Bulblet-productivity of explants from scales, leaves, stems and teplas of Lilium rubellum Baker
TL;DR: The expiants of leaves were excellent for regenerating many bulblets per expiant, while those of stems produced the heaviest bulblet, on average.
About: This article is published in Scientia Horticulturae.The article was published on 1984-03-01. It has received 30 citations till now.
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TL;DR: A method for the regeneration of lily plantlets (Lilium spp.) through different morphogenic pathways is described, with results suggesting that the leaf-segments obtained in this way could be an alternative to the scales as a source of material for propagation.
Abstract: A method for the regeneration of lily plantlets (Lilium spp.) through different morphogenic pathways is described. Plant regeneration was obtained from in vitro cultured leaves of four lily hybrids, cultured on Murashige and Skoog's basal medium supplemented with cytokinins (TDZ and BA) and auxins (NAA and IBA) at different concentrations. Direct shoot regeneration occurred with all tested media for the Asiatic lilies `Elite' and `Pollyanna' and also for the Oriental hybrid `Star Gazer'. Callus developed on TDZ-enriched medium from leaf segments of L. longiflorum cv. `Snow Queen' regenerated by direct organogenesis. This occurred on a medium with auxin/ cytokinin balance which was lower than other genotypes. There were fewer problems of sterilization with leaves from sprouted bulbs than in vitro scale culture. This suggests that the leaf-segments obtained in this way could be an alternative to the scales as a source of material for propagation. A protocol for micropropagation based on bulblets from in vitro shoot-tip-derived stem nodes was also used. The development of pseudo-bulbets is particularly advantageous since it allows for structures characterised by absent or low dormancy. Regenerated shoots have been rooted and successfully acclimatized to greenhouse conditions where they flowered after the second year giving plants with true-to-type shape and colour.
76 citations
Cites background from "Bulblet-productivity of explants fr..."
...Shoot formation by direct or- for L. rubellum (Niimi, 1984). ganogenesis was achieved on all media including growth regulator-free medium....
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...Niimi and Tsuyoshi (1979) and Niimi (1984) reported the induction of adventitious buds from leaf explants in L. rubellum....
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TL;DR: A continuous system of mass propagation of bulblets was achieved through in vitro scale formation (secondary explants) on MS medium supplemented with 23 μM kinetin and 0.5 μM NAA, as well as scale proliferation on MS basal liquid stationary medium.
Abstract: A simple, rapid and cost-effective in vitro scheme has been proposed for mass propagating two cultivars of Asiatic lily hybrids. An average of seven bulblets was formed after 17 d when 1×1 cm2 bulb scale segments (explants) were cultured on Murashige and Skoog (MS) medium with 3% sucrose and 0.5 μM α-naphthaleneacetic acid (NAA). On MS medium containing 0.5 μM NAA and 6 or 9% sucrose, depending on the cultivar, large numbers of bulblets of increased size (3.5–5.0 cm in circumference) were formed under a 16/8 h photoperiod. A continuous system of mass propagation of bulblets was achieved through in vitro scale formation (secondary explants) on MS medium supplemented with 23 μM kinetin and 0.5 μM NAA, as well as scale proliferation on MS basal liquid stationary medium. Upon transplantation all bulblets sprouted, of which 40% flowered in the first season. Under ideal conditions, ca. 9.68×105 bulblets can be produced from a single scale segment in 1 yr by following the systematic propagation steps proposed here.
55 citations
Cites background from "Bulblet-productivity of explants fr..."
...(Takayama and Misawa, 1983); and L. rubellum (Niimi, 1984)....
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...According to Niimi (1985) the increase in the percentage of...
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TL;DR: In vitro bulb scales of Lilium longiflorum ‘Georgia’ were cultured on MS medium with BA (benzyladenine) to induce shoots and the addition of liquid medium stimulated the formation and growth of bulblets remarkably, compared with no addition ofLiquid medium.
46 citations
TL;DR: This procedure has great potential application for micropropagation, genetic transformation, and preparation of shoot tips for cryopreservation and cryotherapy for virus eradication of Lilium.
