Evaluation of responses embryogenic Cycas revoluta thumb., from callus culture obtained in vitro

Processes cell differentiation and dedifferentiation are included in the development of biotechnology protocols to promote somatic embryogenesis as an alternative to the in vitro propagation of plants, somatic embryos may be an excellent strategy for both propagation and conservation of fossil species such as cycads. They were evaluated in vitro with different strategies, morphogenic responses associated with obtaining somatic embryos of C. revolute. calli of megagametophytes, subsequently subcultured in four combinations of basic salts of Murashige and Skoog (1962) MS, with the addition of benzyladenine (BA), and 2, dichlorophenoxyacetic (2,4-D) were used, and kinetin (K), and picloram. In the results, It was possible to characterize potentially embryogenic callus, evaluating the levels of both cellular differentiation, necrosis, texture and color; and increases mass or weight considered to start differentiation or proembryogenic or globular type. Calli were subcultured in a medium containing MS salts, incorporating abscisic acid (ABA) in 0, 0.38, 1.13, 3.78 and 5.67 uM doses influenced both the production and maturation of somatic embryos. Embryonic structures, presented a pink coloration characteristic strongly associated towards maturity. The effect of combinations of BA, Kin, 2,4-D, GA3 and ANA influenced the development and germination of mature somatic embryos. And the combination of 1.36 mM 2,4-D+4.44 uM BA promoted the appearance of calluses with a compact texture, characteristic related to their embryogenic potential. The purpose of this research in Cycas sp was to contribute to the study of the in vitro morphogenic responses of this group of plants. And somatic embryogenesis, will allow the obtaining and multiplication as well as its preservation of Cycas sp. Gender that is evolutionary very important

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

Nusantara Bioscience ◽

10.13057/nusbiosci/n090403 ◽

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 Î¼M 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 (Pâ‰¤ 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 Î¼M CuSO4 + 3% sucrose + 2.75 g/L phytagel) supplemented with 1.6 mg/L of BAP (6-Benzylaminopurine).

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

E-Journal Menara Perkebunan ◽

10.22302/iribb.jur.mp.v88i2.394 ◽

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.

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In vitro propagation of Tilia platyphyllos by axillary shoot proliferation and somatic embryogenesis

Journal of Forest Science ◽

10.17221/4722-jfs ◽

2012 ◽

Vol 49 (No. 12) ◽

pp. 537-543 ◽

Cited By ~ 5

Author(s):

V. Chalupa

Keyword(s):

Somatic Embryogenesis ◽

Somatic Embryos ◽

In Vitro Propagation ◽

Shoot Proliferation ◽

Tree Age ◽

Nodal Segments ◽

Axillary Shoot ◽

Axillary Shoot Proliferation ◽

Tilia Platyphyllos

In vitro propagation of Tilia platyphyllos Scop. has been achieved by axillary shoot proliferation and somatic embryo- genesis. The influence of tree age, explant source, genotype, and phytohormones on micropropagation of juvenile and mature trees of Tilia platyphyllos has been investigated. Nodal segments and shoot tips were used as initial explants for axillary shoot proliferation. Low concentration of cytokinin (BA, BPA, TDZ) plus auxin (IBA) stimulated fast shoot multiplication. Microshoots excised from proliferating cultures were rooted on low salt medium and produced trees were planted in the field. Embryo- genic tissues were initiated from zygotic embryos cultured on MS medium supplemented with 2,4-D. After transfer of  embryogenic tissues with developing embryoids on media lacking 2,4-D and supplemented with low concetration of IBA, the development of somatic embryos was enhanced. Secondary somatic embryogenesis led to the formation of new adventive somatic embryos. Trees produced from somatic embryos were planted in the field and exhibited normal growth and morphology.

