scholarly journals Use of Arabinogalactan Proteins in Coconut (Cocos Nucifera L.) Tissue Culture: An Alternative Approach for Improved Tissue Response

CORD ◽  
2011 ◽  
Vol 27 (2) ◽  
pp. 8
Author(s):  
S C Fernando

Arabinogalactan proteins (AGPs) are extra-cellular proteins involved in plant growth and development. The effects of these proteins on in vitro response of different species are well documented. This study assessed for the first time the role of AGPs on tissue culture of coconut, a highly recalcitrant species in vitro. Coconut (Sri Lanka Tall) plumules were cultured in medium containing 2, 4-dichlorophenoxyaceticacid (2,4-D) for callogenesis. Calli were multiplied by subculturing onto freshly prepared callusing medium. Somatic embryos were induced by transferring of calli to medium with 2/3 concentration of initial 2,4-D and matured by subsequent transferring to medium without any hormones. Embryos were converted in the presence of gibberelic acid. Arabic gum (1-50 mgL-1) was used as the main source of AGP. Alternatively, different AGP sources like Larch wood gum, carrot seeds and defatted coconut kernel were also assessed. Arabic gum in callusing medium resulted in early callogenesis from 20-40% plumules compared to 10% in the control. However, it did not have a significant effect on final callusing efficiency. Its presence in somatic embryogenesis medium showed a significantly higher mass of embryogenic structures per an embryogenic clump (33.3-50.5mg) when compared to the control (26.9mg).The best result was obtained at 10mgL-1 Arabic gum. All AGP sources except Larch wood gum had positive effects on somatic embryogenesis. Among them, defatted coconut kernel (25-50mgL-1) showed the best results giving 70% cultures with embryogenic structures compared to 37% in control. However, so far, the positive effect of AGP did not reflect in plant regeneration efficiency.

2021 ◽  
Vol 28 (1) ◽  
Author(s):  
Renata Orłowska

Abstract Background Somatic embryogenesis is a phenomenon carried out in an environment that generates abiotic stress. Thus, regenerants may differ from the source of explants at the morphological, genetic, and epigenetic levels. The DNA changes may be the outcome of induction media ingredients (i.e., copper and silver ions) and their concentrations and time of in vitro cultures. Results This study optimised the level of copper and silver ion concentration in culture media parallel with the induction medium longevity step towards obtaining barley regenerants via somatic embryogenesis with a minimum or maximum level of tissue culture-induced differences between the donor plant and its regenerants. The optimisation process is based on tissue culture-induced variation evaluated via the metAFLP approach for regenerants derived under varying in vitro tissue culture conditions and exploited by the Taguchi method. In the optimisation and verification experiments, various copper and silver ion concentrations and the different number of days differentiated the tested trials concerning the tissue culture-induced variation level, DNA demethylation, and de novo methylation, including symmetric (CG, CHG) and asymmetric (CHH) DNA sequence contexts. Verification of optimised conditions towards obtaining regenerants with minimum and maximum variability compared to donor plants proved useful. The main changes that discriminate optimised conditions belonged to DNA demethylation events with particular stress on CHG context. Conclusions The combination of tissue culture-induced variation evaluated for eight experimental trials and implementation of the Taguchi method allowed the optimisation of the in vitro tissue culture conditions towards the minimum and maximum differences between a source of tissue explants (donor plant) and its regenerants from somatic embryos. The tissue culture-induced variation characteristic is mostly affected by demethylation with preferences towards CHG sequence context.


2021 ◽  
Vol 6 (3) ◽  
pp. 64403
Author(s):  
Ratna Dewi Eskundari ◽  
Taryono Taryono ◽  
Didik Indradewa ◽  
Yekti Asih Purwestri

Tissue culture is an alternative choice of plant propagation either through somatic embryogenesis or in vitro organogenesis techniques. TRI2025 tea clone has been cultured successfully, however, the scientific information related to morphology, histology, and protein profile at an early event of culturing time has not been reported yet. This study aimed to determine the differences between those pathways, in the context of morphology, histology, and protein profile. The explants were the embryo axis of TRI2025 tea clone cultured on two different induction mediums; somatic embryogenesis and in vitro organogenesis induction medium. The results showed that most of the explants cultured on A medium developed to be a globular-like structure at 11-day after culture (DAC), while all explants cultured on B medium showed the initiation stage of in vitro organogenesis. Histological analysis showed meristem reconstruction at shoot apical meristem (SAM) and root apical meristem (RAM) at 11-DAC at explants cultured on B medium, while explants cultured on A medium showed callusing at 21-DAC. Protein profile analysis using SDS-PAGE showed protein bands of 54 and 81 KDa that only appeared at explants cultured on A medium start from 14-DAC, and those two protein bands thought to be a differentiator at the early stages of the two tissue culture techniques. Thus, these parameters can be used as early detection for plant tissue culture, especially in tea. 