Abstract: Here, we report a widely applicable procedure for direct shoot regeneration via basal leaf segments of Lilium. Leaf segments (0.8–1.0 cm long and 0.4 cm wide) were excised from leaves on shoot nodes 3 to 6 of 4-wk-old in vitro stock shoot cultures. The segments were wounded by three transverse cuts across the midvein on the abaxial side, with 1 mm between cuts, and cultured with the abaxial side in contact with a shoot regeneration medium composed of half-strength Murashige and Skoog medium supplemented with 1 mg/l naphthaleneacetic acid, 0.5 mg/l thidiazuron, 30 g/l sucrose, and 7 g/l agar (pH 5.8). The cultures were incubated for 4 wk under a 16-h photoperiod at 23 ± 2°C for adventitious shoot regeneration. With this procedure, a mean shoot regeneration frequency of 92–100% and mean number of shoots of 4.7–7.0 per segment were obtained in five Lilium species and hybrids, which represent diverse genotypes of Lilium and are commercially popular lilies. Histological studies with Lilium Oriental hybrid “Siberia” revealed that meristemoids initiated from subepidermal cells on the adaxial side of the explant and eventually developed into adventitious buds, without callus formation. In an assessment of genetic stability in the regenerants of “Siberia”, no polymorphic bands were detected by intersimple sequence repeat and only 0.73% polymorphic bands were detected by amplified fragment length polymorphism. The morphologies of the regenerants were identical to those of the control. These results demonstrated that the regenerants were genetically and morphological stable. Thus, this procedure has great potential application for micropropagation, genetic transformation, and preparation of shoot tips for cryopreservation and cryotherapy for virus eradication of Lilium.
40 citations
Cites background from "Bulblet-productivity of explants fr..."
...maximowiczii (Kato and Yoshinori 1977), Lilium rubellum (Niimi 1984), Lilium Asiatic hybrid (Bacchetta et al....
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...2001b; Nhut 2003), and leaf segments (Kato and Yoshinori 1977; Niimi 1984; Bacchetta et al. 2003; Kim et al. 2005; Xu et al. 2009; Liu and Yang 2012); among these, leaf segments are most often used because they are easy to obtain and their availability is independent of the season....
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...A combination of BA and NAA or indole-3-acetic acid (IAA) has been frequently used to induce adventitious shoots from leaf segments in Lilium species (Niimi 1984; Bacchetta et al. 2003; Kim et al. 2005; Liu and Yang 2012)....
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...…has been reported in Lilium longiflorum (Bacchetta et al. 2003), Lilium leichtlinii var. maximowiczii (Kato and Yoshinori 1977), Lilium rubellum (Niimi 1984), Lilium Asiatic hybrid (Bacchetta et al. 2003), Lilium Oriental hybrid (Bacchetta et al. 2003; Kim et al. 2005), Lilium davidii var.…...
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...…stems (Nhut 1998; Nhut et al. 2002; Bacchetta et al. 2003), receptacles (Nhut et al. 2001b; Nhut 2003), and leaf segments (Kato and Yoshinori 1977; Niimi 1984; Bacchetta et al. 2003; Kim et al. 2005; Xu et al. 2009; Liu and Yang 2012); among these, leaf segments are most often used because they…...
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Journal Article•
TL;DR: Histological studies showed that the embryogenic callus was formed at the epidermal cells and near vascular bundles of explants, and can play an important role in the future protection of this rare plant species.
