ヒルガオ科における胞子体型自家不和合性遺伝子座の分子的解析

• Project/Area Number: 11238203
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Mie University
• Principal Investigator: 神山 康夫 三重大学, 生物資源学部, 教授 (80024579)
• Co-Investigator(Kenkyū-buntansha): 土屋 亨 三重大学, 生命科学研究支援センター, 助教授 (30293806) ; 掛田 克行 三重大学, 生物資源学部, 助教授 (50221867)
• Project Period (FY): 1999 – 2003
• Project Status: Completed (Fiscal Year 2003)
• Budget Amount: ¥73,200,000 (Direct Cost: ¥73,200,000); Fiscal Year 2003: ¥10,200,000 (Direct Cost: ¥10,200,000); Fiscal Year 2002: ¥12,800,000 (Direct Cost: ¥12,800,000); Fiscal Year 2001: ¥13,900,000 (Direct Cost: ¥13,900,000); Fiscal Year 2000: ¥18,600,000 (Direct Cost: ¥18,600,000); Fiscal Year 1999: ¥17,700,000 (Direct Cost: ¥17,700,000)
• Keywords: 自家不和合性 / S遺伝子座 / ゲノム解析 / 遺伝子発現解析 / 分子遺伝学 / サツマイモ野生種 / BACライブラリー / 遺伝子地図 / ヒルガオ科植物 / AFLP分析

Research Abstract

ヒルガオ科植物の胞子体型自家不和合性は,1遺伝子座(S遺伝子座)の複対立遺伝子によって支配されている.本研究は,ヒルガオ科のサツマイモ野生種における自家不和合性の分子的機構を明らかにすることを目的として,昨年度までに,(1)DNAマーカーを用いたS遺伝子座周辺の連鎖地図の構築,(2)マップベースクローニング法によるBACとCosmidライブラリーのスクリーニング,(3)S遺伝子座領域をカバーする約300kbpのコンティグ・クローンの単離同定,(4)ショットガン法によるこの約300kbpの塩基配列の解析とORFの推定,というステップを経て研究を進めてきた.今年度は,このゲノム領域において柱頭や葯・花粉で特異的に発現している遺伝子を明らかにするため,ノーザンブロット法による遺伝子の発現解析を行った.その結果,柱頭特異的発現遺伝子が5個(SE-1〜5),葯・花粉特異的発現遺伝子が4個(AB-1〜4)検出され,さらに葯と柱頭で高い発現を示す遺伝子(AS-1),および葯と花弁で高い発現を示す遺伝子(AP-1)も見出された.これら生殖器官特異的な遺伝子について,ゲノムDNAのサザン解析を行った結果,柱頭特異的なSE-1〜5および葯特異的なAB-2〜4遺伝子はシングルコピーでゲノム中に存在すると推定された.次に,これら生殖器官特異的遺伝子に対応するcDNAクローンのスクリーニングを行い,それらの塩基配列と推定アミノ酸配列を明らかにした.データベース検索の結果,これら生殖器官特異的遺伝子については,相同性の高い遺伝子や機能の明らかなドメイン等が見出されていないことから,新規の遺伝子であると推定された.このことは,ヒルガオ科の自家不和合性には,他の植物とは異なりユニークな分子機構が関与していることを示唆している.

