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

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

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An efficient regeneration system through in vitro somatic embryogenesis of barley (Hordeum vulgare L.)

Journal of Bio-Science ◽

10.3329/jbs.v27i0.44674 ◽

2019 ◽

Vol 27 ◽

pp. 89-99

Author(s):

M Haque ◽

SMS Islam

Keyword(s):

Plant Regeneration ◽

Callus Induction ◽

Casein Hydrolysate ◽

Ms Medium ◽

Regeneration System ◽

Embryogenic Calli ◽

Hordeum Vulgare L ◽

The Media ◽

Barley Genotypes

This study was carried out to improve an efficient protocol for in vitro callus induction and plant regeneration using Bangladeshi barley genotypes collected from BARI, Gazipur, Bangladesh. After sterilization embryos were separated carefully from mature seeds of six barley genotypes (BB-1, BB-2, BB-3, BB-4, BB-5 and BB-6) and cultured them in MS medium supplemented with various concentration and combination of PGRs for callus induction and regeneration. Out of six genotypes BB-6 showed highest (38.17%) callus induction in MS + 4.0 mg/l 2,4-D + 200 mg/l L-proline + 300 mg/l casein hydrolysate; whereas, BB-4 and BB-5 showed no callus induction in the same medium. For plant regeneration from embryogenic calli the same genotype (BB-6) also performed the best results (19.25%) in MS medium supplemented with 1.5 mg/l BAP + 30 g/l sucrose. Analysis of variance (ANOVA) showed highly significant differences among the media and the genotypes. J. bio-sci. 27: 89-99, 2019

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Optimization of in vitro regeneration protocol for a popular Indica rice (Oryza sativa L. cv Swarna)

Annals of Plant Sciences ◽

10.21746/aps.2016.08.002 ◽

2016 ◽

Vol 5 (08) ◽

pp. 1395 ◽

Cited By ~ 1

Author(s):

Vijaya Naresh Juturu ◽

Gopala Krishna Mekala ◽

Mallikarjuna Garladinne ◽

Puli Chandra Obul Reddy ◽

Akila Chandra Sekhar*

Keyword(s):

Callus Induction ◽

Oryza Sativa L ◽

Indica Rice ◽

In Vitro Regeneration ◽

Regeneration System ◽

Embryogenic Calli ◽

Rice Varieties ◽

Regenerated Plants ◽

Rooting Medium

Though regeneration system in rice has been very well established compare to other crop plants, the fact remains that, most of the indica rice varieties are still recalcitrant for regeneration and genetic transformation. Therefore, refinement of tissue culture protocol for generation of embryogenic calli and regeneration of the fertile plants from a single cell should be a pre requisite event for development of transgenic plants. Here, in this study we reported high frequency robust regeneration protocols for a popular Indica cultivar Swarna.Mature seeds were used as initial material as explants. Highest callus induction % was observed in MSCIMP medium containing 2.0 mg-1 2,4, D + 0.5 mg-1 Kn as phytohormonal combinations. In addition, maximum regeneration was observed in 2.0 mg-l KN + 0.5 mg-l NAA. Regenerated plants were shifted to rooting medium followed by polyhouse for hardening. The callus induction and regeneration reported in this study were well suited for transformation agronomical important genes or functional genomics studies.

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Regeneration of Pinus halepensis (Mill.) through Organogenesis from Apical Shoot Buds

Forests ◽

10.3390/f12030363 ◽

2021 ◽

Vol 12 (3) ◽

pp. 363

Author(s):

Cátia Pereira ◽

Itziar A. Montalbán ◽

Ana Pedrosa ◽

Jéssica Tavares ◽

Alexey Pestryakov ◽

...

Keyword(s):

