scholarly journals Simplified Regeneration Protocol for Cycas revoluta Thunb. Mature Zygotic Embryos

2015 ◽  
Vol 7 (1) ◽  
pp. 62-65
Author(s):  
Rohangiz NADERI ◽  
Khadije MOHAISENI ◽  
Jaime A. TEIXEIRA DA SILVA ◽  
Mansour OMIDI ◽  
Behjat NADERI
Keyword(s):  
Zygotic Embryo ◽  
Basal Medium ◽  
Adventitious Shoot ◽  
Zygotic Embryos ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Indirect Organogenesis ◽  
Rooted Plantlet ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Cycas Revoluta

Mature zygotic embryos of Cycas revoluta Thunb. were used as explants to investigate direct and indirect organogenesis. Explants were incubated on half-strength Murashige and Skoog (½ MS) basal medium supplemented with various plant growth regulators, singly or in combination (all at 0.5 mg l-1): 6-benzyladenine (BA), kinetin (Kin), 2,4-dichlorophenoxyacetic acid (2,4-D), Kin×2,4-D, BA×Kin and BA×2,4-D. Cultures were placed at a low light intensity (4 µmol m-2 s-1 PPFD). Adventitious shoot regeneration was observed in the presence of 0.5 mg l-1BA after 35 days. The highest number of direct and indirect shoots per zygotic embryo was 3.67 and 29.67, respectively. Roots were induced on indirect shoots by continuous culture on rooting medium (½ MS,‏ 0.1 mg l-1 1-naphthaleneacetic acid) and hardened successfully in perlite. Each rooted plantlet with pinnate leaves and a primary tap root was individually isolated and acclimatized 185 days after the beginning of culture, with a 10% success rate.

Two alternative pathways of somatic embryo origin from polyembryonic mature stored seeds of Pinus koraiensis Sieb et Zucc.

1997 ◽  
Vol 75 (3) ◽  
pp. 509-512 ◽  
Author(s):  
P. V. Bozhkov ◽  
I. S. Ahn ◽  
Y. G. Park
Keyword(s):  
Somatic Embryo ◽  
Somatic Embryos ◽  
Zygotic Embryo ◽  
Pinus Koraiensis ◽  
Zygotic Embryos ◽  
Dichlorophenoxyacetic Acid ◽  
Strong Negative Correlation ◽  
Alternative Pathways ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Embryo Origin

Individual mature stored seeds of Pinus koraiensis sometimes contain several viable zygotic embryos originated through the processes of simple and cleavage polyembryony. To induce the embryonic process, isolated zygotic embryos were cultured on five different media all supplemented with 10 μM 2,4-dichlorophenoxyacetic acid and 5 μM 6-benzyladenine. Two alternative pathways of somatic embryo origin were revealed. The first pathway was associated with the production of a friable, translucent callus in the hypocotyls–cotyledon region of the dominant zygotic embryo. The second pathway was related to the proliferation of a translucent, moist, and mucilaginous tissue (termed embryonal–suspensor mass) in the suspensor region of the dominant zygotic embryo. Both types of tissues contained early somatic embryos. Regression analysis has shown a strong negative correlation between the frequencies of formation of embryogenic callus and embryonal–suspensor mass both at 3 and 8 weeks of culture (r = − 0.85; p = 0.07 and r = −0.71; p = 0.17, respectively). Key words: Pinus koraiensis; polyembryonal seeds; somatic embryogenesis; embryogénie callus; embryonal–suspensor mass.

scholarly journals Shoot Generation and Callus Induction of Dioscorea hispida Dennst by Different Plant Growth Hormones and Basal Media

2020 ◽  
Vol 11 (2) ◽  
pp. 30-38
Author(s):  
Nor Hasima Mahmod ◽  
Zakiah Mustapha ◽  
Ahmad Hilman Ariffin Husni ◽  
Nurul Anisah Ishak ◽  
Hafsah Jaafar
Keyword(s):  
Callus Culture ◽  
Basal Medium ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Plant Growth Hormones ◽  
Efficient Type ◽  
Medicinal Properties ◽  
Tuber Sprouting ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Stem Segments

