Immature embryo is the potential source for in vitro plant regeneration in Jatropha curcas

Context: Jatropha belongs the spurge family Euphorbiaceae. Special interest mounting for its biodiesel which has created enthusiasm in cultivation of the species for oil extraction. Objectives: The study was conducted to develop the protocol for tissue and callus culture in Bangladeshi Jatropha curcus plant particularly to identify the most suitable explants for its wide scale micropropagation. Materials and Methods: Immature embryos taken from four developmental stages of fruits were cultured on growth regulator free MS liquid medium. After fifteen days of germination, elongated hypocotyls and two cotyledonary leaves were used as explants. Results: Embryo derived seedlings acted as the potential source of explants both for callus and plantlets. The immature embryo of size 0.87cm produced highest callus formation (83.33%) on MS medium supplemented with lower concentration of 2, 4-D (0.5 mg/l) and coconut water 2% (v/v). Immature embryos grown on MS basal medium supplemented with 2,4-D (0.2 mg/l, 0.5 mg/l and 1.0 mg/l) alone or in combination with coconut water 2% (v/v) exhibited a wide range of callus induction percentage (26-100%) for hypocotyls and (20 - 40%) for cotyledonary leaves. Conclusion: The age of immature embryo and addition of growth adjuvants and growth additive to the culture medium played the role in promoting better callus and plantlet formation. DOI: http://dx.doi.org/10.3329/jbs.v20i0.17726 J. bio-sci. 20: 125-134, 2012

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The Establishment of an Efficient Callus Induction System for Lotus (Nelumbo nucifera)

Plants ◽

10.3390/plants9111436 ◽

2020 ◽

Vol 9 (11) ◽

pp. 1436

Author(s):

Xianbao Deng ◽

Yaqian Xiong ◽

Jing Li ◽

Dong Yang ◽

Juan Liu ◽

...

Keyword(s):

Callus Induction ◽

Developmental Stages ◽

Callus Formation ◽

Immature Embryo ◽

Basal Medium ◽

Nelumbo Nucifera ◽

Dichlorophenoxyacetic Acid ◽

Highly Efficient ◽

Induction System ◽

2,4 Dichlorophenoxyacetic Acid

The lotus (Nelumbo nucifera) is one of the most popular aquatic plants in Asia, and has emerged as a novel model for studying flower and rhizome development, and primary and secondary metabolite accumulation. Here, we developed a highly efficient callus induction system for the lotus by optimizing a series of key factors that affect callus formation. The highest efficient callus production was induced on immature cotyledon and embryo explants grown on Murashige and Skoog (MS) basal medium containing an optimized combination of 3 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.5 mg/L 6-benzylaminopurine (6-BA). In addition, lotus callus induction was proven to be influenced by lotus genotypes, light conditions, the developmental stages of explants and the time of explant sampling. Collecting immature cotyledons from seeds of the genotype “Shilihe 1”, at 9 days post pollination, and to culture the explants in darkness, are proposed as the optimum conditions for lotus callus induction. Interestingly, highly efficient callus induction was also observed in explants of immature embryo derived aseptic seedlings; and a small amount of lotus benzylisoquinoline alkaloid (BIA) and obvious expression of BIA biosynthetic genes were detected in lotus callus.

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In vitro Callus Induction and Regeneration of Popular Indica Rice Genotypes

International Journal of Agriculture Environment and Biotechnology ◽

10.30954/0974-1712.04.2020.2 ◽

2020 ◽

Vol 13 (4) ◽

Author(s):

Aananthi. N

Keyword(s):

