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Title: Transcriptome Analysis of Self- and Cross-pollinated Pistils of Japanese Apricot (Prunus mume Sieb. et Zucc.)
Other Titles: ウメにおける自家および他家受粉雌ずいのトランスクリプトーム解析
Authors: Habu, Tsuyoshi
Tao, Ryutaro  kyouindb  KAKEN_id
Author's alias: 羽生, 剛
田尾, 龍太郎
Keywords: EST
gametophytic self-incompatibility
next-generation sequencing technology
RNA-Seq
S-RNase
Issue Date: 2014
Publisher: Japanese Society for Horticultural Science(園芸学会)
Journal title: Journal of the Japanese Society for Horticultural Science
Volume: 83
Issue: 2
Start page: 95
End page: 107
Abstract: Solanaceae, Rosaceae, and Plantaginaceae exhibit the S-RNase-based gametophytic self-incompatibility (GSI) system. This type of GSI is controlled by a single polymorphic locus (S locus) containing the pistil S determinant gene, S-ribonuclease (S-RNase), and the pollen S determinant, the S locus F-box gene (SFB/SLF). In addition to these determinant genes, non-S factors, called modifier genes, are required for the GSI reaction. Here, we conducted large-scale transcriptome analysis of unpollinated, self-pollinated, and cross-pollinated pistils of Japanese apricot (Prunus mume Sieb. et Zucc. cv. Nanko) to capture all of the molecular events induced by the GSI reaction in Prunus, using next-generation sequencing technologies. We obtained 40,061 unigenes from 77,521,310 reads from pollinated and unpollinated pistils and pollen grains. Among these unigenes, 29,985 and 27,898 unigene sequences showed at least one hit against the NCBI nr and TAIR10 protein databases, respectively, in BLASTX searches using an E-value cutoff of 1e-6. Digital expression analysis showed that 8,907 and 10,190 unigenes were expressed at significantly different levels between unpollinated (UP) and cross-pollinated (CP) pistils and between UP and self-pollinated (SP) pistils, respectively. The expression of 4,348 unigenes in both CP and SP pollination was commonly and significantly different from that in UP, while the expression of 4,559 and 5,842 unigenes in CP and SP, respectively, was specifically and significantly different from UP. The expression of 2,227 unigenes was up-regulated both in CP and SP compared with UP. Genes supposedly involved in S-RNase-based GSI were included among the unigenes up-regulated by pollination, while no unigenes homologous to the solanaceous pistil modifiers HT-B or 120K were included among the unigenes up-regulated by pollination or in the whole unpollinated/pollinated pistil transcriptome. We discuss the distinct molecular mechanism of S-RNase-based GSI in Prunus.
ナス科, バラ科, オオバコ科の多くの種は S-RNase 依存性配偶体型自家不和合性を示す. このタイプの不和合性は雌ずい側因子 (S-RNase)と花粉側因子 (SFB/SLF)が座乗する S 遺伝子座によって支配されている. また, 自家不和合性共通因子と呼ばれる S 因子以外の因子も配偶体型自家不和合性反応に必要であることが明らかになっている. そこで本研究では, ウメの配偶体型自家不和合性機構解明を目的として, 次世代シークエンス技術を用いた大規模トランスクリプトーム解析を行った. ウメの受粉雌ずい, 未受粉雌ずい, 花粉から合計 77,521,310 個の配列が得られ, それらから 40,061 個の unigene が得られた. これら 40,061 個の unigene を query に, 米国の国立生物工学情報センター (NCBI)の非冗長タンパク質データベース (nr)ならびにシロイヌナズナのタンパク質データベース (TAIR10)に対して, BLASTX 検索したところ, 29,985 個 (全体の 74.8%) の unigene が NCBI の少なくとも1 つ以上のデータに, また 27,898 個 (全体の 69.9%)の unigene が TAIR10 の少なくとも1 つ以上のデータにヒット (E-value cutoff 値 1e-6)した. デジタル発現解析により, 8,907の unigene が未受粉と他家受粉で有意に発現が異なることが, また 10,190 個の unigene が未受粉と自家受粉で有意に発現が異なることが示された. これらのうち, 4,348 個の unigene は両受粉で共通であり, 4,559 個の unigene は他家受粉で特異的に, また 5,842 個の unigene は自家受粉で特異的に発現が変化していた. さらに, 両受粉で発現が変化していた unigene のうち, 2,227 個は両受粉で発現が増加していた. しかし, ナス科で花柱側共通因子であることが示されている HT-B や 120K の相同遺伝子はトランスクリプトーム全体の中に存在しなかった. Prunus 属の配偶体型自家不和合性に特異な認識機構について考察した.
Rights: © 2014 by Japanese Society for Horticultural Science
URI: http://hdl.handle.net/2433/191088
DOI(Published Version): 10.2503/jjshs1.CH-086
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