A Micropropagation, Molecular and Physiochemical Analysis of Canola Brassica napus L. From Seeds (original) (raw)
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AFRICAN JOURNAL OF BIOTECHNOLOGY
The objective of the study was to develop an efficient method for shoot regeneration of canola (Brassica napus L.) and to compare the regeneration capacity of different explants on MS medium with several combinations of plant growth regulators. The experiments showed that the morphogenetical potential of canola depends on genotype, primary explant, hormonal structure and concentration of nutrient medium. Cotyledons possessed higher regeneration ability in comparison to hypocotyls and roots. The best regeneration capability was exhibited by the cultivar 'Quantum'. Its frequency with cotyledonary explants reached 68.8% on all used media. Addition of 3 mg/l ABA in nutrient medium considerably increased the regeneration frequency. The highest shoot regeneration (100%), however, took place when cotyledonary explants were cultivated on medium, containing 1.0 mg/l NAA, 8.0 mg/l BAP and 3.0 mg/l ABA. Precultivation of explants on callus induction medium did not affect the shoot regeneration frequency. Vitrification of regenerants was promoted by increasing the auxin NAA or cytokinin BAP, and ABA in the nutrient medium.
Characterization of seed storage proteins of different varieties of Brassica napus seeds
Brassica napus (Canola), an important cash crop in Pakistan, has rich oil (40%) and protein (15%) content. The present study was designed to investigate nine different varieties of B. napus (Tarch, Altex, Dasi, Dunkled, V- 248, Bulb 98, Rainbow, Asker and Habib-98), for various physiochemical parameters (moisture content, thousand seed weight [TSW] and electrical conductivity [EC]), and also to investigate the genetic relationships among these genotypes using Sodium dodecyl sulphate-polyacralamide gel electrophoresis (SDS-PAGE). The maximum amount of moisture content was 6.31% for variety Dasi and minimum of 4.26% for variety Dunkled. The maximum TSW value was found for V-248 (4.22 g) and the minimum value for Dunkled variety (2.11 g). Electrical conductivity for different varieties was estimated at time intervals of 10 min, 20, 30, 40, 50, 60 and 70 minutes. The EC was determined by conductometer and the mean value ranged from 3.0-6.5 μ sec. Seeds protein analyzed by SDS-PAGE showe...
2007
The objective of the study was to develop an efficient method for shoot regeneration of canola (Brassica napus L.) and to compare the regeneration capacity of different explants on MS medium with several combinations of plant growth regulators. The experiments showed that the morphogenetical potential of canola depends on genotype, primary explant, hormonal structure and concentration of nutrient medium. Cotyledons possessed higher regeneration ability in comparison to hypocotyls and roots. The best regeneration capability was exhibited by the cultivar 'Quantum'. Its frequency with cotyledonary explants reached 68.8% on all used media. Addition of 3 mg/l ABA in nutrient medium considerably increased the regeneration frequency. The highest shoot regeneration (100%), however, took place when cotyledonary explants were cultivated on medium, containing 1.0 mg/l NAA, 8.0 mg/l BAP and 3.0 mg/l ABA. Precultivation of explants on callus induction medium did not affect the shoot regeneration frequency. Vitrification of regenerants was promoted by increasing the auxin NAA or cytokinin BAP, and ABA in the nutrient medium.
Plant regeneration and synthetic seeds production of Brassica oleracea var. italica
Acta Horticulturae, 2012
Agricultural industry has changed tremendously, especially since the end of World War II. Food industry has blossomed due to new technology, addition of preservative usage and government policies that favoured maximizing productions. Thus, sustainable vegetable production has captured the attention of many countries worldwide while at the same time preserving environmental quality and making efficient use of non-renewable resources.
Canola (Brassica napus L.) regeneration and transformation via hypocotyl and hypocotyl derived calli
Sarhad Journal of Agriculture, 2014
Canola (Brassica napus L.) is a leading vegetable oil and protein rich meal crop in the world. Like any other crop plant it is also a target for genetic manipulation. During this study a system of plant tissue culture was established for efficient regeneration and genetic transformation of three cultivars (Wester, Con-1, and Pakola). In vitro regeneration from hypocotyls derived callus and directly from hypocotyls of different cultivars was monitored. A procedure for callus induction, regeneration and selection of explants was also investigated. In case of callus induction more efficient response was observed in Wester and Con-1 compared to Pakola (63.8%). Similarly, high regeneration was noted in Wester (87.6) and Con-1(84.4) while low in Pakola (54.2%). Two Agrobacterium (LBA4404) strains harboring plasmid pCAMBIA2301 and pCAMBIA1391Z (harboring promoter P2F1: GUS) were used for transformation in canola plants using hypocotyls of all three cultivars. PCR and transient GUS expressi...
2012
In this paper, the authors have proposed a study on the variation in efficiency of micropropagation in vitro due to the influence of donor genotype and explant type used red cabbage head. In this test three commercial varieties of red cabbage seeds were tested for the ability of regeneration by organogenesis from hypocotyls, cotyledons, and stem segments. Our results have shown us that the intensity of red cabbage head regeneration depends on the genotype used but also the type of explant. Hipocotil fragments taken from seedlings to "Cabeza Negra 2"had the greatest total capacity to produce shoots than the other two genotypes tested when the combination of phytohormones was used consisting of 2 mg / l 6-benzylaminopurine (BAP) and type auxine ß indolilacetic acid (IBA) and α naftilacetic acid (NAA) at concentrations of 0.2 to 0.4 mg /l. The inoculated cotyledons explants also developed regenerative callus in about 6-8 weeks after initiation of cultures.
