Micropropagation of Pothomorphe umbellata via direct organogenesis from leaf explants
01 Jan 2000-Plant Cell Tissue and Organ Culture (Kluwer Academic Publishers)-Vol. 60, Iss: 1, pp 47-53
TL;DR: An anatomical study confirmed shoot regeneration via direct organogenesis and micropropagation protocol for Pothomorphe umbellata was carried out using leaf segments cultured on 1/4 strength Murashige and Skoog medium.
Abstract: The establishment of a micropropagation protocol for Pothomorphe umbellata was carried out using leaf segments cultured on 1/4 strength Murashige and Skoog medium supplemented with 0.5 mg l-1 6-benzyladenine, 0.1 mg l-1 gibberelic acid added with 10 g l-1 sucrose. Rooting was achieved using MS medium devoid of growth regulators. An anatomical study confirmed shoot regeneration via direct organogenesis.
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TL;DR: This study demonstrated that adventitious shoot regeneration of E. purpurea from leaf explants can be a useful method for the multiplication of this important medicinal plant.
Abstract: Efficient plant regeneration was achieved via organogenesis from callus cultures derived from leaf tissue of Echinacea purpurea. Proliferating shoot cultures were obtained by placing leaf explants on Murashige and Skoog (MS) medium supplemented with 6-benzylaminopurine (BAP) and naphthaleneacetic acid (NAA) combinations. MS medium supplemented with BAP (4.44 μM) and NAA (0.054 μM) was the most effective, providing high shoot regeneration frequencies (100%) associated with a high number of shoots per explant (7.7 shoots/explant). Plantlets were rooted on MS medium alone or in combination with different concentrations of indole-3-butyric acid (IBA), and high rooting and survival was achieved using MS media without plant growth regulators (PGR). All plantlets survived acclimatization producing healthy plants in the greenhouse. This study demonstrated that adventitious shoot regeneration of E. purpurea from leaf explants can be a useful method for the multiplication of this important medicinal plant.
137 citations
Cites background from "Micropropagation of Pothomorphe umb..."
...Reports of auxin and cytokinin combinations supporting organogenic differentiation in other species have been well documented for several species (Lisowska and Wysonkinska, 2000; Pereira et al., 2000, Pretto and Santarém, 2000)....
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TL;DR: Application of this protocol has potential for mass multiplication of the target species in a limited time period and reflected the existence of high inter-explant variability in response to growth regulators.
Abstract: A callus induction and in vitro plantlet regeneration system for the endangered state flower of Uttaranchal (Saussurea obvallata) was optimized by studying the influence of explant type (root, hypocotyl, cotyledon and leaf), age and different concentrations of plant growth regulators. Explants from 10 to 15-day-old seedlings showed maximum callus induction. Callus formation and shoot differentiation was initiated on Murashige-Skoog (MS) medium containing 6-benzyladenine (BA) and α-naphthalene acetic acid (NAA) in all explant types. The best results were obtained using leaf explants: 100% callusing was achieved in MS medium supplemented with 2.5 μM BA and 1.0 μM NAA, and 100% differentiation along with a multiplication rate of 12 shoots per explant with a combination of 5.0 μM BA and 1.0 μM NAA. However, the results reflected the existence of high inter-explant variability in response to growth regulators. In vitro rooting of shoots was achieved at an efficiency of 100% in one-half strength MS medium supplemented with 2.5 μM indole-3-butyric acid. Application of this protocol has potential for mass multiplication of the target species in a limited time period.
103 citations
Cites background from "Micropropagation of Pothomorphe umb..."
...Such variable responses for different explant types have also been reported in other species (Christopher and Rajam 1996; Arockiasamy and Ignacimuthu 1998; Pereira et al. 2000)....
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TL;DR: An efficient and reproducible protocol for high-frequency callus regeneration from immature leaf explants of T. indica was developed, and Histological studies of the regenerative callus revealed that shoot buds were originated from the outermost regions.
Abstract: Tylophora indica (Burm. f.) Merrill is a threatened medicinal climber distributed in the forests of northern and peninsular India. An efficient and reproducible protocol for high-frequency callus regeneration from immature leaf explants of T. indica was developed. Organogenic callus formation from immature leaf pieces was obtained by using Murashige and Skoog (MS) medium supplemented with 7 μM 2,4-dichlorophenoxyacetic acid and 1.5 μM 6-benzyladenine. On this medium 92% explants produced callus. The optimal hormone combination for plantlet regeneration was 8 μM thidiazuron, at which shoot regeneration was obtained from 100% of the cultures, with an average of 66.7 shoots per culture. Histological studies of the regenerative callus revealed that shoot buds were originated from the outermost regions. For root formation, half-strength MS medium supplemented with 3 μM indole-3-butyric acid was used. Plants were transferred to soil, where 92% survived after 3 mo. of acclimatization.
63 citations
Cites background from "Micropropagation of Pothomorphe umb..."
...The promoting effect of auxin and cytokinin combinations on organogenic differentiation has been well documented (Lisowska and Wysokinska, 2000; Pereira et al., 2000; Pretto and Santarém, 2000; Koroch et al., 2002)....
