Efficient regeneration of Brassica napus L. hypocotyls and genetic transformation by Agrobacterium tumefaciens

Jonoubi, Parissa; Mousavi, Amir; Majd, Ahmad; Salmanian, Ali-Hatef; Javaran, Mokhtar Jalali; Daneshian, J

doi:10.1007/s10535-005-5180-2

Cited by 33 publications

(19 citation statements)

References 20 publications

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“…It was observed that 2 d of pre-cultivation and 2 d of co-cultivation were very effective in producing the highest cucumber regeneration frequency and number of shoots in cucumber. These findings of present study are in agreement with those found by Cardoza and Stewart (2003), Jonoubi et al (2005), Khan et al (2010), Wang et al (2013) and Khan et al (2013). The explants co-cultured for 2 d showed the highest regeneration frequency and number of shoots in current study.…”

Section: Discussionsupporting

confidence: 94%

Establishment of a High-efficiency Genetic Transformation System of Cucumber (Cucumis sativus) using Csexpansin 10 (CsEXP10) Gene

Sun¹,

Luo²,

Li³

et al. 2017

IJAB

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Cucumber is one of the most important vegetable crops in the world. To establish an efficient genetic transformation system in cucumber, the role of different factors such as pre-cultivation time, acetosyringone concentration, infection time and cocultivation time that can influence the transformation, was evaluated. In addition, Csexpansin 10 (CsEXP10) gene was transformed into the cucumber genome to produce transgenic lines. The results showed that various optimal parameters such as 100 mg L -1 of kanamycin concentration for selection of transformants, 2 d of pre-cultivation time, 100 μmol L -1 of acetosyringone concentration, 15 min of infection time and 2 d of co-cultivation time were obtained using cotyledonary node explants. The rooting frequency observed on Murashige and Skoog (MS) medium supplemented with 0.2 mg L -1 indole acetic acid and 400 mg L -1 cefotaxime was found to be 100.00%. The positive transgenic cucumber lines were identified using PCR analysis and GUS staining assay. It suggests that the genetic transformation system developed using cotyledonary node explants is efficient and successful in cucumber.

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Section: Discussionsupporting

confidence: 94%

Establishment of a High-efficiency Genetic Transformation System of Cucumber (Cucumis sativus) using Csexpansin 10 (CsEXP10) Gene

Sun¹,

Luo²,

Li³

et al. 2017

IJAB

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“…In earlier studies, 2,4-D was reported to be an effective growth hormone that produced rapid proliferating friable callus in Curcuma amada [15], Gymnema sylvestris [16] and Brassica napus [17] during in vitro cultures.…”

Section: Discussionmentioning

confidence: 99%

A Comparative Study of the Phytochemicals, Antioxidant and Antibacterial Potential of Methanolic Extracts of Trichosanthes Cucumerina (L.) Var. Cucumerina Under in Vitro Culture and Natural Conditions

Bai

Remakanthan

Kumar

et al. 2018

Int J Pharm Pharm Sci

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Objective: To compare the phytochemicals, antioxidative capacity and antibacterial profile of methanolic extracts of callus and naturally propagated plant species-Trichosanthes cucumerina (L.) var. cucumerina and to optimize an ideal protocol for in vitro callus and shoot induction. Methods:The sterilized seeds of Trichosanthes cucumerina (L.) var. cucumerina were inoculated in half Murashige and Skoog (MS) basal medium devoid of growth hormones to raise aseptic seedlings. Explants from aseptic seedlings used for callus induction in MS medium fortified with varying combinations of N 6 -Benzyl amino purine (BAP), 1-Naphthalene acetic acid (NAA) and 2,4-Dichlorophenoxy acetic acid (2,4-D). For in vitro soot induction, MS medium supplemented with different concentrations of 2,4-D, BAP and Kinetin-either alone or in combinations were employed. The callus harvested on 21 st and 45 th days were analyzed for a comparison of the influence of age of callus on the quantity of secondary metabolites. For a comparison with the naturally grown plant, all experiments were carried out with extracts from callus and wild plants. The antioxidant capacity of methanolic extracts was evaluated by 2,2-Diphenyl-1 Picryl Hydrazyl (DPPH) free radical scavenging assay and Ferric Reducing Antioxidant Power (FRAP) analysis. The antibacterial activity of were screened by the agar diffusion method using pathogenic bacteria such as Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus and documented through measurement of the diameter of growth inhibition zone (IZ). Results:The results on in vitro culture indicated that MS medium with BAP (0.5 mg/l) and 2,4-D (1 mg/l) was ideal for callus induction. For shoot induction, supplementation of MS medium with BAP-0.5 mg/l, 2,4,D-1.0 mg/l and Kinetin-0.5 mg/l was found to be most favourable. Direct root induction from the callus was found to occur in medium fortified with BAP-0.5 mg/l, 2,4, D-0.5 mg/l and NAA-1.0 mg/l. The phytoconstituents quantified were alkaloids, flavonoids, tannins, phenols and terpenoids and their levels were higher in wild plant in comparison to callus. Naturally grown plant possesses higher free radical scavenging ability and ferric reducing power than callus. Results of antibacterial activity indicated that the Gram-positive strain (Staphylococcus aureus) was more sensitive than the Gram-negative bacteria (Escherichia coli and Pseudomonas aeruginosa). The highest antibacterial activity recorded for naturally propagated plant extract against S. aureus (IZ = 13 mm) and was quite comparable with standard antibiotic cephatoxim (IZ = 20 mm) at 100µg concentration. Conclusion:Results concluded that this overexploited medicinal plant with lesser seed longevity could be successfully propagated by in vitro methods. The phytoconstituents with antioxidative and antibacterial potential were more abundant in naturally propagated plants than undifferentiated callus tissue. The extracts are potent antibacterial agents.