Dioscorea hispida Dennst produces tuber which possess valuable medicinal properties but unsustainable harvesting has led to its reduction. The plant propagates slowly because of its low tuber sprouting rate. In average, Dioscorea hispida Dennst tubers took approximately 60 d to break dormancy and sprout. Hence, callus culture is proposed as a possible efficient type of culture for manipulation of this species.  In the present study, calli were induced from stem segments to evaluate callus culture potential of Dioscorea hispida Dennst. Results indicate that the combination of 1 mgL-1 naphthaleneacetic acid (NAA), 1 mgL-1 6- benzylaminopurine (BAP) and 0.5 mgL-1 2,4-dichlorophenoxyacetic acid (2, 4-D) in Gamborg (B5) medium improved callus multiplication and differentiation in the stem culture as opposed to those in Murashige and Skoog (MS) medium. The findings from the present study provide the basis of callus culture protocol for stem explant of Dioscorea hispida Dennst with B5 being the more effective basal medium.

scholarly journals Somatic Embryogenesis and Organogenesis in Magnolia dealbata Zucc. (Magnoliaceae), an Endangered, Endemic Mexican Species

HortScience ◽  
2006 ◽  
Vol 41 (5) ◽  
pp. 1325-1329 ◽  
Author(s):  
Martín Mata-Rosas ◽  
Ángel Jiménez-Rodríguez ◽  
Victor M. Chávez-Avila
Keyword(s):  
Somatic Embryogenesis ◽  
Basal Medium ◽  
Direct Somatic Embryogenesis ◽  
Direct Organogenesis ◽  
Limiting Factor ◽  
Zygotic Embryos ◽  
Dichlorophenoxyacetic Acid ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Magnolia Dealbata

Plants of Magnolia dealbata were regenerated from zygotic embryos through somatic embryogenesis and direct organogenesis. Medium and incubation conditions were determinating factors for the development of morphogenetic responses. Photoperiodic exposure was a limiting factor in the general development of the explants, and incubation in darkness allowed their development. The highest formation of shoots per responding explant were obtained on woody plant (WP) medium supplemented with 13.3 μM or 22.2 μM 6-benzylaminopurine (BA) in combination with 2.26 μM or in absence of 2,4-dichlorophenoxyacetic acid (2,4-D) from which 2.5 shoots per explant were induced. Subcultures on WP medium, supplemented with polyvinylpyrrolidone (PUP) 40,000 1 g·L–1) avoided necrosis of explants. Somatic embryos were formed in 85% of explants cultivated on WP medium with 2,4-D (2.3 μM or 4.5 μM); 20% induced indirect embryogenesis and 65% formed direct somatic embryogenesis. The plants were transferred to soil to acclimatize under greenhouse conditions, achieving 90% survival. Somatic embryo conversion to plantlets was obtained with subculture on WP basal medium without growth regulators. In vitro culture can play a key role in the propagation and conservation of this endangered species.

Embryo induction and plant regeneration of Callerya speciosa (Fabaceae) through anther culture

2017 ◽  
Vol 65 (1) ◽  
pp. 80 ◽  
Author(s):  
Bilan Huang ◽  
Li Xu ◽  
Kelie Li ◽  
Yunlu Fu ◽  
Zhiying Li
Keyword(s):  
Anther Culture ◽  
Culture Medium ◽  
Basal Medium ◽  
Dichlorophenoxyacetic Acid ◽  
Anther Wall ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Anther Cultures

