Strategies for overcoming genotypic limitations of in vitro regeneration and determination of genetic components of variability of plant regeneration traits in sorghum (original) (raw)
Related papers
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.
Efficient plant regeneration from shoot apices of sorghum
An efficient and rapid regeneration protocol was developed using shoot apices from germinating seedlings of two cultivars of sorghum, SPV-462 and M35-1, as explants. A vertical slit given from the base of each dissected apex enhanced the efficiency of callusing response by two fold. MS medium containing 0.5 mg dm -3 each of 2,4-D and kinetin was most effective in producing friable and embryogenic calli. Scanning electron microscopy of these calli detected somatic embryogenesis. Calli thus induced gave rise to approximately 42 green shoots per callus in both the genotypes when transferred to regeneration medium containing 1.5 mg dm -3 kinetin.
Industrial Crops and Products, 2017
Ajowan (Carum copticum L.) is an important and endangered industrial medicinal plant that growing in some parts of Iran. Two efficient protocols, without somaclonal variation induction, were developed for indirect somatic embryogenesis and indirect shoot regeneration of three Iranian ecotypes of ajowan. In the first experiment, higher concentration of auxin than cytokinin (1, 1.5 and 2 mg/L of 2,4-dichlorophenoxyacetic acid along with 0.25, 0.5 and 0.75 mg/L kinetin) was used for callus induction in 5, 10 and 15 days old hypocotyl explants. In the second experiment, higher concentration of cytokinin than auxin (1 mg/L of kinetin along with 0.5 mg/L of 2,4-dichlorophenoxyacetic acid) was used for callus induction in 15-d old hypocotyl explants. The higher frequency of somatic embryos was achieved from 15 days old hypocotyl explants of Ghoom ecotype with 28.33 embryos when induced calli from MS medium supplemented with 1.5 mg/L 2,4-dichlorophenoxyacetic acid in combination with 0.5 mg/L kinetin transferred to free plant growth regulator MS medium. Momentary removing of 2,4dichlorophenoxyacetic acid was successful for somatic embryogenesis in Iranian ecotypes of ajowan that it significantly reduce the time of the culture and thus reduce the risk of somaclonal variation. Maximum number of initiated shoots per explant was related to Shiraz ecotype with average 18.33 shoots per callus in MS medium supplemented with 1.5 mg/L of specify type of cytokinin (3-methoxy [-6-benzylamino-9-tetrahydropyran-2-yl] purine) plus 0.25 mg/L naphathalene acetic acid. The survival rate of rooted plantlets was 60.86% and 58.33% for indirect somatic embryogenesis and indirect shoot regeneration derived plants, respectively. The genetic stability of regenerated plants via indirect somatic embryogenesis and indirect shoot regeneration was proved through flow cytometry analysis.
Callus Induction and Plant Regeneration of Commercial Rice (Oryza sativa L.) Cultivars
Journal of the Arkansas Academy of Science, 1995
Manipulation of agronomic traits at the cellular and molecular levels offers an efficient approach to enhance conventional breeding efforts for rice improvement. Plant regeneration protocols, required for biotechnological applications, have not yet been developed for a number of important rice cultivars. This study was conducted to establish a system for plant regeneration of elite rice cultivars adapted to the southern U.S.A. Callus was induced from dehusked grains of cultivars Alan, Katy, and LaGrue, on MS media containing 0.5, 2, and 4 mg L 1 2,4-D, with 0.5 mg L 1 kinetin or without kinetin. Plant regeneration was accomplished by transferring the callus to a hormone-free medium. Callus proliferation was influenced by 2,4-D, kinetin, and genotype in two-way interactions. The effects of these factors on embryogenesis and rhizogenesis was expressed in a three-way interaction. Depending upon the genotype up to 50% plant regeneration was obtained. In most cases treatments consisting of 0.5 to 2 mg L 1 2,4-D plus 0.5 mg L 1 kinetin produced the best callus proliferations with the highest embryogenic capacity. Regenerants grew to maturity in soil and produced viable seeds. The establishment of this regeneration system is essential for the development of a genetic transformation system for the aforementioned commercial rice cultivars.
Improved plant regeneration in callus cultures of Sorghum bicolor (L.) Moench
In Vitro Cellular & Developmental Biology - Plant
Sorghum bicolor is a recalcitrant species for tissue culture regeneration and genetic transformation. Browning of explants is one of the factors limiting organ and tissue cultures. To overcome this, callus tissue was initiated from the shoot tips of in vitro germinating seeds (S. bicolor cv. Róna 1), and then cultured on modified MS media (Murashige and Skoog in Physiol Plant 15:473-497, 1962). In the first experiment, we tested callus induction on several media supplemented with casein hydrolysate, polyvinylpyrrolidone, honey, and sucrose. The best callus induction was recorded for the medium with honey and sucrose (80.0%) and for control medium (79.8%). Shoot regeneration was tested on the MS medium with 6-benzylaminopurine (BAP) supplemented with honey and sucrose at a 1:1 ratio (by weight) or with sucrose only. The highest percentage of calluses regenerating shoots was noted for those induced on the medium with sucrose and honey-approx. four times higher when compared to the control. Rooted plantlets were acclimatized with a 92% survival rate. In the second experiment, we analyzed culture responses to various ways of honey application to the induction media: honey (autoclaved or filtered) in presence or absence of sucrose. Supplementation of the medium with fructose, glucose, and maltose at a proportion typical for honey was also investigated. The explant and callus survival rates were similar to those of the honey-sucrose combination in the first experiment. Only presence of both sucrose and honey in the induction medium improved the total regeneration rate to 37.9% over the control (18.8%). Sucrose and honey appear to act synergistically for shoot regeneration in callus cultures of sorghum.