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TL;DR: Callus cultures were initiated from leaf and nodal explants of V. planifolia and potentially, more than 100,000 plantlets could be produced within eight subcultures from callus obtained from leaf explant through the methods described here.
Abstract: Vanilla planifolia is a tropical orchid mainly known for the aromatic flavor of its cured pods. Callus cultures were initiated from leaf and nodal explants of V. planifolia. Leaf explants showed better callus initiation than the nodal explants with callus biomass production maximal when cultured on Murashige and Skoog (MS) basal medium containing 4.52 mM 2,4-dichlorophenoxy acetic acid and 2.22 mM benzyladenine (BAP). Callus transferred to MS basal medium supplemented with 13.32 μM BAP, and 13.43 μM naphthaleneacetic acid (NAA) showed superior growth response and produced 14.0±1.0 shoots with an average length of 3.6±0.1 cm after 40 d. Subsequent transfer of the proliferated shootlets to MS basal medium supplemented with 8.88 μM BAP and 8.08 μM NAA produced 12.3±0.14 plantlets with an average height of 3.6 cm±0.10 cm. All plantlets produced profuse rooting within 35-40 d after transfer to half- strength MS basal medium supplemented with NAA in combination with indole-3-acetic acid. Rooted plantlets were transferred for hardening, with 80% of the plantlets becoming successfully established in the field. Potentially, more than 100,000 plantlets could be produced within eight subcultures from callus obtained from leaf explant through the methods described here.
62 citations
TL;DR: The effect of vesicular arbuscular mycorrhizae (VAM) association in averting the transplantation shock was tested and proved to be highly beneficial, giving a 100% survival rate after 60 d of transplantation.
Abstract: An efficient regeneration system was developed for Kigelia pinnata L., a multipurpose tree belonging to the family Bignoniaceae. The nodal segments were cultured in vitro, and the optimum concentrations of plant growth regulators for callus induction were determined. The friable organogenic calli were derived from the basal cut end of the nodal segments. The highest yield of morphogenic callus (100%) was observed when nodal segments were cul- tured on Murashige and Skoog (MS) medium supplemented with 3 µM 2,4 dichlorophenoxyacetic acid (2, 4-D). The morphogenic callus maintained high regeneration during the first four subcultures in the callus induction medium. The maximum shoots (28/culture) were regenerated at the highest frequency of 100% when 3 µM thidiazuron (N-phenyl N' 1,2,3-thidiazol-5-yl urea) (TDZ) and 0.5 µM naphthaleneacetic acid (NAA) were added to MS medium. The emergence of multiple shoots from the calli was histologically documented. The regenerated shoots showed maxi- mum rooting on ½ MS medium containing 4 µM indole-3- butyric acid (IBA). The effect of vesicular arbuscular mycorrhizae (VAM) association in averting the transplantation shock was tested and proved to be highly beneficial, giving a 100% survival rate after 60 d of transplantation. This efficient plant regeneration system provides a founda- tion for generating transgenic plants of this multipurpose tree.
57 citations
Cites background from "Micropropagation of Pothomorphe umb..."
...The promoting effect of auxin and cytokinin combinations on organogenic differentiation has been well es tabl ished in several sys tems (Lisowska and Wysonkinska, 2000; Pereira et al., 2000; Pretto and Santarém, 2000; Xie and Hong, 2001; Koroch et al., 2002)....
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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
01 Jan 1962
16,251 citations
"Micropropagation of Pothomorphe umb..." refers methods in this paper
...Disinfested explants were inoculated onto MS semisolid medium (Murashige and Skoog, 1962), contain-...
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...Disinfested explants were inoculated onto MS semisolid medium (Murashige and Skoog, 1962), contain- ing 30 g l−1 sucrose, supplemented with the concentrations and combinations of growth regulators as showed on Table 1....
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Journal Article•
8,559 citations
"Micropropagation of Pothomorphe umb..." refers methods in this paper
...Samples were fixed in Karnovsky solution (Karnovsky, 1965), dehydrated in alcoholic-ethyl series and infiltrated in glycol methacrilate (Reichert-Jung) resin....
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Journal Article•
5,201 citations
TL;DR: Differential staining of paraffin embedded plant material is accomplished without removing theparaffin, and lignified tissue, suberized tissue, and some tannins are stained blue-green, while nonlignified walls are stained red-purple.
Abstract: Differential staining of paraffin embedded plant material is accomplished without removing the paraffin. Material fixed in 3% glutaraldehyde in 0.02 M phosphate buffer is dehydrated, embedded, sectioned, and mounted on glass slides using conventional methods. The slides are placed in 0.05% toluidine blue O in distilled water for 2–30 min, rinsed in water for 1 min, and allowed to air dry. Paraffin is removed with 2 changes of xylene and the cover slip mounted with resin. Lignified tissue, suberized tissue, and some tannins are stained blue-green, while nonlignified walls are stained red-purple. An advantage of the method is the short time required for staining and mounting. Also, critical counterstaining is not required.
447 citations
"Micropropagation of Pothomorphe umb..." refers methods in this paper
...Sections (5 µm) were colored with Toluidine Blue (Sakay, 1973) and set up in synthetic resin (Permount)....
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