An in vitro protocol for Callerya speciosa (Champ.) Schot regeneration through embryogenesis was developed using the anthers as the explants. The late uninucleate stage of the microspore was optimal for the anther culture of C. speciosa. Embryonic callus was induced on a MS basal medium supplemented with 4.4 µM 6-benzylaminopurine (BA) and 9.04 µM 2,4-dichlorophenoxyacetic acid (2,4-D). Embryos were obtained on MS medium supplemented with 2.2 µM BA and 0.5 µM naphthaleneacetic acid (NAA). The highest percentage (16.7%) of embryos was achieved using the culture medium MS + 0.25 µM NAA + 1.1 µM BA. The highest percentage of embryos that developed into plants was 18.3%. However, haploid plants were not observed, which may have been due to the collection of the calli from the anther wall. The results presented here demonstrate the establishment of a highly efficient and rapid system for regenerating C. speciosa using anther cultures.

scholarly journals Somatic embryogenesis from leaf transverse thin cell layer derived-callus of Vietnamese ginseng (Panax vietnamensis Ha et Grushv.)

2016 ◽  
Vol 14 (1) ◽  
pp. 63-73
Author(s):  
Vu Thi Hien ◽  
Nguyen Phuc Huy ◽  
Bui Van The Vinh ◽  
Hoang Xuan Chien ◽  
Hoang Thanh Tung ◽  
...  
Keyword(s):  
Somatic Embryogenesis ◽  
Cell Layer ◽  
Culture Media ◽  
Callus Formation ◽  
Basal Medium ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Transverse Thin Cell Layer ◽  
Cell Layers ◽  
2,4 Dichlorophenoxyacetic Acid

No report on plant regeneration via somatic embryogenesis of P. vietnamensis has been previously published. In the present study, somatic embryogenesis via callus formation from cultures of leaf transverse thin cell layers (tTCLs) of Vietnamese ginseng (Panax vietnamensis Ha et Grushv.) was investigated. α-naphthaleneacetic acid (NAA), 2,4-dichlorophenoxyacetic acid (2,4-D), 6-benzylaminopurine (BA) and thidiazuron (TDZ) were added separately and in combination into the culture media. Explant necrosis or low callogenesis rates were observed when 1-mm wide leaf tTCLs were cultured on media with TDZ, BA, 2,4-D or NAA. On the other hand, calli were successfully induced from the tTCL explants cultured on medium supplemented with either 2,4-D and BA or 2,4-D and TDZ. Callogenesis was observed under both light and dark conditions. The highest callogenesis rate (100%) was obtained on Murashige and Skoog (MS) basal medium supplemented with 1.0 mg l-1 2,4-D in combination with 0.1 mg l-1 TDZ in darkness after eight weeks of culture. White calli were cut into small pieces (1.0 x 1.0 cm dimension) and placed on MS media containing 1.0 mg l-1 2,4-D, 0.5 mg l-1 NAA and TDZ at various concentrations (0.01; 0.1; 0.2; and 0.5 mg l-1), and the best callus proliferation was recorded on medium containing 1.0 mg l-1 2,4-D and 0.2 mg l-1 TDZ. Somatic embryogenesis, with a success rate of 53.3% and 35 embryos per explant, was achieved when calli were subcultured onto MS medium supplemented with 1.0 mg l-1 2,4-D, 0.5 mg l-1 NAA and 0.2 mg l-1 TDZ.

scholarly journals Regeneration of Ornamental Cherry (Prunus) Taxa from Mature Stored Seed

HortScience ◽  
2000 ◽  
Vol 35 (4) ◽  
pp. 745-748 ◽  
Author(s):  
Karen E. Hokanson ◽  
Margaret R. Pooler
Keyword(s):  
Callus Formation ◽  
Adventitious Shoot ◽  
Shoot Formation ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Adventitious Shoot Regeneration ◽  
Potential System ◽  
Regeneration In Vitro ◽  
2,4 Dichlorophenoxyacetic Acid

