scholarly journals 333 Micropropagation of Echinacea angustifolia, E. pallida, and E. purpurea from Vegetative and Seed Explants

HortScience ◽  
2000 ◽  
Vol 35 (3) ◽  
pp. 449C-449
Author(s):  
James F. Harbage

Micropropagation of three Echinacea species, E. angustifolia, E. pallida, and E. purpurea, was investigated as a potential means of germplasm preservation of species faced with overcollection in the wild and rapid clonal propagation of elite individuals with unique medicinal or ornamental properties. Comparison of explant sources indicated vegetative explants resulted in high contamination rates when collected from shoot-tips (100%),but not when collected from nodal explants (11% to 39). Seed coat removal reduced contamination from 100% in intact seeds to near 0% in excised embryos. Removal of seed coats (pericarp and integument layers) also eliminated dormancy requirement for germination. All species responded with shoot multiplication and loss of rooting when BA or thidiazuron was added to culture medium. Medium with thidiazuron resulted in excessive adventitious shoot formation. Shoot multiplication rates were low (one to three shoots/explant) on medium with BA levels low enough to avoid adventitious shoot formation. Medium containing half-strength MS minerals resulted in more shoots with smaller leaves than full-strength MS minerals. Cultures did not perform well on Woody Plant Medium. Increasing subculture frequency from every 4 weeks to every 2 weeks increased shoot multiplication rates from 1.4 to 1.8 shoots per subculture and total shoots produced after 12 weeks of culturing (per initial explant) from 2.8 to 23.9. Rooting occurred readily on shoots isolated from E. purpurea without addition of IBA. Rooting was low or non-existent on shoots from E. angustifolia and E. pallida, respectively, regardless of IBA level, light conditions, or temperature. Methods described in this study allow rapid multiplication of three Echinacea species and subsequent rooting of E. angustifolia and E. purpurea. Future improvements in root induction treatments will allow more effective use of micropropagation for Echinacea germplasm preservation and multiplication. Chemical names used: N-(phenylmethyl)-1H-purine-6-amine (BA), 1H-indole-3-butyric acid (IBA).

HortScience ◽  
2001 ◽  
Vol 36 (2) ◽  
pp. 360-364 ◽  
Author(s):  
James F. Harbage

Micropropagation of three Echinacea species, E. angustifolia DC., E. pallida Nutt., and E. purpurea Moench., was investigated as a potential means of germplasm preservation of species faced with overcollection in the wild and rapid clonal propagation of elite individuals with unique medicinal or ornamental properties. Very high contamination rates occurred with shoot-tip explants but not with nodal segments. Contamination rates for seed explants were inversely related to the number of seedcoat layers removed, ranging from 100% contamination from intact seeds to near 0% contamination from excised embryos. Dormancy of seed explants was also eliminated when the pericarp and integument were removed. Addition of benzyladenine (BA) to the culture medium induced shoot multiplication and inhibited root formation in all three species. Shoot multiplication rates were low (1-3 shoots per culture) when seed explants were placed on a medium with BA levels low enough to avoid adventitious shoot formation (0.45 μm). Shoot count was higher on half-strength Murashige and Skoog (MS) minerals, while leaf size was greater on full-strength MS minerals. Cultures did not perform well in Woody Plant Medium. Reducing subculture frequency from 4 to 2 weeks increased shoot multiplication from 1.4 to 1.8 shoots per subculture and total shoots produced per subculture after 12 weeks from 2.8 to 23.9. Rooting occurred readily on shoots isolated from E. purpurea cultures and was not promoted by addition of IBA to the rooting medium. Rooting was low and nil on shoots from cultures of E. angustifolia and E. pallida, respectively. Methods described in this study allow rapid multiplication of three Echinacea species and subsequent rooting of E. purpurea. Future improvements in root induction treatments will allow these methods to be used effectively for micropropagation and maintenance of disease-free germplasm of Echinacea species. Chemical names used: N-(phenylmethyl)-1H-purine-6-amine (BA); 1H-indole-3-butyric acid (IBA).