Abstract: The effect of two plant growth regulators on the initiation of callus and regeneration of L. martagon was studied on modified MS medium. The cultures were initiated using seeds. The explants were isolated from different parts of seedlings (hypocotyls, seedling bulb, root) and adventitious bulblets. The growth regulators stimulated various types of callus. 4-amino–3,5,6-trichloropyridine–2-carboxylic acid (Picloram) induced a yellow, friable, granular callus, whereas benzyladenine (BA) alone induced a cream-white, compact callus. MS medium containing Picloram plus BA stimulated a yellow, compact, granular callus. The most useful explants for callus initiation were seedling bulbs and adventitious bulblet scales. The most efficient embryogenic callus was obtained on MS medium containing 5 µM Picloram and 5 µM BA. Histological studies showed that the embryogenic callus was formed at the epidermal cells and near vascular bundles of explants. Somatic embryos were solitary, whereas adventitious bulblets were closely connected with vascular tissue. No differences in the amount of DNA between scale tissue and callus were observed. Lilium martagon L. is an endangered species in Poland. In vitro culture techniques can play an important role in the future protection of this rare plant species.
36 citations
Cites background from "Bulblet-productivity of explants fr..."
...…L. regale, L. rubellum, L. speciosum, Lilium × formolongi, and Asiatic and Oriental hybrids (Simmonds and Cumming, 1976; Takayama and Misawa, 1982/83; Nimii, 1984; Priyadarshi and Sen, 1992; Maesato et al., 1994; Mii et al., 1994; Wickremesinhe et al., 1994; Famelaer et al., 1997; Tribulato et…...
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...speciosum, Lilium × formolongi, and Asiatic and Oriental hybrids (Simmonds and Cumming, 1976; Takayama and Misawa, 1982/83; Nimii, 1984; Priyadarshi and Sen, 1992; Maesato et al., 1994; Mii et al., 1994; Wickremesinhe et al., 1994; Famelaer et al., 1997; Tribulato et al., 1997a; Watad et al., 1998; Pelkonen and Kauppi, 1999)....
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References
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TL;DR: In vivo redox biosensing resolves the spatiotemporal dynamics of compartmental responses to local ROS generation and provide a basis for understanding how compartment-specific redox dynamics may operate in retrograde signaling and stress 67 acclimation in plants.
Abstract: In experiments with tobacco tissue cultured on White's modified medium (basal meditmi hi Tnhles 1 and 2) supplemenk'd with kiticthi and hidoleacctic acid, a slrikin^' fourlo (ive-told intTease iu yield was ohtaitu-d within a three to Tour week j^rowth period on addition of an aqtteotis exlrarl of tobacco leaves (Fi^'ures 1 and 2). Subse(iueutly it was found Ihiit this jnoniotiou oi' f^rowih was due mainly though nol entirely to inorj^auic rather than organic con.stitttenls in the extract. In the isolation of Rrowth factors from plant tissues and other sources inorj '̂anic salts are fre(|uently carried along with fhe organic fraclioits. When tissue cultures are used for bioassays, therefore, il is necessary lo lake into account increases in growth which may result from nutrient elements or other known constituents of the medium which may he present in the te.st materials. To minimize interference trom rontaminaitis of this type, an altempt has heen made to de\\eh)p a nieditmi with such adequate supplies of all re(iuired tnineral nutrients and cotntnott orgattic cottslitueitls that no apprecial»le change in growth rate or yield will result from the inlroduclion of additional amounts in the range ordinarily expected to be present in tnaterials to be assayed. As a point of referetice for this work some of the culture media in mc)st common current use will he cotisidered briefly. For ease of comparis4)n Iheir mineral compositions are listed in Tables 1 and 2. White's nutrient .solution, designed originally for excised root cultures, was based on Uspeuski and Uspetiskaia's medium for algae and Trelease and Trelease's micronutrieni solution. This medium also was employed successfully in the original cttltivation of callus from the tobacco Iiybrid Nicotiana gtauca x A', tanijadorffii, atitl as further modified by White in 194̂ ^ and by others it has been used for the
63,098 citations
TL;DR: Tissue cultures of Lilium auratum Lindl.
