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Arabidopsis seed-specific vacuolar aquaporins are involved in maintaining seed longevity under the control of ABSCISIC ACID INSENSITIVE 3.

Mao Z, Sun W - J. Exp. Bot. (2015)

Bottom Line: TIP3;1 and TIP3;2 promoters could be activated by ABI3 in the presence of abscisic acid (ABA) in Arabidopsis protoplasts.TIP3 proteins were detected in the protoplasts transiently expressing ABI3 and in ABI3-overexpressing seedlings when treated with ABA.Furthermore, ABI3 directly binds to the RY motif of the TIP3 promoters.

View Article: PubMed Central - PubMed

Affiliation: Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Fenglin Road 300, Shanghai, 200032, People's Republic of China.

No MeSH data available.


Related in: MedlinePlus

Identification of tip3;1 and tip3;2 T-DNA insertion mutants and three TIP3;1-RNAi transgenic lines (TIP3;1-RNAi/tip3;2) in the tip3;2 mutant background. (A) Schematic representation of the tip3;1 and tip3;2 T-DNA insertion mutant lines. A triangle indicates the position of the T-DNA insertion, and the arrow indicates its orientation. The genomic sequences corresponding to the coding region (black boxes), untranslated region (grey boxes), and introns (black lines) are indicated. The positions of the primers (31LP, 31RP, 32LP, and 32RP) used for PCR analysis of the tip3;1 and tip3;2 T-DNA insertion mutants, respectively, are also indicated. (B) PCR analysis of genomic DNA of Col, tip3;1, tip3;2, and tip3;1/tip3;2. LP, left primer; RP, right primer; LB, T-DNA left border primer. (C) Schematic representation of the construct used for the suppression of TIP3;1 in Arabidopsis seeds. RNAi technology was used with a segment of the TIP3;1 gene driven by the seed-specific TIP3;1 promoter. (D) qRT-PCR analysis of TIP3;1, TIP3;2, and ACT7 transcript abundance in mature seeds of Col, mutants, and RNAi lines. PP2A was used as the endogenous control, and the transcript abundance of TIP3;1, TIP3;2, and ACT7 was quantified by comparisons with that of PP2A. Values are means ±SD, n=3. (E) Immunoblot analysis of TIP3s in mature seeds from Col, tip3 mutants, and three TIP3;1-RNAi/tip3;2 transgenic lines (R3, R7, and R8). HSP17.6, which is expressed in mature seeds, was used as a loading control. (This figure is available in colour at JXB online.)
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Figure 2: Identification of tip3;1 and tip3;2 T-DNA insertion mutants and three TIP3;1-RNAi transgenic lines (TIP3;1-RNAi/tip3;2) in the tip3;2 mutant background. (A) Schematic representation of the tip3;1 and tip3;2 T-DNA insertion mutant lines. A triangle indicates the position of the T-DNA insertion, and the arrow indicates its orientation. The genomic sequences corresponding to the coding region (black boxes), untranslated region (grey boxes), and introns (black lines) are indicated. The positions of the primers (31LP, 31RP, 32LP, and 32RP) used for PCR analysis of the tip3;1 and tip3;2 T-DNA insertion mutants, respectively, are also indicated. (B) PCR analysis of genomic DNA of Col, tip3;1, tip3;2, and tip3;1/tip3;2. LP, left primer; RP, right primer; LB, T-DNA left border primer. (C) Schematic representation of the construct used for the suppression of TIP3;1 in Arabidopsis seeds. RNAi technology was used with a segment of the TIP3;1 gene driven by the seed-specific TIP3;1 promoter. (D) qRT-PCR analysis of TIP3;1, TIP3;2, and ACT7 transcript abundance in mature seeds of Col, mutants, and RNAi lines. PP2A was used as the endogenous control, and the transcript abundance of TIP3;1, TIP3;2, and ACT7 was quantified by comparisons with that of PP2A. Values are means ±SD, n=3. (E) Immunoblot analysis of TIP3s in mature seeds from Col, tip3 mutants, and three TIP3;1-RNAi/tip3;2 transgenic lines (R3, R7, and R8). HSP17.6, which is expressed in mature seeds, was used as a loading control. (This figure is available in colour at JXB online.)

Mentions: To characterize the effects of loss of function of TIP3 genes, two T-DNA insertion mutants were obtained from the ABRC. PCR analysis of genomic DNA from the mutants confirmed the locations of the T-DNA insertions. SALK_053807 has an insertion in the promoter region between the RY2 and RY3 motif of TIP3;1, and SALK_125353c has an insertion in the first intron of TIP3;2 (Fig. 2A, B). The expression level of TIP3;1 in tip3;1 seeds was reduced to 30% of that in WT seeds (Fig. 2D). TIP3;2 transcripts were not detectable in tip3;2 seeds (Fig. 2D), demonstrating that the tip3;2 mutant is transcript .