Author(s):  
Adi Prawoto

Cocoa (Theobroma cacao L.) like most tropical trees is recalcitrant in tissue culture. Somatic embryogenesis is generally efficient micropropagation technique to multiply elite material. However, Somatic embryogenesis in cocoa is difficult and this species is considered as recalcitrant. One of the factors often considered as a component of in vitro recalsitrance is a high phenolic content and oxidation of these compounds. In cocoa tissue culture accumulate large amounts of poliphenolics compounds which probably impair further development. This study was conducted to investigate the composition of phenolic compounds in cocoa flower and leaves, and their changes troughout the somatic embryogenesis process. Calli were induced in cacao floral and leaves explants on a half-strenght Murashige and Skoog medium containing 30 g/L Glucose and combination of 2,4 dichlorophenoxyacetic acid (2,4 D) with kinetin (kin). Total polyphenol content was observed on Sulawesi 1 cocoa clone. Embryogenic and non-embryogenic callus were also compared. The percentage of callus production from flower tissue is 85%, percentage of embryogenic callus 40 %, although  the percentage of somatic embryo production from embryogenic callus callus is 70%. The conservation of callus into somatic embryos followed by decline in phenol content and an increase in peroxidase. The synthesis kinetics for these compounds in calli, under different somatic embryogenesis conditions, revealed a higher concentration under non-embryogenic conditions. So that, phenolic compound can influence the production of calli and an absence the phenolic compound can enhance production of somatic embryo.Kata kunci: Theobroma cacao L., polifenol, embrio somatik, kalus, flavonoid, katekin, in vitro recalcitance


