S-RNases from self-incompatible and -compatible apple cultivars: purification, cloning, enzymic properties, and pollen tube growth inhibitory activity.
Four S-RNases (RNase associated with self-incompatibility) 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 microg/ml (1.0 microM), 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.
Katoh
Naoki
N
Laboratory of Biochemistry, Faculty of Agriculture and Life Sciences, Hirosaki University, Japan.
Goto
Kazunori
K
Asano
Junpei
J
Fukushima
Kiyoe
K
Yamada
Kenji
K
Kasai
Aya
A
Li
Tian Zhong
TZ
Takanoha
Makoto
M
Miyairi
Kazuo
K
Okuno
Toshikatsu
T
D000595:Amino Acid Sequence; D000906:Antibodies; D001483:Base Sequence; D001681:Biological Assay; D015153:Blotting, Western; D003001:Cloning, Molecular; D004795:Enzyme Stability; D035264:Flowers / Q000201:enzymology* / Q000254:growth & development*; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant; D006863:Hydrogen-Ion Concentration; D027845:Malus / Q000145:classification / Q000201:enzymology* / Q000235:genetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010767:Phosphorylcholine / Q000031:analogs & derivatives* / Q000494:pharmacology; D012260:Ribonucleases / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D013696:Temperature
D000906:Antibodies; C437127:O,O-diethylphosphorylcholine iodide; D010767:Phosphorylcholine; D012260:Ribonucleases