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

2016 ◽  
Vol 5 (08) ◽  
pp. 1395 ◽  
Author(s):  
Vijaya Naresh Juturu ◽  
Gopala Krishna Mekala ◽  
Mallikarjuna Garladinne ◽  
Puli Chandra Obul Reddy ◽  
Akila Chandra Sekhar*

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.

2019 ◽  
Vol 7 ◽  
pp. 97-104
Author(s):  
Md. Niuz Morshed Khan ◽  
Md. Monirul Islam ◽  
Dr. Md. Shahidul Islam

Due to growing population, there is an increasing demand of rice production but the productivity of rice is lessened day by day. To overcome this problem various biotechnological tools can be used for developing various rice varieties. However, the lack of a simple and efficient protocol for callus induction, embryogenic callus formation and quick plant regeneration in this cereal crop. In this study embryogenic calli from mature seeds of five indica rice varieties viz. Binadhan-5, Binadhan-6, BRRI dhan-48, BRRI dhan-58 and IR-64 were observed that is done in four different types of media composition. The highest callus induction were observed in media containing Sucrose as a carbon source. Among those varieties Binadhan-6 and BRRI dhan-48 showed highest rate of callus induction respectively. This study will be useful for selecting suitable callus induction medium for callus induction in future that will be useful for not only national but also international plant breeders for producing new variety and so on.


2017 ◽  
Vol 20 (2) ◽  
pp. 81-87
Author(s):  
HN Barman ◽  
ME Hoque ◽  
RK Roy ◽  
PL Biswas ◽  
MAI Khan ◽  
...  

The study was conducted at Biotechnology Division of Bangladesh Rice Research Institute (BRRI) to investigate the effects of plant growing medium and plant growth regulator (PGR) for the callus induction and high frequency plantlets regeneration of indica rice. Ten indica rice varieties viz. BR5, BR11, BRRI dhan28, BRRI dhan29, BRRI dhan33, BRRI dhan41, BRRI dhan47, BRRI dhan48, BRRI dhan49 and BRRI dhan50 were cultured on MS, N6 and LS media. The MS medium was found better for callus induction as compared to N6 and LS media. Among the tested varieties BRRI dhan48 induced the highest percent and best quality callus. Interaction effects of BRRI dhan48 to MS medium yielded 71.55% callus induction. The regeneration efficiency of BRRI dhan48 was tested on MS medium supplemented with different combination of NAA plus BAP and NAA plus kinetin. MS medium supplemented with 2.0 mg L-1 NAA and 2.0 mg L-1 Kn was found the best in respect of percent regenerated (76.67%) plantlet as well as for the growth of plantlets in vitro.Bangladesh Rice j. 2016, 20(2): 81-87


2015 ◽  
Vol 3 (4) ◽  
pp. 680-686 ◽  
Author(s):  
Mohitosh Dey ◽  
Sanjib Kumar Panda ◽  
Lingaraj Sahoo

An efficient plant regeneration system from embryogenic callus of two elite indica rice (Oryza sativa spp. indica) varieties of Northeast India, Ketokijoha and Monoharsali is established. The effect of auxin, 2,4-dicholorophenoxy acetic acid (2,4-D) on callus induction was optimized. Friable, nodular and creamish-white embryogenic calli were induced from mature seeds on NB medium supplemented with 2.5 mg/l 2,4-D. Plants were regenerated from 40-50 days old embryogenic callus on NB medium containing 0.5 mg/l BAP (6-benzylaminopurine) and 0.25 mg/l ABA (abscisic acid). Regenerated plants with multiple tillers were rooted on half strength MS medium and rooted plants were acclimatized with 94% survival rate. Higher frequency of callus induction as well as plant regeneration was recorded in Ketokijoha as compared to Monoharsali. The calli of both the varieties were found amenable to Agrobacterium-mediated transformation as evident from strong GUS (β-glucuronidase) expression. The results may find wide application for genetic improvement for valuable traits these elite indica rice varieties of Northeast India. Int J Appl Sci Biotechnol, Vol 3(4): 680-686


2012 ◽  
Vol 5 (1) ◽  
pp. 173-177 ◽  
Author(s):  
MJ Alam ◽  
M Imran ◽  
L Hassan ◽  
MH Rubel ◽  
M Shamsuddoha