Callus Induction ◽

Callus Formation ◽

Indica Rice ◽

Immature Embryo ◽

Coconut Milk ◽

Rice Genotypes ◽

Mist Chamber ◽

Regenerated Plant ◽

Better Than

Five rice cultivars viz., ASD 16, White Ponni, Pusa Basmati 1, Pusa Sugandh 4 and Pusa Sugandh 5 belonging to subspecies indica were compared for its ability in callus formation and regeneration. In this experiment, the different parameters viz., the effect of hormones (2,4-D and kinetin), organic supplement (coconut milk O1-CM 100 mll-1, O2-CM 75 mll-1, O3-CM 50 mll-1), explants (seed and immature embryo), media (MS and N6), carbon source (sucrose and maltose) using five genotypes on callus response was studied. The effect of hardening methods was also assessed. Results showed that for enhanced callus induction was with MS medium supplemented with 2.0 mgl-1 2, 4-D + 0.5 mgl-1 kinetin + 30 gl-1 maltose irrespective of explants used. Addition of 100 ml l-1 coconut milk was found have improvement in callus response. The performance of immature embryo was better than seed for callus induction, emrbyogenic callus formation, rhizogenic callus formation and regeneration. MS media provided superiority over N6. Among the genotypes Pusa Basmati 1 rendered outstanding performance in callus behavior. The treatment combination MS + 2.5 mgl-1 BAP + 0.5 mgl-1 NAA + 1.0 mgl-1 KN gave the highest organogenesis response and regeneration of plantlets. Hardening in mist chamber was recognized as the best method to give the highest per cent of regenerated plant lets.

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Obtaining calli and regenerated plants in anther cultures of pea

Czech Journal of Genetics and Plant Breeding ◽

10.17221/137/2013-cjgpb ◽

2014 ◽

Vol 50 (No. 2) ◽

pp. 123-129 ◽

Cited By ~ 8

Author(s):

S. Bobkov

Keyword(s):

Developmental Stages ◽

Callus Formation ◽

Naphthaleneacetic Acid ◽

Embryogenic Calli ◽

Nutrient Media ◽

Shoot Morphogenesis ◽

Regenerated Plants ◽

Wide Range ◽

2,4 Dichlorophenoxyacetic Acid ◽

Anther Cultures

Pea (Pisum sativum L.) is a species for which there is no efficient method for the recovery of haploid plants yet. This research investigated the influence of various genotypes, nutrient media, and stress treatments on callus formation, embryogenesis and plant regeneration in anther cultures of pea. A wide range of pea genotypes and nutrient media was studied. Morphogenic calli were initiated on media supplemented with α-naphthaleneacetic acid (NAA), 6-benzyladenine (BA), and 2,4-dichlorophenoxyacetic acid (2,4-D) without application of stress treatments. Embryogenic calli and embryos were regenerated on media with low sucrose content in the presence of 2,4-D or indole-3-butyric acid (IBA) after cold stress (4°C) of isolated buds, alone or in combination with in vitro treatment of isolated anthers at higher temperatures (35–38°C). The efficiency of regeneration via shoot morphogenesis on different nutrient media and the peculiarities of regeneration from embryogenic calli were investigated. Green embryogenic calli initiated on 2,4-D were able to develop through shoot morphogenesis on a medium supplemented with BA and NAA. This process led to regeneration of hypertrophic embryos at various developmental stages. The origin of regenerated plants (i.e. from microspores or somatic anther cells) was estimated using marker alleles determining morphological traits. Almost all R<sub>0</sub> regenerants derived from morphogenic calli originated from anther somatic cells.

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Response of in vitro propagated fig (Ficus carica L.) shoots to the concentrations of benzyl amino purine and coconut water

IOP Conference Series Earth and Environmental Science ◽

10.1088/1755-1315/922/1/012067 ◽

2021 ◽

Vol 922 (1) ◽

pp. 012067

Author(s):

Sophia ◽

M Hayati ◽

E Kesumawati

Keyword(s):

Callus Formation ◽

Formation Rate ◽

Water Concentration ◽

Shoot Formation ◽

Coconut Water ◽

Contamination Rate ◽

Two Factors ◽

Number Of Leaves ◽

Benzyl Amino Purine

Abstract In this study, several concentrations of benzyl amino purine (BAP) and coconut water (CW) were investigated along with the interaction between two factors to the growth of in vitro propagated fig shoots. The investigated factors consisted of BAP concentration: 0, 1, 3, 5 mg L−1 and coconut water concentration: 0, 100, 200, 300 ml L−1. A total of 16 treatment combinations with 6 replications resulting in 96 experimental units consisting of a single fig shoot explant per culture medium. The observed parameters including living explant rate, contamination rate, browning rate, day of first shoot emergence, shoot formation rate, explant height addition, number of leaves, callus formation rate, and number of roots were conducted every week from 1 to 8 weeks after proliferation (WAP). The result indicated that in 8 WAP, the living explant rate reached 23.95%. The combination of concentration 200 ml L−1 CW and 3 mg L−1 BAP + 200 ml L−1 CW-induced early emergence of new shoots at 7 days after proliferation (DAP). The highest shoot formation rate (100%) was observed at a concentration of 300 mL L−1CW. The highest explant height addition (7.10 cm) was observed at a concentration of 200 mL L−1 CW. The highest number of leaves (5.80) was observed at a concentration of 1 mg L−1 BAP + 200 mL L−1 CW. The highest callus formation rate (50%) was observed at a concentration of 100 ml L−1CW and 300 ml L−1 CW. The highest number of roots (17) was observed in the control.

