scholarly journals Genomic Methylated Cytosine Level during the Dedifferentiation and Cellular Competence in Coffea arabica Lines: Insights about the Different In Vitro Responses

Forests ◽  
2021 ◽  
Vol 12 (11) ◽  
pp. 1536
Author(s):  
João Paulo de Morais Oliveira ◽  
Natália Arruda Sanglard ◽  
Adésio Ferreira ◽  
Wellington Ronildo Clarindo
Keyword(s):  
Somatic Embryogenesis ◽  
Somatic Embryos ◽  
Coffea Arabica ◽  
Cytosine Methylation ◽  
Leaf Explants ◽  
Embryogenic Calli ◽  
Ploidy Stability ◽  
Indirect Somatic Embryogenesis ◽  
Cellular Competence

Coffea arabica genotypes present distinct responses in vitro, and somaclonal variation occurrence has been reported. Global cytosine methylation is one of the epigenetic mechanisms that influences the Coffea in vitro responses. We aimed to establish the indirect somatic embryogenesis in C. arabica ‘Catuaí Vermelho’, ‘Caturra’ and ‘Oeiras’, associate the distinct responses to the methylated cytosine genomic level, and check the ploidy stability. Leaf explants were cultured in callus induction and proliferation medium. The resulted calli were transferred to the regeneration medium, and the mature cotyledonary somatic embryos were transferred to the seedling medium. ‘Oeiras’ exhibited the highest number of responsive leaf explants, followed by ‘Caturra’ and ‘Catuaí Vermelho’. Global methylated cytosine level increased over time in the ‘Catuaí Vermelho’ and ‘Caturra’ friable calli, remaining constant in ‘Oeiras’. ‘Oeiras’ did not regenerate somatic embryos, while ‘Catuaí Vermelho’ exhibited the highest number. Somatic embryo regeneration was associated with the increase of the methylated cytosine level. However, the ‘Catuaí Vermelho’ embryogenic calli showed a lower methylated cytosine level than ‘Caturra’. Recovered plantlets exhibited the same 2C value and chromosome number to the explant donors. Therefore, cytosine hypermethylation occurred during C. arabica indirect somatic embryogenesis, influencing cell competence and somatic embryos regeneration.

scholarly journals In vitro somatic embryogenesis of superior clones of robusta coffee from Lampung, Indonesia: Effect of genotypes and callus induction media

2020 ◽  
Vol 21 (8) ◽  
Author(s):  
Dwi Hapsoro ◽  
Rahmadyah Hamiranti ◽  
Yusnita Yusnita
Keyword(s):  
Somatic Embryogenesis ◽  
Callus Induction ◽  
Leaf Explants ◽  
Regeneration Medium ◽  
Embryogenic Calli ◽  
Robusta Coffee ◽  
Primary Callus ◽  
Ascorbic Acids ◽  
Superior Clones

Abstract. Hapsoro D, Hamiranti R, Yusnita Y. 2020. In vitro somatic embryogenesis of superior clones of robusta coffee from Lampung, Indonesia: Effect of genotypes and callus induction media. Biodiversitas 21: 3811-3817. This study aimed to investigate the effects of genotypes and primary callus induction media on somatic embryogenesis of superior robusta coffee clones of Lampung. Leaf explants of clones Tugusari, Komari, Tugino, and Wanto were cultured on two types of primary callus induction media (PCIM). PCIM1 consisted of half-strength MS salts, 30 gL-1 sucrose, added with (mgL-1) 0.1 thiamine-HCl, 0.5 nicotinic acids, 0.5 pyridoxine-HCl, 100 Myo-inositol, 200 ascorbic acids, 150 citric acids, and 1 benzyl adenine. PCIM2 consisted of NPCM salts, 30 gL-1 sucrose, added with (mgL-1) 15 thiamine-HCl, 1 nicotinic acid, 1 pyridoxine-HCl, 2 glycines, 130 Myo-inositol, 200 ascorbic acids, 150 citric acids, 1 2,4-dichlorophenoxyacetic acid, and 2 thidiazuron. The highest percentage (100%) of primary callus formation was found in Komari and Wanto clones. PCIM2 resulted in more primary calli than PCIM1. When subcultured to embryogenic callus induction medium, primary calli of clone Komari and Wanto developed into a high percentage of embryogenic calli, while those of the other two turned brown and died. PCIM2-derived primary calli developed into more embryogenic calli. When subcultured on somatic embryo (SE) regeneration medium, these calli underwent the formation of SE of various stages. When subcultured to plant regeneration medium, these SEs developed into plantlets.