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Somatic Embryogenesis and In vitro Plant Regeneration in Vigna trilobata (L.) Verd. - A Potential Pasture Legume

Plant Tissue Culture and Biotechnology ◽

10.3329/ptcb.v19i1.4990 ◽

1970 ◽

Vol 19 (1) ◽

pp. 89-99

Author(s):

K. Choudhary ◽

M. Singh ◽

M. S. Rathore ◽

N. S. Shekhawat

Keyword(s):

Somatic Embryogenesis ◽

Growth Regulators ◽

Somatic Embryos ◽

Basal Medium ◽

Long Term Study ◽

Continuous Application ◽

First Time ◽

Pasture Legume

This long term study demonstrates for the first time that it is possible to propagate embryogenic Vigna trilobata and to subsequently initiate the differentiation of embryos into complete plantlets. Initiation of callus was possible on 2,4-D. Somatic embryos differentiated on modified MS basal nutrient medium with 1.0 mg/l of 2,4-D and 0.5 mg/l of Kn. Sustained cell division resulted in globular and heart shape stages of somatic embryos. Transfer of embryos on to a fresh modified MS basal medium with 0.5 mg/l of Kn and 0.5 mg/l of GA3 helped them to attain maturation and germination. However, the propagation of cells, as well as the differentiation of embryos, were inhibited by a continuous application of these growth regulators. For this reason, a long period on medium lacking these growth regulators was necessary before the differentiation of embryos occurred again. The consequences for improving the propagation of embryogenic cultures in Vigna species are discussed. Key words: Pasture legume, Vigna trilobata, Globular, Heart shape, somatic embryogenesis D.O.I. 10.3329/ptcb.v19i1.4990 Plant Tissue Cult. & Biotech. 19(1): 89-99, 2009 (June)

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Agrobacterium-mediated transformation of the wild orchid Cattleya maxima Lindl

Universitas Scientiarum ◽

10.11144/javeriana.sc23-1.amto ◽

2018 ◽

Vol 23 (1) ◽

pp. 89

Author(s):

Augusta Yadira Cueva-Agila ◽

Rino Cella

Keyword(s):

Somatic Embryogenesis ◽

Somatic Embryos ◽

Attractive Alternative ◽

Gene Encoding ◽

Anatomical Structures ◽

Large Numbers ◽

Hygromycin Resistance Gene ◽

Growing Conditions ◽

Somatic Embryogenesis Receptor Kinase

Protocorms are unique anatomical structures; they are akin to rhizoids and are formed by young orchid seedlings under physiological conditions. Explanted orchid tissues produce similar structures called protocorm-like bodies (PLBs) when exposed to appropriate in vitro growing conditions. Both the propagative nature of PLBs and the easiness by which they can be generated, make these structures an attractive alternative to seed-mediated production for growing large numbers of plants. To increase somatic embryogenesis and optimize the procedure, PLBs of Cattleya maxima were transformed using the Agrobacterium tumefaciens method. The T-DNA carried a Hygromycin-resistance gene, a visible marker (GFP5-GUSA) and a rice gene encoding the Somatic Embryogenesis Receptor Kinase, deemed to be important for somatic embryogenesis. Treated PLBs generated somatic embryos developing Hygromycin-resistant plantlets. The insertion of T-DNA was confirmed by PCR, and GFP expression was observed using a fluorescent stereomicroscope. Transformed Cattleya maxima PLBs were more efficient in forming somatic embryos (60-80%) than untransformed controls (45-57%), and this contrast was maximized in hormone-free, Murashige and Skoog (MS) medium (80% of the transformed plants compared to 57% of the untransformed ones). This finding supports the notion that SERK plays an important role in Orchid embryogenesis.

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

Indonesian Journal of Agricultural Science ◽

10.21082/ijas.v17n1.2016.p9-16 ◽

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

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.

