Plant tissue culture studies in Sorghum bicolor: immature embryo explants as the source material (original) (raw)
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Adventitious shoot regeneration from immature embryos of sorghum
Plant cell, Tissue and organ culture, 2002
Eleven genotypes of sorghum were examined for their response in tissue culture, and the tissue culture system was optimized. The cultures were initiated from immature embryos taken approximately two weeks after flowering. The response of immature embryos varied with the genotype. 'C. Kafir' and 'PE932 025' showed the highest frequency of callus induction and regenerable callus formation under appropriate culture conditions. Regeneration occurred at high frequencies when cytokinins (kinetin or 6-benzyladenine) had been added in the callus induction medium, followed by regeneration medium devoid of growth regulators. The addition of proline and polyvinylpyrrolidone also enhanced shoot formation, but the addition of cytokinins to regeneration media did not improve shoot formation. On the revised culture medium, plants were regenerated from up to 100% of sorghum immature embryos.
Enhanced shoot regeneration in tissue culture studies of Sorghum bicolor
The effect of combination of benzyl aminopurine (BAP), thidiazuron (TDZ) and indole acetic acid (IAA) was studied on in vitro shoot proliferation from immature embryo explants of Sorghum bicolor (L.) Moench, an economically important cereal. Proliferation of multiple shoots was achieved on MS medium supplemented with 1.5 mgL-1 BAP + 1.5 mgL-1 TDZ + 1.0 mgL-1 IAA + 1000 mgL-1 L-asparagine, L-proline, L-glutamine and serine. Upto 72 plantlets per explant were obtained in the present study. Response of six varieties; IS 3566, SPV 475, CSV 13, CSV 15, CSV 112 and IS 348 to multiple shoot formation was studied. The maximum number of shoots were observed in the variety IS 3566. The in vitro proliferated and elongated shoots were transferred individually to a root induction medium containing 2% sucrose + 1.0 mgL-1 NAA and within 21 days 162 roots per culture which were produced from multiple shoots. The regenerated plantlets were transferred to 1:1 soil and vermiculite mixture and acclimatized with 60 % survival rate. Fully acclimatized plants were grown in garden soil in greenhouse and their morphological and physiological parameters as similar with seedlings.
Regeneration of sorghum from shoot tip cultures and field performance of the progeny1
Plant Cell Tissue and Organ Culture, 2000
A system for rapid plant regeneration through somatic embryogenesis from shoot tip explants of sorghum [Sorghum bicolor (L.) Moench] is described. Somatic embryogenesis was observed after incubation of explants in dark for 6–7 weeks through a friable embryogenic callus phase. Linsmaier and Skoog medium supplemented with 2,4-dichlorophenoxyacetic acid (2 mg l−1) and kinetin (0.1 mg l −1) was used for induction of friable embryogenic calli and somatic embryos. Germination of somatic embryos was achieved about 5 weeks after transfer onto Murashige and Skoog (MS) medium supplemented with 6-benzylaminopurine (2 mg l−1) and indole-3-acetic acid (0.5 mg l −1) under light. Seeds from in vitro-regenerated plants produced a normal crop in a field trial, and were comparable to the crop grown with the seeds of the mother plant used to initiate tissue culture. The simplicity of the protocol and possible advantages of the system for transformation over other protocols using different explants are discussed.
Applied Ecology and Environmental Research, 2019
An effective regeneration system was developed from embryonic callus that was formed by using mature embryos in 6 sorghum cultivars (Gözde 80, Greengo, Leoti, Beydarı, Aldarı and Akdarı). Different auxin (2,4-D and 2,4,5-T) and cytokinin (zeatin and kinetin) combinations on somatic embryogenesis were studied. The highest embryonic callus in all cultivars was derived from cultures in MS medium containing 1 mg/l 2,4-D. The transfer of embryonic callus obtained from medium containing 2,4,5-T + kinetin to the shooting medium (1 or 2 mg/l BA +1.5 mg/l TDZ +1 mg/l IAA) and subsequently rooting medium (½ MS with 1 mg/l NAA) resulted in a higher shooting and rooting. Different concentrations of BA in the shooting medium did not affect shoot formation. Akdarı and Greengo cultivars produced better callus induction and regeneration than the other cultivars as grain and silage types, respectively. Rooting and surviving rates varied between 10.55-68.37% depending on the growth regulators used at the beginning of culture. Growth and survival rates were increased in plants transferred from high-shoot-rate cultivars to the rooting medium.
