In Vitro Flowering of Plantlets Regenerated from Zygotic Embryo-derived Somatic Embryos of Ginseng

Mature zygotic embryos dissected from ginseng (Panax ginseng C.A. Meyer) seeds were cultured on Murashige and Skoog (MS) medium containing various concentrations of 2,4-D and kinetin. Somatic embryos were induced directly from cotyledonary tissue and from intervening callus. The frequency of somatic embryo induction was up to 55% of zygotic embryo explants. Upon transfer onto half-strength MS medium supplemented with 1 mg BA/liter and 1 mg GA3/liter, most somatic embryos developed into plantlets. More than 50% of the plantlets flowered after 4 weeks of culture, and some developed immature fruits in vitro. These results indicate that adulthood of ginseng root explants is not a prerequisite for flowering of plantlets regenerated through somatic embryogenesis. Chemical names used: (2,4 -dichlorophenoxy) acetic acid (2,4-D); N-(2-furanylmethyl) -1H-purin-6-amine(kinetin); N-(phenylmethyl) -1H-purin-6-amine (BA); gibberellic acid (GA3).

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High-frequency Direct Somatic Embryogenesis and Plantlet Regeneration from Leaves Derived from In Vitro-Germinated Seedlings of a Coffea arabica Hybrid Cultivar

HortScience ◽

10.21273/hortsci10771-16 ◽

2016 ◽

Vol 51 (9) ◽

pp. 1148-1152 ◽

Cited By ~ 4

Author(s):

Jane Kahia ◽

Margaret Kirika ◽

Hudson Lubabali ◽

Sinclair Mantell

Keyword(s):

Tissue Culture ◽

High Frequency ◽

Somatic Embryos ◽

Coffea Arabica ◽

Alternative Methods ◽

Direct Somatic Embryogenesis ◽

Ms Medium ◽

Embryogenic Cultures ◽

Half Strength

Breeding work carried out during the period 1971–85 by the Coffee Research Institute, Ruiru, Kenya resulted in the release of a new improved hybrid Coffea arabica named Ruiru 11. The cultivar combines resistance to coffee berry disease (CBD) and leaf rust, with high yield and good cup quality attributes. The propagation by F1 hybrid seeds production, cuttings, and tip grafting do not produce enough planting materials. There was a need to explore alternative methods and tissue culture offers potential options. The objective of the study was to evaluate the effect of explant sources and cytokinins on induction and regeneration of somatic embryos. Eight different explants were cultured on half-strength Murashige and Skoog (MS) medium supplemented with 10 µm benzylaminopurine (BAP). The effect of kinetin, N6-(2-isopentyl) adenine (2iP) evaluated at (0, 0.5, 5, or 25 µm) or thidiazuron (TDZ) (0, 0.5, 1.0, or 5 µm) added in separate experiments was also evaluated. The percentage of embryogenic cultures and the numbers of embryos per explant were determined after 3 months’ culture. The explant type had a significant effect (P > 0.05) on the induction of somatic embryos. Explants from in vitro-germinated seedlings produced the highest embryogenic cultures (90%) and the highest mean number of embryos (19.36) per explant. Cytokinins strongly enhanced induction and regeneration of somatic embryos. TDZ at 1 µm produced the highest embryogenic cultures (100%) and the highest mean number of embryos (24.2). The embryos were germinated on half-strength MS medium without any hormones. A high (98%) survival rate of the regenerated plantlets was recorded over all the treatments in the greenhouse. This is the first report on induction of high-frequency direct somatic embryos from coffee juvenile tissues. This is of great significance in tissue culture and indeed molecular biology manipulations because it allows regeneration of coffee from several explants.

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Coconut Tissue Culture: The Indian Initiatives, Experiences and Achievements

CORD ◽

10.37833/cord.v33i2.48 ◽

2017 ◽

Vol 33 (2) ◽

pp. 11

Author(s):

Anitha Karun

Keyword(s):

