Norway spruce (Picea abies) genetic transformation with modified Cry3A gene of Bacillus thuringiensis.

Modified versions of the Cry3A gene of Bacillus thuringiensis (Bt) were transferred into Norway spruce (Picea abies). Both the biolistic approach and Agrobacterium tumefaciens mediated procedure were employed for transformation of embryogenic tissue (ET) cultures. The latter method proved to be more efficient yielding 70 transgenic embryogenic tissue lines compared with 18 lines obtained by biolistics. The modified Cry3A genes were driven by a 35S promoter and the nptII screenable selection marker gene was used in all vectors. The transgenic ETs were molecularly characterized and converted into mature somatic embryos. Germinating embryos formed plantlets which were finally planted into perlite and their Cry3A gene transcription activities were demonstrated by RT-PCR.

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Transformation of HBsAg (hepatitis B surface antigen) gene into tomato mediated by Agrobacterium tumefaciens

Czech Journal of Genetics and Plant Breeding ◽

10.17221/5/2011-cjgpb ◽

2011 ◽

Vol 47 (No. 2) ◽

pp. 69-77 ◽

Cited By ~ 5

Author(s):

T. Li ◽

J.K. Sun ◽

Z.H. Lu ◽

Q. Liu

Keyword(s):

Agrobacterium Tumefaciens ◽

Hepatitis B ◽

Surface Antigen ◽

Hepatitis B Surface Antigen ◽

Marker Gene ◽

Selection Marker ◽

35S Promoter ◽

Antigen Gene ◽

Plant Expression ◽

Plant Expression Vector

The plant expression vector pBRSAg was constructed as suitable for transformation via Agrobacterium-mediated approach. It contains all elements for plant expression, such as CaMV 35S promoter, both left and right border sequence for transferred DNA (T-DNA) in Agrobacterium, plant reporter gene gus, and plant selection marker gene hpt. The recombinant binary vector pBRSAg was transformed into Agrobacterium tumefaciens strains by using the freeze-thaw method. Tomato cotyledon explants were transformed by A. tumefaciens and plants were regenerated on selection medium. GUS staining, PCR and PCR-Southern analysis of the plants were positive. It was for the first time shown that the HBsAg (hepatitis B surface antigen) gene was introduced into tomato plants. The expression of transgene is under investigation.

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Polyamines during somatic embryo development in Norway spruce (Picea abies [L.])

Journal of Forest Science ◽

10.17221/84/2008-jfs ◽

2009 ◽

Vol 55 (No. 2) ◽

pp. 75-80 ◽

Cited By ~ 9

Author(s):

J. Malá ◽

M. Cvikrová ◽

P. Máchová ◽

O. Martincová

Keyword(s):

Somatic Embryogenesis ◽

Picea Abies ◽

Norway Spruce ◽

Somatic Embryos ◽

Developmental Stages ◽

Embryogenic Tissue ◽

Growth Media ◽

Mature Embryos ◽

Proliferation Medium ◽

Maturation Medium

Contents of free polyamines (putrescine, spermidine and spermine) were determined in different developmental stages of Norway spruce (<I>Picea abies</I> [L.] Karst.) somatic embryos by means of HPLC. Determinations were performed embryogenic tissue after 4 weeks of the growth on proliferation medium, after 2 and 5 weeks of the culturing on maturation medium, and 2 weeks after desiccation. Maturation of somatic embryos (after 5 weeks) was accompanied by increase of concentrations of putrescine (2.3 times) and spermidine (3.2 times). In comparison with above mentioned polyamines, spermine concentrations were significantly lower (4.3 times). Two weeks after desiccation, the concentrations of putrescine decreased 5.4 times and spermidine 2.2 times in comparison with mature embryos. To improve the efficiency of somatic embryogenesis of less responsive genotypes, the supplementation of growth media by polyamines is discussed.

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A tonoplast intrinsic protein (TIP) is present in seeds, roots and somatic embryos of Norway spruce (Picea abies)

Physiologia Plantarum ◽

10.1034/j.1399-3054.1995.950217.x ◽

1995 ◽

Vol 95 (2) ◽

pp. 288-295

Author(s):

Peter Oliviusson ◽

Inger Hakman

Keyword(s):

Picea Abies ◽

Norway Spruce ◽

Somatic Embryos ◽

Tonoplast Intrinsic Protein ◽

Intrinsic Protein

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A Tonoplast Intrinsic Protein, α-TIP, is Present in Seeds and Somatic Embryos of Norway Spruce (Picea abies)

Current Issues in Plant Molecular and Cellular Biology - Current Plant Science and Biotechnology in Agriculture ◽

10.1007/978-94-011-0307-7_90 ◽

1995 ◽

pp. 647-652

Author(s):

P. Oliviusson ◽

I. Hakman

Keyword(s):

Picea Abies ◽

Norway Spruce ◽

Somatic Embryos ◽

Tonoplast Intrinsic Protein ◽

Intrinsic Protein

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INTRODUCTION OF β-GLUCURONIDASE GENE INTO GINSENG (PANAX CINSENG) BY A TI-PLASMID VECTOR SYSTEM AND ITS EXPRESSION

HortScience ◽

10.21273/hortsci.27.6.621e ◽

1992 ◽

Vol 27 (6) ◽

pp. 621e-621

Author(s):

Jang R. Liu ◽

Haeng S. Lee ◽

Suk W. Kim ◽

Hyo W. Lee

Keyword(s):

Somatic Embryos ◽

Binary Vector ◽

Zygotic Embryo ◽

Plasmid Vector ◽

Vector System ◽

Selection Marker ◽

35S Promoter ◽

Dichlorophenoxyacetic Acid ◽

Gus Gene ◽

2,4 Dichlorophenoxyacetic Acid

β-Glucuronidase (GUS) gene of Escherichia coli was introduced into ginseng cells by an Agrobacterium binary vector system and expressed in somatic embryos derived from the cells. A binary vector pBI121 carrying CaMV 35S promoter-GUS gene fusion and a neomycin phosphotransferase gene as selection marker was transferred into Agrobacterium tumefaciens LBA4404. Zygotic embryo cotyledonary segments were co-cultivated with A. tumefaciens and transferred to the medium containg 1 mg 2,4-dichlorophenoxyacetic acid/liter, 0.5 mg kinetin/liter, and 100 mg kanamycin/liter. Kanamycin-resistant calli were formed after 3 to 4 weeks of culture. Southern analysis confirmed the resistant calli were transformed with GUS gene. High GUS activities were detected in somatic embryos developed from the calli.

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