scholarly journals Calli induction in leaf explants of coffee elite genotypes

2011 ◽  
Vol 41 (3) ◽  
pp. 384-389 ◽  
Author(s):  
Juliana Costa de Rezende ◽  
Carlos Henrique Siqueira de Carvalho ◽  
Moacir Pasqual ◽  
Ana Carolina Ramia Santos ◽  
Stephan Malfitano de Carvalho
Keyword(s):  
Coffea Arabica ◽  
Block Design ◽  
Leaf Explants ◽  
Production Potential ◽  
Embryogenic Calli ◽  
The Third ◽  
Randomized Block Design ◽  
Indirect Somatic Embryogenesis ◽  
Growth Room ◽  
Opposite Behavior

Three experiments were carried out with the objective of achieving high effectiveness in calli induction from high heterozygosis leaf explants of Coffea arabica through indirect somatic embryogenesis. A randomized-block design in a 2x5 factorial arrangement made up of two media [BOXTEL & BERTHOULY (1996) and TEIXEIRA et al. (2004)] and five C. arabica genotypes were used in the first experiment. In the second experiment the embryogenic calli production potential was evaluated in ten genotypes. Each of them was considered as a treatment. In the third experiment the variations in both 2.4-D (2.5 e 20µM) and 2-iP (2.5 e 20µM) concentrations in TEIXEIRA et al. (2004) medium and secondary media were evaluated. Crops were kept in a growth room under darkness, at 25±2oC. The medium described by TEIXEIRA et al (2004) was found to be superior when compared to that described by BOXTEL & BERTHOULY (1996) in the 2.2 and 7.2 genotypes. An opposite behavior was noticed in 4.2 genotype, that is, BOXTEL & BERTHOULY (1996) had medium superiority. Both 3.0 and 5.0 genotypes had the same behavior in both media studied, which shows that the somatic embryo production depends on the genotype. Calli induction depends on the 2-iP and 2.4 D ratio. The 20.0µM of 2.4-D and 20.0µM of 2-iP combination caused the highest embryogenic calli induction rate.

scholarly journals Propagasi in vitro tanaman kurma (Phoenix dactylifera L.) pada bioreaktor dengan perendaman sesaat

2020 ◽  
Vol 88 (2) ◽  
Author(s):  
Rizka Tamania SAPTARI ◽  
Masna Maya SINTA ◽  
Imron RIYADI ◽  
. PRIYONO ◽  
. SUMARYONO
Keyword(s):  
Somatic Embryogenesis ◽  
Somatic Embryos ◽  
In Vitro Propagation ◽  
Date Palm ◽  
Phoenix Dactylifera ◽  
Embryogenic Calli ◽  
Temporary Immersion Bioreactor ◽  
Phoenix Dactylifera L ◽  
Immersion Period

The cultivation of date palm in Indonesia has increased since the last decade. However, the superior date palm seedlings are still limited and most of them are imported from other countries. The mass supply of superior date palm seedlings can be provided by in vitro propagation in the bioreactor. Therefore, the research was conducted to develop a protocol of date palm in vitro propagation by using Temporary Immersion Bioreactor (TIB). The in vitro propagation was carried out through somatic embryogenesis technique using meristematic tissues isolated from offshoots of date palm female clone cv. Zambli as explants. The explants were sterilized and then cultured to produce embryogenic calli and somatic embryos. Afterwards, somatic embryos germination and plantlets formation were conducted in TIB with treatments of immersion period: 3, 10, and 30 minutes every 6 hours, with 8 replications, The results showed that the optimal somatic embryo germination in TIB was with the immersion period of 30 min every 6 h, resulting in the most formation of shoots and fresh biomass weight increment up to nearly threefold in 6 weeks. Thereafter, plantlets formation in TIB with immersion period of 10 min and 30 min every 6 h exhibited similar performances in producing more plantlets with higher total fresh weight and better vigor than those of 3 min every 6 h. However, there were more rooted plantlets in the TIB with immersion period of 10 min every 6 h. Based on the results, an in vitro propagation protocol via somatic embryogenesis in TIB has been successfully developed for mass propagation of date palm cv. Zambli, which produced plantlets with good vigor and rooting.