Arabidopsis seed-specific vacuolar aquaporins are involved in maintaining seed longevity under the control of ABSCISIC ACID INSENSITIVE 3.

Mao Z, Sun W - J. Exp. Bot. (2015)

Identification of tip3;1 and tip3;2 T-DNA insertion mutants and three TIP3;1-RNAi transgenic lines (TIP3;1-RNAi/tip3;2) in the tip3;2 mutant background. (A) Schematic representation of the tip3;1 and tip3;2 T-DNA insertion mutant lines. A triangle indicates the position of the T-DNA insertion, and the arrow indicates its orientation. The genomic sequences corresponding to the coding region (black boxes), untranslated region (grey boxes), and introns (black lines) are indicated. The positions of the primers (31LP, 31RP, 32LP, and 32RP) used for PCR analysis of the tip3;1 and tip3;2 T-DNA insertion mutants, respectively, are also indicated. (B) PCR analysis of genomic DNA of Col, tip3;1, tip3;2, and tip3;1/tip3;2. LP, left primer; RP, right primer; LB, T-DNA left border primer. (C) Schematic representation of the construct used for the suppression of TIP3;1 in Arabidopsis seeds. RNAi technology was used with a segment of the TIP3;1 gene driven by the seed-specific TIP3;1 promoter. (D) qRT-PCR analysis of TIP3;1, TIP3;2, and ACT7 transcript abundance in mature seeds of Col, mutants, and RNAi lines. PP2A was used as the endogenous control, and the transcript abundance of TIP3;1, TIP3;2, and ACT7 was quantified by comparisons with that of PP2A. Values are means ±SD, n=3. (E) Immunoblot analysis of TIP3s in mature seeds from Col, tip3 mutants, and three TIP3;1-RNAi/tip3;2 transgenic lines (R3, R7, and R8). HSP17.6, which is expressed in mature seeds, was used as a loading control. (This figure is available in colour at JXB online.)
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Figure 2: Identification of tip3;1 and tip3;2 T-DNA insertion mutants and three TIP3;1-RNAi transgenic lines (TIP3;1-RNAi/tip3;2) in the tip3;2 mutant background. (A) Schematic representation of the tip3;1 and tip3;2 T-DNA insertion mutant lines. A triangle indicates the position of the T-DNA insertion, and the arrow indicates its orientation. The genomic sequences corresponding to the coding region (black boxes), untranslated region (grey boxes), and introns (black lines) are indicated. The positions of the primers (31LP, 31RP, 32LP, and 32RP) used for PCR analysis of the tip3;1 and tip3;2 T-DNA insertion mutants, respectively, are also indicated. (B) PCR analysis of genomic DNA of Col, tip3;1, tip3;2, and tip3;1/tip3;2. LP, left primer; RP, right primer; LB, T-DNA left border primer. (C) Schematic representation of the construct used for the suppression of TIP3;1 in Arabidopsis seeds. RNAi technology was used with a segment of the TIP3;1 gene driven by the seed-specific TIP3;1 promoter. (D) qRT-PCR analysis of TIP3;1, TIP3;2, and ACT7 transcript abundance in mature seeds of Col, mutants, and RNAi lines. PP2A was used as the endogenous control, and the transcript abundance of TIP3;1, TIP3;2, and ACT7 was quantified by comparisons with that of PP2A. Values are means ±SD, n=3. (E) Immunoblot analysis of TIP3s in mature seeds from Col, tip3 mutants, and three TIP3;1-RNAi/tip3;2 transgenic lines (R3, R7, and R8). HSP17.6, which is expressed in mature seeds, was used as a loading control. (This figure is available in colour at JXB online.)
Mentions: To characterize the effects of loss of function of TIP3 genes, two T-DNA insertion mutants were obtained from the ABRC. PCR analysis of genomic DNA from the mutants confirmed the locations of the T-DNA insertions. SALK_053807 has an insertion in the promoter region between the RY2 and RY3 motif of TIP3;1, and SALK_125353c has an insertion in the first intron of TIP3;2 (Fig. 2A, B). The expression level of TIP3;1 in tip3;1 seeds was reduced to 30% of that in WT seeds (Fig. 2D). TIP3;2 transcripts were not detectable in tip3;2 seeds (Fig. 2D), demonstrating that the tip3;2 mutant is transcript .

Bottom Line: TIP3;1 and TIP3;2 promoters could be activated by ABI3 in the presence of abscisic acid (ABA) in Arabidopsis protoplasts.TIP3 proteins were detected in the protoplasts transiently expressing ABI3 and in ABI3-overexpressing seedlings when treated with ABA.Furthermore, ABI3 directly binds to the RY motif of the TIP3 promoters.

View Article: PubMed Central - PubMed

Affiliation: Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Fenglin Road 300, Shanghai, 200032, People's Republic of China.

No MeSH data available.


Related in: MedlinePlus