Establishment of Regeneration Protocol for Canola (Brassica napus L.)
Global Journal of Science Frontier Research, 2013
Abstrac t- The stu dy was conducted with an aim of developing a genotype independent efficient in vitr o regen eration system for Canola (Brassic a napus L.) th at is not grown in Bangladesh. Plant regeneration was achieved through embryogenic callus production from hypocotyls and cotyledonary leaves with petioles. The optimum medium for callus induction was found with 0.5 mgl -1 2, 4dic hlorophenoxyacetic acid (2, 4-D). The best shooting medium for canola contained 3.0 mgl-1 benzyl amino purine (BAP), 0.1 mgl -1 naphtal ene acetic acid (NAA), and 5.0 mgl -1 AgNO3 and pr oduced maximum number of shoots when kinetin was used at a concentration of 0.5 mgl - 1 . The p rofound positive effect of AgNO3 for Can ola was found for callogenesis at 0.5 mgl -1 and for regeneration at 5.0 mgl -1 . In vitr o regen erated shoots of canola produced well developed root system on both the media with 2.5 mgl -1 NAA and 1.0 mgl -1 indol e 3-butyric acid (IBA). Keywor ds: canola, plant regeneration, an...
A reproducible protocol for regeneration and transformation in canola (Brassica napus L
2006
The objective of the present study is to develop an efficient protocol for shoot and plant regeneration using five commercial canola cultivars grown under the Egyptian agricultural conditions. The regeneration efficiency from hypocotyl explants was examined. The data indicated that embryonic calli were formed within two weeks in the presence of 1 mgl -1 2,4-D. Adventitious shoots emerged from the embryonic callus in the presence of 4.5 mgl -1 BA. The cultivars showed a varied response to shoot regeneration. Regeneration frequency was high in the cultivar Sarow-4 (68%) followed by Masrri L-16 (64%) compared with the other cultivars tested. Hypocotyl explants from the cultivars Sarow-4 and Semu-249 were inoculated and co-cultivated with Agrobacterium tumefaciens strain LBA4404 harboring a binary vector pBI-121 containing the neomycin phosphotransferase-II gene (NPT-II). The resulted putative transgenic plantlets were able to grow under knanamycin containing medium. The stable integration of the NPT-II gene into the plant genomes was tested by PCR using NPT-II -specific primers. The GUS gene expression can be detected only in the transgenic plants. The reported protocol in the present study is repeatable and can be used to regenerate transgenic canola plants expressing the genes present in A. tumifaciens binary vectors.
ESTABLISHMENT OF REGENERATION AND TRANSFORMATION OF Brassica napus
Egyptian Journal of Genetics and Cytology, 2012
Production of genetically enhanced canola varieties with desirable new traits requires the establishment of efficient regeneration and transformation protocols. In the current work, experiments were conducted using two commercial cultivars Sarow-4 and Bactool, hypocotyl explants and Agrobacterium strain LBA4404 (harboring pISV2678 plasmid, which contains both gus and bar genes as reporter and selectable marker, respectively). Six days old hypocotyls segment were co-cultivated on callus induction medium (CIM) containing MS basal salt mixture and supplemented with 1mg/l 2,4-D for 3 days, Agrobacterium inoculation was conducted and cultures were incubated for 3 days before being placed on selection medium for a further 11 days. Two weeks old cultures were transferred to shoot induction RG3 medium supplemented with 4 mg/l BAP, 0.2 mg/l NAA, 5 mg/l AgNo3, 500 mg cefotaxime and 3 mg/l bialaphos. Shoots were rooted on rooting R4 medium containing 0.3 mg/l IBA. Regenerated plantlets were su...
Sinai Journal of Applied Sciences, 2016
This study was designed to investigate the effect of some factors affecting on in vitro growth enhancement of Topas canola genotype. During establishment stage, MS medium strengths (full and half), explant types (shoot tip and axillary node) and cytokinin types (BA and Kin) were investigated. Using shoot tip explants on full MS medium that supplemented with 1.0 mgl-1 BA proved to be the best treatment during this stage. In addition, different BA concentrations (0.0, 0.1, 0.5 and 2.5 mgl-1) and the same previous concentrations of IBA or NAA in the presence of 1.0 mgl-1 of BA or Kin were examined during multiplication stage. Results indicated that BA proved to be the best cytokinin especially at 2.5 mgl-1 or at 1.0 mgl-1 in the presence of 0.5 mgl-1 IBA since it produced the maximum number of shoots/explant. During rooting stage, effects of auxin type (IBA and NAA) at different concentrations (0.0, 0.1, 0.5 and 2.5 mgl-1) and the same previous concentrations of IAA, NAA and 2,4-D in the presence of 0.5 mgl-1 IBA were evaluated. Generally, using of half MS medium supplemented with 2.5 mgl-1 IBA individual or 0.5 mgl-1 IBA in combination with 0.1 mgl-1 IAA stated to be the most suitable treatments for root induction and growth. Obtained plantlets were successfully acclimatized (70-80% survivability) in peat moss and sand (1:1, v/v) medium in the greenhouse.