scholarly journals Improvement of the in vitro regeneration and Agrobacterium-mediated genetic transformation of Medicago sativa L.

2019 ◽  
Vol 43 (1) ◽  
pp. 96-104
Author(s):  
Farzad NOFOUZI ◽  
Muhammet Çağrı OĞUZ ◽  
Saber Delpasand KHABBAZI ◽  
Ali ERGÜL
Euphytica ◽  
1988 ◽  
Vol 39 (1) ◽  
pp. 3-9 ◽  
Author(s):  
Y. Wan ◽  
E. L. Sorensen ◽  
G. H. Liang

Genome ◽  
1992 ◽  
Vol 35 (3) ◽  
pp. 474-477 ◽  
Author(s):  
G. A. Kielly ◽  
S. R. Bowley

The genetic control of somatic embryogenesis in alfalfa (Medicago sativa L.) was studied using one nonembryogenic and three embryogenic genotypes: A70-34, a selection from 'Rangelander'; RA3, a selection from 'Regen-S'; and C2-4, a selection from a breeding population that had A70-34 in its pedigree. Crosses of embryogenic × embryogenic and embryogenic × nonembryogenic and S1 and BC1 testcrosses were evaluated for in vitro regeneration. Selfing reduced the expression of the trait. Somatic embryogenesis was dominant and explained by two loci. All three regenerating genotypes shared a common genetic system.Key words: alfalfa, Medicago sativa, somatic embryogenesis, tissue culture.


2020 ◽  
Vol 16 (6) ◽  
pp. 937-941
Author(s):  
Sharad Vats ◽  
Preeti Mehra

Background: Vector-borne diseases are quite prevalent globally and are one of the major causes of deaths due to infectious diseases. There is an availability of synthetic insecticides, however, their excessive and indiscriminate use have resulted in the emergence of resistant varieties of insects. Thus, a search for novel biopesticide has become inevitable. Methods: Rotenoids were isolated and identified from different parts of Medicago sativa L. This group of metabolites was also identified in the callus culture, and the rotenoid content was monitored during subculturing for a period of 10 months. Enhancement of the rotenoid content was evaluated by feeding precursors in a tissue culture medium. Results: Four rotenoids (elliptone, deguelin, rotenone and Dehydrorotenone) were identified, which were confirmed using spectral and chromatographic techniques. The maximum rotenoid content was found in the seeds (0.33±0.01%), followed by roots (0.31±0.01%) and minimum in the aerial parts (0.20±0.05%). A gradual decrease in the rotenoid content was observed with the ageing of subcultured tissue maintained for 10 months. The production of rotenoids was enhanced up to 2 folds in the callus culture using amino acids, Phenylalanine and Methionine as precursors as compared to the control. The LC50 value of the rotenoids was found to be 91 ppm and 162 ppm against disease vectors of malaria and Dracunculiasis, respectively. Conclusion: The study projects M. sativa as a novel source of biopesticide against the disease vectors of malaria and Dracunculiasis. The use of precursors to enhance the rotenoid content in vitro can be an effective venture from a commercial point of view.


1972 ◽  
Vol 52 (6) ◽  
pp. 941-948 ◽  
Author(s):  
L. B. MacLEOD ◽  
H. T. KUNELIUS ◽  
F. W. CALDER

The effects of early summer and fall cutting managements on two alfalfa (Medicago sativa L.) cultivars were studied over a 3-year period. Removal of the first crop of Saranac and Narrangansett alfalfas at prebud stage, followed by two cuts at the early bloom stage before early September, and a late fall harvest on October 25, produced seasonal dry matter (DM) yields of 7.5–8.6 metric T/ha in the 1st year. Stand deterioration, however, was severe and by the 3rd year, practically all alfalfa had disappeared. Average percent crude protein (CP) in cut 1 was 17.7–21.9 and percent in vitro dry matter digestibility (IVD) 67.4–73.8. Removal of the first cut at bud stage with two subsequent harvests at the early bloom stage also allowed a total of three harvests before early September and resulted in seasonal DM yields of 8.6–8.8 in the 1st year. Although the survival of alfalfa was increased over the managements where cut 1 was at prebud stage, a considerable decrease of alfalfa was still observed in the 3rd year. Under this management, average CP in cut 1 was 15.8–16.3 and IVD 64.1–68.4. Harvesting the first crop at early to midbloom stage and the second crop at early bloom in early to mid-August, followed by third harvest in late October, resulted in good survival with seasonal DM yields of 6.1–10.6. Lower CP and IVD were recorded in cut 1 than in the more intensive managements. Under all managements, average CP in cuts 2 and 3 was 14.9–17.3 and 13.3–19.4, respectively. IVD was 54.5–60.7 in cut 2 and 56.0–64.4 in cut 3.


