Pectin engineering:: Modification of potato pectin by in vivo expression of an endo-1,4-β-D-galactanase

被引:125
作者
Sorensen, SO [1 ]
Pauly, M
Bush, M
Skjot, M
McCann, MC
Borkhardt, B
Ulvskov, P
机构
[1] Danish Inst Agr Sci, Biotechnol Grp, DK-1871 Copenhagen, Denmark
[2] Royal Vet & Agr Univ, Dept Plant Biol, DK-1871 Copenhagen, Denmark
[3] John Innes Ctr Plant Sci Res, Dept Cell Biol, Norwich NR4 7UH, Norfolk, England
关键词
D O I
10.1073/pnas.130568297
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Potato tuber pectin is rich in galactan (oligomer of beta-1,4-linked galactosyl residues). We have expressed a fungal endo-galactanase cDNA in potato under control of the granule bound starch synthase promoter to obtain expression of the enzyme in tubers during growth. The transgenic plants displayed no altered phenotype compared with the wild type. Fungal endo-galactanase activity was quantified in the transgenic tubers, and its expression was verified by Western blot analysis. The effect of the endo-galactanase activity on potato tuber pectin was studied by Fourier transform infrared microspectroscopy. immune-gold labeling, and sugar analysis. All analyses revealed alterations in pectin composition. Monosaccharide composition of total cell walls and isolated rhamnogalacturonan I fragments showed a reduction in galactosyl content to 30% in the transformants compared with the wild type. Increased solubility of pectin from transgenic cell walls by endo-polygalacturonase/pectin methylesterase digestion points to other changes in wall architecture.
引用
收藏
页码:7639 / 7644
页数:6
相关论文
共 35 条
[1]   PECTINS AS MEDIATORS OF WALL POROSITY IN SOYBEAN CELLS [J].
BARONEPEL, O ;
GHARYAL, PK ;
SCHINDLER, M .
PLANTA, 1988, 175 (03) :389-395
[2]   NEW METHOD FOR QUANTITATIVE-DETERMINATION OF URONIC ACIDS [J].
BLUMENKR.N ;
ASBOEHAN.G .
ANALYTICAL BIOCHEMISTRY, 1973, 54 (02) :484-489
[3]   Pectic epitopes are differentially distributed in the cell walls of potato (Solanum tuberosum) tubers [J].
Bush, MS ;
McCann, MC .
PHYSIOLOGIA PLANTARUM, 1999, 107 (02) :201-213
[4]   TOMATO EXO-(1-]4)-BETA-D-GALACTANASE - ISOLATION, CHANGES DURING RIPENING IN NORMAL AND MUTANT TOMATO FRUIT, AND CHARACTERIZATION OF A RELATED CDNA CLONE [J].
CAREY, AT ;
HOLT, K ;
PICARD, S ;
WILDE, R ;
TUCKER, GA ;
BIRD, CR ;
SCHUCH, W ;
SEYMOUR, GB .
PLANT PHYSIOLOGY, 1995, 108 (03) :1099-1107
[5]   STRUCTURAL MODELS OF PRIMARY-CELL WALLS IN FLOWERING PLANTS - CONSISTENCY OF MOLECULAR-STRUCTURE WITH THE PHYSICAL-PROPERTIES OF THE WALLS DURING GROWTH [J].
CARPITA, NC ;
GIBEAUT, DM .
PLANT JOURNAL, 1993, 3 (01) :1-30
[6]   In vitro synthesis and properties of pectin/Acetobacter xylinus cellulose composites [J].
Chanliaud, E ;
Gidley, MJ .
PLANT JOURNAL, 1999, 20 (01) :25-35
[7]   A rapid method to screen for cell-wall mutants using discriminant analysis of Fourier transform infrared spectra [J].
Chen, LM ;
Carpita, NC ;
Reiter, WD ;
Wilson, RH ;
Jeffries, C ;
McCann, MC .
PLANT JOURNAL, 1998, 16 (03) :385-392
[8]   EXPRESSION CLONING, PURIFICATION AND CHARACTERIZATION OF A BETA-1-4-GALACTANASE FROM ASPERGILLUS-ACULEATUS [J].
CHRISTGAU, S ;
SANDAL, T ;
KOFOD, LV ;
DALBOGE, H .
CURRENT GENETICS, 1995, 27 (02) :135-141
[9]   OLIGOSACCHARINS - STRUCTURES AND SIGNAL-TRANSDUCTION [J].
COTE, F ;
HAHN, MG .
PLANT MOLECULAR BIOLOGY, 1994, 26 (05) :1379-1411
[10]  
DATLA RSS, 1993, GENE, V129, P311