Tissue Culture ◽

Somatic Embryos ◽

Culture Media ◽

Zygotic Embryo ◽

Shoot Meristem ◽

Direct Organogenesis ◽

Zygotic Embryos ◽

Immature Inflorescence ◽

High Yielding Varieties

Coconut is one of the principal crops of India cultivated in over 35 districts mainly in the southern states. The productivity of the crop is declining in many of the traditionally cultivated regions owing to ageing plantations as well as biotic and abiotic stresses. These plantations are to be replanted with high yielding varieties/hybrids for which adequate quantity of quality planting material is not available. Even though tissue culture research was initiated in many laboratories in the country, the work was eventually phased out in most of the laboratories for want of a repeatable protocol. At ICAR-CPCRI, coconut tissue culture programs have been continuing for the past three decades. The attempts made include experimentation with different explants viz., immature inflorescence, plumular tissues, mature palm shoot meristem, ovary and anthers and different culture media supplemented with varying levels and types of hormones. Some of the successful protocols developed at the Institute include coconut zygotic embryo culture for collection and exchange of germplasm, cryopreservation and retrieval of zygotic embryos and pollen and plantlet regeneration from plumular tissues. Even though ICAR-CPCRI has succeeded in obtaining plantlets via direct organogenesis from inflorescence explants, the absence of friable calli formation from explants, the low rate of somatic embryo formation, large number of cultures turning to abnormal shoot development, non conversion of somatic embryos into plantlets, and formation of abnormal somatic embryos remain the major bottlenecks. Gene expression studies are being currently undertaken to decipher the molecular basis of in vitro recalcitrance.

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Enhanced Somatic Embryo Induction of a Tree Peony, Paeonia ostii ‘Fengdan’, by a Combination of 6-benzylaminopurine (BA) and 1-naphthylacetic Acid (NAA)

Plants ◽

10.3390/plants9010003 ◽

2019 ◽

Vol 9 (1) ◽

pp. 3 ◽

Cited By ~ 2

Author(s):

Xiuxia Ren ◽

Ya Liu ◽

Byoung Ryong Jeong

Keyword(s):

Somatic Embryogenesis ◽

Somatic Embryos ◽

Zygotic Embryo ◽

Zygotic Embryos ◽

Ms Medium ◽

Hypocotyl Explants ◽

Cotyledon Explants ◽

Embryo Induction ◽

Naphthylacetic Acid ◽

Better Than

Somatic embryogenesis is a preferred method for vegetative propagation due to its high propagation efficiency. In this study, zygotic embryos, cotyledons, and hypocotyls of Paeonia ostii ‘Fengdan’ were used as the explant to induce somatic embryogenesis. The results showed that a combination of 0.5 mg·L−1 thidiazuron (TDZ) and 0.5 mg·L−1 2,4-dichlorophenoxyacetic acid (2,4-D) was effective in inducing somatic embryos from the zygotic embryo and cotyledon explants. Hypocotyls only formed somatic embryos on Murashige and Skoog (MS) medium supplemented with both 0.5 mg·L−1 TDZ and 0.5 mg·L−1 1-naphthylacetic acid (NAA). Moreover, the compact callus was effectively produced from zygotic embryo, cotyledon, and hypocotyl explants in medium supplemented with a combination of 3.0 mg·L−1 6-benzylaminopurine (BA) and 1.0 mg·L−1 NAA, and then converted into somatic embryos in the same medium, and the ratio of the explants with embryo induction and number of embryos induced per explant were much higher than those induced by 0.5 mg·L−1 TDZ and either 0.5 mg·L−1 2,4-D or 0.5 mg·L−1 NAA. The MS medium was better than the woody plant medium (WPM) for inducing somatic embryos from zygotic embryo and hypocotyl explants, whereas the WPM was better than the MS medium for somatic embryogenesis induction from cotyledon explants. All of the somatic embryos developed well into mature embryos on their respective media supplemented with both 3.0 mg·L−1 BA and 1.0 mg·L−1 NAA. Overall, the protocols for indirect somatic embryogenesis from zygotic embryo, cotyledon, and hypocotyl of P. ostii ‘Fengdan’ were successfully established, which can greatly facilitate their propagation and breeding processes.

