S複合遺伝子座の構造と葯発現遺伝子のS特異性の解析

• Project/Area Number: 11238202
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Tohoku University
• Principal Investigator: 西尾 剛 東北大学, 大学院・農学研究科, 教授 (30301039)
• Co-Investigator(Kenkyū-buntansha): 鳥山 欽哉 東北大学, 大学院・農学研究科, 助教授 (20183882)
• Project Period (FY): 1999 – 2003
• Project Status: Completed (Fiscal Year 2003)
• Budget Amount: ¥58,300,000 (Direct Cost: ¥58,300,000); Fiscal Year 2003: ¥9,100,000 (Direct Cost: ¥9,100,000); Fiscal Year 2002: ¥9,100,000 (Direct Cost: ¥9,100,000); Fiscal Year 2001: ¥10,700,000 (Direct Cost: ¥10,700,000); Fiscal Year 2000: ¥13,200,000 (Direct Cost: ¥13,200,000); Fiscal Year 1999: ¥16,200,000 (Direct Cost: ¥16,200,000)
• Keywords: 自家不和合性 / アブラナ科野菜 / 生殖 / 自己・非自己認識 / 花粉 / 柱頭 / S遺伝子 / 進化 / Sハプロタイプ / 花粉側認識分子 / 塩基配列 / 多様性 / 種分化 / 認識特異性 / 不和合性強度 / ウブラナ科植物 / S複合遺伝子座 / 塩基配列変異

Research Abstract

R.sativusの10種類のSP11と9種類のSRKの塩基配列を決定した。R.sativusのS-21はB.rapaのS-9とSP11が極めて類似し、SRKも推定アミノ酸配列で91%の相同性を示した。これらSハプロタイプは属分化前は同一のSハプロタイプであることが示唆された。
SP11タンパク質において、B.oleraceaとB.rapa同一認識特異性をもつSハプロタイプ間で異なるアミノ酸が少ないRegion IIIとRegion Vは認識特異性に重要と考え、その領域を類似SP11間で組み換えた変異SP11遺伝子を導入した形質転換体を作成し、認識特異性の変化を検定中である。
Brassica napusの内、最も多くの品種が有するBnS-1とBnS-13についてSP11とSRKのゲノムクローンを単離し、全長の塩基配列を決定したところ、BnS-1はB.rapaのS-47と極めて類似しており、SRKは正常に機能していることを見出した。また、BnS-13もB.oleraceaのS-15に酷似し、このSRKも正常に機能していることを明らかにした。
種分化前は同一のSハプロタイプであったと推定されるB.rapaのS-47とB.oleraceaのS-12のS遺伝子座の塩基配列を決定し、B.oleraceaのS-12のS遺伝子座の方がB.rapaのS-47よりも大きいことを見出し、劣性グループのクラスIIハプロタイプであるB.oleraceaのS-15とB.rapaのS-60の比較においてもB.oleraceaの方がS遺伝子座が大きいことを明らかにした。B.oleraceaのS遺伝子座が大きい原因としてレトロトランスポゾンの挿入が大きく関わっていることが分かった。S遺伝子座に存在するレトロトランスポゾン様配列は、B.rapaよりもB.oleraceaのゲノム中により多いコピー数で検出された。