1992 ◽  
Vol 22 (5) ◽  
pp. 750-755 ◽  
Author(s):  
Ben A. Bergmann ◽  
Anne-Marie Stomp

Embryos of Pinusechinata Mill., Pinustaeda L., Pinusserotina Michx., Pinuseldarica Medwed., Pinuscaribaea Morelet, Pinusoocarpa Scheide, Pinustecunumanii (Schwd.) Equiluz & Perry, Pinusstrobus L., and Pinusradiata D. Don were cultured following the protocol for Pinusradiata to determine if a perpetual meristem culture could be produced. Two subsequent experiments were done that included modifications to the medium's mineral composition, strength (modified LePoivre and Murashige and Skoog at half-strength and full strength), cytokinin concentration (1.0, 2.5, and 5.0 mg/L benzyladenine), and auxin concentration (0 and 0.1 mg/L indolebutyric acid). Pines in subsection Australes performed poorly in culture relative to those species in subsection Oocarpae. Pinusradiata and Pinuseldarica were the only species to produce long-term subculturable meristematic tissue, although shoots were obtained with seven of the species. Half-strength modified LePoivre medium containing 2.5 mg/L benzyladenine but no auxin gave the best results with most species. Significant differences in shoot production were found among Pinusoocarpa provenances.


2014 ◽  
Vol 20 (1-2) ◽  
Author(s):  
N. M. Ali Bacha ◽  
M. Batha ◽  
A. M. Abdul-Kader

In the present study, g2ps1 gene from Gerbera hybrida coding for 2-pyrone synthase which contribute for fungal and insect resistance was used. The aim was to work out an efficient approach of genetic transformation for apple cvs. ‘Golden Delicious’, ‘Royal Gala’ and ‘MM111’, ‘M26’ rootstocks for improving their fungal resistance using genetic engineering techniques. Adventitious shoot formation from leaf pieces of apples studied was achieved using middle leaf segments taken from the youngest leaves from in vitro-grown plants.Optimum conditions for ‚direct’ shoot organogenesis resulted in high regeneration efficiency of  0%, 95%, 92%, 94% in the studied apples respectively. Putative transgenic shoots could be obtained on MS media with B5 Vitamins, 5.0 mg l-1 BAP, or 2.0 mg l-1 TDZ with 0.2 mg l-1 NAA in the presence of the selection agent “PPT” at 3.0-5.0 mgl-1. Shoot multiplication of transgenic shoots was achieved on: MS + B5 vitamins + 1.0 mg l-1 BAP + 0.3 mg l-1 IBA, 0.2 mg l-1 GA3+1.0 g/l MES+ 30 g/l sucrose + 7.0 g/l Agar, with the selection agent PPT at 5.0 mg l-1 and were subcultured every 4 weeks in order to get sufficient material to confirm transformation of the putative shoots obtained. Six, seven, one and six transgenic clones of the apples studied respectively have been obtained and confirmed by selection on the media containing the selection agent “PPT” and by PCR analysis using the suitable primers in all clones obtained for the presence of the selection” bar gene (447 bp) and the gene-of- interest “g2PS1” (1244 bp), with transformation efficiency of 0.4%, 0.6%, 0.1% and 0.3% respectively. These transgenic clones were multiplied further in vitro in the presence of the selection agent ‘PPT’ and rooted in vitro. Rooted transgenic plantlets were successfully acclimatized and are being kept under-containment conditions according to the biosafety by-law in Syria to evaluate their performance for fungal resistance .


2020 ◽  
Vol 51 (3) ◽  
pp. 976-981
Author(s):  
Al- Dabagh & Salih

 Technique of tissue culture for Chia (Salvia hispanica) micropropagation was achieved, this study investigated the impact of various concentrations of plant growth regulators on shoot multiplication and root induction with the Chia’s mature seed as a source explant. The highest percentage of shoot formation (80%), shoots number per explant(3.20) and shoot length(3.26 cm), were recorded on MS medium enriched with BAP(1.0 mgl-1) after eight weeks of seed culture. The optimal medium for the rhizogenesis was achieved on half strength MS medium fortified with 1.0 mgl-1 IBA after four weeks of culture, which had the highest rooting percentage (100%) with highest mean of roots number (5.6 roots per shoot) with (3.40 cm root length). The rooted plants were successfully adapted ex vitro with a survival rate of 85%.