The experiment was conducted to observe the regeneration potential and also to establish a suitable in vitro plantlet regeneration protocol from mature seed derived embryogenic calli of four indica rice varieties viz BRRI dhan28, BRRI dhan29, BRRI dhan47 and Binadhan-7 after partial desiccation treatment. Different concentrations and combinations of growth regulators were used in MS medium to observe the callus induction ability using mature embryo as explants. The percentage of callus induction frequency  was highest (86.00) in BRRI dhan47 and the lowest (56.50) in Binadhan-7 Among the culture media the performance of MS +500 mgl-1 L- Proline + 2.0 mgl-12, 4-D + 0.8 mgl-1BAP was better than any other media for callus induction frequency (%), rapid  callusing, size of the callus (mm), texture of callus and color of callus. Among the four varieties, shoot regeneration was highest in BRRI dhan29 (84.33%) which required minimum (14.80) days to and the lowest was in Binadhan-7 (39.67%) which required maximum (15.47) days. Among the treatments, the highest (65.75%) shoot regeneration was observed with MS + 6.0 mgl-1Kn +0.5 mgl-1 NAA which required minimum days (13.75) to develop green bud formation and the lowest shoot regeneration (56.50) was observed with MS + 2.0 mgl-1Kn + .05 mgl-1 NAA. BRRI dhan29 produced more number of shoots (4.67) per callus while Binadhan-7 showed minimum number of shoots (2.87) per callus. The highest number of shoot producing roots (3.66) was observed in BRRI dhan29 which showed maximum number of root per plant and the lowest (3.11) in Binadhan-7. Among the three treatments MS + 0.6 mgl-1IBA showed highest percentage 86.67 of root followed by MS+ 0.6 mgl-1IBA 70% and the lowest (66.7%) was in MS+ 0.4 mgl-1IBA. The establishment rate of the plantlet in the pot was the highest (67.67%) in BRRI dhan29 and Binadhan-7 showed lowest establishment rate (51.22%) in pot. DOI: http://dx.doi.org/10.3329/jesnr.v5i1.11573J. Environ. Sci. & Natural Resources, 5(1): 173 - 177, 2012


1970 ◽  
pp. 41-45 ◽  
Author(s):  
Md. Niuz Morshed Khan ◽  
Dr. Md. Monirul Islam ◽  
Dr. Md. Shahidul Islam

Due to growing population, there is an increasing demand of rice production but the productivity of rice is lessened day by day. To overcome this problem various biotechnological tools can be used for developing various rice varieties. However, the lack of a simple and efficient protocol for callus induction, embryogenic callus formation and quick plant regeneration in this cereal crop. In this study embryogenic calli from mature seeds of five indica rice varieties viz. Binadhan-5, Binadhan-6, BRRI dhan-48, BRRI dhan-58 and IR-64were observed that is done in four different concentration of growth regulators (2,4-D). The highest callus induction were observed in media containing 2 mg/L 2,4-D. Among those varieties BRRI dhan-48 showed highest rate of callus induction. In this experiment Agrobacterium mediated gene transfer is also tried although plants were found nut need to be tested by molecular methods and tested the desired characters for selecting those plantlets as transgenic plant. This study will be useful for selecting suitable callus induction medium for callus induction in future.


1970 ◽  
Vol 19 (2) ◽  
pp. 185-197
Author(s):  
T.L. Aditya

An efficient protocol was developed for in vitro morphogenic ability along with plantlet regeneration of two Bangladeshi indica rice varieties (BR24 and BR26) via somatic embryogenesis by applying 50 mM NaCl stress in callus induction and suspension initiation media. Osmotic stress was induced by NaCl (50, 100, 150, 200 and 250 mM) on the cell growth in suspension maintenance media. In viability test stress adapted cells showed 85 - 95% viability up to 200 mM NaCl compared with stress shocked (MS1-50) and control (MS1-0) treatments. Higher stress adapted cells showed growth retardation and the induction of plasmolysis. For both genotypes somatic embryos were obtained in both MS based liquid and semisolid media with or without 50 and 100 mM NaCl. Cell suspension-derived micro-calli were partially desiccated (6 - 12 hr) and subsequently maintained in MS1 callus induction media supplemented with proline (12 mM), ABA (2 mg/l) and 0.6% phytagel in the presence or absence of 50 and 100 mM NaCl. Subsequently, desiccated somatic embryos were transferred in MS based regeneration media with or without 50 and 100 mM NaCl. Proline mediated callus was found to be more effective in embryo differentiation than ABA. Partial desiccation dramatically enhanced callus growth and partially increased regeneration percentage. BR24 showed a better regeneration response producing plantlets in presence of proline in control media while BR26 restored regeneration potential in the presence of ABA and 100 mM NaCl. Plantlets regenerated from salt stressed callus cultures were then grown in compost in a glasshouse and produced normal, fertile plants.  Key words: Indica rice, Cell suspension, Morphogenic, Regeneration D.O.I. 10.3329/ptcb.v19i2.5436 Plant Tissue Cult. & Biotech. 19(2): 185-197, 2009 (December)


2006 ◽  
Vol 86 (1) ◽  
pp. 63-69
Author(s):  
Seedhabadee Ganeshan ◽  
Brian J Weir ◽  
Monica Båga ◽  
Brian G Rossnagel ◽  
Ravindra N Chibbar

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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