Effect of experimental conditions and genotypic variability on somatic embryogenesis in Coffea arabica

1993 ◽  
Vol 71 (11) ◽  
pp. 1496-1502 ◽  
Author(s):  
D. Bieysse ◽  
A. Gofflot ◽  
N. Michaux-Ferrière
Keyword(s):  
Somatic Embryogenesis ◽  
Coffea Arabica ◽  
Culture Media ◽  
Leaf Explants ◽  
Subsequent Development ◽  
Culture Conditions ◽  
Genotypic Variability ◽  
Experimental Conditions ◽  
Embryogenic Cells

The somatic embryogenic potential of leaf explants from greenhouse-grown plants of eight Coffea arabica genotypes was investigated on three different gelose-gelled culture media (Dublin, Pierson, and Yasuda). Four of these genotypes were reactive. Optimal somatic embryogenesis was obtained when the explants were taken from microcutting and cultured on gelrite-gelled Yasuda's medium. Under these culture conditions, somatic embryos and plantlets were obtained in two previously recalcitrant genotypes. The histocytological callus development was found to be identical in responsive or recalcitrant genotypes. Embryogenic cells formed at two successive points during callogenesis and their subsequent development varied according to culture conditions. Cells initiated in 10- to 15-day-old calli either degenerated or developed directly into embryos. Cells initiated in 60-day-old calli became isolated and developed into embryos or their development was arrested. Embryos obtained in these conditions were able to develop into plantlets. Key words: Coffea arabica, genotypic variability, histocytology, in vitro culture, somatic embryogenesis.

scholarly journals Somatic Embryogenesis and Plant Regeneration from Cotyledon and Hypocotyl Explants of Fagopyrum esculentum Moench lpls Mutant

Agronomy ◽  
2019 ◽  
Vol 9 (11) ◽  
pp. 768
Author(s):  
Yue Fei ◽  
Lan-Xiang Wang ◽  
Zheng-Wu Fang ◽  
Zhi-Xiong Liu
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Somatic Embryos ◽  
Basal Medium ◽  
Fagopyrum Esculentum ◽  
Cereal Crop ◽  
Embryogenic Calli ◽  
Hypocotyl Explants ◽  
Indirect Somatic Embryogenesis ◽  
Fagopyrum Esculentum Moench

Buckwheat (Fagopyrum esculentum, Family Polygonaceae) is an annual pseudo-cereal crop with healing benefits. However, the genetic improvement of common buckwheat has achieved only limited success, mainly due to buckwheat’s dimorphic flowers and heteromorphic self-incompatibility. Here, we develop a useful protocol for indirect somatic embryogenesis and subsequent plant regeneration from hypocotyl explants of F. esculentum. Firstly, the initial calli of hypocotyl explants were induced on Murashige and Skoog (MS) basal medium containing 2.0 mgL−1 2,4-D and 1.5 mgL−1 6-BA for 30 days culture, and then the yellowish white friable embryogenic calli were developed when the initial calli were transferred to fresh MS basal medium supplemented with 1.0 mgL−1 6-BA and 0.5 mgL−1 thidiazuron (TDZ)two to three times subculture at 40–60 days intervals. Subsequently, the somatic embryos were able to germinate from embryogenic callus sub-cultured on MS basal medium containing 1.0 mgL−1 6-BA and 0.5 mgL−1 TDZ with 15% potato puree for 20 days subculture. Finally, maximum mean percentage (75.75%) of somatic embryo-derived plants were obtained when the mature somatic embryos were transferred to MS basal medium without growth regulators for 40 days culture. Our result provides a useful protocol for plant regeneration and SE from hypocotyl explants of F. esculentum.

scholarly journals Effect of Plant Growth Regulators on Somatic Embryogenesis and Plantlet Development of Turkey Berry (Solanum torvum SW)