2017 ◽  
Vol 17 (2) ◽  
pp. 91
Author(s):  
Meutia Zahara

Abstract Phalaenopsis orchids are recognized as the most popular orchid genus in the world, especially in horticultural industry due to their large, colorful, and durable flowers as well as their wider adaptability to room conditions. The characteristics of seedling propagated by vegetative means are not uniform; therefore, propagation through tissue culture is desirable. Although the micro propagation of Phalaenopsis has shown very good development, but the wide spread of micro propagation still limited due some problems such as the exudation of phenolic compounds, the PGR concentration, the media used, somaclonal variation, the chosen explants, etc. This paper endeavor to include some important investigations based on the common explants used; leaf and flower stalk. Keywords: Micropropagation, Phalaenopsis, leaf explant, flower stalk ReferencesAnonymous. Orchid (Orchidaceae). Diakes tanggal 13 Januari 2013 dari http://www.rainforest-alliance.org/kids/species-profiles/orchid. Rainforest Alliance. 2002.Pillon, Y.; Chase, M. W.Taxonomic exaggeration and its effects on orchid conservation. Conservation Biology. 2007, 21, 263–265.Thengane, S. R.; Deodhar, S. R.; Bhosle, S. V.; Rawal, S. K. Direct somatic embryogenesis and plant regenaration in Garciniaindica Chois’. Current Science. 2006, 91(8), 1074-1078.Yuswanti, H.; Dharma, I. P.; Utama. ; Wiraatmaja, I. W. Mikropropagasi anggrek Phalaenopsis dengan menggunakan eksplan tangkai bunga. AGROTROP. 2015, 5(2): 161-166.Raynalta, E.; Sukma, D.  Pengaruh komposisi media dalam perbanyakan protocorm like bodies, pertumbuhan plantlet, dan aklimatisasi Phalaenopsis amabilis. J. Hort. Indonesia. 2013, 4(3): 131-139.Kosir, P.; Skof, S.; Luthar, Z. Direct Shoot Regeneration from Nodes of Phalaenopsis of Orchids. Acta Agriculturae Slovenica. 2004, 83, 233–242.Arditti, J. R. ; Ernst. Micropropagation of Orchids. Wiley-Interscience. New York, 1993.Park, Y. S.;Kakuta, S.; Kano, A.; Okabe, M.Efficient propagation of protocorm-like bodies of Phalaenopsis in liquid medium. Plant Cell, Tissue and Organ Culture. 1996, 45, 79–85.Park, S. Y. ; Yeung, E. C.; Chakrabarty, D. ; Paek, K. Y. An efficient direct induction of protocorm-like bodies from leaf subepidermal cells of Doritaenopsis hybrid using thin-section culture. Plant Cell Reports. 2002, 21, 46–51.Zahara, M.; Datta, A.; Boonkorkaew, P. Effects of sucrose, carrot juice and culture media on growth and net CO2 exchange rate in Phalaenopsis hybrid ‘Pink’. ScientiaHorticulturae. 2016,205, 17–24.Hee, K. H.; Loh, C. S.; Yeoh, H. H. In vitro flowering and rapid in vitro embryo production in Dendrobium Chao Praya Smile (Orchidaceae). Plant Cell Reports. 2007, 26, 2055–2062.Kannan, N. An in vitro study on micropropagation of Cymbidium orchids. Current Biotica. 2009, 3, 244–250.Steward, Jr. N. C. Plant Biotechnology and Genetics. Willey, A john Willey & Sons, INC., Publication. 2008.George, E. F.; Sherington, P. D.Biotechnology by tissue culture. Exegetics Ltd. 1994.Nursyamsi. Teknik kultur jaringan sebagai alternatif perbanyakan tanaman untuk mendukung rehabilitasi lahan. Makalah pada ekspose hasil-hasil penelitian balai penelitian kehutanan makasar. Makasar, 2010.Aditi, J. F. L. S.; Krikorian, A. D. Orchid mircropropagation: the path from laboratory to commercialization and an account of several unappreciated investigators. Botanical Journal of of the Linnean Society. 1996, 122: 183-241.Gunawan, L. W. Teknik Kultur Jaringan Tanaman. Pusat Antar Universitas (PAU) Bioteknologi IPB. 1998. Bogor.Chugh, S. Guha, S.; Rao, I. U. Micropropagation of orchids: A review on the potential of different explants. Scientia Horticulturae. 2009, 122, 507–520.Ramdan. Kultur daun dan pangkal batang in vitro anggrek bulan raksasa (Phalaenopsis gigantea J.J.Smith) pada beberapa media kultur jaringan. Departemen agronomi dan hortikultura, Fakultas pertanian IPB. 2011.Latip, M. A. R.; Murdad, Z. A.; Aziz, L. H.; Ting, L. M.; Govindasamy.; R. Pipin. Effects of N6-Benzyladenine and Thidiazuron on Poliferation of Phalaenopsis gigantea Protocorm. AsPac J. Mol. Biol. Biotechnol. 2010, 18(1): 217-220 p.Niknejad, A.; Kadir, M. A.; Kadzimin, B. S. In vitro plant regeneration from protocorms-like bodies (PLBs) and callus of Phalaenopsis gigantea (Epidendroidaceae: Orchidaceae). African Journal of Biotechnology.2010, 10, 11808–11816.Chen, J. T.; Chang, W. C. Direct somatic embryogenesis and plant regeneration from leaf explants of Phalaenopsis amabilis. Biologia Plantarum. 2006, 50, 169–173.Zahara, M. Disertasi doktor: The Effects of Plant Growth Regulators and Natural Additives on Direct Shoot Regeneration and Plantlet Growth of Phalaenopsis hybrid ‘Pink’. Asian Institute of Technology, Pathumthani. Thailand. 2016.Xu, C. J.; Li, H.; Zhang, M. G. Preliminary studies on the elements of browning and the changes in cellular texture of leaf explant browning in Phalaenopsis. Acta Horticulturae Sinica. 2005, 32, 1111–1113.Tokuhara, K; Mii, M. Induction of embryonic callus and cell suspension culture from shoot tips excised from flower stalk buds of Phalaenopsis (Orchidaceae). In Vitro Cellular & Developmental Biology–Plant. 2001, 37, 457–461Balilashaki, K.; Naderi, R.; Kalantari, S.; Soorni, A. Mircropropagation of Phalaenopsis amabilis cv Cool ‘Breeze’ with using flower stakl nodes and leaves of sterile obtained from node cultures. IJFAS, 2014.Semiarti, E.; Indrianto, A.; Purwanto, A. Agrobacterium-Mediated transformation of Indonesian orchids for  micropropagation, genetic transformation, Prof. MarÃa Alvarez (Ed.), ISBN: 978-953-307-364-4, InTech, 2011. Available from: http://www.intechopen.com/books/ genetic-transformation/agrobacterium-mediated-transformation-ofindonesian-orchids-for-micropropagation.


