scholarly journals Development of a protocol for the micropropagation of mature Eucalyptus grandis clones through somatic embryogenesis

2001 ◽  
Author(s):  
◽  
Andiswa Tsewana
Keyword(s):  
Somatic Embryogenesis ◽  
Embryo Development ◽  
Callus Induction ◽  
Embryogenic Callus ◽  
Somatic Embryos ◽  
Casein Hydrolysate ◽  
Induction Medium ◽  
Mature Trees ◽  
The Media ◽  
Callus Induction Medium

Biotechnology techniques such as micropropagation VIa somatic embryogenesis offer potential significant advances in the improvement of forest species, which could sustain forest production in South Africa, as well as globally, without increased use of land. In order to apply such techniques to commercial breeding and clonal programmes of E. grandis species, it is necessary to develop reliable and efficient protocols applicable to explants of proven superior genotypes. Most of the research on E. grandis somatic embryogenesis has used the genetically variable embryos or seedlings as explant sources, which results in the propagation of material of unproven genetic value. In order to exploit somatic embryogenesis maximally for cloning of superior trees, somatic embryos have to be induced from highly selected and, hence, mature trees. The aim of this investigation was to develop such a protocol for E. grandis and to test its applicability to various E. grandis hybrids. Somatic embryos were induced from buds, stems, leaves and petioles, with petioles and buds giving the best results. Thus, these were selected for further studies which involved testing the effect of medium composition on embryogenic callus induction. Media used for this purpose contained MS or B5 nutrients, 1 mg.l' 2,4-D, 0.5 g.r! glutamine, 0.5 g.r! casein hydrolysate, 4 g.r! Gelrite and 30 or 50 g.rl sucrose. All the media tested were able to support induction of embryogenic callus, although the number of explants producing embryogenic calli was affected significantly by the media composition (10-91 %). Callus induction media with B5 nutrients seemed to have a significant effect onn the developmental stage of embryos in the callus induction medium. Presence of 50 g.r! sucrose in the callus induction medium reduced the embryo yield, but the progress of embryo development was enhanced. The callus induction medium containing B5, 1 mg.l' 2,4-D, 0.5 g.rl glutamine, 0.5 g.r! casein hydrolysate, 4 g.r! Gelrite and 30 g.l' sucrose was chosen for subsequent studies. Of all the media tested for embryo development, the medium with B5, 2.5 mg.l' 2iP, 0.5 g.r! glutamine, 0.5 g.r! casein hydrolysate, 4 g.r! Gelrite and 50 g.r! sucrose was found to be the most suitable for embryo development to the cotyledonary stage. Experiments involving incorporation of both ABA and 2iP aiming at maturation of E. grandis somatic embryos led to an increase in size of the cotyledonary embryos formed but not to germination.

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 The Effect of Daminozide, Dark/Light Schedule and Copper Sulphate in Tissue Culture of Triticum timopheevii

Plants ◽  
2021 ◽  
Vol 10 (12) ◽  
pp. 2620
Author(s):  
Dmitry Miroshnichenko ◽  
Anna Klementyeva ◽  
Sergey Dolgov
Keyword(s):  
Tissue Culture ◽  
Somatic Embryogenesis ◽  
Callus Induction ◽  
Copper Ions ◽  
Induction Medium ◽  
Tissue Cultures ◽  
Triticum Timopheevii ◽  
Dark Light ◽  
Green Plants ◽  
Callus Induction Medium

Triticum timopheevii Zhuk. is a tetraploid wheat that is utilized worldwide as a valuable breeding source for wheat improvement. Gene-based biotechnologies can contribute to this field; however, T. timopheevii exhibits recalcitrance and albinism in tissue cultures, making this species of little use for manipulation through genetic engineering and genome editing. This study tested various approaches to increasing in vitro somatic embryogenesis and plant regeneration, while reducing the portion of albinos in cultures derived from immature embryos (IEs) of T. timopheevii. They included (i) adjusting the balance between 2,4-D and daminozide in callus induction medium; (ii) cultivation using various darkness/illumination schedules; and (iii) inclusion of additional concentrations of copper ions in the tissue culture medium. We achieved a 2.5-fold increase in somatic embryogenesis (up to 80%) when 50 mg L−1 daminozide was included in the callus induction medium together with 3 mg L−1 2,4-D. It was found that the dark cultivation for 20–30 days was superior in terms of achieving maximum culture efficiency; moreover, switching to light in under 2 weeks from culture initiation significantly increased the number of albino plants, suppressed somatic embryogenesis, and decreased the regeneration of green plants. Media containing higher levels of copper ions did not have a positive effect on the regeneration of green plants; contrarily, the elevated concentrations caused albinism in plantlets. The results and relevant conclusions of the present study might be valuable for establishing an improved protocol for the regeneration of green plants in tissue cultures of T. timopheevii.

