Abstract
Japanese pear (Pyrus serotina Rehd.) exhibits gametophytic self-incompatibility. Following our previous findings that basic ribonucleases in the styles of Japanese pear are associated with self-incompatibility genes (S-RNases), stylar proteins with high pI values were analyzed by two-dimensional gel electrophoresis further to characterize S-RNases. A group of basic proteins of about 30 kDa associated with self-incompatibility genes were identified. These proteins contained sugar chains which reacted with concanavalin A and wheat germ agglutinin, and thus were designated as S-glycoproteins of Japanese pear. The fact that the S-glycoprotein was expressed at a much lower level in a self-compatible mutant than in the original variety suggested a role of S-glycoproteins in mediating self-incompatibility of Japanese pear. Immunoblot analysis indicated that S-glycoproteins are identical to previously identified S-RNases. The S-glycoproteins were predominantly expressed in the style, in the ovary in trace amounts, and not in leaf, pollen or germinated pollen. The N-terminal amino acid sequences of the S-glycoproteins showed homology not only with each other but also with those of the S-allele-associated proteins from plants of the family Solanaceae at levels of about 30–50%.
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Ai Y, Singh A, Coleman CE, Ioerger TR, Kheyr-Pour A, Kao T-H (1990) Self-incompatibility in Petunia inflata. isolation and characterization of cDNAs encoding three S-allele associated proteins. Sex Plant Reprod 3:130–138
Anderson MA, Cornish EC, Mau S-L, Williams EG, Hoggart R, Atkinson A, Bonig I, Grego B, Simpson R, Roche PJ, Haley JD, Penschow JD, Niall HD, Tregear GW, Coghlan JP, Crawford RJ, Clarke AE (1986) Cloning of cDNA for a stylar glycoprotein associated with expression of self-incompatibility in Nicotiana alata. Nature 321:38–44
Clarke RE, Okuley JJ, Collins PD, Sims TL (1990) Sequence variability and developmental expression of S-alleles in self-incompatible and pseudo-self-compatible Petunia. Plant Cell 2:815–826
Damerval C, de Vienne D, Zivy M, Thiellement H (1986) Technical improvements in two-dimensional electrophoresis increase the level of genetic variation detected in wheat-seedling proteins. Electrophoresis 7:52–54
de Nettancourt D (1977) Incompatibility in angiosperms. Springer-Verlag, Berlin
Franklin-Tong VE, Franklin FCH (1992) Gametophytic self-incompatibility in Papaver rhoeas L. Sex Plant Reprod 5:1–7
Haring VH, Gray JE, McClure BA, Anderson MA, Clarke AE (1990) Self-incompatibility: A self-recognition system in plants. Science 250:937–941
Hirano H, Watanabe T (1990) Microsequencing of proteins electroblotted onto immobilizing matrices from polyacrylamide gel electrophoresis: Application to an insoluble protein. Electrophoresis 11:573–580
Hirano H, Komatsu S, Nakamura A, Kikuchi F, Kajiwara H, Tsunasawa S, Sakiyama F (1991) Structural homology between semidwarfism-related proteins and glutelin seed protein in rice (Oryza sativa L.) Theor Appl Genet 83:153–158
Hirata N (1989) Self-compatible mutant in Japanese pear. Gamma Field Symposia 28:71–80
Hiratsuka S (1992) Detection and inheritance of a stylar protein associated with a self-incompatibility genotype of Japanese pear. Euphytica 61:55–59
Hiratsuka S, Takahashi E, Hirata N (1987) Crossability of selfcompatible mutant “Osa-Nijisseiki” (Pyrus serotina Rehd.). J Jpn Soc Hort Sci 56:166–172
Horiuchi H, Yanai K, Takagi M, Yano K, Wakabayashi E, Sanda A, Mine S, Ohgi K, Irie M (1988) Primary structure of a base non-specific ribonuclease from Rhizopus niveus. J Biochem 103:408–418
Kajiura M, Kanato K, Machida Y, Kozaki I (1967) New Japanese pear variety “Shinsui”. Bull Hort Res Station 6:69–76 (in Japanese)
Kaufmann H, Salamini F, Thompson RD (1991) Sequence variability and gene structure at the self-incompatibility locus of Solanum tuberosum. Mol Gen Genet 226:457–466
Kawata Y, Sakiyama F, Tamaoki H (1988) Amino-acid sequence of ribonuclease T2 from Aspergillus oryzae. Eur J Biochem 176:683–697