Callus formation and adventitious shoot regeneration in vitro from mature stored seed were evaluated in eight ornamental cherry (Prunus) taxa: P. campanulata Maxim., P. maackii Rupr., P. sargentii Rehd., P. serrula Franch., P. serrulata Lindl., P. subhirtella Miq., P. virginiana L., and P. yedoensis Matsum. Several portions of the embryo (cotyledons and hypocotyl sections) and nine combinations of growth regulators (BA, 2,4-D, IBA, NAA, and TDZ) were compared. Effects of embryo portions and growth regulator treatments were generally small within taxa, but shoot formation differed among taxa. About 20% to 50% of the embryos from P. virginiana and P. serrula and ≈5% to 30% of those from P. maackii produced shoots. The other taxa generally did not produce shoots. Regeneration from mature stored seed in the responsive taxa represents a potential system for genetic transformation. Chemical names used: 6-benzyladenine (BA); 2,4-dichlorophenoxyacetic acid (2,4-D); indole-3-butyric acid (IBA); α-naphthaleneacetic acid (NAA); thidiazuron (TDZ).

Callus formation and shoot bud differentiation in anther culture of Solanum surattense

1979 ◽  
Vol 57 (22) ◽  
pp. 2524-2527 ◽  
Author(s):  
S. Sinha ◽  
R. P. Roy ◽  
K. K. Jha
Keyword(s):  
Anther Culture ◽  
Callus Formation ◽  
Low Frequency ◽  
Basal Medium ◽  
Pollen Grains ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Cytological Examination ◽  
Shoot Bud ◽  
2,4 Dichlorophenoxyacetic Acid

In anther culture of Solatium surattense, the Murashige and Skoog's medium supplemented with 2,4-dichlorophenoxyacetic acid (2.2 mg/L), indoleacetic acid or naphthaleneacetic acid (1.9 mg/L), and kinetin (2.2 mg/L) served as “callus-producing medium.” Histological and cytological observations indicated that the callus originated from the pollen grains. Synergistic action of kinetin (5.0 mg/L) and coconut milk (15%) in basal medium was able to induce differentiation of shoot buds either from the anthers directly or from the callus. Directly differentiating buds were formed by whole shoot bud morphogenesis of pollen. They were produced at a low frequency and showed presence of well-developed radicular and plumular regions. But the buds originating from callus lacked radicular ends. Root initiation in such buds was achieved by transferring them to basal medium. Cytological examination of the androgenic plantlets revealed a chromosomal series ranging from the haploid to the hexaploid with a few aneuploids.

The Effect of Auxins and Antiauxins on Shoot-Bud Induction and Morphology in the Moss, Bryum atrovirens Will ex Brid

1990 ◽  
Vol 38 (2) ◽  
pp. 177 ◽  
Author(s):  
RN Chopra ◽  
BD Vashistha
Keyword(s):  
Basal Medium ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Simultaneous Application ◽  
Shoot Bud ◽  
Cultural Conditions ◽  
Bud Induction ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Indole 3 Acetic Acid ◽  
Do So

The protonema of the moss Bryum atrovirens remains bud-free under ordinary cultural conditions on Nitsch's basal medium. Exogenously applied auxins (Indole-3-acetic acid; 2-4-dichlorophenoxyacetic acid; α-naphthaleneacetic acid and β-naphthoxyacetic acid) induced buds on protonemata, whereas antiauxins (Maleic hydrazide and 2,3,5-triiodobenzoic acid) failed to do so. Morphology of the gametophores depended upon the concentration of auxin in the medium. In general, normal leafy gametophores resulted at lower concentrations, and at higher levels of auxins morphology was adversely affected. Simultaneous application of 6-benzylaminopurine and 2,4-dichlorophenoxyacetic acid advanced bud formation as well as increased bud number, but had no significant effect on the improvement of shoot morphology.

scholarly journals 099 Indirect and Direct Regeneration of Kalmia latifolia

HortScience ◽  
1999 ◽  
Vol 34 (3) ◽  
pp. 458D-458
Author(s):  
Mark H. Brand
Keyword(s):  
Shoot Regeneration ◽  
Leaf Explants ◽  
Adventitious Shoot ◽  
Direct Regeneration ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Kalmia Latifolia ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Optimal Medium ◽  
Silver Dollar

