scholarly journals Strategies for Fast Multiplication and Conservation of Forest Trees by Somatic Embryogenesis and Cryopreservation: a Case Study with Cypress (Cupressus sempervirens L.)

2018 ◽  
Vol 46 (1) ◽  
pp. 32-38 ◽  
Author(s):  
Maurizio LAMBARDI ◽  
Elif Aylin OZUDOGRU ◽  
Sara BARBERINI ◽  
Roberto DANTI
Keyword(s):  
Somatic Embryogenesis ◽  
Embryogenic Callus ◽  
In Vitro Regeneration ◽  
Pathogenic Fungus ◽  
Cupressus Sempervirens ◽  
Slow Cooling ◽  
Callus Line ◽  
Widespread Species ◽  
Rural Landscapes

Common cypress (Cupressus sempervirens L.) is one of the most widespread species in the Mediterranean area. It has been traditionally cultivated for its ornamental value, becoming a typical feature of urban and rural landscapes, and high timber quality. In the last 30 years, cypress has been subjected to important breeding programmes, aimed to select clones tolerant to the widespread canker, caused by the pathogenic fungus Seiridium cardinale, leading to various patented varieties today available on the market, as well as for genotypes producing null or low amount of allergenic pollen. Somatic embryogenesis is a suitable in vitro regeneration method for fast cloning of conifer trees, and the cryopreservation of embryogenic callus is a significant tool for the safe long-term conservation of valuable cell lines. Recently, a complete protocol for the production of cypress plants from somatic embryogenesis was developed for the patented clone ‘Mediterraneo’. Here, the coupling of somatic embryogenesis and cryopreservation may offer a superior tool to propagate and maintain selected genotypes of cypress by overcoming repetitive subculturing of selected embryogenic callus lines. For the above, this study aimed to compare different cryopreservation techniques (PVS2-based vitrification and slow cooling) with the ‘Mediterraneo’ embryogenic callus line. Best results were obtained after the optimization of a slow cooling procedure, based on the 30-min treatment of embryogenic masses with a cryoprotective solution containing 180 g l-1 sucrose and 7.5% DMSO, followed by the reduction of the temperature at a rate of -1 °C min-1 up to -40 °C and the subsequent immersion in liquid nitrogen  (“two-step freezing”).

scholarly journals EFFECT OF DIFFERENT SOURCES OF PLANT GROWTH REGULATOR ON THE INDUCTION AND DEVELOPMENT OF MANGOSTEEN SOMATIC EMBRYOS

2017 ◽  
Vol 17 (1) ◽  
pp. 9
Author(s):  
Yosi Zendra Joni ◽  
Riry Prihatini ◽  
Darda Efendi ◽  
Ika Roostika
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Growth ◽  
Embryogenic Callus ◽  
Plant Growth Regulator ◽  
Somatic Embryos ◽  
Casein Hydrolysate ◽  
Malt Extract ◽  
Mass Propagation ◽  
Embryogenic Callus Induction

<p>Somatic embryogenesis is a technique for regenerating embryos derived from somatic cells of various plant species. This technique along with the utilization of plant growth regulator (PGR) might benefit for mass propagation and improvement of plant species through biotechnological tools. The study aimed to determine the effect of different plant growth regu-lators, namely 6-benzyladenine (BA) and thidiazuron (TDZ) on the embryogenic callus induction as well as casein hydrolysate and malt extract on the somatic embryo development of mangosteen. The explants used were in vitro young stems of mangosteen clone Leuwiliang. This study consisted of two experiments, namely induction of embryogenic callus and formation of somatic embryo. The first experiment was arranged as factorial in a completely randomized design with BA (0 and 0.7 mg l-1) as the first factor and TDZ (0, 0.1, 0.5 and 1.0 mg l-1) as the second factor. The second experiment consisted of four treatments, i.e. casein hydrolysate and malt extract at the rate of 500 and 1,000 mg l-1. The results showed that the best medium for embryogenic callus induction was MS supplemented with 0.1 mg l-1 TDZ, which resulted semifriable calli. Casein hydrolysate and malt extract could not induce the formation of somatic embryos. After two times subcultures on the same MS medium supplemented with 0.5 mg l-1 TDZ and 0.7 mg l-1 BA, a total of 33.8 somatic embryos per explant was induced. The successful somatic embryogenesis would support mangosteen breeding and in vitro mass propagation program.</p>

scholarly journals In vitro Regeneration of Dryland Kenyan Maize Genotypes Through Somatic Embryogenesis

2006 ◽  
Vol 2 (2) ◽  
pp. 146-151 ◽  
Author(s):  
R.O. Oduor ◽  
E.N.M. Njagi ◽  
S. Ndung` u ◽  
J.S. Machuka
Keyword(s):  
Somatic Embryogenesis ◽  
In Vitro Regeneration ◽  
Maize Genotypes

In vitro Regeneration of Irvingia gabonensis by Somatic Embryogenesis

2008 ◽  
Vol 11 (5) ◽  
pp. 726-732 ◽  
Author(s):  
Fotso . ◽  
Oumar . ◽  
Niemenak Nicolas ◽  
Donfagsiteli Tchinda Ne ◽  
Omokolo Ndoumou De
Keyword(s):  
Somatic Embryogenesis ◽  
In Vitro Regeneration ◽  
Irvingia Gabonensis