Kheyr-Pour A, Bintrim SB, Ioerger TR, Remy R, Hammond SA, Kao T-H (1990) Sequence diversity of pistil S-proteins associated with gametophytic self-incompatibility in Nicotiana alata. Sex Plant Reprod 3:88–97
Kijimoto-Ochiai S, Katagiri YU, Ochiai H (1985) Analysis of N-linked oligosaccharide chains of glycoproteins on nitrocellulose sheets using lectin-peroxidase reagents. Anal Biochem 147:222–229
Kirch HH, Uhrig H, Lottspeich F, Salamini F, Thompson RD (1989) Characterization of proteins associated with self-incompatibility in Solanum tuberosum. Theor Appl Genet 78:581–588
Machida Y (1972) In: Sato K, Mori H, Matsui O, Kitajima H, Chiba T (eds) Dictionary of pomology. Yokendo, Tokyo pp 527–529 (in Japanese)
McClure BA, Haring V, Ebert PR, Anderson MA, Simpson RJ, Sakiyama F, Clarke AE (1989) Style self-incompatibility gene products of Nicotiana alata are ribonucleases. Nature 342:955–957
McClure BA, Gray JE, Anderson MA, Clarke AE (1990) Self-incompatibility in Nicotiana alata involves degradation of pollen rRNA. Nature 347:757–760
Nakanishi TM, Saeki N, Maeno M, Ozaki T, Kawai T, Ichii T (1991) Ultrastructural study on the stylar transmitting tissue in Japanese pear. Sex Plant Reprod 4:95–103
Nakanishi T, Yamazaki T, Funadera K, Tomonaga H, Ozaki T, Kawai Y, Ichii T, Sato Y, Kurihara A (1992) Isoelectric focusing analysis of stylar proteins associated with self-incompatibility alleles in Japanese pear. J Jpn Soc Hort Sci 61:239–248
Nasrallah JB, Nishio T, Nasrallah ME (1991) The self-incompatibility genes of Brassica: Expression and use in genetic ablation of floral tissue. Annu Rev Plant Physiol Plant Mol Biol 42:393–422
O'Farrell PH (1975) High resolution two-dimensional electrophoresis of proteins. J Biol Chem 250:4007–4021
O'Farrell PZ, Goodman HM, O'Farrell PH (1977) High resolution two-dimensional electrophoresis of basic as well as acidic proteins. Cell 12:1133–1142
Ogaki C (1958) The sterility-factors in new varieties of the Japanese pear (Pyrus serotina Rehd. var. culta Rehd.). Res Rep Kanagawa Agric Inst Hort 5:23–26 (in Japanese)
Ramagli LS, Rodriguez LV (1985) Quantitation of microgram amounts of protein in two-dimensional polyacrylamide gel electrophoresis sample buffer. Electrophoresis 6:559–563
Rubin RW, Warren RW (1977) Quantitation of microgram amounts of protein in SDS-mercaptoethanol-Tris electrophoresis sample buffer. Anal Biochem 83:773–777
Sassa H, Hirano H, Ikehashi H (1992) Self-incompatibility-related RNases in styles of Japanese pear (Pyrus serotina Rehd.). Plant Cell Physiol 33:811–814
Sato Y, Kurihara A, Abe K, Ogata T, Kajiura I, Kotobuki K, Machida Y (1988) Studies on the mode of inheritance of self-compatibility in Japanese pear. Abstracts of the Japanese Society of Horticultural Science, Autumn Meeting, pp 76–77 (in Japanese)
Sato Y, Abe K, Saito T, Kotobuki K (1992) Selection of S-gene homozygotes and its utilization for S-genotype analysis of “Osa-Nijisseiki”. Abstracts of the Japanese Society of Horticultural Science Autumn Meeting, pp 16–17 (in Japanese)
Terami H, Torikawa H, Shimazu Y (1946) Analysis of the sterility factors existing in varieties of the Japanese pear (Pyrus serotina Rehd.). Studies Hort Inst Kyoto Imp Univ 3:267–271 (in Japanese)
Tsai D-S, Lee H-S, Post LC, Kreiling KM, Kao TH (1992) Sequence of an S-protein of Lycopersicon peruvianum and comparison with other solanaceous S-proteins. Sex Plant Reprod 5:256–263
Umbach AL, Lalonde BA, Kandasamy MK, Nasrallah JB, Nasrallah ME (1990) Immunodetection of protein glycoforms encoded by two independent genes of the self-incompatibility multigene family of Brassica. Plant Physiol 93:739–747
Xu B, Mu J, Nevins DL, Grun P, Kao T-H (1990) Cloning and sequencing of cDNAs encoding two self-incompatibility associated proteins in Solanum chacoense. Mol Gen Genet 224:341–346
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Sassa, H., Hirano, H. & Ikehashi, H. Identification and characterization of stylar glycoproteins associated with self-incompatibility genes of Japanese pear, Pyrus serotina Rehd. Molec. Gen. Genet. 241, 17–25 (1993). https://doi.org/10.1007/BF00280196
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DOI: https://doi.org/10.1007/BF00280196