Abstract: Tissue cultures of Lilium auratum Lindl and L speciosum Thunb, which were derived from bulbscales, all appeared to differentiate organs The effect of cultural conditions on the differentiation of bulblets and roots was examined
The best material for bulblet formation was bulbscales of intact or in vitro produced bulblets The optimum temperature was 20°C and optimum pH was 6 Effect of irradiance on organ formation was not obvious but leaf emergence was stimulated
Higher kinetin concentrations stimulate the formation of numerous bulbscalcs High NAA concentrations induce roots On the other hand kinetin inhibits the NAA effect on root formation A high sucrose concentration stimulated organ formation, but the number of bulblets was at a constant level in the medium containing between 10 and 90 g/l of sucrose The formation of bulblets and their growth were stimulated at increasing strength of Murashige-Skoog's (MS) medium, but the length of roots was inhibited Inter action of strength of MS medium and sucrose concentration was examined High concentration of both components stimulated bulb lei growth, but the second strength of MS medium containing 90 or 120 g/l sucrose stimulated callus induction and inhibited the growth of bulblets
Maximum growth took 100 days for bulblets and about 50 days for roots The change of fresh weight/dry weight ratio during differentiation is also discussed
123 citations
TL;DR: Differences in responses of the twelve species tend to cut across the three families and no simple relation is evident between the natural rate of vegetative increase and the in vitro behaviour.
Abstract: In vitro responses of twelve species of bulbs and corms were compared. Plantlets could be induced directly without intervening callus on stem tissue in nine species, on ovary tissue in five species, and on leaf tissue in four species. In Gladiolus, Hyacinthus, Muscari, Ornithogalum, and Scilla plantlets were formed without growth factors added to the Murashige and Skoog medium. In Hippeastrum, Schizostylis, Sparaxis, and Ipheion auxin was required. No plantlets could be induced directly on expiants of growing tissue of Freesia, Tulipa, or Narcissus. Adventitious plantlets could be induced on pieces of bulb or corm from ten species but such material was difficult to free from contamination. Callus was obtained from all species except Tulipa and Hippeastrum. Plantlets could be regenerated from callus except that of Gladiolus, Sparaxis, and Schizostylis. Differences in responses of the twelve species tend to cut across the three families and no simple relation is evident between the natural rate of vegetative increase and the in vitro behaviour.
95 citations
TL;DR: This communication reports the successful establishment in both agar and liquid shake cultures of callus of Lilium longiflorum Thunb and the repeated subculturing of the callus on growth factor free media, and the production of large numbers of plantlets from callus under certain growth conditions in liquid suspension cultures.
Abstract: The monocot Lilium, possessing very large nuclei and chromosomes, is a favorable material for studying problems of development, parti cularly with regard to the proteins of the nucleus (Sheridan and Stern, 1967). Since it would broaden the possible approaches to such study if successful, an attempt was made to establish Lilium in tissue culture. This communication reports (1) the successful establishment in both agar and liquid shake cultures of callus of Lilium longiflorum Thunb ; (2) the repeated subculturing of the callus on growth factor free media; and (3) the production of large numbers of plantlets from callus under certain growth conditions in liquid suspension cultures. Callus was established by placing tissue expiants from steBi apices onto the basal medium supplemented with 2 mg/1 of indoleacetic acid (IAA). The terminal 10 cm of stems which were approximately 25 cm in height were removed and all but the smallest leaves were stripped off. The apices were washed for 5 min in a solution of the detergent Heikol, 10 min in 10% Clorox, and 5 min in sterile distilled water. Each apex was then placed in a sterile petri dish and the outer portion in cluding all the leaves and primordia was removed with a sc^pel leaving a cylinder which was free of any surface tissue. The terminal 2 cm were removed and placed on the medium. The basal medium was that of Linsmaier and Skoog (1965) con taining major and minor salts, organic supplements of thiamin and inositol, and 40 g of sucrose per liter. Basal agar medium refers to the above medium solidified with 0.8% agar. Fresh weight was used for determining growth response. Within three weeks of explantation, callus appeared on the edge of the tissue block in contact with the medium. The presence of IAA in the culture medium stimulated the production of callus by the stem tissue but was not essential for it. When expiants were placed on medium lacking IAA, buds appeared on the surface of the tissue blocks and shoots and roots were formed producing new plants; on occasion callus was also formed, but usually only after the formation of roots and shoots.
70 citations