HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Abstract Full Text Free Full Text (PDF) Free Alert me when this article is cited Alert me if a correction is posted Services Related articles in Crop Science Similar articles in this journal Similar articles in ISI Web of Science Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via ISI Web of Science (5) Citing Articles via Google Scholar Google Scholar Articles by Cao, M. X. Articles by Lu, J. A. Search for Related Content PubMed Articles by Cao, M. X. Articles by Lu, J. A. Agricola Articles by Cao, M. X. Articles by Lu, J. A. Related Collections Rice Crop Genetics

Engineering Higher Yield and Herbicide Resistance in Rice by Agrobacterium-Mediated Multiple Gene Transformation

^a National Lab. of Plant Molecular Genetics, Inst. of Plant Physiology and Ecology, Shanghai Inst. for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China
^b Agrobiotech Research Center, Shanghai Academy of Agricultural Sciences, Shanghai 201106, China
^c Crop Breeding and Cultivation Research Institute, Shanghai Academy of Agricultural Sciences, Shanghai 201106, China

View larger version (7K): [in a new window]   Fig. 1. Schematic representation of the T-DNA region in the pYP1203E plasmid. 5'NOS, 5'-untranslated promoter region of the Agrobacterium tumefaciens nopaline synthase gene. VHb, homodimeric hemoglobin gene; 3'rbcs, 3'-untranslated terminator region of the Arabidopsis rbcs gene; SAR68, chromosome matrix-attachment region (MAR) of tobacco; 5'STS14, the pistil-specific promoter derived from potato; tzs, trans-zeatin secretion gene; 3'nos, 3'-untranslated terminator region of the nopaline synthase gene; EPSPS, 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene; RBCS, the chloroplast-targeting transit peptide RBCS from Arabidopsis at the 5' region of the EPSPS expression cassette; 5'35S, CaMV35S promoter; gus (intron), ß-glucuronidase gene with an intron; 5'D35S, double CaMV35S promoter; hpt, hygromycin phosphotransferase gene; LB, left border; RB, right border; BglII cut the plasmid DNA at a single site between 5'35S and GUS gene sequence.

View larger version (153K): [in a new window]   Fig. 2. Rice transformation mediated by Agrobaterium tumefaciens EHA105/pYP1203E and the results of GUS (ß-D-glucuronidase) histochemical analysis and herbicide-resistance test of transgenic rice plants. (A) Regenerated resistant shoots on RE2–CH50 medium. (B) Regenerated shoots and roots on 1/2MSNH50 medium (Left is the nontransformed control shoots). (C) Transgenic plants carrying the five transgenes putatively growing in soil in the greenhouse. (D) T1 progeny of transgenic Qiufeng carrying the five transgenes. (E) The roots and leaf of transgenic rice plants and the hygromycin-resistent calli (right) compared with the nontransformed control (left). (F) The leaves of transgenic rice plants (right) and the nontransformed control (left) after daubing with 5.0 mM glyphosate solution. (G) Transgenic plants (right) and the nontransformed control (left) after spraying with the equivalent of 13 g ha–1 glyphosate.

View larger version (68K): [in a new window]   Fig. 3. Polymerase chain reaction analysis of VHb (Vitreoscilla hemoglobin gene), tzs (trans-zeatin secretion gene), and EPSPS (5-enolpyruvylshikimate-3-phosphate synthase) in T0 rice plant. Lanes: M, molecular weight marker (A) and (C) DL2000, (B) X174 HaeIII digest; P, positive control (amplified fragments from plasmid pYP1203E); C, a nontransformed control plant; X-1 and X-2, putative T0 plants of ‘Xiushui-11’; Q-1 and 2, putative T0 plants of ‘Qiufeng’; H-1, putative T0 plants of ‘Hanfeng’; Y-1, putative T0 plants of ‘Youfeng’.

View larger version (96K): [in a new window]   Fig. 4. Southern blot analysis of T1 transgenic rice plants. Lane: P, 5 ng plasmid pYP1203E DNA cut by BglII at a single site was loaded as a positive control; C, a nontransformed control plant; X-1-1, X-2-9, Q-1-5, Q-2-1, H-1-1, and Y-1-1, representative T1 transgenic plants from their T0 progenies respectively (Plant genomic DNA digested with BglII. X stands for ‘Xiushui-11’, Q for ‘Qiufeng’, H for ‘Hanfeng’, and Y for ‘Youfeng’). The three polymerase chain reaction products were used as probes and labeled with DIG-Prime (digoxigenin; Roche Applied Science, Penzberg, Germany). The same southern filter was rehybridized with different probes. (A) VHb, Vitreoscilla hemoglobin gene; (B) tzs, trans-zeatin secretion gene; (C) EPSPS, 5-enolpyruvylshikimate-3-phosphate synthase gene.

View larger version (95K): [in a new window]   Fig. 5. Northern analysis of the expression of VHb (Vitreoscilla hemoglobin gene) and tzs (trans-zeatin secretion gene) in T1 plants. Lanes: X-1-1(k), Q-2-1(k), H-1-1(k), and Y-1-1(k) in (A) and (B) were loaded with 20 µg total RNA from kernels of T1 plants; Lanes: X-1-1(p), Q-2-1(p), and H-1-1(p) in (B) were loaded with 20 µg total RNA from pistils of T1 plants; Lanes: C(k) and C(p) were loaded with 20 µg total RNA from kernels and pistils of nontransformed control respectively. X stands for ‘Xiushui-11’, Q for ‘Qiufeng’, H for ‘Hanfeng’, and Y for ‘Youfeng’.