scholarly journals Efficient Plant Regeneration from Cotyledon and Midrib Derived Callus in Eggplant (Solanum melongena L.)

1970 ◽  
Vol 14 ◽  
pp. 31-38 ◽  
Author(s):  
M Rahman ◽  
M Asaduzzaman ◽  
N Nahar ◽  
MA Bari
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Key Words ◽  
Somatic Embryo ◽  
Callus Induction ◽  
Somatic Embryos ◽  
Solanum Melongena ◽  
Induction Medium ◽  
Root Induction ◽  
Ms Medium

Somatic embryos were obtained from cotyledon and midrib explants of Solanum melongena L., cultivar Loda. For callus induction, medium was supplemented with different concentrations of auxin singly or in combination with BAP. The best callusing 83-85% was obtained from both of the explants cultured on MS medium containing 2.0 mgl-1NAA + 0.05 mgl-1BAP. Somatic embryogenesis and shoot regeneration was achieved after transferring the calli to MS medium supplemented with BAP, GA3, NAA and Zeatin. Cotyledon derived calli showed better performance (87%) for regeneration than that of midrib (82%) when sub cultured on MS medium having 2.0 mgl-1 Zeatin + 1.0 mgl-1 BAP. For root induction, MS + 3.0 mgl-1 IBA was proved to be better treatment for average number (14-15) and mean length (12 cm) of roots than those of other treatments. Key words: Eggplant; cotyledon; midrib; callus induction; somatic embryo J. bio-sci. 14: 1-9, 2006

scholarly journals REGENERATION OF Pimpinella pruatjan THROUGH SOMATIC EMBRYOGENESIS

2016 ◽  
Vol 8 (2) ◽  
pp. 60
Author(s):  
I. Roostika ◽  
R. Purnamaningsih ◽  
I. Darwati ◽  
I. Mariska
Keyword(s):  
Somatic Embryogenesis ◽  
Embryo Development ◽  
Callus Formation ◽  
Natural Habitat ◽  
Casein Hydrolysate ◽  
Shoot Multiplication ◽  
Dry Weight ◽  
Embryogenic Calli ◽  
The Media ◽  
Medicinal Properties

Pruatjan (Pimpinella pruatjan Molk.) is an Indonesian endangered plant which has various medicinal properties such as aphrodisiac, diuretic, and tonic. The plant is commonly harvested from its natural habitat, therefore it becomes endangered. Regeneration of pruatjan through organogenesis has been studied, but its shoot multiplication was very low (5 shoots per explant). The study aimed to investigate the best regeneration technique of pruatjan through somatic embryogenesis. This research was conducted at the tissue culture laboratory, Indonesian Center for Agricultural Biotechnology and Genetic Resources Research and Development in 2004-2005. Callus formation of pruatjan was induced from the petioles and leaves in Driver and Kuniyaki’s (DKW) based medium containing 2,4-D combined with picloram at the level of 0.5, 1.0, 1.5, and 1.5 ppm. Embryogenic calli were then transferred into embryo development medium in two ways. First, they were directly transferred into media containing IBA/NAA at the level of 0.5, 1.0, and 1.5 ppm. Second, they were indirectly transferred into media containing 2.0 ppm 2,4-D and 0.3% casein hydrolysate prior to the IBA/ NAA media. Parameters evaluated were fresh weight, dry weight, time initiation of embryogenic callus formation, and total number of embryos. The result showed that calli of pruatjan were successfully induced from the petioles and leaves. The best calli were induced from the leaves in the DKW medium containing 2.0 ppm 2,4-D and 0.5 ppm picloram. Embryo development of the calli was best if they were first grown in the media containing 2.0 ppm 2,4-D and 0.3% casein hydrolysate then transferred to the IBA/NAA media. The total number of somatic embryos was counted up to 103 on the medium containing 1.5 ppm IBA. This study indicated that pruatjan somatic embryogenesis regeneration required three different media, i.e. for callus induction, development and maturation, and for germination.

