An acidic class III chitinase in sugar beet: induction by Cercospora beticola, characterization, and expression in transgenic tobacco plants.
An acidic chitinase (SE) was found to accumulate in leaves of sugar beet (Beta vulgaris) during infection with Cercospora beticola. Two isoforms, SE1 and SE2, with MW of 29 kDa and pI of approximately 3.0 were purified to homogeneity. SE2 is an endochitinase that also exhibits exochitinase activity, i.e., it is capable of hydrolyzing chito-oligosaccharides, including chitobiose, into N-acetyl-glucosamine. Partial amino acid sequence data for SE2 were used to obtain a cDNA clone by polymerase chain reaction. The clone was used to isolate a cDNA clone encoding SE2. The deduced amino acid sequence for SE2 is 58-67% identical to the class III chitinases from cucumber, Arabidopsis, and tobacco. A transient induction of SE2 mRNA during the early stages of infection with C. beticola is much stronger in tolerant plants than in susceptible plants. Transgenic tobacco (Nicotiana benthamiana) plants constitutively accumulate SE2 protein in the intercellular space of their leaves. In a preliminary infection experiment, the transgenic plants did not show increase in resistance against C. nicotianae.
Nielsen
K K
KK
Maribo Seed Biotechnology, DANISCO A/S, Copenhagen K, Denmark.
Mikkelsen
J D
JD
Kragh
K M
KM
Bojsen
K
K
D000595:Amino Acid Sequence; D001483:Base Sequence; D002688:Chitinases / Q000096:biosynthesis* / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004591:Electrophoresis, Polyacrylamide Gel; D004790:Enzyme Induction; D015151:Immunoblotting; D007527:Isoenzymes / Q000096:biosynthesis* / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D003904:Mitosporic Fungi / Q000502:physiology*; D008969:Molecular Sequence Data; D010935:Plant Diseases / Q000382:microbiology; D010940:Plant Proteins / Q000096:biosynthesis* / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D030821:Plants, Genetically Modified; D012333:RNA, Messenger / Q000096:biosynthesis; D017386:Sequence Homology, Amino Acid; D014675:Vegetables / Q000201:enzymology* / Q000382:microbiology
D018076:DNA, Complementary; D007527:Isoenzymes; D010940:Plant Proteins; D012333:RNA, Messenger; D002688:Chitinases