1984 ◽  
Vol 3 (6) ◽  
pp. 222-225 ◽  
Author(s):  
P. K. Gupta ◽  
S. V. Kendurkar ◽  
V. M. Kulkarni ◽  
M. V. Shirgurkar ◽  
A. F. Mascarenhas

1992 ◽  
Vol 72 (2) ◽  
pp. 483-485 ◽  
Author(s):  
A. R. McElroy ◽  
D. C. W. Brown

A transplant plug technique was developed that uses in vitro somatic embryogenesis techniques to mass-multiply alfalfa plants in a form suitable for direct transplanting. The plug contains potting media covered with an agar nutrient cap. Plants develop from embryos placed on the cap and then establish in the potting media.Key words: Hybrid alfalfa, Medicago sativa L., tissue culture, transplant plug, somatic embryogenesis


Author(s):  
Sulistyani Pancaningtyas

Cocoa (Theobroma cacao L.) like most tropical trees is recalcitrant in tissue culture. Somatic embryogenesis is generally efficient micropropagation technique to multiply elite material. However, Somatic embryogenesis in cocoa is difficult and this species is considered as recalcitrant. One of the factors often considered as a component of in vitro recalsitrance is a high phenolic content and oxidation of these compounds. In cocoa tissue culture accumulate large amounts of poliphenolics compounds which probably impair further development. This study was conducted to investigate the composition of phenolic compounds in cocoa flower and leaves, and their changes troughout the somatic embryogenesis process. Calli were induced in cacao floral and leaves explants on a half-strenght Murashige and Skoog medium containing 30 g/L Glucose and combination of 2,4 dichlorophenoxyacetic acid (2,4 D) with kinetin (kin). Total polyphenol content was observed on Sulawesi 1 cocoa clone. Embryogenic and non-embryogenic callus were also compared. The percentage of callus production from flower tissue is 85%, percentage of embryogenic callus 40 %, although  the percentage of somatic embryo production from embryogenic callus callus is 70%. The conservation of callus into somatic embryos followed by decline in phenol content and an increase in peroxidase. The synthesis kinetics for these compounds in calli, under different somatic embryogenesis conditions, revealed a higher concentration under non-embryogenic conditions. So that, phenolic compound can influence the production of calli and an absence the phenolic compound can enhance production of somatic embryo.Kata kunci: Theobroma cacao L., polifenol, embrio somatik, kalus, flavonoid, katekin, in vitro recalcitance


2017 ◽  
Vol 10 (1) ◽  
pp. 46-54 ◽  
Author(s):  
H.D.D. Bandupriya ◽  
W.W.M.A. Iroshini ◽  
S A C N Perera ◽  
V.R.M. Vidhanaarachchi ◽  
S.C. Fernando ◽  
...  

Background: In vitro culture techniques provide an excellent platform for the multiplication of recalcitrant species such as coconut and thereby increase the homogeneity of the plantations. Clonal fidelity is one of the most important pre-requisites in a micropropagation protocol of crop species especially those with long life spans. Objective: The present study was conducted in order to determine the genetic homogeneity of coconut plantlets derived from unfertilized ovaries through somatic embryogenesis. Method: Twenty randomly selected plantlets at acclimatization stage, from two mother palms were subjected to Simple Sequence Repeats analysis. Thirteen highly polymorphic microsatellite primers were used for the detection of genetic fidelity in the clonal plantlets and their respective parent. Results: These plantlets showed no apparent differences among themselves and were comparable with the respective mother palm in the Simple Sequence Repeats analysis. The results obtained from this study suggest that there is no somaclonal variation or genetic instability occurring in plantlets that are regenerated from ovary explants. Conclusion: The absence of any sign of somaclonal variation suggests that somatic embryogenesis protocol did not induce the changes in gene structure, which had remained stable throughout the period that had been maintained in vitro. Determination of genetic fidelity of in vitro plants proved the suitability of regeneration protocol for large scale micropropagation applications for coconut.


Genome ◽  
1992 ◽  
Vol 35 (3) ◽  
pp. 474-477 ◽  
Author(s):  
G. A. Kielly ◽  
S. R. Bowley

The genetic control of somatic embryogenesis in alfalfa (Medicago sativa L.) was studied using one nonembryogenic and three embryogenic genotypes: A70-34, a selection from 'Rangelander'; RA3, a selection from 'Regen-S'; and C2-4, a selection from a breeding population that had A70-34 in its pedigree. Crosses of embryogenic × embryogenic and embryogenic × nonembryogenic and S1 and BC1 testcrosses were evaluated for in vitro regeneration. Selfing reduced the expression of the trait. Somatic embryogenesis was dominant and explained by two loci. All three regenerating genotypes shared a common genetic system.Key words: alfalfa, Medicago sativa, somatic embryogenesis, tissue culture.


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