2021 ◽  
pp. 1-8
Author(s):  
Ghan Singh Maloth ◽  
Rajinikanth Marka ◽  
Rama Swamy Nanna
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Somatic Embryos ◽  
Leaf Explants ◽  
Direct Somatic Embryogenesis ◽  
Ms Medium ◽  
Solanum Torvum ◽  
Plantlet Formation ◽  
Regenerated Plants

In the present study it was reported on direct somatic embryogenesis and plant regeneration from cotyledon and leaf explants of Turkey berry/pea egg plant (Solanum torvum SW), a medicinally important plant. Somatic embryogenesis has several advantages over other routes of in vitro plant regeneration. Somatic embryogenesis was induced directly from cotyledon and leaf explants on MS medium fortified with BAP (0.5 mg/L)+NAA (0.5-6.0 mg/L). High percentage of somatic embryogenesis (90%), maximum number of somatic embryos formation (62±0.18)  along with high percentage (76%) conversion of somatic embryos into bipolar embryos was observed on cotyledon explants in 0.5 mg/L BAP+2.5 mg/L NAA. At the same concentration of BAP (0.5 mg/L)+NAA (2.5 mg/L) also resulted  on the maximum percentage of somatic embryogenesis (92%), the highest number of somatic embryos formation (88±0.15) and the highest percentage (76%) of somatic embryos conversion into bipolar embryos in leaf explants. A mixture of globular, heart and torpedo-shaped embryos were germinated on MS medium supplemented with 0.5 mg/L IAA+1.0-4.0 mg/L BAP. Maximum germination frequency (75±0.14) of somatic embryos and plantlet formation was found in 0.5 mg/L IAA+2.0 mg/L BAP, but they didn’t germinate on ½ MSO and MSO media. The survival rate of regenerated plants after field transfer was recorded to be 75%. These regenerated plants were found morphologically similar to donor plants. The present protocol can be used for conservation of the species and also for genetic transformation experiments in S. torvum.

scholarly journals (291) Plant Regeneration through Direct Somatic Embryogenesis in Homalomena `Emerald Gem'

HortScience ◽  
2006 ◽  
Vol 41 (4) ◽  
pp. 1078A-1078
Author(s):  
Qian Zhang ◽  
Jianjun Chen ◽  
Richard J. Henny
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Somatic Embryo ◽  
Somatic Embryos ◽  
Leaf Explants ◽  
Multiple Shoot ◽  
Direct Somatic Embryogenesis ◽  
Induction Medium ◽  
Ms Medium

Homalomena `Emerald Gem' is an important ornamental foliage plant and widely used for interior plantscaping. Current propagation of this cultivar has been primarily carried out through in vitro culture by organogenesis; regeneration through somatic embryogenesis has not been documented. This report describes successful plant regeneration via direct somatic embryogenesis from explants of different organs. Somatic embryos formed at and around the cut surface of petiole, spathe, and peduncle explants. Embryos also appeared at the base between expanded ovaries of the spadix segment, and around midrib of leaf explants. The optimal treatments for somatic embryo occurrence from petiole, spathe, and peduncle explants were MS medium containing 0.2 mg/L NAA or 0.5 mg/L 2, 4-D with 2.0 mg/L CPPU, and for spadix explants were MS medium with 0.5 mg/L PAA and 2.5 mg/L TDZ. Somatic embryos appeared 6 to 8 weeks after culture and formed large embryo clumps in 3 to 4 months. Somatic embryos produced more secondary embryos and geminated on induction medium. Multiple shoot development and plant regeneration occurred from somatic embryo clusters on MS medium without hormone or with 2 mg/L BA and 0.2 mg/L NAA. The regenerated plants grew vigorously after transplanting to a soilless container substrate in a shaded greenhouse.

scholarly journals 2- Isopentenyladenine in the induction of direct somatic embryogenesis capacity of Coffea arabica L.

