Journal article
Characterization of chitinases able to rescue somatic embryos of the temperature-sensitive carrot variant ts 11
To characterize the acidic endochitinase EP3, able to rescue somatic embryos of the carrot cell line ts11, the enzyme was purified from the medium of wild-type suspension cultures. Peptide sequences, deduced amino acid sequences of corresponding PCR-generated cDNA clones, serological relation and biochemical properties showed that there were at least five closely related chitinases, four of which could be identified as class IV EP3 chitinases with an apparent size of 30 kDa.
Two other proteins were identified as a serologically related class I acidic chitinase (DcChitI) of 34 kDa, and a serologically unrelated 29 kDa class II acidic chitinase (DcChitII), respectively. Additional cDNA sequences, Western and Southern analysis showed the presence of a least two, but possibly more, highly homologous class IV EP3 genes in the carrot genome.
Two class IV EP3 chitinases were tested and found to be able to increase the number of ts11 globular embryos formed under non-permissive conditions. One of the class IV EP3 chitinases as well as the class I chitinase DcChitI promoted the transition from globular to heart-stage ts11 embryos. The class II endochitinase and a heterologous class IV chitinase from sugar-beet were not active on ts11.
This suggests that there are differences in the specificity of chitinases in terms of their effect on plant somatic embryos.
Language: | English |
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Publisher: | Kluwer Academic Publishers |
Year: | 1996 |
Pages: | 631-45 |
Journal subtitle: | An International Journal on Molecular Biology, Molecular Genetics and Biochemistry |
ISSN: | 15735028 and 01674412 |
Types: | Journal article |
DOI: | 10.1007/BF00042235 |
Amino Acid Sequence Base Sequence Biochemistry, general Cells, Cultured Chitinases Chromatography, DEAE-Cellulose Chromatography, Gel Cloning, Molecular DNA Primers DNA, Complementary Daucus carota Fabaceae Genetic Variation Isoenzymes Kinetics Life Sciences Molecular Sequence Data Plant Pathology Plant Sciences Plants, Medicinal Polymerase Chain Reaction Recombinant Fusion Proteins Seeds Sequence Homology, Amino Acid Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization carrot class I acidic endochitinase class II acidic endochitinase plant class IV endochitinase (EC 3.2.1.14) somatic embryos,ts11