scholarly journals Regeneration of Bigleaf Magnolia by Somatic Embryogenesis

HortScience ◽  
1993 ◽  
Vol 28 (6) ◽  
pp. 672-673 ◽  
Author(s):  
S.A. Merkle ◽  
B.A. Watson-Pauley
Keyword(s):  
Acetic Acid ◽  
Somatic Embryogenesis ◽  
Somatic Embryos ◽  
Casein Hydrolysate ◽  
Basal Medium ◽  
Induction Medium ◽  
Immature Seeds ◽  
Dichlorophenoxy Acetic Acid ◽  
Adventive Embryos

Bigleaf magnolia (Magnolia macrophylla Michx.) cultures were initiated from immature seeds on an induction medium containing 9.0 μm 2,4-D, 1.1μm BA, and 1 g casein hydrolysate/liter. After 2 months on induction medium, one culture produced adventive embryos. Clumps of embryos transferred to liquid induction medium proliferated as nodules, which grew in diameter, but failed to produce embryos while maintained in induction medium. Nodules transferred to basal medium produced clumps of somatic embryos, which continued to produce repetitive embryos with monthly transfer to fresh basal medium. Individual embryos transferred to basal medium lacking casein hydrolysate germinated and leaves expanded. Plantlets derived from these embryos were transferred to potting mix and acclimatized to greenhouse conditions. Chemical names used: (2,4-dichlorophenoxy)acetic acid (2,4-D); N -(phenylmethyl)-lH-purin-6-amine (BA).

scholarly journals Robust in vitro propagation and regeneration of ubi kuning high beta carotene cassava genotype through somatic embryogenesis

2017 ◽  
Vol 9 (4) ◽  
pp. 352-360
Author(s):  
SUPATMI SUPATMI ◽  
HANI FITRIANI ◽  
NURHAMIDARR RAHMAN ◽  
N. SRI HARTATI ◽  
ENNY SUDARMONOWATI
Keyword(s):  
Somatic Embryogenesis ◽  
Embryogenic Callus ◽  
Somatic Embryos ◽  
In Vitro Propagation ◽  
Light Condition ◽  
Beta Carotene ◽  
Induction Medium ◽  
Dark Light ◽  
High Beta

Supatmi, Fitriani H, Rahman N, Hartati NS, Sudarmonowati E. 2017. Robust in vitro propagation and regeneration of ubi kuning high beta carotene cassava genotype through somatic embryogenesis. Nusantara Bioscience 9: 352-360. Ubi kuning is a local genotype of cassava with high beta carotene content but the development of this genotype is still low because of plant disease susceptibility. Objectives of this study were to robust induce and regenerate somatic embryos of ubi kuning in vitro as well as to define a protocol of cyclic somatic embryogenesis of ubi kuning. Different size of leaf lobes, various concentration of picloram and different light conditions were tested to produce an effective and efficient somatic embryos (SEs).The best response of the induction of embryogenic callus was observed in leaf lobes explant with range size of 1-3 mm and >5mm cultured on induction medium (MS + 4% sucrose + 4 μM CuSO4 + 0.1 mM Glutamine + 0.8% Microagar) supplemented with either 10 or 18 mg/L picloram grown under dark light for 4 weeks. Retransferring embryogenic callus to the same medium supplemented with 16 mg/L picloram gave the advanced development of primary somatic embryos (PSEs) after 70 d grown under both dark and light condition treatments. A positive correlation between globular and cotyledon stages was obtained in all treatments (P≤ 0.01). The highest shoot and root growth (30% and 25%) was achieved in the regeneration of cotyledonary like-tissues cultured on callus embryogenic media (CEM) (MS basal+ 2.5 μM CuSO4 + 3% sucrose + 2.75 g/L phytagel) supplemented with 1.6 mg/L of BAP (6-Benzylaminopurine).

scholarly journals Response to Callus Induction and Regeneration of Newly Released BRRI Rice Varieties