Evaluation of the Enhanced Regeneration System for in vitro regeneration in barley

2006 ◽  
Vol 86 (1) ◽  
pp. 63-69
Author(s):  
Seedhabadee Ganeshan ◽  
Brian J Weir ◽  
Monica Båga ◽  
Brian G Rossnagel ◽  
Ravindra N Chibbar
Keyword(s):  
Hordeum Vulgare ◽  
Embryogenic Callus ◽  
Culture Media ◽  
In Vitro Regeneration ◽  
Cold Treatment ◽  
Regeneration System ◽  
Embryogenic Calli ◽  
Hordeum Vulgare L ◽  
Best Response

A simple two-step model for evaluation of in vitro regeneration protocols is proposed based on callus induction and regeneration from immature scutella of two Canadian barley (Hordeum vulgare L.) genotypes, AC Metcalfe and SB92559 using the Enhanced Regeneration System (ERS). The number of explants producing embryogenic callus, the number of plants per embryogenic callus and the number of plants per explant were considered. Tissue culture parameters included three combinations of growth regulators, two carbon sources in culture media, and three cold treatment regimes of spikes prior to scutella isolation. Culture medium containing 5 µM 2,4-dichlorophenoxy acetic acid (2,4-D) and 0.5 µM benzyl adenine (BA) induced the highest percent of embryogenic calli and the highest number of shoots per embryogenic callus from AC Metcalfe. Medium containing 3.75 µM 2,4-D and 0.75 µM BA gave the best response for SB92559. Both genotypes produced more shoots on maltose than on sucrose medium. A 2-d treatment of spikes at 4°C resulted in best response for SB92559. Regeneration response from AC Metcalfe scutella from spikes was unaffected by being subjected to 2, 4 or 6 d of cold. Conditions resulting in best responses from both genotypes were tested on four commercial barley varieties. However, these lines showed inferior regeneration compared to SB92559 and AC Metcalfe. Key words: Hordeum vulgare, scutella, embryogenic callus, shoot production

scholarly journals In vitro regeneration and somatic embryogenesis in citrus

2015 ◽  
Vol 24 (2) ◽  
pp. 247-262 ◽  
Author(s):  
El Sawy A Mohamed ◽  
Amina Gomaa ◽  
Nancy Danial
Keyword(s):  
Somatic Embryogenesis ◽  
Genetic Stability ◽  
Somatic Embryos ◽  
Rapd Analysis ◽  
In Vitro Regeneration ◽  
Rt Pcr ◽  
Mother Plants

Better results were obtained when stigma explants of variegated lemon and citron were used. After ten months, somatic embryos developed into plantlets at a frequency ranged from 13.3 for lime to 66.7% for lemon. Virus presence was tested by ELISA and RT?PCR. The results indicated that the plantlets regenerated through somatic embryogenesis are CTV?free. RAPD analysis was used to asses the genetic stability of plantlets as compared to the mother plants. The results indicated that most plantlets belong to the respective mother plants and the polymorphism percentage was genotype and explant?dependant.Plant Tissue Cult. & Biotech. 24(2): 247-262, 2014 (December

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

scholarly journals The effect of 2,4 dichlorophenoxyacetic acid on in vitro callogenesis of cocoa (Theobroma cacao L.)

2015 ◽  
Vol 31 (2) ◽  
pp. 90-98
Author(s):  
Sulistyani Pancaningtyas
Keyword(s):  
Acetic Acid ◽  
Somatic Embryogenesis ◽  
Embryogenic Callus ◽  
Theobroma Cacao ◽  
Randomized Design ◽  
Significant Difference ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Dichlorophenoxy Acetic Acid ◽  
Theobroma Cacao L

Cocoa (Theobroma cacao L.) development using modern breeding techniques can be facilitated by propagation of planting material through somatic embryogenesis. Various factors that may affect embryogenesis are the composition of culture medium and culture condition. Hormone commonly used to initiate the formation of callus is auxin with type 2.4-D (2.4 Dichlorophenoxy acetic acid). The aim of this study was to determine the effect of the addition of 2.4 -D hormoneson the process of cocoa embryogenesis. The treatments were arragged in factorial combination in completely randomized design, which consisted of two factors. Thefirst factor was the concentration of auxin 2,4-D 25 %, 50 %, 75 %, and 100 %; and the second factor was cocoa clones; Sulawesi 01 and Sulawesi 02. The resultshowed that the addition of 2.4-D hormone up to 100% on somatic embryogenesis of cocoa for Sulawesi 01 clone was not significantly different from Sulawesi 02 clone for all parameters. While on the addition of 2.4-D, there was significant difference between Sulawesi 01 and 02. Cocoa embryogenic callus using the addition of 2.4-D (25%-100%) was significantly different from control. Increased concentrations of 2,4-D hormone which is applied onto media would inhibit the formation of the somatic embryo. Addition of 2.4 D 25%, encouraged towards non-embryogenic callus. Keywords: 2.4 Dichlorophenoxy acetic acid, embryogenic callus, somatic embryos, cocoa, medium culture, hormone

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