Agrobacterium tumefaciens -mediated creeping bentgrass ( Agrostis stolonifera L.) transformation using phosphinothricin selection results in a high frequency of single-copy transgene integration

Luo, Hong; Hu, Qian; Nelson, Kimberly; Longo, Chip; Kausch, Albert P.; Chandlee, Joel M.; Wipff, J K; Fricker, C. R.

doi:10.1007/s00299-003-0734-2

Cited by 79 publications

(47 citation statements)

References 32 publications

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“…4b). The low copy numbers obtained are consistent with the results from other grass species transformed using A. tumefaciens-mediated gene delivery (Bettany et al 2003;Luo et al 2004;Dong and Qu 2005;Han et al 2005;Wang and Ge 2005;Bajaj et al 2006;Ge et al 2006;Gao et al 2008). …”

Section: Molecular Characterization Of Transgenic Annual Ryegrass Plantssupporting

confidence: 79%

Agrobacterium-mediated transformation of Lolium rigidum Gaud.

Ran

Patron

et al. 2014

Plant Cell Tiss Organ Cult

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Lolium rigidum Gaud. is an annual grass grown for forage but also an economically damaging crop weed. A single genotype somatic embryogenic callus line, VLR1-60, was identified from a herbicide susceptible L. rigidum population, VLR1, and proved to be amenable to Agrobacterium tumefaciens-mediated transformation. Somatic embryogenic calli were continuously induced from the meristematic region of VLR1-60 plants multiplied in vitro and the basic tolerance level of VLR1-60 to hygromycin B was determined. A hygromycin phosphotransferase gene was used as a selectable marker for hygromycin B selection. Somatic embryogenic calli derived from in vitro grown vegetative tillers were co-cultivated with the A. tumefaciens strain EHA105 harbouring binary vector carrying reporter genes and selectable marker in the presence of acetosyringone for 3 days. Inoculated calli were recovered on callus proliferation medium containing Timentin TM but lacking hygromycin and were then subcultured onto media with hygromycin concentrations increased progressively through time for selection of transformed plant cells. Putative transgenic plants were recovered and integration of transgenes was confirmed by Southern hybridization analysis and by detection of DsRed or GUS activity in transgenic plants. The frequency of plant transformation was 1.3 %. The ability to transform L. rigidum will provide opportunities for functional characterization of genes to improve forage quality and increase our understanding of the evolution of herbicide resistance and of the basic genetics underlying traits that make L. rigidum a damaging crop weed.

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Section: Molecular Characterization Of Transgenic Annual Ryegrass Plantssupporting

confidence: 79%

Agrobacterium-mediated transformation of Lolium rigidum Gaud.

Ran

Patron

et al. 2014

Plant Cell Tiss Organ Cult

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“…Transgenic plants overexpressing Osa-miR319a were produced and maintained as described previously (Luo et al, 2004a(Luo et al, , 2004b.…”

Section: Plant Materials and Transformationmentioning

confidence: 99%

Constitutive Expression of a miR319 Gene Alters Plant Development and Enhances Salt and Drought Tolerance in Transgenic Creeping Bentgrass

et al. 2013

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MicroRNA319 (miR319) is one of the first characterized and conserved microRNA families in plants and has been demonstrated to target TCP (for TEOSINTE BRANCHED/CYCLOIDEA/PROLIFERATING CELL FACTORS [PCF]) genes encoding plant-specific transcription factors. MiR319 expression is regulated by environmental stimuli, suggesting its involvement in plant stress response, although experimental evidence is lacking and the underlying mechanism remains elusive. This study investigates the role that miR319 plays in the plant response to abiotic stress using transgenic creeping bentgrass (Agrostis stolonifera) overexpressing a rice (Oryza sativa) miR319 gene, Osa-miR319a. We found that transgenic plants overexpressing Osa-miR319a displayed morphological changes and exhibited enhanced drought and salt tolerance associated with increased leaf wax content and water retention but reduced sodium uptake. Gene expression analysis indicated that at least four putative miR319 target genes, AsPCF5, AsPCF6, AsPCF8, and AsTCP14, and a homolog of the rice NAC domain gene AsNAC60 were down-regulated in transgenic plants. Our results demonstrate that miR319 controls plant responses to drought and salinity stress. The enhanced abiotic stress tolerance in transgenic plants is related to significant down-regulation of miR319 target genes, implying their potential for use in the development of novel molecular strategies to genetically engineer crop species for enhanced resistance to environmental stress.