Proceedings of the National Academy of Sciences, India Section B: Biological Sciences, 2017
An efficient indirect in vitro plant regeneration protocol for Dianthus caryophyllus L. cv. 'Master' was developed using leaf explants. This study revealed the morphogenetic potential of leaf explant as a source for micropropagation. Murashige and Skoog (MS) medium supplemented with 2.0 mg/l 2,4-dichlorophenoxyacetic acid and 0.5 mg/l Naphthalene acetic acid (NAA) resulted in maximum (94.44%) callus induction. MS medium supplemented with 1.5 mg/l thidiazuron, 0.25 mg/l Kinetin and 0.25 mg/l NAA was found to be highest for average shoot regeneration (80.56%), average number of shoots (6.01) and average shoot length (1.93 cm). For in vitro multiplication of shoots, MS medium supplemented with 2.0 mg/l Kinetin and 0.25 mg/l NAA was found to be the best which resulted in 14.64 average number of microshoots. It was observed that the half strength MS basal medium supplemented with 1.5 mg/l Indole-3-butyric acid and 0.02% activated charcoal showed maximum rooting (98.19%) with 9.60 average number of roots per microshoot having 4.24 cm root length. The in vitro rooted plantlets were hardened gradually and were successfully acclimatized under ex vitro conditions. The genetic variation in the in vitro raised plants was confirmed using DNA based markers [Random Amplified Polymorphic DNA and Inter Simple Sequence Repeats] for assessment of genetic stability of plants raised through indirect regeneration.
AFRICAN JOURNAL OF BIOTECHNOLOGY
Optimization of tissue culture conditions for Sorghum bicolor L. through somatic embryogenesis from immature embryos is important for the genetic manipulation and improvement of this agronomically valuable crop. In an attempt to develop a successfully reproducible in vitro regeneration protocol for a group of diverse sorghum genotypes, 10 sorghum lines including locally adapted and commercially important elite genotypes were assessed for their regeneration potential on different culture media–containing adequate growth regulators combinations. The maximum response of embryogenic callus induction was obtained from explants cultured on Murashige and Skoog (MS) medium supplemented with 1.5 mgL-1 of 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.7 mgL-1 L-proline. The addition of kinetin to the MS-based culture media had a negative effect on the formation of embryogenic calli. The results reveal that embryogenic callus formation and regeneration were highly genotype dependent. The line LG...
Scientific World, 2010
Response of genotypes to culture media for callus induction and subsequent regeneration from rice anthers were investigated at Biotechnology Unit, Khumaltar, Lalitpur, Nepal. Boots of several rice genotypes viz. Bindeshwari, Hardinath- 1, Prabhat, Khumal-4, Chhomrong local and Chandnath-3 were cold pretreated at 8±1°C for seven days. Anthers from these boots were aseptically cultured on three different medium designated as Medium A: N6 mineral salts and vitamins (2 mg/l each) + myoinositol 100 mg/l + 2,4-D 2.5 mg/l + kinetin 0.5 mg/l + AgNO3 10 mg/l + maltose 50 gm/l; Medium B: N6 mineral salts and MS vitamins + NAA 4 mg/l + Kinetin 2 mg/l + AgNO3 5 mg/l and sucrose 60 gm/l; and Medium C: Medium B without AgNO3. Results revealed that the response of genotype to various media compositions were highly significant for response of anthers for callus induction and embryogenic calli formation. The interaction between genotypes and media were also significant. Among media, the frequency of...
Advance Journal of Food Science and Technology
The objective of the present study is to develop callus induction and plant regeneration; an experiment was conducted to investigate the effect of growth regulators on callus induction in rice mature embryo culture. In this experiment, callus was initiated from the mature seed scutellumof two Malaysian indica rice, MRQ74 and MR269. Callus was induced using different concentrations of 2, 4-D and BAP on MS medium under dark condition or under 16/8 h photoperiod conditions. For complete plant regeneration, the calli of both varieties were planted for shoot regeneration on MS media supplemented with different concentrations of BAP and NAA, for root regeneration on IBA. The results revealed that the maximum callus inductions in MRQ74 and MR269 were observed in the MS medium containing 3 mg/L 2, 4-D with 0.1 mg/L BAP under dark conditions. Furthermore, our protocol uses mature seeds as the explants in both rice varieties in terms of callus induction. Additionally, green plant production of plants was also observed in all of the media tested for shoot regeneration and MS media supplemented with 3 mg/L of BAP and 0.1 mg/L of NAA was found to be responsible for the recovery of more highly regenerated shoots. Effective root induction (100%) was achieved in a medium containing 1mg/LIBA. The described protocol can be useful in the establishment of an in vitro system for rice that can be a major technique for a genetic transformation system and crop improvement.