To introduce desirable trait genes into Kalmia latifolia, efficient adventitious shoot regeneration methods are needed. Silver Dollar (S$) callus induction and growth in the dark was compared on Woody Plant (WP) medium containing 2,4-dichlorophenoxyacetic acid (2,4-D) (1, 5, 10, 20 μM) or naphthaleneacetic acid (NAA) (1, 10, 20, 40 μM) with and without 5 μM isopentenyladenine (2iP). Both 2,4-D and NAA produced >450 mg of callus from leaf explants in 8 weeks. The addition of 2iP tripled growth for 2,4-D and doubled growth for NAA. Greatest callus growth was obtained on 20-40 μM NAA or 5-20 μM 2,4-D. Shoot regeneration on callus was achieved on WP medium containing 30 μM 2iP or 1 μM thidiazuron (TDZ), but a combination of the two was best, with 68% of dark-grown calli regenerating shoots in 4 weeks. 26% more dark-grown calli regenerated shoots than light-grown calli. The type of auxin (2,4-D or NAA) used to grow the calli did not affect shoot regeneration. For direct shoot regeneration, S$ leaf explants were tested on WP medium containing 5, 15, 30, 45 and 60 μM 2iP. The addition of 1 μM indole-3-butyric acid (IBA) doubled the percentage of leaves that regenerated shoots. 2iP concentrations between 15 and 45 μM supported excellent shoot regeneration, but optimal regeneration (95% of explants, 5.1 shoots/leaf) occurred on 30 μM 2iP+1 μM IBA. Leaf explants of six cultivars were grown on optimal medium with shoot regeneration ranging from 17% to 93% of leaves and 1.8 to 8.2 shoots per leaf, depending on the cultivar.

scholarly journals (282) In Vitro Indirect Organogenesis of Leucocoryne purpurea, an Ornamental Geophyte Species Endemic to Chile

HortScience ◽  
2005 ◽  
Vol 40 (4) ◽  
pp. 1051B-1051
Author(s):  
Luis Humberto Escobar Torres ◽  
Eduardo Alejandro Olate Muñoz ◽  
Miguel Jordan ◽  
Marlene Gebauer
Keyword(s):  
Callus Induction ◽  
Basal Medium ◽  
Average Frequency ◽  
Dichlorophenoxyacetic Acid ◽  
Fresh Weight ◽  
Root Tips ◽  
Indirect Organogenesis ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Different Sources

Callus induction (CI) and later shoot induction (SI) were studied in Leucocoryne purpurea, a native and endemic Chilean geophyte species. Basal leaf portions (BL), bulb basal plate (BP), and root tips (RT) from in vitro plants were used as explants. Treatments for CI included all three explants and media containing different sources and concentrations of auxins and cytokinins as plant growth regulators (PGRs). Plant material was initiated on MS basal medium (Murashige and Skoog, 1962), supplemented with vitamins, 30 g·L-1 sucrose, 6.0 g·L-1 agar and pH adjusted to 5.7 before autoclaving. The experiments were carried on a growth chamber at 24 ± 1.5 °C. CI cultures were maintained in darkness for 16 weeks, and SI for 12 weeks in a 16-hour photoperiod. BL and RT explants did not respond to any of the CI treatments. BP explants cultured on MS basal medium without PGRs also did not produce any callus. The average frequency of callus induction for BP was 78% and the average fresh weight of callus was 10.06 g/explant after 16 weeks of culture. Best treatment for CI was BP cultured on 4.52 μm 2,4-dichlorophenoxyacetic acid (2,4-D) in combination with 0.45 μm 6-benzyladenine (BA), when they were compared to 2,4-D alone or picloram as auxin source. After 16 weeks of culture, calli were transferred to SI medium, supplemented with three different concentrations of thidiazuron (TDZ), either intact or subdivided (150 mg/explant). SI treatments had a greater and significant response when the callus came from a CI medium containing auxin and cytokinin combined, in comparison to those coming from a CI medium containing auxins only.

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