+ Site Statistics
+ Search Articles
+ Subscribe to Site Feeds
EurekaMag Most Shared ContentMost Shared
EurekaMag PDF Full Text ContentPDF Full Text
+ PDF Full Text
Request PDF Full TextRequest PDF Full Text
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter
Follow on Google+Follow on Google+
Follow on LinkedInFollow on LinkedIn

+ Translate

The use of the Emu promoter with antibiotic and herbicide resistance genes for the selection of transgenic wheat callus and rice plants

Australian Journal of Plant Physiology 21(1): 95-112

The use of the Emu promoter with antibiotic and herbicide resistance genes for the selection of transgenic wheat callus and rice plants

Coding sequences of selectable marker genes were introduced into a plasmid containing the Emu promoter sequence which drives high levels of expression in cereal cells. The selectable markers were those coding for the enzymes neomycin phosphotransferase (NPT*) for kanamycin resistance, hygromycin phosphotransferase (HPT) for hygromycin resistance, phosphinothricin N-acetyltransferase (PAT) for resistance to the herbicidal compound phosphinothricin, and a mutant acetolactate synthase (ALS) for resistance to sulfonylurea herbicides. Plasmid DNAs were introduced into wheat and rice protoplasts by electroporation. Wheat protoplast-derived callus lines producing NPT and PAT were selected with kanamycin and phosphinothricin respectively. Wheat callus transformed with the gene coding for the mutant ALS was selected with sulfometuron methyl following the bombardment of suspension cultures with DNA-coated microparticles. With rice, transformed plants were regenerated from protoplast-derived cultures selected with kanamycin and hygromycin. Integration of the introduced DNA in transformed rice plants was confirmed by gel blot hybridisation, and significant levels of the NPT enzyme were measured by ELISA in leaves of the primary transformants as well as in leaves, seeds and roots of progeny plants. The results confirm that efficient selection of transformed cereal cultures and plants can be achieved by using selectable genes linked to the Emu promoter.

Accession: 002530641

DOI: 10.1071/PP9940095

Related references

Carlson, A.R.; Kaeppler, H.F., 2000: Comparison of visual screening, herbicide and antibiotic selection of transgenic oat plants. In Vitro Cellular & Developmental Biology Animal 36(3 Part 2): 47 A, March

Mandal, N.; Dana, I., 1998: Herbicide resistant transgenic rice (Oryza sativa L.) plants obtained by microprojectile bombardment of regenerable embryogenic callus. Transgenic rice plants were produced by the direct delivery of plasmid DNA, containing the uidA and bar genes, to young embryogenic calluses using high velocity microprojectiles. Only 2 independent callus lines were selected on varying concentrati...

Christou, P.; Ford, T.L., 1995: The impact of selection parameters on the phenotype and genotype of transgenic rice callus and plants. Transgenic rice embryogenic callus and plants were recovered from experiments involving electric discharge particle acceleration (Accell technology). Critical parameters influencing successful delivery and stable integration of exogenous DNA into...

Endo, M.; Shimizu, T.; Toki, S., 2012: Selection of transgenic rice plants using a herbicide tolerant form of the acetolactate synthase gene. Acetolactate synthase (ALS) is an enzyme in the biosynthetic pathway for branched-chain amino acids, and bispyribac-sodium (BS), a pyrimidinyl carboxy herbicide, is a well-known inhibitor of ALS activity. However, it appears that a mutated form of...

Kawahigashi, H.; Hirose, S.; Ohkawa, H.; Ohkawa, Y., 2006: Herbicide resistance of transgenic rice plants expressing human CYP1A1. Cytochrome P450 monooxygenases (P450s) metabolize herbicides to produce mainly non-phytotoxic metabolites. Although rice plants endogenously express multiple P450 enzymes, transgenic plants expressing other P450 isoforms might show improved herbic...

Y.H.ngXiu; Zhao ZhiPeng; Wang Ling; Liu QiaoQuan; Gong ZhiYun; G.M.ngHong, 2007: Breeding of transgenic rice lines with GNA and Bar genes resistance to both brown planthopper and herbicide. Acta Phytophylacica Sinica 34(5): 555-556

Joersbo, M., 2001: Advances in the selection of transgenic plants using non-antibiotic marker genes. Production of transgenic plants started more than a decade ago, but it is still a time-consuming operation. One of the critical points is the selection procedure used for the recovery of transgenic shoots after transformation. Moreover, as more tr...

Inui, H.; Ohkawa, H., 2005: Herbicide resistance in transgenic plants with mammalian P450 monooxygenase genes. Transgenic potato and rice plants were generated by the introduction of human P450 species, CYP1A1, CYP2B6, CYP2C9 and CYP2C19, which metabolized a number of herbicides, insecticides and industrial chemicals. The transgenic potato plant T1977 co-e...

Wang XiaoJun; Liu YuLe; Huang Yong; Tian Bo, 1996: Self-fertile transgenic wheat plants expressing bromoxynil herbicide resistance. A reproducible protocol for rapid production of transgenic wheat via microprojectile bombardment was developed. Immature embryos excised from caryopses 14 to 18 days after anthesis were used for bombardment with tungsten particles coated with a pl...

Wang Xiao Jun, L.Y.L.; Huang Yong; Tian Bo, 1996: Self-fertile transgenic wheat plants expressing herbicide bromoxynil resistance. Genetic manipulation of wheat (Triticum aestivum L.) by biotechnological approaches is currently limited by a lack of efficient and reliable transformation method. The authors report a reproducible protocol for rapid production of transgenic wheat...