2020 ◽  
Vol 23 (2) ◽  
pp. 17-25
Author(s):  
SD Joya ◽  
S Sultana ◽  
J Ferdous ◽  
MA Qayum ◽  
ME Hoque
Keyword(s):  
Callus Induction ◽  
Casein Hydrolysate ◽  
The Other ◽  
Induction Medium ◽  
Regeneration Ability ◽  
Regeneration System ◽  
Rice Varieties ◽  
Green Plants ◽  
Induction Frequency ◽  
The Media

A study was carried out for developing an efficient callus induction and regeneration system for three newly developed BRRI varieties namely BRRI dhan86, BRRI dhan87 and BRRI dhan89. Dehusked seeds were plated onto MS and N6 media with two hormone combinations for callus induction. Calli obtained from each callus induction medium were transferred to four different regeneration media. Callus induction frequency and regeneration ability were significantly influenced by rice varieties, and interactions of variety and media. Among the media compositions, the highest callus (59.44%) were obtained from C1 (MS+2mg/l 2,4-D) followed by C2 ( MS+2 mg/l 2,4-D+0.5 mg/l kinetin) , C3 ( N6+2 mg/l 2,4-D) and C4 (N6+2 mg/l 2,4-D+0.5 mg/l kinetin) medium. The highest regeneration (45.74%) was obtained from R2 (MS+4 mg/ml BAP+1.2 mg/ml kinetin+0.5 mg/ml NAA), followed by R3 (1 mg/ml BAP+1 mg/ml Kinetin+1 mg/ml NAA), R4 (2 mg/ml kinetin+1 mg/ml NAA+300 mg casein hydrolysate) and R1 (2 mg/ml BAP+1 mg/ml kinetin+1 mg/ml NAA). BRRI dhan86 showed the highest regeneration ability (53.06%) than the other two varieties. It is observed that all varieties performed better in C1 medium for callus induction and R2 medium for regeneration. This study also revealed that BRRI dhan86 was more responsive to callus induction and regeneration of green plants than the other two varieties. Bangladesh Rice j. 2019, 23(2): 17-25

scholarly journals REGENERATION OF Pimpinella pruatjan THROUGH SOMATIC EMBRYOGENESIS

2016 ◽  
Vol 8 (2) ◽  
pp. 60
Author(s):  
I. Roostika ◽  
R. Purnamaningsih ◽  
I. Darwati ◽  
I. Mariska
Keyword(s):  
Somatic Embryogenesis ◽  
Embryo Development ◽  
Callus Formation ◽  
Natural Habitat ◽  
Casein Hydrolysate ◽  
Shoot Multiplication ◽  
Dry Weight ◽  
Embryogenic Calli ◽  
The Media ◽  
Medicinal Properties

Pruatjan (Pimpinella pruatjan Molk.) is an Indonesian endangered plant which has various medicinal properties such as aphrodisiac, diuretic, and tonic. The plant is commonly harvested from its natural habitat, therefore it becomes endangered. Regeneration of pruatjan through organogenesis has been studied, but its shoot multiplication was very low (5 shoots per explant). The study aimed to investigate the best regeneration technique of pruatjan through somatic embryogenesis. This research was conducted at the tissue culture laboratory, Indonesian Center for Agricultural Biotechnology and Genetic Resources Research and Development in 2004-2005. Callus formation of pruatjan was induced from the petioles and leaves in Driver and Kuniyaki’s (DKW) based medium containing 2,4-D combined with picloram at the level of 0.5, 1.0, 1.5, and 1.5 ppm. Embryogenic calli were then transferred into embryo development medium in two ways. First, they were directly transferred into media containing IBA/NAA at the level of 0.5, 1.0, and 1.5 ppm. Second, they were indirectly transferred into media containing 2.0 ppm 2,4-D and 0.3% casein hydrolysate prior to the IBA/ NAA media. Parameters evaluated were fresh weight, dry weight, time initiation of embryogenic callus formation, and total number of embryos. The result showed that calli of pruatjan were successfully induced from the petioles and leaves. The best calli were induced from the leaves in the DKW medium containing 2.0 ppm 2,4-D and 0.5 ppm picloram. Embryo development of the calli was best if they were first grown in the media containing 2.0 ppm 2,4-D and 0.3% casein hydrolysate then transferred to the IBA/NAA media. The total number of somatic embryos was counted up to 103 on the medium containing 1.5 ppm IBA. This study indicated that pruatjan somatic embryogenesis regeneration required three different media, i.e. for callus induction, development and maturation, and for germination.

