Promoters of Rice Seed Storage Protein Genes Direct Endosperm-Specific Gene Expression in Transgenic Rice

Wu, Chuanyin; Adachi, Takahiro; Hatano, Tetsuya; Washida, Haruhiko; Suzuki, Akihiro; Takaiwa, Fumio

doi:10.1093/oxfordjournals.pcp.a029449

Cited by 71 publications

(64 citation statements)

References 31 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To visualize prolamine RNA movement in developing rice endosperm cells, the promoters from the rice glutelin Gt1 (Zheng et al, 1993) and prolamine pProl (Wu et al, 1998) were inserted upstream of NLS-MS2-GFP and GUS-MS6X-prolamine, respectively (Figure 1). These two genes then were cloned into the T-DNA vector pCAMBIA1301, which was used to transform rice via Agro-bacterium tumefaciens .…”

Section: Prolamine Rnas Form Particles In Developing Rice Endospermmentioning

confidence: 99%

“…The RNA transcriptional gene fusion was driven by the prolamine Pprol promoter (Wu et al, 1998) and was prepared as follows. A SacIHindIII fragment containing the nos terminator (Tnos) was cloned into pSL1180.…”

Section: Plasmid Dna Construction and Plant Transformationmentioning

confidence: 99%

“…A SacIHindIII fragment containing the nos terminator (Tnos) was cloned into pSL1180. To form the Pprol-MS6X fusion, the MS6X fragment (BamHISacI) from pSL-MS2-6 was cloned into the BamHI-SacI of pProl (Wu et al, 1998). The resulting plasmid was digested with EcoRI and NotI, and the Pprol:MS6X fragment was cloned upstream of Tnos.…”

Section: Plasmid Dna Construction and Plant Transformationmentioning

confidence: 99%

See 2 more Smart Citations

The Transport of Prolamine RNAs to Prolamine Protein Bodies in Living Rice Endosperm Cells[W]

et al. 2003

View full text Add to dashboard Cite

RNAs that code for the major rice storage proteins are localized to specific subdomains of the cortical endoplasmic reticulum (ER) in developing endosperm. Prolamine RNAs are localized to the ER and delimit the prolamine intracisternal inclusion granules (PB-ER), whereas glutelin RNAs are targeted to the cisternal ER. To study the transport of prolamine RNAs to the surface of the prolamine protein bodies in living endosperm cells, we adapted a two-gene system consisting of green fluorescent protein (GFP) fused to the viral RNA binding protein MS2 and a hybrid prolamine RNA containing tandem MS2 RNA binding sites. Using laser scanning confocal microscopy, we show that the GFP-labeled prolamine RNAs are transported as particles that move at an average speed of 0.3 to 0.4 m/s. These prolamine RNA transport particles generally move unidirectionally in a stop-and-go manner, although nonlinear bidirectional, restricted, and nearly random movement patterns also were observed. Transport is dependent on intact microfilaments, because particle movement is inhibited rapidly by the actin filament-disrupting drugs cytochalasin D and latrunculin B. Direct evidence was obtained that these prolamine RNA-containing particles are transported to the prolamine protein bodies. The significance of these results with regard to protein synthesis in plants is discussed.

show abstract

Section: Prolamine Rnas Form Particles In Developing Rice Endospermmentioning

confidence: 99%

Section: Plasmid Dna Construction and Plant Transformationmentioning

confidence: 99%

Section: Plasmid Dna Construction and Plant Transformationmentioning

confidence: 99%

See 1 more Smart Citation

The Transport of Prolamine RNAs to Prolamine Protein Bodies in Living Rice Endosperm Cells[W]

et al. 2003

View full text Add to dashboard Cite

show abstract

“…Tissue-specific expression of seed protein genes from legumes (Beachy et al 1985;Sengupta-Gopalan et al 1985;Chen et al 1986;Okamuro et al 1986), wheat (Colot et al 1987), Rice (Wu et al 1998;Patti et al 2012) and maize (Schernthaner et al 1988;Russell and Fromm 1997) have been shown previously. The zein promoter consisting 933 bp of 23 kDa zein gene and 1,100 bp of 27 kDa zein gene was expressed in tobacco (Schernthaner et al 1988) and maize (Russell and Fromm 1997) and endosperm-specific expression was achieved by them.…”