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Can Agricultural Biotechnology Help Guava Growing in Temperate Climate

HortScience ◽

10.21273/hortsci.39.4.861b ◽

2004 ◽

Vol 39 (4) ◽

pp. 861B-861

Author(s):

Bipul K. Biswas* ◽

Nirmal Joshee ◽

Anand K. Yadav

Keyword(s):

Tissue Culture ◽

Somatic Embryos ◽

Adventitious Shoots ◽

Multiple Shoots ◽

Cold Climate ◽

Nodal Explants ◽

Temperate Climate ◽

Zygotic Embryos ◽

Ms Medium

Guava (Psidium guajava L.), also called `apple of tropics,' is immensely nutraceutical and horticulturally important. Being a tropical plant, it cannot stand temperatures below 25° F and needs frost protection to grow in temperate regions. To adapt in cold climate, cold hardy guava cultivars are needed. Conventional ways are uneconomic in time and efforts. Still, transgenic plants developed using biotechnological approaches of tissue culture and rDNA technology, appear to have great potential. Thus, protocols for in vitro propagation of guava were developed via organogenesis and somatic embryogenesis using nodal explants from mature trees and young zygotic embryos, respectively. Nodal explants induced multiple shoots when cultured on MS medium fortified with KIN, BAP and Ad.S. Adding a (NO3)2 to medium was useful to prevent in vitro shoot tip browning of adventitious shoots. Rocker liquid culture greatly increased growth of multiple shoots compared to the agar-based medium. It appears to be a good tool for woody plant tissue culture. Induction of somatic embryos in guava was also achieved on MS medium supplemented with IAA auxin. About 80% to 90% somatic embryos germinated normally. To achieve Agro-bacterium-mediated gene transfer in guava, on-going co-cultivation of organogenic tissues of guava is to optimize protocols for freeze tolerance gene (CBF1, CBF2, CBF3) transfer. Plasmid vectors containing selectable markers (nptII gene for antibiotic selection and GUS reporter gene as scorable gene mediated selection), with CaMV 35S promoter gene has been introduced into guava tissues and the resultant plants showed antibiotic resistance. Details of the experimental procedures and up-to-date results will be discussed.

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Somatic embryogenesis from zygotic embryos of Euterpe oleracea Mart.

Revista Brasileira de Fruticultura ◽

10.1590/s0100-29452002000300004 ◽

2002 ◽

Vol 24 (3) ◽

pp. 601-603 ◽

Cited By ~ 12

Author(s):

Ana da Silva Ledo ◽

Osmar Alves Lameira ◽

Abdellatif Kemaleddine Benbadis ◽

Ilmarina Campos de Menezes ◽

Maria do Socorro Padilha de Oliveira ◽

...

Keyword(s):

Somatic Embryogenesis ◽

Somatic Embryos ◽

Zygotic Embryos ◽

Ms Medium ◽

Euterpe Oleracea ◽

Mineral Salts ◽

Vegetable Material ◽

Morphogenetic Responses ◽

Euterpe Oleracea Mart

The aim of this work was to study the morphogenetic responses of zygotic embryos of açai palm (Euterpe oleracea Mart.) submitted to several conditions of in vitro culture. Several research experiments were conducted, in laboratory, using vegetable material collected from açai palm plants at Embrapa Amazon Oriental, Belém-PA, Brazil. It was possible to verify the expression of a direct, repetitive and no-synchronized model of somatic embryogenesis in mature zygotic embryos cultivated in primary MS medium supplemented with 2,4-D (339.36 muM) and transferred to a secondary MS medium in the presence of NAA (0.537 muM) and 2iP (12.30 muM). The conversion of somatic embryos into seedlings was reached after 210 days with the transfer of the cultures to a third medium with sucrose and mineral salts concentrations reduced to a half, without growth regulators.

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Somatic Embryogenesis and Plantlet Regeneration in the Carica papaya L. cv. Eksotika

Plants ◽

10.3390/plants9030360 ◽

2020 ◽

Vol 9 (3) ◽

pp. 360

Author(s):

Baker Al-Shara ◽

Rosna Mat Taha ◽

Jamaludin Mohamad ◽

Hashimah Elias ◽

Asif Khan

Keyword(s):

