scholarly journals Morphological, Histological, and Protein Profiling of Tea Embryo Axis at Early Stage of Culture

2021 ◽  
Vol 6 (3) ◽  
pp. 64403
Author(s):  
Ratna Dewi Eskundari ◽  
Taryono Taryono ◽  
Didik Indradewa ◽  
Yekti Asih Purwestri
Keyword(s):  
Tissue Culture ◽  
Somatic Embryogenesis ◽  
Apical Meristem ◽  
Protein Profile ◽  
Scientific Information ◽  
Protein Profiling ◽  
Root Apical Meristem ◽  
Embryo Axis ◽  
In Vitro Organogenesis

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. 

scholarly journals Protein Profile of Tissue Culture of TRI2025 Tea Clone

2019 ◽  
Vol 11 (1) ◽  
pp. 8-14
Author(s):  
Ratna Dewi Eskundari ◽  
Taryono Taryono ◽  
Didik Indradewa ◽  
Yekti Asih Purwestri
Keyword(s):  
Tissue Culture ◽  
Somatic Embryogenesis ◽  
Protein Profile ◽  
Scientific Information ◽  
Tea Plant ◽  
Embryonic Axes ◽  
Sds Page ◽  
The World ◽  
The Difference ◽  
Almost All

Tea is well known as favourite healthy drink for almost all people over the world. Tea propagation using conventional and modern ways are now developing rapidly. However, information regarding the protein profile of tissue culture of tea plant has not been revealed yet. This study aimed to determine the difference of protein profile of tea’s tissue culture using SDS-PAGE. This study was conducted using embryonic axes of TRI2025 tea clone cultured on MS media supplemented with 2,4-D for inducing somatic embryogenesis and globular-like structure (GLS) regeneration, and MS media supplemented with BAP for inducing shoot via organogenesis. The results revealed that proteins in the size of 37.69; 54.89; 60.77; 71.35; 87.34; and 92.99 KDa might be involved at somatic embryogenesis, and about 38.69 KDa, 69.27 KDa, and 55.76 KDa respectively for GLS, initiation of shoot, and initiation of GLS derived leaf. Predicted key protein for leaf initiation both directly or through GLS was about 31-33 KDa, while for callusing were about 27.56 and 52.73 KDa, and constitutive protein was about 22.75 KDa. This study provides the first report of protein profile of tea’s tissue culture. The information obtained can be beneficial as a marker for explant for somatic embryogenesis, GLS, or organogenesis pathway and as a scientific information for further biotechnology development.

scholarly journals Barley somatic embryogenesis-an attempt to modify variation induced in tissue culture

2021 ◽  
Vol 28 (1) ◽  
Author(s):  
Renata Orłowska
Keyword(s):  
Tissue Culture ◽  
Somatic Embryogenesis ◽  
Taguchi Method ◽  
Culture Conditions ◽  
Dna Demethylation ◽  
Silver Ion ◽  
Donor Plant ◽  
In Vitro Tissue Culture ◽  
Induced Variation

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.

scholarly journals Coffee Yield and Mineral Cycle in Intercropping of Coffea canephora and Some Species of Timber Shade Trees

2008 ◽  
Vol 24 (1) ◽  
Author(s):  
Adi Prawoto
Keyword(s):  
Tissue Culture ◽  
Somatic Embryogenesis ◽  
Phenolic Compound ◽  
Embryogenic Callus ◽  
Theobroma Cacao ◽  
Coffea Canephora ◽  
Coffee Yield ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Theobroma Cacao L

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

Abstract 11072: Profiling of a Platelet Proteome Discriminates Between Subjects and the Effects of Antiplatelet Agents Alone and in Combination

Circulation ◽  
2014 ◽  
Vol 130 (suppl_2) ◽  
Author(s):  
Simone Marcone ◽  
Desmond J Fitzgerald
Keyword(s):  
Platelet Aggregation ◽  
Protein Profile ◽  
Drug Effects ◽  
Antiplatelet Agents ◽  
Protein Profiling ◽  
Antiplatelet Drugs ◽  
Label Free ◽  
Functional Protein ◽  
Platelet Proteome

