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

Plants ◽  
2020 ◽  
Vol 9 (3) ◽  
pp. 360
Author(s):  
Baker Al-Shara ◽  
Rosna Mat Taha ◽  
Jamaludin Mohamad ◽  
Hashimah Elias ◽  
Asif Khan

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.

2013 ◽  
Vol 7 (2) ◽  
pp. 5-12
Author(s):  
Ragad M. Abdullah ◽  
Mazahim K. AL-Mallah

Calli cultures of stems and leaves explants excised from field -grown rubber, Ficus elastica Decora, plants were formed on agar-solidified Murashige and Skoog (MS) medium.The results proved that MS medium supplemented with 1.0 mg L-1 benzyl adenin (BA) and 0.8mg L-1 2,4-dichloro-phenoxy acetic acid (2,4-D) was suitable to stimulate stem’s callus at ratio 87.5% . Whereas supplementation of MS medium with 0.5 mg L-1 of both BA and Indole -3-butyric acid (IBA)encouraged callus formation to reach76.6%. Leaves showed responses for callus initiation up to 75% at the same media. Stem calli showed limited ability to regenerate shoots on both agar solidified MS medium containing 1.0 mg L-1 2,4-D and kinetin (kin) 0.5 mg L-1 and MS medium supplemented with 0.5 mg L-1 of both BA and IBA. Transferring of shoots to differentiation medium (MS+0.5 mg L-1 BA+ 0.1 mg L-1 IBA) stimulated the growth and elongation of these shoots. Shoots were transferred to agar-solidified MS medium free from growth regulators failed to form roots. Because of the less number of shoots, other rooting media were not tested. The data showed, that shoot tips succeeded to regenerate shoots when they were cultured in different MS. The results proved clear increase in chlorophyll and protein content of the rubber shoots as compared with content of field grown rubber plants. It was noticed that agar-solidified MS medium supplemented with 4.0 mg L-1 BA was considered the optimum medium for shoots regeneration. All plants regenerated from shoot tips were readily rooted in agar-solidified MS medium with increasing Potassium Nitrate KNO3 from 1900 mg L-1 to 2000 mg L-1, and at the same medium supplemented with 3.0 mg L-1 IBA and 1.0 mg L-1 BA. All these plants were successfully acclimated and transferred to peat moss.


HortScience ◽  
1995 ◽  
Vol 30 (4) ◽  
pp. 783E-783
Author(s):  
S.K. Dhir ◽  
U.L. Yadava

An efficient protocol has been developed for the in vitro multiplication of papaya (Carica papaya L.) through somatic embryogenesis utilizing immature zgotic embryos. Somatic embryos were initiated on MS basel media supplemented with 5 mg·liter–1 2,4-D, 400 mg·liter–1 glutamine, and 6% sucrose. After culturing for 2 months, 65% of the explants became highly embryogenic. Each explant produced 50 to 80 embryos in 4 months on culture induction medium. Frequency of embryogenesis was increased (75 to 150 somatic embryos on 80% explants) upon supplementing medium with 4% maltose as a carbon source and 100 mg·liter–1 L-asparagine. The embryogenic callus appeared yellow and embryos at different stages of development were well-organized. On regular subculturing, these cultures continued to produce secondary embryos. Following their transfer to the hormone-free medium supplemented with 4% maltose, these embryos germinated. The somatic embryogenesis system is rapid, repetitive, and highly proliferative. Thus, this system may have a potential use in the development of synthetic seed and transgenic papaya plants. Details of important factors affecting somatic embryogenesis will be discussed.


2011 ◽  
Vol 183-185 ◽  
pp. 1462-1466
Author(s):  
Ling Yang ◽  
Yu Hua Li ◽  
Hai Long Shen

Somatic embryogenesis was obtained by using immature zygotic embryos of S. pohuashanesis as explants and emblings were obtained. For induction of somatic embryos, immature zygotic embryos which 30 days old after pollination were cultured on solid MS medium with 1.0 mg•L-1 NAA, 0.1 mg•L-1 6-BA, 500 mg•L-1casein hydrolysate (CH) and 40 g•L-1 sucrose . Inducted somatic embryos were cultured in solid MS medium containing 500 mg•L-1CH and 40 g•L-1 sucrose. After 30 days of culture, many normal cotyledonary embryos were produced. Plantlets were regenerated when somatic embryos were transferred to MS medium with 30 g•L-1 sucrose. The somatic embryos germinated at a germination frequency of approximately 80%, but rate of the plantlets that successfully acclimated and continued growing was 40% in the greenhouse.


HortScience ◽  
2004 ◽  
Vol 39 (4) ◽  
pp. 861B-861
Author(s):  
Bipul K. Biswas* ◽  
Nirmal Joshee ◽  
Anand K. Yadav

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.