Research Products

• [Publications] Tomita, R.N.: "Genetic mapping of AFLP/AMF-derived DNA markers in the vicinity of the self-incompatibility locus in Ipomoea trifida."Sexual Plant Reproduction. 16. 265-272 (2004)
• [Publications] Tomita, R.N.: "Molecular characterization of a 313-kb genomic region containing the self-incompatibility locus of Ipomoea trifida, a diploid ralative of sweet potato."Breeding Science. 54(印刷中). (2004)
• [Publications] Kowyama.Y.: "Map-based cloning of S-locus genes in a wild diploid species of sweet potato, Ipomoea trifida."Abstract of the 7th International Congress of Plant Molecular Biology. 434 (2003)
• [Publications] O'Brien, M.: "The Solanum chacoense HT modifier of gametophytic self-incompatibility works through delayed abscission of the floral organs."Abstract of the 7th International Congress of Plant Molecular Biology. 435 (2003)
• [Publications] Kakeda, K.: "Two-locus self-incompatibility in wild barley, Hordeum bulbosum."Abstract of International Symposium on Plant Self-incompatibility. 17 (2003)
• [Publications] Tsuchiya, T.: "Molecular analysis of the S-locus region of Ipomoea trifida."Abstract of International Symposium on Plant Self-incompatibility. 19 (2003)
• [Publications] 日向康吉: "植物育種学各論"文永堂. 330 (2003)
• [Publications] Kondo, K.: "Cultivated tomato has defects in both S-RNase and HT genes required for stylar function of self-incompatibility"The Plant Journal. 29(5). 627-636 (2002)
• [Publications] Kondo, K.: "Insights into the evolution of self-compatibility in Lycopersicon from a study of stylar factors"The Plant Journal. 30(2). 143-153 (2002)
• [Publications] O'Brien, M.: "Molecular analysis of the stylar-expressed Solanum chachoense small asparagine-rich protein family related to the HT modifier"The Plant Journal. 32(6). 985-996 (2002)
• [Publications] Kowyama, Y.: "Evolutionary aspect of self-incompatibility in the genus Lycopersicon"Abstract of 18th International Congress on Sexual Plant Reproduction. 81 (2002)
• [Publications] Tsuchiya, T.: "Sexual reproductive organ-specific genes : Surveying with cDNA microarray"Abstract of the 8th International Congress of Ecology. 58 (2002)
• [Publications] 掛田克行: "Agrobacteriumによるサツマイモ野生二倍体種の形質転換"育種学研究. 4(1)(印刷中). (2003)
• [Publications] Kowyama Y.: "Self-incompatibility reactions and map-based cloning of self-incompatibility locus in a wild sweet potato, Ipomoea trifida"Abstract of 17th international conrerence on Plant Growth Substances. 160 (2001)
• [Publications] Tomita, R.N.: "サツマイモ野生種におけるS遺伝子座周辺のBACクローンのスクリーニングと解析"育種学研究. 3(1). 213 (2001)
• [Publications] 掛田克行: "サツマイモ野生種からのARC1(Arm Repeat Containing)相同遺伝子の単離と解析"育種学研究. 3(1). 212 (2001)
• [Publications] 渡瀬智子: "サツマイモ野生種の自家不和合性遺伝子座近傍に座乗する遺伝子の探索"育種学研究. 3(2). 198 (2001)
• [Publications] Kondo, K.: "Cultivated tomato has defects in both S-RNase and HT genes required for stylar function or self-compatibility"The Plant Journal. 29(5)(印刷中). (2002)
• [Publications] Kondo, K.: "Insights into the evolution of self-compatibility in Lycopersicon from a study of stylar Factors"The Plant Journal. 29(5)(印刷中). (2002)
• [Publications] Kowyama,Y.: "Sporophytic self-incompatibility in Ipomoea trifida, a close relative of sweet potato."Annals of Botany. 85. 191-196 (2000)
• [Publications] Kakeda,K.: "A self-compatible mutant S allele conferring a dominant negative effect on the functional S alelle in Ipomoea trifida."Sexual Plant Reproduction. 13. 119-125 (2000)
• [Publications] Kakeda,K.: "Molecular genetic analysis of self-incompatibility in Hordeum bulbosum."Barley Genetics. 8. 157-159 (2000)
• [Publications] Takei,N.: "Isolation and expression of a pistil-specific chitinase gene in rice (Oryza sativa L.)"Breeding Science. 50. 225-228 (2000)
• [Publications] Guerin,J.: "A DEFICIENS homologue is down regulated during apomictic initiation in ovules of Hieracium."Planta. 210. 914-920 (2000)
• [Publications] Kowyama,Y.: "Self-incompatibility and reproductive-organ specific gene products in a wild diploid species of the sweet potato."Proceeding of 12th symposium of the International Society for Tropical Root Crops. (in press). (2000)
• [Publications] Jordan, N. D.: "S-Protein mutants indicate a functional role for SBP in the self-incompatibility reaction of Papaver rhoeas"Plant Journal. 20. 119-125 (1999)
• [Publications] Tomimoto, Y.: "A pistil-specific PR-1 like protein of Camellia, its expression sequence and genealogical position"Breeding Science. 49. 97-104 (1999)
• [Publications] Hashizume, F.: "Efficient Agrobacterium-mediated transformnation and the usefulness of a synthetic GFP reporter gene in Japonica rice"Plant Biotechnology. 16. 397-401 (1999)
• [Publications] Hobo, T.: "ACGT-containing abscisic acid response element(ABRE) and coupling element 3(CE3) are functionally equivalent"Plant Journal. 19. 679-689 (1999)
• [Publications] Hobo, T.: "A bZIP factor, TRAB1, interacts with VP1 and mediates abscisic acid-induced transcription"Proceeding of National Academy of Sciences, USA. 96. 15348-15353 (1999)
• [Publications] Kowyama, Y.: "Sporophytic self-incompatibility of Ipomoea trifida, a close relative to sweet potapo"Annals of Botany. (in press). (1999)
• [Publications] 神山康夫: "生物の生殖と遺伝的多様性,IGEシリーズ25(分担執筆)"東北大学遺伝生態研究センター. 131 (1999)