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“…Our results indicate that four key factors, seedling age, Agrobacterium concentration, infection time, and co-cultivation time, affect Agrobacterium-mediated transformation as revealed by GUS expression and hygromycin resistance. Previous studies have reported the use of seedlings of different ages from various plant species as donor material for transgenic experiments, including 8-day-old canola seedlings (Maheshwari et al, 2011) and 21-day-old B. napus seedlings (Jonoubi et al, 2005). In the present study, GUS expression and hygromycin resistance were monitored to evaluate the effect of explant age on the transformation frequency of the B. napus 'Precocity'.…”

Section: Optimization Of Factors Affecting Gus Expression and Hygromymentioning

confidence: 97%

Improved Agrobacterium-mediated transformation and high efficiency of root formation from hypocotyl meristem of spring Brassica napus ‘Precocity’ cultivar

Liu

Lang

et al. 2015

Genet. Mol. Res.

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ABSTRACT. Rape seed (Brassica napus L.) is one of the most important oil seed crops in the world. Genetic manipulation of rapeseed requires a suitable tissue culture system and an efficient method for plant regeneration, as well as an efficient transformation procedure. However, development of transgenic B. napus has been problematic, and current studies are limited to cultivated varieties. In this study, we report a protocol for regeneration of transgenic rape after Agrobacterium-mediated transformation of hypocotyls from the spring B. napus 'Precocity' cultivar. We analyzed the effects of plant growth regulators in the medium on regeneration. Additionally, factors affecting the transformation efficiency, including seedling age, Agrobacterium concentration, infection time, and co-cultivation time, were assessed by monitoring GUS expression. Results from these experiments revealed that transformation was optimized when the meristematic parts of the 16841 Transformation and root formation of Brassica napus ©FUNPEC-RP www.funpecrp.com.br Genetics and Molecular Research 14 (4): 16840-16855 (2015) hypocotyls were taken from 8 day-old seedlings, cultured on Murashinge and Skoog basal media containing 0.1 mg/L 1-naphthaleneacetic acid and 2.5 mg/L 6-benzylaminopurine, and incubated in Agrobacterium suspension (OD 600 = 0.5) for 3 to 5 min, followed by 2 days of cocultivation. Integration of T-DNA into the plant genome was confirmed by polymerase chain reaction (PCR), b-glucuronidase histochemical staining, and quantitative real-time PCR. The protocols developed for regeneration, transformation, and rooting described in this study could help to accelerate the development of transgenic spring rape varieties with novel features.

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Efficient regeneration of Brassica napus L. hypocotyls and genetic transformation by Agrobacterium tumefaciens

Cited by 33 publications

References 20 publications

Establishment of a High-efficiency Genetic Transformation System of Cucumber (Cucumis sativus) using Csexpansin 10 (CsEXP10) Gene

Establishment of a High-efficiency Genetic Transformation System of Cucumber (Cucumis sativus) using Csexpansin 10 (CsEXP10) Gene

A Comparative Study of the Phytochemicals, Antioxidant and Antibacterial Potential of Methanolic Extracts of Trichosanthes Cucumerina (L.) Var. Cucumerina Under in Vitro Culture and Natural Conditions

Improved Agrobacterium-mediated transformation and high efficiency of root formation from hypocotyl meristem of spring Brassica napus ‘Precocity’ cultivar

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