Transgenic rice as a system to study the stability of transgene expression: multiple heterologous transgenes show similar behaviour in diverse genetic backgrounds

Gahakwa, Daphrose; Maqbool, Shahina; Fu, Xiang‐Dong; Sudhakar, D.; Christou, Paul; Kohli, Ajay

doi:10.1007/s001220051495

Cited by 50 publications

(20 citation statements)

References 74 publications

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“…(ii) normal single gene Mendelian inheritance of hph gene has occurred in the progeny of transformed tobacco plants. Our report of normal Mendelian inheritance of antibiotic resistance gene is an addition to the growing literature such as petunia transformed with pMON120 type vectors (see Horsch et al, 1983), in tomato transformed with pBin19 carrying the nptII gene (Cortina and Culianez-Macia, 2004), in peanut transformed with pBI121 carrying nptII gene (Cheng et al, 1997), in rice transformed with hpt, bar, gna and gusA genes (Gahakwa et al, 2000) and in barley plants transformed with bar gene (Bregitzer and Tonks, 2003). Transgene inherited in Mendelian ratio (present study) showed that there is single copy insertion in transgenic plants of tobacco.…”

Section: Discussionmentioning

confidence: 66%

Expression of a fungal endochitinase gene in transgenic tomato and tobacco results in enhanced tolerance to fungal pathogens

Shah

Mukherjee

Eapen

2010

Physiol Mol Biol Plants

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Development of transgenic Nicotiana tabacum and Lycopersicon esculentum expressing an endochitinase (ech42) gene from biocontrol fungus Trichoderma virens using Agrobacterium-mediated genetic transformation is reported in this paper. Integration of transgene in the genome of transgenic plants was demonstrated using polymerase chain reaction and Southern-blot hybridization, while expression was ascertained by reverse transcription polymerase chain reaction. Histochemical analysis confirmed the expression of GUS enzyme in transformed shoots. Levels of endochitinase enzyme in transgenic plants were found to be up to 10 fold higher compared to control plants. Endochitinase enzyme of 42 kDa was also visualized on SDS-PAGE gel using fluorimetric zymogram in transgenic plants. Endochitinase activity was found to be higher in leaf and stem than the root tissue in transgenic tomato plants. Transgenic lines of both plants showed enhanced resistance to fungal pathogens and a strong negative correlation was found between expression level of endochitinase enzyme and size of disease lesions. Inheritance of transgene, expression and resistance to fungal pathogens of T 1 transgenic tobacco lines was also analysed. The results of the present studies show that ech42 is a promising candidate gene for developing fungal disease resistance in tomato plants.

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Section: Discussionmentioning

confidence: 66%

Expression of a fungal endochitinase gene in transgenic tomato and tobacco results in enhanced tolerance to fungal pathogens

Shah

Mukherjee

Eapen

2010

Physiol Mol Biol Plants

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“…Although multiple copies of genes and promoter sequences could result in the inhibition of gene expression in plants (Matzke et al 1994;Meyer 1998) a number of studies have indicated that this does not always occur (Hobbs et al 1990;Maqbool et al 1999;Gahakwa et al 2000). Kohli et al (1999) showed that lines containing up to five copies of gusA and/or bar could express the gene stably at high levels through the R 3 generation.…”

Section: Discussionmentioning

confidence: 99%

Broccoli plants with pyramided cry1Ac and cry1C Bt genes control diamondback moths resistant to Cry1A and Cry1C proteins

et al. 2002

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This study was undertaken to determine the effects of pyramiding two Bacillus thuringiensis (Bt) genes in the same plant on the production of Bt proteins and the control of diamondback moths (DBM, Plutella xylostella) resistant to one or the other protein. Broccoli lines carrying both cry1Ac and cry1C Bt genes were produced by sexual crosses of cry1Ac- and cry1C-transgenic plants. Plants containing both genes were selected by tests for resistance to kanamycin and hygromycin, and confirmed by PCR analysis for the Bt genes. Both cry1Ac and cry1C mRNAs were detected in the hybrid lines, and Cry1Ac and Cry1C proteins were stably produced at levels comparable to the parental plants. Plants producing both Cry1Ac and Cry1C proteins caused rapid and complete mortality of DBM larvae resistant to Cry1A or Cry1C, and suffered little or no leaf damage. These plants, in combination with the resistant DBM populations available, will allow greenhouse or field studies of resistance management strategies involving gene pyramiding.

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“…Evidence from many transformation experiments indicates that there is no simple correlation between transgene copy number and expression level, with the exception of certain carefully controlled experiments using boundary elements. In some cases, higher copy numbers have suppressed overall expression levels (e.g., Cannell et al 1999;Spencer et al 1992) whereas in others higher copy numbers have enhanced expression (Stoger et al 1998;Gahakwa et al 2000). Where suppression effects have occurred, it has been suggested that ''runaway expression'' resulting in the generation of aberrant RNAs lacking polyadenylate tails has triggered potent silencing through the post-transcriptional silencing pathway (Schubert et al 2004).…”

Section: Repetitious Promoters and Transcriptional Gene Silencingmentioning

confidence: 99%

Promoter diversity in multigene transformation

et al. 2010

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Multigene transformation (MGT) is becoming routine in plant biotechnology as researchers seek to generate more complex and ambitious phenotypes in transgenic plants. Every nuclear transgene requires its own promoter, so when coordinated expression is required, the introduction of multiple genes leads inevitably to two opposing strategies: different promoters may be used for each transgene, or the same promoter may be used over and over again. In the former case, there may be a shortage of different promoters with matching activities, but repetitious promoter use may in some cases have a negative impact on transgene stability and expression. Using illustrative case studies, we discuss promoter deployment strategies in transgenic plants that increase the likelihood of successful and stable multiple transgene expression.

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Transgenic rice as a system to study the stability of transgene expression: multiple heterologous transgenes show similar behaviour in diverse genetic backgrounds

Cited by 50 publications

References 74 publications

Expression of a fungal endochitinase gene in transgenic tomato and tobacco results in enhanced tolerance to fungal pathogens

Expression of a fungal endochitinase gene in transgenic tomato and tobacco results in enhanced tolerance to fungal pathogens

Broccoli plants with pyramided cry1Ac and cry1C Bt genes control diamondback moths resistant to Cry1A and Cry1C proteins

Promoter diversity in multigene transformation

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