2012 ◽  
Vol 40 (2) ◽  
pp. 140 ◽  
Author(s):  
Hafiz Mamoon REHMAN ◽  
Iqrar Ahmad RANA ◽  
Siddra IJAZ ◽  
Ghulam MUSTAFA ◽  
Faiz Ahmad JOYIA ◽  
...  

Dalbergia sissoo Roxb. ex DC. (Sissoo) is a native forest tree species in Pakistan. Many ecological and economical uses are associated with this premier timber species, but dieback disease is of major concern. The objective of this study was to develop a protocol for in vitro regeneration of Sissoo that could serve as target material for genetic transformation, in order to improve this species. Callus formation and plantlet regeneration was achieved by culturing cotyledons, immature seeds, and mature embryos on a modified Murashige and Skoog (1962) (MS) medium supplemented with plant growth regulators. Callus induction medium containing 2.71 ?M 2, 4-dichlorophenoxyacetic acid (2,4-D) and 0.93 ?M kinetin produced better callus on all explants tested compared to other treatments, such as 8.88 ?M 6-benzylaminopurine (BA) and 2.69 ?M ?-naphthalene acetic acid (NAA), or 2.71 ?M 2, 4-D and 2.69 ?M NAA. Shoot regeneration was best on MS medium containing 1.4 ?M NAA and 8.88 ?M BA compared to other treatments, such as 1.4 ?M NAA and 9.9 ?M kinetin, or 2.86 ?M indole-3-acetic acid and 8.88 ?M BA. Murashige and Skoog medium containing 1.4 NAA ?M and 8.88 ?M BA was better in general for regeneration regardless of callus induction medium and the type of explant used. Rooting was best on half-strength MS medium with 7.35 ?M indole-3-butyric acid. Regenerated plantlets were acclimatized for plantation in the field. Preliminary genetic transformation potential of D. sissoo was evaluated by particle bombardment of callus explants with a pUbiGus vector. The bombarded tissue showed transient Gus activity 1week after bombardment. Transformation of this woody tree is possible provided excellent regeneration protocols. The best combination for regeneration explained in this study is one of such protocols.


1970 ◽  
Vol 20 (2) ◽  
pp. 145-155 ◽  
Author(s):  
Rita Sarah Borna ◽  
M. I. Hoque ◽  
R. H. Sarker

Genetic transformation using nodal and internodal segments from three economically important potato (Solanum tuberosum L.) varieties namely, Diamant, Cardinal and Granola was conducted using an Agrobacterium tumefaciens strain LBA4404 harbouring binary plasmid pBI12 containing the GUS and nptII genes. Node and internodal segments were used for direct regeneration as well as regeneration with the intervention of callus. best responses were  obtained for direct regeneration of shoots when the explants were cultured on MS supplemented with 4.0 mg/l BAP +1.0 mg/l IAA, 1.5 mg/l BAP  + 0.5 mg/l IAA and 5.0 mg/l BAP +1.0 mg/l IAA in Diamant, Cardinal  and  Granola, respectively. In Diamant spontaneous in vitro microtuberization was obtained from these proliferated shoots. Further culturing of these in vitro grown green microtubers regenerated a large number of shoots on MS containing 4.0 mg/l BAP +1.0 mg/l IAA. By combining the best treatments, this protocol yielded an average transformation rate of 87% of treared explants. Stable expression of GUS gene was visualized in the various parts of transformed shoots through histochemical assay. Genomic DNA was isolated from transformed shoots and stable integration of the GUS and nptII genes was confirmed by PCR analysis.   Key words:  Potato, in vitro regeneration, transformation   D.O.I. 10.3329/ptcb.v20i2.6894   Plant Tissue Cult. & Biotech. 20(2): 145-155, 2010 (December)


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