2018 ◽  
Vol 48 (11) ◽  
Author(s):  
Ivanilda dos Santos Alves ◽  
Valéria Cristina Barbosa Carmazini ◽  
Cosme Damião dos Santos ◽  
Julieta Andrea Silva de Almeida
Keyword(s):  
Somatic Embryogenesis ◽  
Salt Concentration ◽  
Somatic Embryos ◽  
Coffea Arabica ◽  
Leaf Explants ◽  
Direct Somatic Embryogenesis ◽  
Coffea Arabica L

ABSTRACT: This study evaluated the effect of different concentrations of 2-isopentenyladenine (2-iP) on the direct somatic embryogenesis capacity of the Mundo Novo cultivar of Coffea arabica. Leaf explants were cultivated with half the MS salt concentration and the addition of sucrose (20gL-1) and 2-iP (0; 2.5; 5; 7.5 and 10µM). The 2-iP doses of 7.5 and 10µM produced the greatest responses with respect to the percentage of explants with embryogenic structures and the size of the embryogenic structures. However, the greatest production of somatic embryos occurred on the explants treated with 10µM of 2-iP, followed by 7.5µM, whereas their production was absent or reduced with 0 and 5µM, respectively.

scholarly journals Regeneration of Asparagus officinalis L. Through Embryogenic Callus

2017 ◽  
Vol 27 (1) ◽  
pp. 21-31 ◽  
Author(s):  
Mustafa Abul Kalam Azad ◽  
Muhammad Nurul Amin
Keyword(s):  
Somatic Embryogenesis ◽  
Somatic Embryos ◽  
Shoot Proliferation ◽  
Asparagus Officinalis ◽  
Garden Soil ◽  
Ms Medium ◽  
Regeneration System ◽  
Embryo Germination ◽  
Embryogenic Calli

A plant regeneration system was established from hypocotyl explants of in vitro grown seedlings of A. officinalis and in vitro proliferated shoots, respectively through somatic embryogenesis and embryogenic calli. Somatic embryogenesis was significantly influenced by the types of plant growth regulators. Embryogenic calli with somatic embryos developed well in MS supplemented with 2.0 ‐ 4.0 μM BAP and 1.0 ‐ 4.0 μM 22,4‐D, NAA or IBA. The highest frequency (95.3%) of embryogenic calli and 55.2 somatic embryos formation were obtained when the MS was amended with 4.0 μM BAP and 2.0 μM 2,4‐D. The best embryo germination occurred in 1.0 μM BAP supplemented MMS. The highest 97.2% of shoot proliferation was observed in embryogenic calli in MS medium containing 2.0 μM BAP and 1.0 μM IBA. In vitro grown shoots were rooted in MMS with 1.0 ‐ 2.0 μM IBA. Regenerants were transferred to vermicompost and successfully established under an ex vitro environment in garden soil with 80% survival rate.Plant Tissue Cult. & Biotech. 27(1): 21-31, 2017 (June)

scholarly journals Somatic Embryogenesis and Plant Regeneration from in-vitro-grown Leaf Explants of Rose

HortScience ◽  
2004 ◽  
Vol 39 (6) ◽  
pp. 1378-1380 ◽  
Author(s):  
C.K. Kim ◽  
J.Y. Oh ◽  
J.D. Chung ◽  
A.M. Burrell ◽  
D.H. Byrne
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Somatic Embryos ◽  
Leaf Explants ◽  
Basal Medium ◽  
Shoot Formation ◽  
Ms Medium ◽  
Regeneration Frequency

Somatic embryogenesis was initiated from in vitro-grown leaf explants of rose using an induction period of 4 weeks on MS basal medium supplemented with auxin followed by several subcultures on MS basal medium with cytokinin. `4th of July' showed the highest regeneration frequency (24.4%) on 5.3 μm NAA followed by culture on medium containing 18.2 μm zeatin. `Tournament of Roses' produced somatic embryos when cultured for 4 weeks on medium containing dicamba, 2.3 μm followed by three subcultures on medium containing 18.2 μm zeatin. Embryogenic callus matured on MS media containing 0.5 μm NAA, 6.8 μm zeatin, and 2.9 μm GA3. Long-term cultures were established for both cultivars. Somatic embryos germinated on MS medium containing IBA and BA. Silver nitrate (58.8 μm) enhanced shoot formation and germination of somatic embryos. Plants derived from somatic embryos were acclimatized and successfully established in the greenhouse.

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