2002 ◽  
Vol 24 (3) ◽  
pp. 601-603 ◽  
Author(s):  
Ana da Silva Ledo ◽  
Osmar Alves Lameira ◽  
Abdellatif Kemaleddine Benbadis ◽  
Ilmarina Campos de Menezes ◽  
Maria do Socorro Padilha de Oliveira ◽  
...  

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.


2014 ◽  
Vol 26 (2) ◽  
pp. 155-162 ◽  
Author(s):  
Jaime A. Teixeira da Silva

ABSTRACT The mid- to long-term preservation of papaya (Carica papaya L.) would allow for the safeguarding of important germplasm. In this study, soft friable callus (SFC) and hard callus (HC) were induced from the first two true leaves of 10-day-old seedlings containing a midrib derived from the germinated seed of two cultivars (‘Rainbow’ and ‘Sunrise Solo’). Following germination on a Murashige and Skoog (MS) medium that contained 3% sucrose and was free of plant growth regulators (PGRs), sections of the first true leaves from 10-day-old seedlings were exposed to seven published callus or somatic embryogenesis protocols for zygotic embryos, leaves or hypocotyls. Optimal SFC and HC induction was carried out on a half-strength MS medium following the Fitch (1993) or the Ascêncio-Cabral et al. (2008) protocol, respectively. SFC formed shoots that could then convert to plants when transferred to a full-strength MS medium devoid of PGRs. Plantlets 10-cm tall were acclimatised in two steps: first by in vitro acclimatisation in aerated vessels, the Vitron, under CO2-enriched (3000 ppm CO2), then by the transfer of individually rooted plantlets in Rockwool® blocks to a substrate of soil: pine bark : perlite (1:1:1, v/v/v). SFC and HC were then encapsulated in alginate beads, which were exposed to low temperature storage (LTS) at 10°C and 15°C, and also cryopreserved for 30 days. All encapsulated alginate beads that contained SFC, HC or leaf tissue that had been stored under LTS or cryopreserved were able to regenerate callus when placed on an optimal callus induction medium. Plants derived from the control, LTS and cryopreservation protocols, either from SFC or HC, were successfully acclimatised.


Author(s):  
C. K. Rajesh ◽  
D. Sudhakar ◽  
K. K. Kumar ◽  
C. Kavitha ◽  
G. Karthikeyan ◽  
...  

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.


2012 ◽  
Vol 12 (3) ◽  
pp. 171-178 ◽  
Author(s):  
Cleber Witt Saldanha ◽  
Maisa Pimentel Martins-Corder

This study evaluated different aspects of in vitro germination and embryogenic competence of immature zygotic embryos of E. edulis. The embryos germinated on full or half-strength MS (MS or MS/2) medium combined with sucrose (20, 30 and 40 g L-1). The effect of calcium chloride concentrations (0, 2, 4, 8, and 12 mM) on the induction of somatic embryogenesis was tested. The embryos were germinated on MS or MS/2. Germination of zygotic embryos and the number of roots per plantlet were not affected by the culture medium and sucrose concentration. Plantlet height and fresh weight were influenced by both; the difference was greatest in MS medium with 40 g L-1 sucrose. The induction of somatic embryogenesis was not influenced by doses of calcium chloride, whereas the number of somatic embryos formed was affected. The germination capacity of somatic embryos of heart-of-palm tree was not influenced by the media tested.


HortScience ◽  
1990 ◽  
Vol 25 (12) ◽  
pp. 1652-1654 ◽  
Author(s):  
Haeng S. Lee ◽  
Jang R. Liu ◽  
Seung G. Yang ◽  
Young H. Lee ◽  
Kwang-W. Lee

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).


2008 ◽  
Vol 43 (10) ◽  
pp. 1325-1330 ◽  
Author(s):  
Lucymeire Souza Morais-Lino ◽  
Janay Almeida dos Santos-Serejo ◽  
Sebastião de Oliveira e Silva ◽  
José Raniere Ferreira de Santana ◽  
Adilson Kenji Kobayashi

The objective of this study was to establish cell suspension culture and plant regeneration via somatic embryogenesis of a Brazilian plantain, cultivar Terra Maranhão, AAB. Immature male flowers were used as explant source for generating highly embryogenic cultures 45 days after inoculation, which were used for establishment of cell suspension culture and multiplication of secondary somatic embryos. Five semisolid culture media were tested for differentiation, maturation, somatic embryos germination and for plant regeneration. An average of 558 plants per one milliliter of 5% SCV (settled cell volume) were regenerated in the MS medium, with 11.4 µM indolacetic acid and 2.2 µM 6-benzylaminopurine. Regenerated plants showed a normal development, and no visible somaclonal variation was observed in vitro. It is possible to regenerate plants from cell suspensions of plantain banana cultivar Terra using MS medium supplemented with 11.4 µM of IAA and 2.2 µM of BAP.


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