Protein profiling of biological samples provides an integrated measurement of genetic, environmental and drug effects. We examined the response to a number of antiplatelet drugs alone and in combination on the human platelet secreteome following stimulation with thrombin. The drugs included a P2Y12 antagonist (PSB0739) and a PAR-1 antagonist (RWJ56110) at concentration (10nM) that blocked the platelet responses to their respective agonists, and aspirin (10microM). Thrombin (1U/ml) was used to stimulate aggregation and secretion of washed platelets despite the presence of the drugs. Platelets were obtained from healthy donors and drugs added in vitro. The proteome profile was determined by label-free quantitative mass spectrometry (Q Exactive and MaxQuant software). The profile was markedly different between individuals (n=3) but it was similar for the same individual over 3 separate samples obtained weeks apart. The addition of the antiplatelet agents showed a similar strong individual clustering of all proteins (>1,000). Cluster analysis of the 270 proteins that changed with drug showed marked differences between drugs although there was no impact on platelet aggregation. Similarly, combinations of 2 or 3 drugs demonstrated distinctive clustering of changes in the protein profile. Functional ontology of the proteome showed marked differences between drugs with very distinctive patterns. Similarly, drugs differed clearly in their effects on discrete functional protein clusters, such as ‘coagulation proteins’. Our findings suggest that individuals have distinct platelet proteomes (‘proteome barcode’) that remain evident in the presence of drug. Despite this, antiplatelet drugs exhibit distinct effects on the platelet proteome, when applied alone and in combination. The profiling of the platelet proteome better discriminates the effects of antiplatelet drugs than is possible with platelet aggregation and may help tailor therapy more effectively.

In vitro organogenesis and somatic embryogenesis in Seseli lehmannii Degen inflorescence culture

2019 ◽  
Vol 305 ◽  
pp. S29-S30 ◽  
Author(s):  
O. Mitrofanova ◽  
T. Kuzmina ◽  
I. Mitrofanova ◽  
M. Rudenko
Keyword(s):  
Somatic Embryogenesis ◽  
In Vitro Organogenesis

scholarly journals A REVIEW: MICROPROPAGATION OF PHALAENOPSIS sp FROM LEAF AND FLOWER STALK EXPLANTS

2017 ◽  
Vol 17 (2) ◽  
pp. 91
Author(s):  
Meutia Zahara
Keyword(s):  
Tissue Culture ◽  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Plant Cell ◽  
Direct Somatic Embryogenesis ◽  
Direct Shoot Regeneration ◽  
Flower Stalk ◽  
Micro Propagation ◽  
Direct Shoot

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.

scholarly journals In vitro Organogenesis of Stevia rebaudiana Bert. with Different Explants and Growth Regulators

2021 ◽  
Vol 14 (3) ◽  
Author(s):  
Juan Francisco Aguirre-Medina ◽  
Ana Laura Gálvez-López ◽  
Juan Francisco Aguirre-Cadena
Keyword(s):  
Growth Regulators ◽  
Apical Meristem ◽  
Stevia Rebaudiana ◽  
Leaf Number ◽  
Naphthaleneacetic Acid ◽  
Nodal Segment ◽  
In Vitro Organogenesis ◽  
Meristem Explants ◽  
Induction Stage

Objective: To evaluate various explants and growth regulators in order to improve invitro propagation of Stevia rebaudiana through organogenesis.Design/Methodology/Approach: Explants and growth regulators in two differentconcentrations were evaluated. The explants were nodal segment, axillary bud, andapical meristem; while the growth regulators were benzylaminopurine (BAP) at 1.125 mgL -1 and 3.0 mg L -1 , naphthaleneacetic acid (NAA) at 1.5 mg L -1 and 3.0 mg L -1 , andCIDEF-4 brassinosteroids (BRs) at 1.0 mg L -1 and 1.5 mg L -1 . In total 18 treatments withseven repetitions. Contamination, oxidation, and survival were recorded duringinduction; while leaf number, regrowth height, and root presence were recorded duringmultiplication.Results: At the induction stage there was a differential response between explantsaccording to their ontogenetic age; during multiplication, the morphological componentsshowed differences between concentrations of growth regulators and explants, withhigher effectiveness when adding BAP to apical meristems.Study Limitations/Implications: Both the origin and the age of explants can inducedifferential growth while interacting with growth regulators. Findings/Conclusions: Apical meristem explants showed better advantages for in vitroreproduction of S. rebaudiana since they present less contamination and higher survivalat the induction stage, even when exhibiting the highest oxidation among explants,which did not influence the decrease in their survival. At the multiplication stage withapical meristem, height, leaf number, and root presence were increased. Values werehigh when interacting with BAP.

A transplant plug technique for production of alfalfa (Medicago sativa L.) plants from somatic embryos

1992 ◽  
Vol 72 (2) ◽  
pp. 483-485 ◽  
Author(s):  
A. R. McElroy ◽  
D. C. W. Brown
Keyword(s):  
Tissue Culture ◽  
Somatic Embryogenesis ◽  
Medicago Sativa ◽  
Somatic Embryos ◽  
Potting Media ◽  
Medicago Sativa L

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

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