scholarly journals Adaptation of in vitro regeneration protocol for Brazilian wheat genotypes

2018 ◽  
Vol 48 (11) ◽  
Author(s):  
Eduardo André Roesler ◽  
Ernandes Manfroi ◽  
Andréa Morás ◽  
Dielli Aparecida Didoné ◽  
Magali Ferrari Grando ◽  
...  
Keyword(s):  
Callus Induction ◽  
Somatic Embryos ◽  
In Vitro Regeneration ◽  
Induction Medium ◽  
Regeneration Capacity ◽  
Target Region ◽  
Wheat Genotypes ◽  
Regeneration Efficiency ◽  
Callus Induction Medium

ABSTRACT: The availability of an efficient protocol for in vitro regeneration is imperative for genetic transformation. Using genotypes adapted to the target region as a transgenic platform accelerates the development of cultivars. Therefore, this study aimed to adapt an in vitro regeneration protocol for Brazilian wheat genotypes. For this purpose, the in vitro regeneration capacity of immature embryos from six Brazilian wheat genotypes using two protocols of regeneration of somatic embryos was analysed. Furthermore, combinations of 2,4-D and picloram in the callus induction medium were tested in order to improve regeneration efficiency. Genotypes with higher regeneration efficiency were BR18-Terena and PF020037, yielding 0.42 and 1.13 shoots per explant using the Hu and the Wu protocol, respectively. Adding 1mgL-1 2,4-D in the callus induction medium was the most favourable, producing 3.73 and 3.07 shoots per explant for PF020037 and BR18-Terena, respectively. In conclusion, a protocol for regeneration for two Brazilian wheat genotypes recommended and developed to be cultivated at the Cerrado region has been adapted.

scholarly journals Indirect Organogenesis and Somatic Embryogenesis of Pineapple Induced by Dichlorophenoxy Acetic Acid

2016 ◽  
Vol 8 (1) ◽  
pp. 8
Author(s):  
Ika Roostika ◽  
Ika Mariska ◽  
Nurul Khumaida ◽  
Gustaaf A. Wattimena
Keyword(s):  
Somatic Embryogenesis ◽  
Embryogenic Callus ◽  
Somatic Embryos ◽  
Callus Formation ◽  
Induction Medium ◽  
Ms Medium ◽  
Regeneration Medium ◽  
Embryogenic Calli ◽  
Indirect Organogenesis ◽  
Basal Media

<p>This research aimed to study the effect of 2,4-D,<br />AdS, and basal media to the regeneration of pineapple<br />through indirect organogenesis and somatic embryogenesis,<br />and to study the complete event of somatic embryogenesis.<br />Callus formation was induced by 21, 41, and 62 μM 2,4-D<br />with addition of 9 μM TDZ. The non embryogenic calli were<br />transferred onto 4.65 μM Kn containing medium.<br />Embryogenic callus formation was induced on MS or Bac<br />basal media consisted of N-organic compounds with<br />addition of AdS (0, 0.05 and 0.1 μM). The embryogenic calli<br />were regenerated on modified MS medium with addition of<br />0.9 μM IBA, 1.1 μM BA, 0.09 μM GA3 or MS medium<br />supplemented with 0.018 mM BA. The result proved that the<br />single auxin of 2,4-D was not enough to induce embryogenic<br />cells. Therefore the non embryogenic calli were regenerated<br />through organogenesis. The 21 μM 2,4-D yielded high level of<br />callus formation (80%), higher fresh weight (0.2 g/explant)<br />and higher number of shoot (25 shoots/explant in two<br />months). Embryogenic calli were produced on N-organic<br />compounds enriched media. The regeneration medium<br />significantly affected the level of browning, where the MS<br />medium with addition of 0.018 mM BA yielded lower level of<br />browning. There was an interaction of embryogenic callus<br />induction medium and regeneration medium to the number<br />of mature somatic embryos. The embryogenic callus<br />induction on MS medium enriched with N-organic<br />compounds and 0.05 μM AdS followed by the regeneration<br />of somatic embryos on MS medium with addition of 0.018<br />mM BA was the best treatment which yielded 17 mature<br />somatic embryos/explant.</p>

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