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Propagation of Dendrobium aggregatum by green capsule culture

Lankesteriana ◽

10.15517/lank.v0i0.11763 ◽

2013 ◽

Cited By ~ 1

Author(s):

S. Vijayakumar ◽

G. Rajalkshmi ◽

K. Kalimuthu

Keyword(s):

Root Formation ◽

Coconut Water ◽

Immature Embryos ◽

Ms Medium ◽

Medium Supplement ◽

In Vitro Shoots ◽

Immature Seeds ◽

Benzyl Amino Purine ◽

Number Of Shoots

An efficient protocol for propagation of Dendrobium aggregatum using the axenic immature seeds, derived from green capsule, was developed. The immature embryos from 120 days old capsules after pollination were germinated on Murashige and Skoog (MS) medium supplement with various concentration of BAP alone or in combination with NAA along with coconut water, and the same media were used for induction, multiplication, elongation and rooting in vitro shoots. MS medium with the addition of 3% sucrose 1.5 mg L-1 Benzyl amino purine (BAP) and 15% coconut water (CW) favoured the higher rate of germination, more number of protocorm like bodies, production of maximum number of shoots, elongation of shoots, as well as root formation. During acclimatization, 95% of the plantlets survived after one month.

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THE EFFICIENCY OF SHOOT AND PLANTLET FORMATION OF Cephaelis ipecacuanha AFTER THREE SUBCULTURES IN VITRO

Ciência Rural ◽

10.1590/s0103-84781994000300013 ◽

1994 ◽

Vol 24 (3) ◽

pp. 523-526 ◽

Cited By ~ 2

Author(s):

Osmar Alves Lameira ◽

Marly Pedroso da Costa ◽

José Eduardo Brasil Pereira Pinto

Keyword(s):

Gibberellic Acid ◽

Butyric Acid ◽

Growth Regulators ◽

Activated Charcoal ◽

Basal Medium ◽

Adventitious Shoot ◽

Shoot Elongation ◽

Plantlet Formation ◽

Cephaelis Ipecacuanha

Multiple adventitious shoot formed from internodal segments of Cephaelis ipecacuanha cultured 25 days on Gamborg basal medium (GAMBORG et al., 1968) supplemented with 6.66mM 6-benzylaminopurine there was a maximum of nine shoots per segment and an average of five shoots per segment formed. The presence of gibberellic acid in the subculture media promoted shoot elongation in all treatments. The shoots attained 3cm in height and rooting of 100% after 35 days of culturing upon Murashige and Skoog's basal medium (MS), added with 4.92mM indole-3-butyric acid, 0.87m gibberellic acid and 0.1% activated charcoal. Further growth was accelerated after the transfer to 1/2 MS without growth regulators. Rooted plantlets transferred to potting soil could be successfully established.

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Somatic Embryogenesis in Centella asiatica, a Valuable Medicinal Plant

HortScience ◽

10.21273/hortsci.39.4.859c ◽

2004 ◽

Vol 39 (4) ◽

pp. 859C-859

Author(s):

Nirmal Joshee* ◽

Bipul K. Biswas ◽

Anand K. Yadav

Keyword(s):