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“…Transgenic plants constitutively expressing Osa-miR528 were produced via A. tumefaciens-mediated transformation of embryonic callus induced from mature seeds as described previously (Luo et al, 2004).…”

Section: Plant Materials and Transformationmentioning

confidence: 99%

Constitutive Expression of Rice MicroRNA528 Alters Plant Development and Enhances Tolerance to Salinity Stress and Nitrogen Starvation in Creeping Bentgrass

et al. 2015

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MicroRNA528 (miR528) is a conserved monocot-specific small RNA that has the potential of mediating multiple stress responses. So far, however, experimental functional studies of miR528 are lacking. Here, we report that overexpression of a rice (Oryza sativa) miR528 (Osa-miR528) in transgenic creeping bentgrass (Agrostis stolonifera) alters plant development and improves plant salt stress and nitrogen (N) deficiency tolerance. Morphologically, miR528-overexpressing transgenic plants display shortened internodes, increased tiller number, and upright growth. Improved salt stress resistance is associated with increased water retention, cell membrane integrity, chlorophyll content, capacity for maintaining potassium homeostasis, CATALASE activity, and reduced ASCORBIC ACID OXIDASE (AAO) activity; while enhanced tolerance to N deficiency is associated with increased biomass, total N accumulation and chlorophyll synthesis, nitrite reductase activity, and reduced AAO activity. In addition, AsAAO and COPPER ION BINDING PROTEIN1 are identified as two putative targets of miR528 in creeping bentgrass. Both of them respond to salinity and N starvation and are significantly down-regulated in miR528-overexpressing transgenics. Our data establish a key role that miR528 plays in modulating plant growth and development and in the plant response to salinity and N deficiency and indicate the potential of manipulating miR528 in improving plant abiotic stress resistance.Abiotic stresses, especially drought, salt, and nitrogen (N) deficiency, are limiting factors for plant growth, development, and agricultural productivity. To cope with drought and salt stresses, plants have evolved similar strategies of osmotic adjustment (Munns, 2002;Zhu, 2002). Plants also evolved salinity-specific adjustments against ionic disequilibrium, which encompass excluding salt entry into plants and compartmentalizing ions into vacuoles or old leaves (Munns, 1993;Yeo, 1998). Another worldwide limiting factor for crop yields is N deficiency, which triggers reduced leaf growth rate and photosynthetic rate (Chapin et al., 1988). Due to the sessile nature of plants, abiotic stresses are unavoidable. Therefore, it is critical to develop reliable procedures to genetically modify plants for improved performance under environmental stresses, thereby enhancing agricultural productivity to meet the ever-growing demands in food production.Genetic engineering plays an increasingly important role in agronomic trait modifications in crop species. Currently, many genes encoding functional proteins, transcription factors, and proteins involved in signaling pathways have been identified as abiotic stressresponsive genes (Shinozaki and Yamaguchi-Shinozaki, 2007;Masclaux-Daubresse et al., 2010;Turan et al., 2012). Constitutive expression of some of these genes in transgenic plants has been demonstrated to lead to enhanced salt or drought tolerance (Golldack et al., 2011;Kim et al., 2013;Lu et al., 2013;Li et al., 2014). However, details of the regulatory network in the plant...

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Agrobacterium tumefaciens -mediated creeping bentgrass ( Agrostis stolonifera L.) transformation using phosphinothricin selection results in a high frequency of single-copy transgene integration

Cited by 79 publications

References 32 publications

Agrobacterium-mediated transformation of Lolium rigidum Gaud.

Agrobacterium-mediated transformation of Lolium rigidum Gaud.

Constitutive Expression of a miR319 Gene Alters Plant Development and Enhances Salt and Drought Tolerance in Transgenic Creeping Bentgrass

Constitutive Expression of Rice MicroRNA528 Alters Plant Development and Enhances Tolerance to Salinity Stress and Nitrogen Starvation in Creeping Bentgrass

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