Butyric Acid ◽

Somatic Embryos ◽

Carica Papaya ◽

Callus Formation ◽

Peat Moss ◽

Zygotic Embryos ◽

Ms Medium ◽

Immature Zygotic Embryos ◽

Adenine Sulphate

A highly efficient protocol for regeneration of Carica papaya L. cv. Eksotika somatic embryos from immature zygotic embryos was developed. This study was designed to overcome the obstacles in regeneration of somatic embryos from immature zygotic embryos of “Eksotika”, especially problems associated with formation of better root quality and callus formation at the base of somatic embryos. Somatic embryos were generated by incubation of immature zygotic embryos in half-strength salt Murashige and Skoog (MS) medium with full-strength vitamins supplemented with 7.5 mg L−1 2,4-D, 100 mg L−1 L-glutamine, 50 mg L−1 myo-inositol, 45 mg L−1 adenine sulphate, 0.33% gelrite, and 6% sucrose, followed by transfer to maturation medium consisting of ½ MS medium supplemented with 5 mg L−1 phloroglucinol, 100 mg L−1 L-glutamine, 100 mg L−1 myo-inositol, 68 mg L−1 adenine sulphate, 0.38% gelrite, and 3% sucrose. After that, well-formed somatic embryos were transferred to MS medium containing 3% sucrose and 0.8% agar for shoot production. The embryos were elongated in MS medium supplemented with 1 mg L−1 gibberellic acid, 0.5 mg L−1 indole-3-butyric acid, 100 mg L−1 myo-inositol, and 3.76 mg L−1 riboflavin. Root regeneration was achieved on MS medium containing 7.9 mg L−1 phloroglucinol and supported with vermiculite after 4 days of cultivation on ½ MS medium with 2 mg L−1 indole-3-butyric acid. After the rooting phase, in vitro plantlets were acclimatized in peat moss soil.

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Regeneration potential of different explants during micropropagation of neem tree (Azadirachta indica A. Juss.)

Journal of Phytology ◽

10.25081/jp.2021.v13.7352 ◽

2021 ◽

pp. 171-177

Author(s):

Bhavadharani Dhandapani ◽

Gnanam Ramasamy ◽

Senthil Natesan ◽

Kumaran Kalayanasundaram

Keyword(s):

Azadirachta Indica ◽

Zygotic Embryo ◽

Casein Hydrolysate ◽

Nodal Segments ◽

Naphthalene Acetic Acid ◽

Zygotic Embryos ◽

Ms Medium ◽

Regeneration System ◽

Regeneration Potential ◽

Half Strength

Azadirachta indica A. Juss., (Neem), a prodigious multipurpose tree, has immense potential to benefit mankind and to protect the environment. In order to investigate the effects of three different explants for its regeneration potential, de embryonated cotyledon, immature zygotic embryo and nodal segments from a 30 year old neem plus tree were used. Half strength MS medium with benzyl amino purine (3 mg/L) and naphthalene acetic acid (0.5 mg/L) and casein hydrolysate (1 g/L) was effective in shoot bud sprouting from both nodes and cotyledons. Half strength MS medium fortified with TDZ (0.2 mg/L) was effective for induction of somatic embryogenesis from zygotic embryos. Shoot buds initiated from the cotyledons produced a maximum number of shoots per explants (4.33) which on further sub culturing induced maximum multiple shoots (15) on half strength MS medium fortified with BAP (1.5 mg/L), NAA (0.5 mg/L) and CH (400 g /L) and the nodal explants induced only 4-5 axillary shoots on further sub culturing. Even though immature zygotic embryos produced more number of somatic embryos per explant (24.97) within a short time (30-45 days), the plantlet conversion was poor (25.52 %). In vitro rooting was observed in half strength MS medium supplemented with IBA (2 mg/L). The regeneration potential of de embryonated cotyledons through a simple regeneration system may be beneficial for efficient mass propagation of selected plus trees of neem.

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Somatic Embryogenesis in Papaya (Carica papaya L. cv. TNAU Papaya CO.8)

Current Journal of Applied Science and Technology ◽

10.9734/cjast/2020/v39i330510 ◽

2020 ◽

pp. 18-26

Author(s):

C. K. Rajesh ◽

D. Sudhakar ◽

K. K. Kumar ◽

C. Kavitha ◽

G. Karthikeyan ◽

...

Keyword(s):

