Production of Hairy Root Cultures and Transgenic Plants by <I>Agrobacterium rhizogenes</I>-Mediated Transformation
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Agrobacterium rhizogenes-mediated transformation results in the development of hairy roots at the site of infection. The production of hairy roots involves cocultivation of explants with A. rhizogenes and the subsequent selection of hairy roots on hormone-free medium. Hairy roots have many applications for research including secondary product production and for the study of biochemical pathways. In addition, transgenic plants regenerated from hairy roots often show an altered phenotype due to the presence of the rol genes. In this chapter we describe how to produce and grow hairy root cultures, how to regenerate shoots from these hairy roots, and how to conduct molecular analysis of these cultures.Keywords:
Hairy Root Culture
Hairy Root Culture
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Agrobacterium rhizogenes strain RPB13 induced highest survival percentage (100%) along with the highest no. of roots per explant, whereas, in hormone treated micro-propagated explants the survival percentage was 94%.The maximum number. of roots were 11 and 6 due to the treatment with A. rhizogenes and hormones respectively per explant. Furthermore, the induction of rooting in explant by strain RPB13 required less days than that of hormonal treatment. Thus, A. rhizogenes strain RPB13 has opened new avenue for rooting in micro-propagated explants without hormonal use in strawberry plants.
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Agrobacterium rhizogenes-transformed hairy roots from Catharanthus roseus have been widely used in research and in life. Consequently, methods for preservation are essential to maintain valuable hairy root lines. Our results showed that Gamborg’B5 was the most suitable medium for hairy roots from VIN002 and VIN005 while solid White media was more comfortable for the hairy lines from VIN022 and VIN077. All hairy root lines must be preserved in the dark at 25–27 oC. Under suitable conditions, the rate of lines growing normally reached 69.3, 67.0, 57.3 and 60.7 % for VIN002, VIN005, VIN072, and VIN077, respectively. There were 90.0, 93.3, 80.0 and 93.3 % of lines could preserve rolB, a gene which has important effect on the growth of hairy roots.
Catharanthus roseus
Hairy Root Culture
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Regeneration of whole viable plants from hairy root cultures, which were established from transformation with Agrobacterium rhizogenes, has been reported in a number of plant species. Such transgenic plants frequently show a very characteristic phenotype, different from their normal counterparts. Therefore, scientists have been studying on production of transformed roots for creating transgenic ornament plants. In this study, several factors affecting A. rhizogenes C26 mediated genetic transformation in Catharanthus roseus VIN077, i.e. cell density of Agrobacterium rhizogenes C26 suspension, infection time, co-cultivation time, illumination conditions and culture media, were investigated. The results indicated that cell density of bacterial suspension, infection time, co-cultivation time, and culture media were important factors that had positive effects on the formation of hairy root at appropriate conditions. It was suggested that leaves of C. roseus VIN077 soaked in A. rhizogenes C26 suspension at cell density of 0.2 (OD600 nm) for 10 minutes and co-cultivated on ½ White medium in the dark condition for 6 days resulted in the most effective hairy root formation. Illumination with fluorescent white light was found to inhibit hairy root formation in co-cultivation period but able to induce this process during the bacterial elimination step. The transgenic hairy root lines were confirmed by polymerase chain reaction (PCR) using rolB specific primers.
Catharanthus roseus
Hairy Root Culture
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Hairy Root Culture
Plant Physiology
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Acetosyringone
Strain (injury)
clone (Java method)
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Abstract Agrobacterium rhizogenes Conn. causes hairy root disease in plants. Hairy root‐infected A. rhizogenes is characterized by a high growth rate and genetic stability. Hairy root cultures have been proven to be an efficient means of producing secondary metabolites that are normally biosynthesized in roots of differentiated plants. Furthermore, a transgenic root system offers tremendous potential for introducing additional genes along with the Ri plasmid, especially with modified genes, into medicinal plant cells with A. rhizogenes vector systems. The cultures have turned out to be a valuable tool with which to study the biochemical properties and the gene expression profile of metabolic pathways. Moreover, the cultures can be used to elucidate the intermediates and key enzymes involved in the biosynthesis of secondary metabolites. The present article discusses various applications of hairy root cultures in plant genetic engineering and potential problems associated with them. (Managing editor: Wei Wang)
Hairy Root Culture
Gene engineering
Plant cell
Metabolic Engineering
Root (linguistics)
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Hairy roots, transformed with Agrobacterium rhizogenes, have been found to be suitable for the production of secondary metabolites because of their stable and high productivity in hormone-free culture conditions. A number of plant species including many medicinal plants have been successfully transformed with Agrobacterium rhizogenes. Transformed root cultures have also been found to be a potential source of high-value pharmaceuticals. In this article the most important alkaloids produced by hairy roots are summarised. Several different methods have been used to increase the alkaloid accumulation in hairy root cultures. The selection of high productive root lines based on somaclonal variation offers an interesting option to enhance the productivity. Elicitors and modification of culture conditions have been shown to increase the growth and the alkaloid production in some cases. Genetic engineering is a modern tool to regulate the secondary metabolism also in hairy roots. However, our knowledge on biosynthesis of many alkaloids is still poor. Only a limited number of enzymes and their respective genes which regulate the biosynthetic pathways are fully characterised.
Hairy Root Culture
Secondary metabolism
Somaclonal variation
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Hairy Root Culture
Plant Roots
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Agrobacterium rhizogenes-mediated transformation results in the development of hairy roots at the site of infection. The production of hairy roots involves cocultivation of explants with A. rhizogenes and the subsequent selection of hairy roots on hormone-free medium. Hairy roots have many applications for research including secondary product production and for the study of biochemical pathways. In addition, transgenic plants regenerated from hairy roots often show an altered phenotype due to the presence of the rol genes. In this chapter we describe how to produce and grow hairy root cultures, how to regenerate shoots from these hairy roots, and how to conduct molecular analysis of these cultures.
Hairy Root Culture
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Citations (77)