Cell and Tissue Culture Approaches in Relation to Crop Improvement
Biotechnologies of Crop Improvement, Volume 1, 2018
Plant cell and tissue culture involves the growing of cells, tissues and organs on synthetic medium under closely controlled and aseptic conditions. Plant cell and tissue culture methods offer a rich scope for the creation, conservation and utilization of genetic variability for the improvement of field, horticultural and forest plant species. Micropropagation of selected plant species is one of the best and most successful examples of the commercial application of tissue culture technology. Micropropagation ensures true-to-type, rapid and large-scale multiplication. Now scores of multimillion-dollar industries around the world propagate a variety of plant species through micropropagation. Tissue culture technology offers environmental-friendly industries to flourish. It is likely that automation of multiplication systems will be commercially feasible within the next few years for several species including potato microtubers, lily bulblets and gladiolus corms. Meristem culturing and in vitro grafting help in developing disease-free plants. Improvement of somatic embryogenesis, coupled with embryo desiccation and encapsulation technology, may lead to the utilization of 'artificial seeds' for mass cloning of plants. Further induction of somatic embryogenesis in plants helps in cloning and transformation. Somaclonal variation is a potent emerging aspect for broadening the genetic base and thus obtaining incremental improvement in the commercial cultivars, more particularly, in the vegetatively propagated plant species. Using the technique of in vitro selection, many million cells/protoplasts can be screened against various biotic and abiotic stress factors in a single Petri dish which is more efficient as compared to the screening of similar number of plants in the field which requires more time and space as well. Production of haploids through bulbosum, anther/pollen culture and embryo rescue from wide hybrids has been exploited for the produc
In vitro regeneration and morphogenesis studies in common bean
Plant Cell Tissue and Organ Culture, 2010
An efficient protocol for high frequency in vitro regeneration of multiple shoots and somatic embryos from the embryonic axis of common bean (Phaseolus vulgaris) was developed. Ten common bean cultivars representing a wide range of diversity among current commercial market classes were used for in vitro regeneration evaluation in our study. These cultivars were tested on 63 different media formulations consisting of combinations of cytokinins, namely benzyladenine (BA) and thidiazuron (TDZ) at concentration levels of 0.0, 1.0, 2.5, 5.0 and 10.0 mg l−1 and auxin, namely naphthalene acetic acid (NAA) and indole-3-acetic acid (IAA) at concentration levels of 0.0, 0.05, 0.1 and 1.0 mg l−1. P. vulgaris cv. Olathe pinto bean performed the best producing over 20 multiple shoots per explant while cv. Condor black bean was the poorest with nine multiple shoots per explant. The optimum media for regeneration of multiple shoots was 4.4 mg l−1 Murashige and Skoog (MS) containing 2.5 mg l−1 BA and 0.1 mg l−1 IAA supplemented with 30 mg l−1 silver nitrate. Adventitious shoots and somatic embryos were regenerated on 4.4 mg l−1 MS medium containing 1 mg l−1 TDZ and 0.05 mg l−1 NAA supplemented with 30 mg l−1 silver nitrate or activated charcoal. Efficient and effective rooting of plantlets was achieved by dipping the cut end base of in vitro regenerated shoots in 1.0 mg l−1 indole-3-butyric acid (IBA) solution and culturing on media containing 4.4 mg l−1 MS supplemented by 0.1 mg l−1 IAA, NAA or IBA.
African Journal of Biotechnology
In the present study, an efficient protocol for somatic embryogenesis and plant regeneration was established in six finger millet varieties (GBK-043137, GBK-043128, GBK-043124, GBK-043122, GBK-043094 and GBK-043050). Shoot tips from 3 days in vitro grown plants were inoculated on MS supplemented with various concentrations and combinations of α-naphthaleneacetic acid (NAA), 2,4-Dichlorophenoxyacetic acid (2,4-D), benzylaminopurine (BAP) and kinetin for callus induction and somatic embryogenesis. For shoot regeneration, somatic embryos were cultured on various concentrations of BAP, while root induction was done using different concentrations and combinations of NAA, kinetin, BAP and 2,4-D. Acclimatization of regenerated plants was tested using forest soil, cocopeat, manure, sand and fertilizer either singly or in combination. Best callus formation was achieved on 2.5 mg/l of 2,4-D and 1.5 mg/l BAP with a mean of 12.33±0.33 on variety GBK-043128 while shooting and rooting were best on 1.75 mg/l BAP with a mean of 25.07±0.64 and 1.0 BAP+0.25 NAA with a mean of 15.00±2.2, respectively. Best acclimatization was attained using soil, sand and fertilizer on GBK-043094. Plants regenerated were morphologically similar to in vivo plants with 97% survival rate. Moreover, they were fertile and able to set viable seeds. This efficient protocol has the potential for crop improvement and genomic studies.