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Genotype and transcriptome effects on somatic embryogenesis in Cryptomeria japonica

PLoS ONE ◽

10.1371/journal.pone.0244634 ◽

2020 ◽

Vol 15 (12) ◽

pp. e0244634

Author(s):

Ayako Izuno ◽

Tsuyoshi E. Maruyama ◽

Saneyoshi Ueno ◽

Tokuko Ujino-Ihara ◽

Yoshinari Moriguchi

Keyword(s):

Gene Expression ◽

Somatic Embryogenesis ◽

Tree Species ◽

Somatic Embryos ◽

Cryptomeria Japonica ◽

Expression Profiles ◽

Gene Expression Profiles ◽

Normal Embryo ◽

Conifer Species ◽

Embryogenic Potential

Somatic embryogenesis (SE), which is in vitro regeneration of plant bodies from somatic cells, represents a useful means of clonal propagation and genetic engineering of forest trees. While protocols to obtain calluses and induce regeneration in somatic embryos have been reported for many tree species, the knowledge of molecular mechanisms of SE development is still insufficient to achieve an efficient supply of somatic embryos required for the industrial application. Cryptomeria japonica, a conifer species widely used for plantation forestry in Japan, is one of the tree species waiting for a secure SE protocol; the probability of normal embryo development appears to depend on genotype. To discriminate the embryogenic potential of embryonal masses (EMs) and efficiently obtain normal somatic embryos of C. japonica, we investigated the effects of genotype and transcriptome on the variation in embryogenic potential. Using an induction experiment with 12 EMs each from six genotypes, we showed that embryogenic potential differs between/within genotypes. Comparisons of gene expression profiles among EMs with different embryogenic potentials revealed that 742 differently expressed genes were mainly associated with pattern forming and metabolism. Thus, we suggest that not only genotype but also gene expression profiles can determine success in SE development. Consistent with previous findings for other conifer species, genes encoding leafy cotyledon, wuschel, germin-like proteins, and glutathione-S-transferases are likely to be involved in SE development in C. japonica and indeed highly expressed in EMs with high-embryogenic potential; therefore, these proteins represent candidate markers for distinguishing embryogenic potential.

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In vitro ROOTING OF TENERA HYBRID OIL PALM (Elaeis guineensis Jacq.) PLANTS1

Revista Árvore ◽

10.1590/1806-90882017000400014 ◽

2018 ◽

Vol 41 (4) ◽

Cited By ~ 1

Author(s):

Marlúcia Souza Pádua ◽

Raíssa Silveira Santos ◽

Luciano Vilela Paiva ◽

Vanessa Cristina Stein ◽

Luciano Coutinho Silva

Keyword(s):

Somatic Embryogenesis ◽

Growth Regulators ◽

Oil Palm ◽

Somatic Embryos ◽

Large Scale ◽

Elaeis Guineensis ◽

Germination Percentage ◽

Shoot Length ◽

Indole Butyric Acid

ABSTRACT Oil palm is a woody monocot of economic importance due to high oil production from its fruits. Currently, the conventional method most used to propagate oil palm is seed germination, but success is limited by long time requirements and low germination percentage. An alternative for large-scale propagation of oil palm is the biotechnological technique of somatic embryogenesis. The rooting of plants germinated from somatic embryos is a difficult step, yet it is of great importance for later acclimatization and success in propagation. The aim of this study was to evaluate the effect of the auxins indole acetic acid (IAA) and indole butyric acid (IBA) on the rooting of somatic embryos of Tenera hybrid oil palm. Plants obtained by somatic embryogenesis were inoculated in modified MS medium with 10% sucrose and 0.6% agar and supplemented with IAA or IBA at concentrations of 5 µM, 10 µM, and 15 µM, and the absence of growth regulators. After 120 days, the presence of roots, root type, length of the longest root, number of roots, number of leaves, and shoot length were analyzed. Growth regulators were favorable to rooting; plants cultivated with IBA growth regulator at 15 µM showed higher rooting percentage (87%) and better results for the parameters of number of roots (1.33) and shoot length (9.83).

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