S-RNases from self-incompatible and -compatible apple cultivars: Purification, cloning, enzymic properties, and pollen tube growth inhibitory activity

• Published in: Bioscience, biotechnology, and biochemistry Vol. 66; no. 6; pp. 1185 - 1195
• Main Authors: Katoh, N. (Hirosaki Univ., Aomori (Japan). Faculty of Agriculture and Life Science), Goto, K, Asano, J, Fukushima, K, Yamada, K, Kasai, A, Li, T.Z, Takanoha, M, Miyairi, K, Okuno, T
• Format: Journal Article
• Language: English
• Published: Tokyo Japan Society for Bioscience, Biotechnology, and Agrochemistry 01.06.2002; Japan Society for Bioscience Biotechnology and Agrochemistry
• Subjects: Agronomy. Soil science and plant productions; Amino Acid Sequence; amino acids; Antibodies; apple; apples; Base Sequence; bioassays; Biological and medical sciences; Biological Assay; biotechnology; Blotting, Western; Cloning, Molecular; cultivars; Economic plant physiology; Enzyme Stability; Flowering, floral biology, reproduction patterns; Flowers - enzymology; Flowers - growth & development; Fundamental and applied biological sciences. Psychology; Gene Expression Regulation, Enzymologic; Gene Expression Regulation, Plant; GROWTH; Growth and development; Hydrogen-Ion Concentration; INHIBITION; MALUS; Malus - classification; Malus - enzymology; Malus - genetics; Malus domestica; MOLECULAR CLONING; Molecular Sequence Data; Molecular Weight; nucleotide sequences; Phosphorylcholine - analogs & derivatives; Phosphorylcholine - pharmacology; Plant physiology and development; pollen; pollen growth inhibition; POLLEN TUBES; PURIFICATION; RIBONUCLEASES; Ribonucleases - chemistry; Ribonucleases - genetics; Ribonucleases - isolation & purification; Ribonucleases - metabolism; S-RNase; SELF COMPATIBILITY; SELF STERILITY; self-incompatibility; Sexual reproduction; Temperature; thermal stability; VARIETIES; Vegetative and sexual reproduction, floral biology, fructification; Western blotting; Bioassay; Purification; Genetic variability; Nucleotide sequence; Enzyme; Rosaceae; Homology; Esterases; Malus domestica; Ribonuclease; Sexual reproduction; Pollen tube; Gynoecium; Characterization; Selfincompatibility; Complementary DNA; Dicotyledones; Angiospermae; Development; Hydrolases; Spermatophyta; Fruit tree; Aminoacid sequence
• ISSN: 0916-8451; 1347-6947; 1347-6947
• DOI: 10.1271/bbb.66.1185

Four S-RNases (RNase associated with self-incom-patibility) were purified from the styles of two apple cultivars (Malus domestica), a self-incompatible cv., Starking Delicious (SD), and a self-compatible cv., Megumi (MG). Each cultivar produced two S-RNases and their enzymatic properties such as specific activity, pH optimum, thermal stability, and molecular mass, were characterized. The four S-RNases inhibited the tube growth of apple pollen in an in vitro bioassay at 25 micro g/ml (1.0 micro M), but did not distinguish self from non-self pollen. The cDNAs of four S-RNases were cloned, and the nucleotide and deduced amino acid sequences were analyzed. The nucleotide sequence of SD-Se RNase was a new one and the other was identical to that of Sc-RNase of cv. Fuji. In MG one was identical to the sequence of SD-Sc RNase and the other to that of Sa-RNase of cv. Golden Delicious except for one base. From results of the isolation amounts and the Western blot analysis for stylar crude extracts the amount of S-RNases in MG was apparently less than that in SD.