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 Similar articles in this journal Similar articles in Web of Science Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Web of Science (19) Citing Articles via Google Scholar Google Scholar Articles by Saruul, P. Articles by Samac, D. A. Search for Related Content PubMed Articles by Saruul, P. Articles by Samac, D. A. Agricola Articles by Saruul, P. Articles by Samac, D. A. Related Collections Germplasm Enhancement Alfalfa Cell Biology & Molecular Genetics

Production of a Biodegradable Plastic Polymer, Poly-ß-Hydroxybutyrate, in Transgenic Alfalfa

^a Dep. of Plant Pathology, 495 Borlaug Hall
^b Biological Process Technol. Inst., 240 Gortner Laboratories
^c Dept. of Agronomy and Plant Genetics, 411 Borlaug Hall
^d USDA-ARS-Plant Science Research Unit and Dep. of Plant Pathology, 495 Borlaug Hall, Univ. of Minnesota, St. Paul, MN 55108-6030

View larger version (72K): [in a new window]   Fig. 1. DNA Southern blot analysis of transgenic alfalfa plants. Restriction enzyme-digested genomic DNA was hybridized with 32P-labeled phbA (Panel B) or phbB (Panel C). (A) Restriction map of the T-DNA from pMON25948. Bars underneath the map indicate sizes of restriction fragments containing phbA or phbB. R1, EcoR1 site; RV, EcoRV site; RB and LB, right and left T-DNA border, respectively; P-35S, CaMV 35S promoter; SS-TP, transit peptide of the small subunit of Rubisco. E9-3' and nos 3', terminator region of soybean E9 and Agrobacterium nopaline synthase, respectively. (B and C) Lane P, EcoRV-digested pMON25948; Lane 1, Line 863 digested with EcoR1; Lane 2, Line 863 digested with EcoRV; Lane 3, Line 864 digested with EcoR1; Lane 4, Line 864 digested with EcoRV; Lane 5, Line 916 digested with EcoR1; Lane 6, Line 916 digested with EcoRV; Lane 7, untransformed control digested with EcoR1; Lane 8, untransformed control digested with EcoRV.

View larger version (76K): [in a new window]   Fig. 2. RNA northern blot analysis of transgenic alfalfa plants. Total RNA from mature leaves of transgenic lines and untransformed alfalfa were hybridized with individual phb genes. RNA loaded in each lane is shown by ethidium bromide staining of 28S rRNA. Lane 1, Line 864; Lane 2, untransformed alfalfa; Lane 3, Line 913.

View larger version (23K): [in a new window]   Fig. 3. Accumulation of 3-hydroxybutyrate (3-HB) in alfalfa plants. (A) Plants transformed with Vector pMON25948 (phbABC). (B) Plants transformed with Vector pMON25949 (bktB, phbBC).

View larger version (22K): [in a new window]   Fig. 4. Gas chromatography–mass-spectrometry analysis of PHB produced in transgenic alfalfa. (A) Electron impact mass fragmentation spectrum of Line 864. (B) Poly-ß-hydroxybutyrate standard.

View larger version (13K): [in a new window]   Fig. 5. Proton-nuclear magnetic resonance spectrometry (1H-NMR) analysis of alfalfa leaf extracts for poly-ß-hydroxybutyrate (PHB). (A) 500-MHz 1H-NMR spectrum of PHB standard; (B) Line 864 transformed with phbABC genes; (C) Control, untransformed alfalfa. Note the expanded spectrum containing PHB peaks (2.5 and 5.2 ppm) present in transgenic plants.

View larger version (106K): [in a new window]   Fig. 6. Visualization of poly-ß-hydroxybutyrate (PHB) granule accumulation in alfalfa Line 864 transformed with phbABC genes. Leaf tissues stained with Nile Blue A and viewed by confocal microscopy. (A) Untransformed alfalfa; (B) Line 864. Chloroplasts from mature leaves of alfalfa viewed by transmission electron microscopy. (C) Untransformed alfalfa, bar = 1 µm; (D) Transgenic alfalfa with agglomerations of electron-lucent PHB granules (indicated with arrows), bar = 1 µm; (E) Immuno-gold labeling of granules with antibodies against PHB synthase, bar = 0.5 µm.