Section: Discussionmentioning

confidence: 67%

A maize α-zein promoter drives an endosperm-specific expression of transgene in rice

Joshi

Geetha

Singh

et al. 2014

Physiol Mol Biol Plants

View full text Add to dashboard Cite

An alpha-zein promoter isolated from maize containing P-box, E motif sequence TGTAAAGT, opaque-2 box and TATA box was studied for its tissuespecific expression in rice. A 1,098 bp promoter region of alpha-zein gene, fused to the upstream of gusA reporter gene was used for transforming rice immature embryos (ASD 16 or IR 64) via the particle bombardmentmediated method. PCR analysis of putative transformants demonstrated the presence of transgenes (the zein promoter, gusA and hpt). Nineteen out of 37 and two out of five events generated from ASD 16 and IR 64 were found to be GUS-positive. A histological staining analysis performed on sections of mature T 1 seeds revealed that the GUS expression was limited to the endosperm and not to the pericarp or the endothelial region. GUS expression was observed only in the following seed development stages : milky (14-15 DAF), soft dough (17-18 DAF), hard dough (20-23 DAF), and mature stages (28-30 DAF) of zein-gusA transformed (T 0 ) plants. On the contrary a constitutive expression of GUS was evident in CaMV35S-gusA plants. PCR and Southern blotting analyses on T 1 plants demonstrated a stable integration and inheritance of transgene in the subsequent T 1 generation. GUS assay on T 2 seeds revealed that the expression of gusA gene driven by alphazein promoter was stable and tissue-specific over two generations. Results suggest that this alpha-zein promoter could serve as an alternative promoter to drive endosperm-specific expression of transgenes in rice and other cereal transformation experiments.

show abstract

“…To make a marker-free vector, we first inserted GluB1 promoter, which regulates seed storage protein genes in the endosperm of rice seeds [21] into the to the pCAMBIA1300 binary vector. The HPTII expression cassette (CaMV 35S promoter-HPTII gene-CaMV 35S terminator) in the pCAMBIA1300 binary vector was removed by XhoI and EcoRI restriction enzyme treatment.…”

Section: Dna Constructsmentioning

confidence: 99%

Generation of Transgenic Rice without Antibiotic Selection Marker through Agrobacterium-mediated Co-transformation System

Park¹,

Kwon²,

Lee³

et al. 2012

Journal of Life Science

View full text Add to dashboard Cite

Development of transgenic plant increasing crop yield or disease resistance is good way to solve the world food shortage. However, the persistence of marker genes in crops leads to serious public concerns about the safety of transgenic crops. In the present paper, we developed marker-free transgenic rice inserted high molecular-weight glutenin subunit (HMW-GS) gene (Dx5) from the Korean wheat cultivar 'Jokyeong' using Agrobacterium-mediated co-transformation method. Two expression cassettes comprised of separate DNA fragments containing only the Dx5 and hygromycin resistance (HPTII) genes were introduced separately into Agrobacterium tumefaciens EHA105 strain for co-infection. Each EHA105 strain harboring Dx5 or HPTII was infected into rice calli at a 3: 1 ratio of EHA105 with Dx5 gene and EHA105 with HPTII gene expressing cassette. Then, among 66 hygromycin-resistant transformants, we obtained two transgenic lines inserted with both the Dx5 and HPTII genes into the rice genome. We reconfirmed integration of the Dx5 and HPTII genes into the rice genome by Southern blot analysis. Wheat Dx5 transcripts in T1 rice seeds were examined with semi-quantitative RT-PCR. Finally, the marker-free plants containing only the Dx5 gene were successfully screened at the T1 generation. These results show that a co-infection system with two expression cassettes could be an efficient strategy to generate marker-free transgenic rice plants.

show abstract

Promoters of Rice Seed Storage Protein Genes Direct Endosperm-Specific Gene Expression in Transgenic Rice

Cited by 71 publications

References 31 publications

The Transport of Prolamine RNAs to Prolamine Protein Bodies in Living Rice Endosperm Cells[W]

The Transport of Prolamine RNAs to Prolamine Protein Bodies in Living Rice Endosperm Cells[W]

A maize α-zein promoter drives an endosperm-specific expression of transgene in rice

Generation of Transgenic Rice without Antibiotic Selection Marker through Agrobacterium-mediated Co-transformation System

Contact Info

Product

Resources

About