Activity of a chimeric promoter with the doubled CaMV 35S enhancer element in protoplast-derived cells and transgenic plants in maize

Omirulleh, S.; Ábrahám, Mariann; Golovkin, Maxim; Stefanov, I.; Karabaev, M K; Mustárdy, László; Mórocz, S.; Dudits, Dénes

doi:10.1007/bf00028800

Cited by 97 publications

(32 citation statements)

References 37 publications

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“…In rice, the CaMV 35S promoter is widely used as a constitutive strong promoter (Terada and Shimamoto, 1990). Likewise, the CaMV 35S enhancer can elevate gene expression in monocotyledonous plants, including maize (Omirulleh et al, 1993) and rice (Mitsuhara et al, 1996). We found in our rice study that the CaMV 35S enhancers were functional in both orientations, either upstream or downstream of nearby genes.…”

Section: Discussionsupporting

confidence: 54%

T-DNA Insertional Mutagenesis for Activation Tagging in Rice

et al. 2002

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We have developed a new T-DNA vector, pGA2715, which can be used for promoter trapping and activation tagging of rice (Oryza sativa) genes. The binary vector contains the promoterless ␤-glucuronidase (GUS) reporter gene next to the right border. In addition, the multimerized transcriptional enhancers from the cauliflower mosaic virus 35S promoter are located next to the left border. A total of 13,450 T-DNA insertional lines have been generated using pGA2715. Histochemical GUS assays have revealed that the GUS-staining frequency from those lines is about twice as high as that from lines transformed with the binary vector pGA2707, which lacks the enhancer element. This result suggests that the enhancer sequence present in the T-DNA improves the GUS-tagging efficiency. Reverse transcriptase-PCR analysis of a subset of randomly selected pGA2715 lines shows that expression of the genes immediately adjacent to the inserted enhancer is increased significantly. Therefore, the large population of T-DNA-tagged lines transformed with pGA2715 could be used to screen for promoter activity using the gus reporter, as well as for creating gain-of-function mutants.Recent completion of the draft sequence for the rice (Oryza sativa) genome has resulted in an explosion of information on rice genes (Goff et al., 2002; Yu et al., 2002). The challenge for the post-sequencing era is to identify the biological functions for these genes. Of all the approaches used to discover gene function, the most direct is to disrupt the genes and analyze the consequences. Various methods have been developed in plants for this purpose. These include using ethyl methanesulfonate, fast-neutron treatment, or insertion of an element, such as a transposable element or T-DNA (Koornneef et al., 1982;Sundaresan, 1996;Krysan et al., 1999). Insertional mutagenesis has the advantage that the inserted element acts as a tag for gene identification. However, all gene disruption approaches also have some limitations. For example, it is difficult to identify the function of redundant genes, or of genes required in early embryogenesis or gametophyte development.To overcome those limitations, modified insertional elements have been developed. One of these modified designs is the gene trap system that involves creating fusions between the tagged genes and a reporter gene, such as ␤-glucuronidase (gus) or green fluorescent protein (gfp; Sundaresan et al., 1995;Springer, 2000). This system provides a way of identifying novel genes based on their expression patterns. Insertion of the promoterless reporter not only destroys normal gene function but also activates expression of the reporter gene. Because expression levels can be monitored in heterozygote plants, the gene trap system is useful for studying the patterns of most plant genes, including essential genes that cause lethal mutations. This system is convenient for observing mutant phenotypes because reporter activation indicates the location, condition, and time of expression for the disrupted gene. In Arabidopsis, ...

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Section: Discussionsupporting

confidence: 54%

T-DNA Insertional Mutagenesis for Activation Tagging in Rice

et al. 2002

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“…For Brassica, the most commonly used is neomycin phosphotransferase II (nptll) from transposon Tn5 (Bevan et al, 1983;Carrer et al, 1993;Murry et al, 1993;Omirulleh et al, 1993). This gene confers resistance toward aminoglycosides such as kanamycin, neomycin, geneticin (G418), and paromycin.…”

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confidence: 99%

A non-antibiotic selection system uses the phosphomannose-isomerase (PMI) gene forAgrobacterium-mediated transformation of Chinese cabbage

Park

2006

J. Plant Biol.

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To establish a non-antibiotic selection system that utilizes the phosphomannose-isomerase (PMI) gene for Chinese cabbage transformation, we first determined the optimum mannose concentration for selecting transformed cells. Hypocotyl and cotyledon explants that were grown on media containing more than 5 g L 1 mannose did not induce green calli but, rather became chlorotic and withered before dying. In contrast, media containing 20 g I. 1 sucrose plus 5 g L -1 mannose proved suitable for selection. We then used this particular level of mannose to transform hypocotyl tissues. Within 6 weeks, shoots were regenerated from some of the calli; subsequently, these plants were transplanted to pots and grown in the greenhouse. A 514-bp PCR fragment was obtained from most transformants but not from the non-transformed plants. Southern blot analysis also revealed the expected PMI gene in those PCR-confirmed transgenic plants. RT-PCR of total RNA was performed to confirm P&fl expression. We have now demonstrated that this gene does not inhibit the growth of transgenic plants, and that this selection system can be applied to Chinese cabbage transformation.

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“…The first successful transgenic maize was generated from protoplasts of embryogenic suspension cultures of the inbred maize line A188 (Rhodes et al, 1988), but none of the 38 plants from 10 transformed lines could set seed. In 1993, the first fertile transgenic maize was reported from protoplasts that had been isolated from embryogenic suspension cultures of genotype HE/89 Omirulleh et al, 1993). More than 60% of the transgenic plants could set seed.…”

Section: Tissue Culturementioning

confidence: 99%

Genetic transformation of major cereal crops

Ji¹,

Xu²,

Wang³

2013

Int. J. Dev. Biol.

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Of the more than 50,000 edible plant species in the world, at least 10,000 species are cereal grains. Three major cereal crops, rice (Oryza sativa), maize (Zea mays), and wheat (Triticum sp.), provide two-thirds of the world's food energy intake. Although crop yields have improved tremendously thanks to technological advances in the past 50 years, population increases and climate changes continue to threaten the sustainability of current crop productions. Whereas conventional and marker-assisted breeding programs continue to play a major role in crop improvement, genetic engineering has drawn an intense worldwide interest from the scientific community. In the past decade, genetic transformation technologies have revolutionized agricultural practices and millions of hectares of biotech crops have been cultured. Because of its unique ability to insert well-characterized gene sequences into the plant genome, genetic engineering can also provide effective tools to address fundamental biological questions. This technology is expected to continue to be an indispensable approach for both basic and applied research. Here, we overview briefly the development of the genetic transformation in the top seven cereals, namely maize, rice, wheat, barley (Hordeum vulgare), sorghum (Sorghum sp.), oat (Avena sativa), and millets. The advantages and disadvantages of the two major transformation methods, Agrobacterium tumefaciens-mediated and biolistic methods, are also discussed.

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Activity of a chimeric promoter with the doubled CaMV 35S enhancer element in protoplast-derived cells and transgenic plants in maize

Cited by 97 publications

References 37 publications

T-DNA Insertional Mutagenesis for Activation Tagging in Rice

T-DNA Insertional Mutagenesis for Activation Tagging in Rice

A non-antibiotic selection system uses the phosphomannose-isomerase (PMI) gene forAgrobacterium-mediated transformation of Chinese cabbage

Genetic transformation of major cereal crops

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