 or 1.0 mg L-1. Among the media used for shoot regeneration, the highest frequency of shoot organogenesis was obtained from MS medium containing 2.0 mg L-1 TDZ and 0.5 mg L-1 IAA, producing 11.0 shoots per explant with 100% regeneration frequency from hypocotyl explants after 4 wk. For successful rooting, IAA was the most effective at 1.0 or 2.0 mg L-1, producing 16.3 or 17.3 roots per shoot, respectively, after 4 wk. Regenerants from several medium types were grown in a greenhouse for 6 mo, and all displayed similar cardenolide contents, i.e., the lanatoside C level was around 0.3% (w/w) while the levels of lanatosides A and B, digoxin, and digitoxin were less than 0.08% (w/w), similar to plants grown from seeds collected from Trabzon district.Öğe In vitro regeneration and cardenolide determination of an endemic foxglove, Digitalis cariensis (Aegean Foxglove)(Springer, 2015) Mohammed, Aliyu; Yücesan, Buhara; Demir-Ordu, Öznur; Cihangir, Cansu; Eker, İsmail; Gürel, EkremThe genus Digitalis L. is one of the most pharmacologically important plant genera because many Digitalis species produce cardenolides, which are commonly used for cardiac insufficiencies. In this study, different pretreatments were employed to increase germination of Digitalis cariensis seeds, which germinate poorly because of extensive dormancy. Seed germination was improved when seeds were pretreated by scarification followed by soaking in sterile distilled water overnight. In addition, the regeneration capacity of four different types of explants (cotyledonary leaf, root, hypocotyl, or flamingo-bill-type [FBT]) was tested on Linsmaier and Skoog (LS), Murashige and Skoog (MS) and Gamborg's B5 (B5) medium containing no plant growth regulators or supplemented with 0.5 mg L-1 thidiazuron (TDZ) alone or in combination with 0.25 mg L-1 indole-3-acetic acid (IAA). Among the explants tested, only FBT explants regenerated. The highest mean number of shoots was produced on LS medium supplemented with a combination of 0.5 mg L-1 TDZ and 0.25 mg L-1 IAA, producing 3.9 shoots per FBT explant after 4 wk of incubation. Regenerated shoots were rooted on LS medium containing different concentrations of IAA or indole-3-butyric acid (IBA) (0.1, 0.5, or 1.0 mg L-1). The greatest number of roots developed on medium supplemented with 1.0 mg L-1 IAA, which produced 5.6 roots per shoot after 3 wk of culture. Cardenolides were profiled by HPLC analysis from basal leaves of D. cariensis plants from natural populations and from in vitro-derived plantlets. There was no significant difference in lanatoside contents (A, B, and C) between the two sources. Digoxin and digitoxin were not detected in either source.Öğe Plantlet regeneration from young leaf segments of Curaua (Ananas Erectifolius), an Amazon species(2016) Moreira, Carolina Mariane; Andrade, Helena Botelho De; Bertolucci, Suzan Kelly Vilela; Lameira, Osmar Alves; Mohammed, AliyuThe present study reports the first efficient in vitro regeneration of Ananas erectifolius via indirect organogenesis. Leaf segments (leaf base, middle, and apex) excised from 3- or 5-week-old in vitro plantlets were cultured on 1/4 strength MS medium supplemented with different concentrations and combinations of plant growth regulators. The explants were also exposed to pulse treatment with thidiazuron (TDZ) and 2,4-dichlorophenoxyacetic acid (2,4-D) for different periods of time. The results showed that using juvenile rather than old explants enhanced the frequency of callus induction (35.0% and 16.0%, respectively). Among the explant types tested, only leaf base segments induced calli; the highest frequency occurred via culture treatment containing 4-amino-3,5,6-trichloropicolinic acid (picloram; 48.57%), 2,4-D (40%), or TDZ (35.71%). However, only calli generated in treatments containing TDZ or N6-isopentenyladenine (2ip) were able to develop shoots (maximum 35.71% and 14.28%, respectively). A combination of TDZ and 2,4-D improved callus induction (60.0%) but did not increase shooting response. However, both callusing and shooting response increased when 10 days of pulse treatment with TDZ and 2,4-D was applied (66.83% and 48.7%, respectively). Rooted plantlets that exhibited normal growth and development were acclimatized in a greenhouse and had a survival rate of 95%.Öğe Shoot-tip cultivation and cardenolide content analysis of natural populations of Digitalis Lanata Ehrh. Subsp. Lanata (Wooly Foxglove) in Thrace Region(2018) Yücesan, Bahtiyar Buhara; Eker, İsmail; Lazzarini, Luiz Eduardo Santos; Aslam, Noreen; Mohammed, Aliyu; Gürel, EkremDigitalis lanata, due to the presence of cardenolides in the leaves, is widely used in drug industry for the treatments of congestive heart diesases. In this present study, natural populations of D. lanata in Thrace region were collected. Thereafter, cardiac glycoside analysis of D. lanata subsp. lanata grown in natural habitats and micropropagation from shoot tips were established as a model work for a consistent and a sustainable plant production. Of the cardenolides, Lan A (ranging between 24.8 and 300.4 mg 100g-1) and Lan C (42.1 and 258 mg 100 g-1 were predominantly found in the samples as compared to lanatoside B, digoxin and digitoxin. After seed germination, shoot-tip cultivation was achieved after a subsequent shoot and root formation on MS medium. Indole-3-acetic acid (IAA) and indole-3-butyric acid (IBA) depending on their concentrations were found to be effective in shoot and root formation accordingly. Maximum two shoots from shoot tips were produced after 8 weeks of cultivation, and at the end of 12th weeks of cultivation each shoots produced an average number of 6.4 roots in 8.1 cm length. Acclimatization was also achieved successfully with a maximum survival rate (95 %) for 4 weeks. This study is the first report on cardenolide profiling of Thracian D. lanata at different locations, and shoot tip cultivation of the population selected under in vitro conditions.
