Somatic embryo an alternative target tissue for agrobacterium-mediated transformation in Hevea brasiliensis

Li Y. T., . and Li J., . and Huang H. S., . and Hua Y. W., . and Huang T. D., . Somatic embryo an alternative target tissue for agrobacterium-mediated transformation in Hevea brasiliensis. pp. 171-188. ISSN 1511-1768

Traditionally embryogenic callus has been used as the only receptor source of genetic transformation in H. brasiliensis. In this study we showed that somatic embryos were a viable alternative to embryogenic callus. The appropriate concentration of both decontamination and selection antibiotics was firstly screened. Visual and cytological observations found both epidermal and subepidermal cotyledon cells could be infected indicating that somatic embryos were susceptible to antibiotics and Agrobacterium. Subsequently five isolated consecutive transformation experiments were carried out to produce the transgenic plants using a total of 1158 somatic embryos of which 47 (4.06 ) GUS-positive embryos were produced after cell differentiation and embryogenesis. Six were successfully multiplied and produced 33 plants. PCR and Southern blot analysis confirmed that the T-DNA was integrated into the recovered plants. Finally three additional cycles of secondary embryogenesis were carried out for transgenic embryo proliferation efficiency assessment. The rate of proliferation tended to slightly increase with each proliferation generation and one initial transgenic embryo-derived GUS-negative embryo were observed in the fifth multiplication cycle indicating that transgenic plants could be propagated through secondary embryogenesis and some initial transgenic embryos might be chimeric. This is the first attempt on producing transgenic plants using somatic embryos as the target tissue in H. brasiliensis. It is demonstrated that somatic embryos can be used as an alternative target tissue for Agrobacterium-mediated transformation in Hevea brasiliensis.

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Abstract

Traditionally embryogenic callus has been used as the only receptor source of genetic transformation in H. brasiliensis. In this study we showed that somatic embryos were a viable alternative to embryogenic callus. The appropriate concentration of both decontamination and selection antibiotics was firstly screened. Visual and cytological observations found both epidermal and subepidermal cotyledon cells could be infected indicating that somatic embryos were susceptible to antibiotics and Agrobacterium. Subsequently five isolated consecutive transformation experiments were carried out to produce the transgenic plants using a total of 1158 somatic embryos of which 47 (4.06 ) GUS-positive embryos were produced after cell differentiation and embryogenesis. Six were successfully multiplied and produced 33 plants. PCR and Southern blot analysis confirmed that the T-DNA was integrated into the recovered plants. Finally three additional cycles of secondary embryogenesis were carried out for transgenic embryo proliferation efficiency assessment. The rate of proliferation tended to slightly increase with each proliferation generation and one initial transgenic embryo-derived GUS-negative embryo were observed in the fifth multiplication cycle indicating that transgenic plants could be propagated through secondary embryogenesis and some initial transgenic embryos might be chimeric. This is the first attempt on producing transgenic plants using somatic embryos as the target tissue in H. brasiliensis. It is demonstrated that somatic embryos can be used as an alternative target tissue for Agrobacterium-mediated transformation in Hevea brasiliensis.

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Item Type:	Article
AGROVOC Term:	Somatic embryos
AGROVOC Term:	Agrobacterium
AGROVOC Term:	Hevea brasiliensis
AGROVOC Term:	Rubber
AGROVOC Term:	Cytological analysis
AGROVOC Term:	Antibiotics
AGROVOC Term:	Cotyledons
AGROVOC Term:	Transgenic plants
AGROVOC Term:	Embryogenesis
AGROVOC Term:	DNA
Depositing User:	Ms. Suzila Mohamad Kasim
Last Modified:	24 Apr 2025 06:28
URI:	http://webagris.upm.edu.my/id/eprint/22943

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