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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

TIP3 genes are specifically expressed during seed maturation. (A and B) Expression analysis of TIP3;1 and TIP3;2 during seed development (A) and seed germination (B) in Arabidopsis. qRT-PCR analysis of TIP3;1 and TIP3;2 transcript abundance during seed development and seed germination. The relative expression level of each gene was normalized with four reference genes, and calculated according to the geNorm 3.5 manual. Values are means ±SD, n=3. DPA, days post-anthesis. (C and D) Immunoblot analysis of TIP3s during seed development (C) and seed germination (D). The same amounts of proteins separated by SDS–PAGE were stained with Coomassie Brilliant Blue and used as a loading control.
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Figure 1: TIP3 genes are specifically expressed during seed maturation. (A and B) Expression analysis of TIP3;1 and TIP3;2 during seed development (A) and seed germination (B) in Arabidopsis. qRT-PCR analysis of TIP3;1 and TIP3;2 transcript abundance during seed development and seed germination. The relative expression level of each gene was normalized with four reference genes, and calculated according to the geNorm 3.5 manual. Values are means ±SD, n=3. DPA, days post-anthesis. (C and D) Immunoblot analysis of TIP3s during seed development (C) and seed germination (D). The same amounts of proteins separated by SDS–PAGE were stained with Coomassie Brilliant Blue and used as a loading control.

Mentions: The Arabidopsis genome contains two TIP3 genes, namely TIP3;1 and TIP3;2. Using qRT-PCR, the temporal expression patterns of TIP3 genes were investigated in a precise manner. Transcripts of TIP3;1 and TIP3;2 began to be detectable in siliques at 12 days post-anthesis (DPA) (Fig. 1A). TIP3 transcript levels increased sharply throughout the maturation phase. Immunoblot analysis also indicated that TIP3s began to accumulate at the same time point (Fig. 1C). Since the antibody raised against the C-terminal peptide of TIP3;1 cannot discriminate between TIP3;1 and TP3;2 (Supplementary Fig. S1 at JXB online), the detected signals represented both TIP3 isoforms. In germinating seeds, the levels of TIP3 transcripts decreased to <1% during the first 3h after germination (Fig. 1B). Interestingly, the protein levels of TIP3s did not decrease significantly within 24h, but they started to decrease sharply 48h after germination (Fig. 1D). At the same time, TIP1s (TIP1;1 and TIP1;2) began to be detectable (Fig. 1D). Fluorescent signals were detected in the seeds of ProTIP3;1:GFP transgenic plants, but not in WT seeds (Supplementary Fig. S2). These data suggest that TIP3;1 and TIP3;2 are specifically expressed in seeds, and the TIP3;1 promoter is active in seeds.


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)

TIP3 genes are specifically expressed during seed maturation. (A and B) Expression analysis of TIP3;1 and TIP3;2 during seed development (A) and seed germination (B) in Arabidopsis. qRT-PCR analysis of TIP3;1 and TIP3;2 transcript abundance during seed development and seed germination. The relative expression level of each gene was normalized with four reference genes, and calculated according to the geNorm 3.5 manual. Values are means ±SD, n=3. DPA, days post-anthesis. (C and D) Immunoblot analysis of TIP3s during seed development (C) and seed germination (D). The same amounts of proteins separated by SDS–PAGE were stained with Coomassie Brilliant Blue and used as a loading control.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

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Figure 1: TIP3 genes are specifically expressed during seed maturation. (A and B) Expression analysis of TIP3;1 and TIP3;2 during seed development (A) and seed germination (B) in Arabidopsis. qRT-PCR analysis of TIP3;1 and TIP3;2 transcript abundance during seed development and seed germination. The relative expression level of each gene was normalized with four reference genes, and calculated according to the geNorm 3.5 manual. Values are means ±SD, n=3. DPA, days post-anthesis. (C and D) Immunoblot analysis of TIP3s during seed development (C) and seed germination (D). The same amounts of proteins separated by SDS–PAGE were stained with Coomassie Brilliant Blue and used as a loading control.
Mentions: The Arabidopsis genome contains two TIP3 genes, namely TIP3;1 and TIP3;2. Using qRT-PCR, the temporal expression patterns of TIP3 genes were investigated in a precise manner. Transcripts of TIP3;1 and TIP3;2 began to be detectable in siliques at 12 days post-anthesis (DPA) (Fig. 1A). TIP3 transcript levels increased sharply throughout the maturation phase. Immunoblot analysis also indicated that TIP3s began to accumulate at the same time point (Fig. 1C). Since the antibody raised against the C-terminal peptide of TIP3;1 cannot discriminate between TIP3;1 and TP3;2 (Supplementary Fig. S1 at JXB online), the detected signals represented both TIP3 isoforms. In germinating seeds, the levels of TIP3 transcripts decreased to <1% during the first 3h after germination (Fig. 1B). Interestingly, the protein levels of TIP3s did not decrease significantly within 24h, but they started to decrease sharply 48h after germination (Fig. 1D). At the same time, TIP1s (TIP1;1 and TIP1;2) began to be detectable (Fig. 1D). Fluorescent signals were detected in the seeds of ProTIP3;1:GFP transgenic plants, but not in WT seeds (Supplementary Fig. S2). These data suggest that TIP3;1 and TIP3;2 are specifically expressed in seeds, and the TIP3;1 promoter is active in seeds.

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