Somatic Embryogenesis ◽

Pinus Halepensis ◽

Culture Media ◽

In Vitro Regeneration ◽

Naphthaleneacetic Acid ◽

Embryogenic Calli ◽

Shoot Buds ◽

Apical Shoot ◽

Adult Trees

Organogenesis and somatic embryogenesis have been widely applied as the two main regeneration pathways in plant tissue cultures. However, recalcitrance is still the main restriction in the clonal propagation of many woody species, especially in conifers. They undergo a “phase change” that leads to significant loss of vegetative propagation capacity, reducing the aptitude of tissues and organs to be regenerated in vitro beyond this point. In line with this, the in vitro regeneration of mature conifer trees has been a long-cherished goal in many laboratories worldwide. Based on previous works in Pinus species regeneration from adult trees, we now present data about the culture of apical shoot buds in an attempt to induce organogenesis and somatic embryogenesis to clone mature trees of Aleppo pine (Pinus halepensis). Reinvigorated axillary shoots were submitted to conditions usually applied to induce somatic embryogenesis through the manipulation of culture media, including the use of auxins such as 2,4-Dichlorophenoxyacetic acid and 1-Naphthaleneacetic acid, cytokinins (6-benzyladenine and kinetin), and phytosulfokine (50, 100, and 200 nM). Although somatic embryos could not be obtained, an embryogenic-like tissue was produced, followed by the emergence of actively proliferating non-embryogenic calli. Variations in the consistence, texture, and color of non-embryogenic calli were observed; especially those arising in the media containing phytosulfokine. Reinvigorated shoots, induced by 22 or 44 µM 6-benzyladenine, were obtained through organogenesis and acclimatized, and phenotypically normal plants were obtained.

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In vitro Plant Regeneration of Four Local Varieties of Chickpea (Cicer arietinum L.) Grown in Bangladesh

Bangladesh Journal of Scientific and Industrial Research ◽

10.3329/bjsir.v46i3.9047 ◽

1970 ◽

Vol 46 (3) ◽

pp. 379-384 ◽

Cited By ~ 2

Author(s):

TA Banu ◽

RH Sarker ◽

MI Hoque

Keyword(s):

In Vitro Regeneration ◽

Multiple Shoots ◽

Mature Embryo ◽

Multiple Shoot ◽

Direct Organogenesis ◽

Ms Medium ◽

Regeneration System ◽

Best Response ◽

Multiple Shoot Regeneration

In vitro regeneration system was developed through direct organogenesis from decapitated mature embryo explants of locally grown four chickpea varieties, namely, Barichhola-4, Hyprochhola, Binachhola-3 and Binachhola-4. Best response towards multiple shoot regeneration was obtained on MS medium supplemented with 0.5 mg/l BAP, 0.5 mg/l Kn, 0.2 mg/l NAA along with double concentrations of CaCl2 and NH4NO3. However good shoot health and expanded leaf was found on MS medium containing 1.0 mg/l kn. Apart from this, few experiments were conducted with decapitated embryo attached cotyledon. Using this explants highest number of multiple shoots were obtained on MSB medium containing 4× micronutrients of MS medium with 3.0 mg/l BAP and 0.04 mg/l NAA in all four varieties. Shoots regenerated on 1.0 mg/l kn supplemented medium showed good response towards rooting on MS medium supplemented with 0.2 mg/l IBA in all four varieties. It was observed that micrografting is an alternative technique to in vitro rooting in chickpea. Key words: In vitro regeneration; Decapitated embryo; Chickpea. DOI: http://dx.doi.org/10.3329/bjsir.v46i3.9047 BJSIR 2011; 46(3): 379-384

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In vitro regeneration of three varieties of Brassica campestris L. grown in Bangladesh

Bangladesh Journal of Scientific and Industrial Research ◽

10.3329/bjsir.v55i3.49391 ◽

2020 ◽

Vol 55 (3) ◽

pp. 181-188

Author(s):

B Goswami ◽

MI Hoque ◽

S Khan ◽

RH Sarker

Keyword(s):

Shoot Regeneration ◽

Brassica Campestris ◽

In Vitro Regeneration ◽

Shoot Formation ◽

Multiple Shoot ◽

Root Induction ◽

Regeneration System ◽

Cotyledonary Leaf ◽

Best Response

A reproducible in vitro regeneration system was developed for three varieties of Brassica campestris L. namely Agrani, BINA Sarisha-10 and BINA Sarisha-6 using hypocotyl and cotyledonary leaf with petiole as explants. MS medium supplemented with 2.0 mg/l BAP and 0.2 mg/l IAA was found to be the best for the multiple shoot formation for all the three varieties. Among three varieties, BINA Sarisha-6 showed best response in terms of shoot regeneration as well as number of shoots per explant (9.0) using hypocotyls as explants. In case of Agrani and BINA Sarisha-10 highest number of shoot per explants were found 8.2 and 7.0, respectively. Interestingly in vitro regenerated shoots of BINA Sarisha-6 and BINA Sarisha-10 were produced in vitro flowers on shoot regeneration media. Best root induction in BINA Sarisha-6, Agrani and BINA Sarisha-10 was achieved on MS media supplemented with 0.5 mg/l IBA. After proper hardening, the in vitro regenerated plantlets were successfully transplanted into soil. Bangladesh J. Sci. Ind. Res.55(3), 181-188, 2020

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