Research Products

• [Publications] Fukai E, Fujimoto R, Nishio T: "Genomic organization of the S core region and the S flanking regions of a class-I S haplotype in Brassica rapa."Mol Gen Genomics. 269. 361-369 (2003)
• [Publications] Matsui K, Tetsuka T, Nishio T, Hara T: "Heteromorphic incompatibility retained in self compatible plants produced by a cross between common and wild buckwheat."New Phytologist. 159. 701-708 (2003)
• [Publications] Sato Y, Fujimoto R, Toriyama K, Nishio T: "Commonalty of self recognition specificity of S haplotypes between Brassica oleracea and Brassica rapa."Plant Mol Biol. 52. 617-626 (2003)
• [Publications] Fujimoto R, Nishio T: "Identification of S haplotypes in Brassica by dot blot analysis of SP11 alleles."Theor Appl Genet. 106. 1433-1437 (2003)
• [Publications] Okamoto S, Sato Y, Sakamoto K, Nishio T: "Distribution of similar self-incompatibility (S) haplotypes in different genera, Raphanus and Brassica."Sex Plant Reprod. (in press).
• [Publications] Sato Y, Fujimoto R, Toriyama K, Nishio T: "Commonality of self-recognition specificity of S haplotypes between Bracssica oleracea and Brassica rapa"Plant Mol.Biol. (In press). (2003)
• [Publications] Fujimoto R, Nishio T: "Identification of S haplotypes in Brassica by dot blot analysis of SP11 alleles"Theor Appl Genet. (In press). (2003)
• [Publications] Inaba R, Nishio T: "Phylogenetic analysis of Brassiceae based on the nucleotide sequences of the S-locus related gene, SLR1"Theor Appl Genet. 105. 1159-1165 (2002)
• [Publications] Sato K, Nishlo T, Kimura R, Kusaba M, Suzuki T, Hatakeyama K, Ockendon DJ, Satta Y: "Coevolution of the S-locus genes SRK, SLG, and SP11/SCR in Brassica oleracea and B.rapa"Genetics. 162. 931-940 (2002)
• [Publications] Sato K, Nishlo T: "Efficient detection of DNA polymorphism in cabbage and rice cultivars by PCR-RF-SSCP (PRS)"Plant Cell Rep. 21. 276-281 (2002)
• [Publications] Matsushita Y, Suzuki T.Mori M, Shimosato H, Kuota R, Watanabe M, Kayano T, Nishio T, Nyunoya H: "Isolation of a cDNA for a nucleoside-diphosphate kinase capable of phosphorylating the kinase domain of the self-incompatibility factor SRK of Brassica campestris"J.Exp.Bot.. 53. 765-767 (2002)
• [Publications] Kimura R, Sato K, Fujimoto R, Nishio T: "Recognition specificity of self-incompatibility maintained after the divergence of Brassica oleracea and Brassica rapa"Plant J.. 29. 215-223 (2002)
• [Publications] Matsushita Y, Suzuki T, Mori M, Shimosato H, Kubota R, Watanabe M, Kayano T, Nishio T, Nyunoya H: "Isolation of a cDNA for a nucleoside-diphosphate kinase capable of phosphorylating the kinase domain of the self-incompatibility factor SRK of Brassica campestris."J. Exp. Bot.. (In press). (2002)
• [Publications] Fukai E, Nishio T, Nasrallah ME: "Molecular genetic analysis of the candidate gene for MOD, a locus required for self-incompatibility in Brassica rapa"Mol. Genet. Genomics. 265. 519-525 (2001)
• [Publications] Hatakeyama K, Takasaki T, Suzuki G, Nishio T, Watanabe M, Isogai A, Hinata K: "The S receptor kinase gene determines dominance relationships in stigma expression of self-incompatibility in Brassica"Plant J.. 26. 155-161 (2001)
• [Publications] Sakamoto K, Nishio T: "Distribution of S haplotypes in commercial cultivars of Brassica rapa"Plant Breeding. 120. 155-161 (2001)
• [Publications] Takasaki T, Hatakeyama K, Watanabe M, Toriyama K, Hinata K: "Homology-dependent suppression of stigma phenotype by an antisense S-locus glycoprotein (SLG) gene in Brassica rapa L"Breeding Science. 51. 89-94 (2001)
• [Publications] Okada T, Toriyama K: "Pollen vegetative cell-specific expression of Bra r 1 : useful tool for observation of the vegetative nucleus and identification of transgenic pollen by nuclear-targeted GFP"Sex Plant Reprod. 13. 301-307 (2001)
• [Publications] Katashiba H, Ota R, Toriyama K: "Analyses of the genomic sequence and promoter activity of a gene for a protein similar to Tat binding protein isolated from Brassica rapa"Plant Biotech. 18. 33-38 (2001)
• [Publications] Suzuki T,Kusaba M,Matsushita M,Okazaki K,Nishio T: "Characterization of Brassica S-haplotypes lacking S-locus glycoprotein."FEBS Letters. 482. 102-108 (2000)
• [Publications] Sakamoto K,Kusaba M,Nishio T: "Single seed PCR-RFLP analysis for identification of S haplotypes in commercial F1 hybrid cultivars of broccoli and cabbage."Plant Cell Rep.. 19. 400-406 (2000)
• [Publications] Suzuki G,Watanabe M,Nishio T: "Physical distances between S-locus genes in various S haplotypes of Brassica rapa and B.oleracea."Theor.Appl.Genet.. 101. 80-85 (2000)
• [Publications] Nishio T,Kusaba K: "Sequence diversity of SLG and SRK in Brassica oleracea L."Ann.Bot.. 85 Sup.A. 141-146 (2000)
• [Publications] Fukai E,Nishio T,Nasrallah M E: "Molecular genetic analysis of the candidate gene for MOD, a locus required for self-incompatibility in Brassica rapa."Mol.Gen.Genet.. (In press).
• [Publications] Okada T,Sasaki Y,Ohta R,Onozuka N,Toriyama K: "Expression of Bra r 1 gene in transgenic tobacco and Bra r 1 promoter activity in pollen of vari ous plant species."Plant Cell Physiol.. 41. 757-766 (2000)
• [Publications] Toriyama K,Tanabe M,Rashid H: "Biotechnology in Agriculture and Forestry 48 Transgenic Crops III"Springer. 352-358 (2001)
• [Publications] Kusaba M.,Matsushita M.,Okazaki K.,Satta Y.,Nishio T.: "Sequence and structural diversity of the S locus genes from different lines with the same self-recongnition specificities in Brassica oleracea"Genetics. 154. 413-420 (2000)
• [Publications] Nishio T.,Kusaba K.: "Sequence diversity of SLG and SRK in Brassica oleracea L."Ann.Bot.. (in press). (2000)
• [Publications] Suzuki G.,Watanabe M.,Nishio T.: "Physical distances between S-locus genes in various S haplotypes of Brassica rapa and B.oleracea"Theor.Appl.Genet.. (in press). (2000)
• [Publications] Sakamoto K.,Kusaba M.,Nishio T.: "Single seed PCR-RFLP analysis for identification of S haplotypes in commercial F,hybrid cultivars of broccoli and cabbage"Plant Cell Rep.. (in press). (2000)
• [Publications] Okazaki K.,Kusaba,M.,Ockendon DJ,Nishio T.: "Characterization of S tester lines in Brassica oleracea : polymorphism of restriction fragment length of SLG homologues and isoelectric points of S-locus glycoproteins"Theor.Appl.Genet.. 98. 1329-1334 (1999)
• [Publications] Suzuki G.,Kai N.,Hirose T.,Fukui K.,Nishio T.,Takayama S.,Isogai A.,Watanabe M.,Hinata K.: "Genomic organization of the S locus : Identification and characterization of genes in SLG/SRK region of S9 haplotype of Brassica campestris(syn.rapa)"Genetics. 153. 391-400 (1999)
• [Publications] Kusaba M.,Nishio T.: "Plant Biotech."1)The molecular mechanism of self-recongnition in Brassica self-incompatibility. 10 (1999)