Somatic Embryos ◽

Bacterial Contamination ◽

Developmental Stages ◽

Basal Medium ◽

Centella Asiatica ◽

Perennial Herb ◽

Culture Techniques ◽

Nodular Callus ◽

Further Development

Centella asiatica L. (Apiaceae family), also called `Indian Pennywort,' is a prostrate, faintly aromatic, and stoloniferous perennial herb with long petiolated leaves. In the Ayurvedic medicine, it is reputed as a nervine tonic along with antibacterial, antifeedant, antileprotic and wound healing properties. Centella contains glycosides, indocentelloside, brahmoside, and asiaticoside. Its leaves are rich in carotenoids and vitamins B and C. In vitro culture techniques which offer a viable tool for mass propagation of plants have recently become increasingly popular for conservation of rare, endangered and threatened medicinal plants germplasm. Centella tissue culture has been reported to experience high incidences of microbial contamination which drastically reduces survival of explants. Thus, the main purpose of this study was to develop an efficient micropropagation technique for Centella asiatica to reduce explant contamination and rapidly disseminate superior clones for research and production. Here we present induction and further development of somatic embryos, using Centella stolons as explants. Somatic embryos were induced in response to 2,4-D shock on MS medium. Initially, somatic embryos appeared as highly nodular callus and eventually developed into somatic embryos that exhibited globular, heart shaped and cotyledonary stages. After auxin shock, cultures were regularly transferred to MS basal medium where somatic embryos completed various developmental stages and then germinated to give rise to new plantlets. In this presentation, we will demonstrate complete protocols for the successful sterilization of Centella explants prepared from plants that had abundance of fungal and bacterial contamination.

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Micropropagationof Male and Female Trees of Andaleh (MorusmacrouraMiq.) through In-vitro Culture using Several Compositionsof Basal Medium

JERAMI Indonesian Journal of Crop Science ◽

10.25077/jijcs.1.1.32-38.2018 ◽

2018 ◽

Vol 1 (1) ◽

pp. 32-38

Author(s):

Aswaldi Anwar

Keyword(s):

Callus Formation ◽

Basal Medium ◽

Shoot Multiplication ◽

Bud Break ◽

Shoot Induction ◽

Male And Female ◽

West Sumatra ◽

Parental Material

Andalehis the local name of MorusmacrouraMiq. in West Sumatra, Indonesia. Nowadays, this dioeciously speciesis in endangered situation. The aim of the research is to find out the appropriate combination of plant growth regulator to induce shoot multiplication of explants from male and female trees of andaleh. The plantlets from this research will be used in the next future to conserve this endangered species in vitro and in vivo, especially in preparing parental material in breeding program. Young buds from male and female trees were used as an explants in basal medium Murashige and Skoog supplemented with BAP (0.5, 1.0, 1.5 and 2.0 mg.L-1) in combination with NAA (1.0 mg.L-1 for each). The frequency of bud break was 50 % in MS medium supplemented with BAP (0.5 mg.L-1) and NAA 1.0 mg.L-1 for both source of explants (female and female trees of andaleh) after 3 weeks of culture. Generally, the number of shoot induction was very low. On the other hand, the rate of callus formation was high (100%) in highest BAP concentration (2.0 mg.L-1).

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Anther Culture of Potato and Molecular Analysis of Anther-derived Plants as Laboratory Exercises for Plant Breeding Courses

HortTechnology ◽

10.21273/horttech.9.4.585 ◽

1999 ◽

Vol 9 (4) ◽

pp. 585-588 ◽

Cited By ~ 2

Author(s):

Richard E. Veilleux

Keyword(s):

Plant Breeding ◽

Anther Culture ◽

Molecular Analysis ◽

Simple Procedure ◽

Extraction Procedure ◽

Basal Medium ◽

Root Tips ◽

Wide Range ◽

Simple Medium

Anther culture has been one of the most successful techniques for generating haploid plants over a wide range of species. It is a reasonably simple procedure that can be accomplished successfully without sophisticated laboratory facilities; yet, the plants generated through anther culture can be used to demonstrate the application of many modern methods that have direct applicability to plant breeding. Anthers of diploid potato clones that have been selected for competence in anther culture can be cultured in a simple medium to yield androgenic embryos after 5 weeks. Plant regeneration requires an additional 3 to 4 weeks. Regenerated plants should be large enough 2 weeks after transfer to basal medium for ploidy determination by any of three methods depending on available facilities: chromosome counts in root tips; chloroplast counts in stomatal guard cells; or flow cytometry of nuclei released from in vitro plantlets. DNA can be extracted from anther-derived plantlets using a rapid extraction procedure to demonstrate segregation of PCR (polymerase chain reaction)-based markers such as RAPD (randomly amplified polymorphic DNA), RAMPs (randomly amplified microsatellite polymorphisms), or microsatellites. Microsatellite markers that were heterozygous in the anther donor can be used to verify haploidy in anther-derived plants. If an anther culture laboratory is scheduled early in a semester, such molecular analysis can be planned for late in the same semester.

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