Elucidation of the roles of GABA in Tomato Fruit 060727 Takashi AKIHIRO Food for specified health use （FOSHU） Permission from Ministry of Health, Labour and Welfare billion € 47.1 € 40.4 € 33.7 €27.0 € 20.3 €13.4 € 6.7 In vertebrates, GABA is an inhibitory neurotransmitter in the brain GABA Gamma-aminobutyric acid\ Fig. 1. Sequence alignment of GADs from various plants. Identical and similar amino acid residues are indicated by black and gray boxes, respectively. Gaps are shown by hyphens. the C-terminal peptide extension that is unique to plant GADs is shown by a dashed line. Table 1. Homology comparisons between tomato GADs Fig. 2. Alignment of the amino acid sequences of the C-terminus of plant GADs. OsGAD1 LeGADA OsGAD2 Petunia LeGADB Lotus LeGADF Brassica juncea Fig. 3. Structural features of C-terminal portions of GADs from tomato, rice, lotus Indian mustard and petunia. GABAT Fig. 4. Sequence alignment of GABATs from tomato and Arabidopsis. Identical and similar amino acid residues are indicated by black and gray boxes, respectively. Gaps are shown by hyphens. Table 2. Prediction of the subcellular location of GABATs using TargetP, MitoProII,PSORT and PSORTII program. SSADH Fig. 5. Sequence alignment of SSADHs from tomato and Arabidopsis. Identical and similar amino acid residues are indicated by black and gray boxes, respectively. Gaps are shown by hyphens. Table 3. Prediction of the subcellular location of SSADHs using TargetP SSR Fig. 6. Sequence alignment of SSRs from tomato and Arabidopsis. Identical and similar amino acid residues are indicated by black and gray boxes, respectively. Gaps are shown by hyphens. Table 4. Prediction of the subcellular location of SSRs using TargetP Fig. 7. Isolation of total RNAs from tomato 3.5 Leaf Stem Root Flower Trichome Columella Pericarp Loculus Seed Jelly 3 2 1.5 1 0.5 DAF45 DAF42 DAF39 DAF36 DAF33 DAF30 DAF27 DAF24 DAF21 DAF18 DAF15 DAF12 DAF9-2 DAF9-1 DAF6 DAF3 0 Control ug/ul 2.5 OK OK OK OK Failed OK OK OK OK OK Conclusion GAD GABAT SSADH SSR X3 X3 X2 X2 Isolated from MicroTom Future works 優先順位１ (8・25まで） GABAT,SSRおよびSSADHのGenome配列を決定する →TillingしてGABA代謝系の変異株をスクリーニングする 優先順位2 (8・２５まで） 果実における発現解析を行う →RNAi、Tillingのターゲット遺伝子の絞り込み 優先順位3 発現ベクターへの組み込み →単離したクローンがファンクショナルであることを確かめる 優先順位4 Isoformの数を確定する →サザン解析
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