Somatic Embryogenesis ◽

Growth Regulators ◽

Callus Induction ◽

Somatic Embryos ◽

Carica Papaya ◽

Naphthalene Acetic Acid ◽

Zygotic Embryos ◽

Ms Medium ◽

Half Strength ◽

Cotyledonary Stage

An efficient indirect somatic embryogenesis protocol for Carica papaya var TNAU Papaya CO.8 was developed using immature zygotic embryos as an explant. Two growth regulators namely 2,4-D and picloram each at 1, 2, 3 mg/L were tested for callus induction and the highest callus induction frequency (83.33%) was observed in MS medium supplemented with 3 mg/L 2,4-D. However the rate of conversion into somatic embryos was highest (63.33%) on MS medium supplemented with 2 mg/L 2,4-D. Maturation of somatic embryos was studied by using MS medium with different concentrations of abscisic acid (ABA) and benzyl amino purine (BAP) along with glutamine (400 mg/L). The maturation of globular embryos was observed to be higher in the combination of ABA (1.5 mg/L), BAP (0.4 mg/L) along with glutamine (400 mg/L). Even though regeneration was observed from cotyledonary stage embryos in presence of different growth regulators like BAP, α-naphthalene acetic acid (NAA), phloridzin dehydrate kinetin and gibberellic acid, further growth was not observed due to abnormal regenerative structures. Regeneration of cotyledonary stage somatic embryos were highest (77.4%) in half strength MS medium without growth regulators. The well-developed plantlets with shoots and roots were subsequently transferred for hardening.

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Somatic Embryogenesis in Members of the Plumbaginaceae Ornamental Statice Limonium and Sea Thrift Armeria maritima

HortScience ◽

10.21273/hortsci.37.7.1122 ◽

2002 ◽

Vol 37 (7) ◽

pp. 1122-1123 ◽

Cited By ~ 1

Author(s):

Mohammed A.M. Aly ◽

Bala Rathinasabapathi ◽

Sheevani Bhalsod

Keyword(s):

Acetic Acid ◽

Somatic Embryogenesis ◽

Somatic Embryos ◽

Ms Medium ◽

Cotyledon Explants ◽

Poor Seed ◽

Armeria Maritima ◽

Dichlorophenoxy Acetic Acid ◽

Somatic Embryo Induction ◽

Better Than

Many members of the Plumbaginaceae are important flower crops wherein propagation is hindered by poor seed germination. Micropropagation via organogenesis is commercially practiced for certain Limonium species. However, somatic embryogenesis was not reported for members of the Plumbaginaceae until recently for L. bellidifolium Durmort. The induction of somatic embryogenesis from cotyledon explants in a modified Murashige and Skoog (MS) medium was examined in four other members of this family, Limonium aureum O. Kuntze, L. latifolium O. Kuntze, L. sinuatum Mill., and Armeria maritima Willd. Induction of embryogenic callus was achieved in all the species examined on MS medium supplemented with 4.5 μm 2,4-D and 88 or 118 mm sucrose. Species of the genus Limonium responded better than A. maritima Willd. in somatic embryo induction and maturation. Somatic embryos of L. aureum O. Kuntze matured readily on MS medium supplemented with 0.93 μm kinetin and 88 mm mannitol. Chemical name used: 2,4-Dichlorophenoxy acetic acid (2,4-D).

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Influence of growth regulators and explants on shoot regeneration in carnation

Horticultural Science ◽

10.17221/1/2009-hortsci ◽

2009 ◽

Vol 36 (No. 4) ◽

pp. 140-146 ◽

Cited By ~ 4

Author(s):

J.K. Kanwar ◽

S. Kumar

Keyword(s):

Acetic Acid ◽

Shoot Regeneration ◽

Growth Regulators ◽

Dianthus Caryophyllus ◽

Naphthalene Acetic Acid ◽

Ms Medium ◽

Shoot Bud ◽

Dichlorophenoxy Acetic Acid ◽

Number Of Shoots

The influence of growth regulators, explants and their interactions on in vitro shoot bud formation from callus was studied in <I>Dianthus caryophyllus</I> L. The leaf and internode explants were cultured on Murashige and Skoog (MS) medium containing different concentrations of growth regulators. The highest callus induction was observed with 2 mg/l 2,4-dichlorophenoxy acetic acid (2,4-D) and 1 mg/l benzyl adenine (BA). Out of twenty seven shoot regeneration media tested, only 2 mg/l thidiazuron (TDZ) and zeatin alone or in combination with naphthalene acetic acid (NAA) and/or indole acetic acid (IAA) could differentiate calli. The highest average number of shoots was observed with 2 mg/l TDZ and 1 mg/l IAA. Significant differences were observed in calli producing shoots and number of shoots per callus in the explants of leaf and internode. The shoots were elongated and multiplied on MS medium supplemented with 1 mg/l BA and solidified with 1% agar. The shoots were rooted and hardened with 76% survival success in pots after six weeks of transfer to the pots.

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