HortScience ◽  
1998 ◽  
Vol 33 (4) ◽  
pp. 749-750 ◽  
Author(s):  
María Victoria González ◽  
Manuel Rey ◽  
Raffaela Tavazza ◽  
Stefano La Malfa ◽  
Luigi Cuozzo ◽  
...  

Plant regeneration was obtained from adventitious buds induced in isolated cotyledons of Italian stone pine (Pinus pinea L.). The best results for bud induction were obtained by using half-strength LePoivre medium with 4.5 μM 6-benzyladenine for 30 days. Shoot elongation was achieved in the same medium without growth regulators but with the addition of 0.5% activated charcoal. The induction medium was the best also for shoot multiplication, but it was necessary to include subcultures on elongation medium. The slow elongation rate of adventitious shoots remains the greatest obstacle to multiplication. Root formation (15%) after 5 months was observed when shoots were cultured on elongation medium for long periods.


1970 ◽  
Vol 18 (2) ◽  
pp. 173-179 ◽  
Author(s):  
T. Mallikadevi ◽  
P. Senthilkumar ◽  
S. Paulsamy

The in vitro regeneration of Plubago zeylanica exhibited that the callus was initiated in the basal medium containing BAP, NAA, 2, 4-D, and IBA.  The high amount (90%) of organic calli was induced in the basal medium supplemented with 2, 4-D, alone at 2.0 mg/l. In the subculture the adventitious shoot formation was prominently higher (83%) in the basal medium containing BAP, and NAA at 3.5 and 0.3 mg/l, respectively. IAA (1.0 mg/l)effectively produced higher percen-tage (90) of roots and root growth. After sequential hardening, survivability rate was observed to be significantly higher (80%) in the hardening medium containing garden soil, sand and vermicompost in the ratio of 1 : 1 : 1 by volume under greenhouse condition.  Key words: Plumbago zeylanica, In vitro regeneration, Medicinal plant D.O.I. 10.3329/ptcb.v18i2.3648 Plant Tissue Cult. & Biotech. 18(2): 173-179, 2008 (December)


1993 ◽  
Vol 118 (1) ◽  
pp. 151-157 ◽  
Author(s):  
Michael E. Compton ◽  
D.J. Gray

Adventitious shoots were obtained from watermelon [Citrullus lanatus (Thunb.) Matsun. & Nakai] cotyledons incubated on a modified Murashige and Skoog medium containing BA. Initial experiments comparing the effects of BA (0, 5, 10, or 20 μm) and IA4 (0, 0.5, or 5 μm) demonstrated that BA was required for adventitious shoot formation but its concentration in the medium was not critical. The addition of IAA to medium with BA increased callus production and inhibited shoot formation. However, the percentage of responding explants in the best treatment was <30%. Therefore, the manner in which cotyledon explants were prepared and seedling age at the time of explantation was examined to improve the organogenic response. The percentage of explants with shoots was improved by using explants that consisted of cotyledon bases (43%) or cotyledons cut in half longitudinally (39%). A lower percentage (16%) of cotyledons cut longitudinally into four pieces produced shoots. Explants taken from the apical half of cotyledons failed to regenerate shoots. Shoot formation was improved further by using explants from young seedlings. The percentage of explants with shoots was >90% for `Minilee', 64% for S86NE, and 50% for `Jubilee II' when explants were prepared from 5-day-old seedlings. Explants from nongerminated embryos or seedlings germinated for 10, 15, or 20 days produced fewer shoots. The effect of several cytokinins on shoot organogenesis was then examined using the optimized protocol. The percentage of explants with shoots and the number of shoots per explant were about two to four times higher when 5 to 10 μm BA was used compared to the most effective kinetin (20 μm) or thidiazuron (0.1 μm) concentration. The percentage of explants with shoots and the number of shoots per explant were greater for diploid (57% and 2.2, respectively) than for triploid (22% and 0.6, respectively) or tetraploid (20% and 0.8, respectively) lines. Chemical names used: N -(phenylmethyl)-1 H -purin-6-amine (BA); 6-furfurylaminopurine (kinetin); N -phenyl-N' -1,2,3-thiadiazol-5-ylurea (thidiazuron); 1 H -indole3-acetic acid (IAA).


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