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Overexpression of PwTUA1, a pollen-specific tubulin gene, increases pollen tube elongation by altering the distribution of alpha-tubulin and promoting vesicle transport.

Yu Y, Li Y, Li L, Lin J, Zheng C, Zhang L - J. Exp. Bot. (2009)

Bottom Line: Here, a TUA1 gene from Picea wilsonii, which is specifically expressed in pollen, was isolated.Transient expression analysis in P. wilsonii pollen indicated that PwTUA1 improved pollen germination and pollen tube growth.The possible role of PwTUA1 in microtubule dynamics and organization was discussed.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Crop Biology, College of Life Sciences, Shandong Agricultural University, Taian, Shandong 271018, PR China.

ABSTRACT
Tubulin genes are intimately associated with cell division and cell elongation, which are central to plant secondary cell wall development. However, their roles in pollen tube polar growth remain elusive. Here, a TUA1 gene from Picea wilsonii, which is specifically expressed in pollen, was isolated. Semi-quantitative RT-PCR analysis showed that the amount of PwTUA1 transcript varied at each stage of growth of the pollen tube and was induced by calcium ions and boron. Transient expression analysis in P. wilsonii pollen indicated that PwTUA1 improved pollen germination and pollen tube growth. The pollen of transgenic Arabidopsis overexpressing PwTUA1 also showed a higher percentage of germination and faster growth than wild-type plants not only in optimal germination medium, but also in medium supplemented with elevated levels of exogenous calcium ions or boron. Immunofluorescence and electron microscopy showed alpha-tubulin to be enriched and more vesicles accumulated in the apex region in germinating transgenic Arabidopsis pollen compared with wild-type plants. These results demonstrate that PwTUA1 up-regulated by calcium ions and boron contributes to pollen tube elongation by altering the distribution of alpha-tubulin and regulating the deposition of pollen cell wall components during the process of tube growth. The possible role of PwTUA1 in microtubule dynamics and organization was discussed.

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Expression of PwTUA1 in different tissues of Picea wilsonii. (A) Tissue-specific expression of PwTUA1 in P. wilsonii. Total RNA was isolated from needles (N), stems (S), roots (R), and pollen (P) (incubated for 0, 6, 12, 18, and 24 h). (B) Expression of PwTUA1 in pollen at different growth stages. (C) PwTUA1 gene expression induced by 0.1% (w/v) Ca2+ and 0.1% (w/v) H3BO3 at different time points.
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fig3: Expression of PwTUA1 in different tissues of Picea wilsonii. (A) Tissue-specific expression of PwTUA1 in P. wilsonii. Total RNA was isolated from needles (N), stems (S), roots (R), and pollen (P) (incubated for 0, 6, 12, 18, and 24 h). (B) Expression of PwTUA1 in pollen at different growth stages. (C) PwTUA1 gene expression induced by 0.1% (w/v) Ca2+ and 0.1% (w/v) H3BO3 at different time points.

Mentions: To obtain a profile of PwTUA1 expression patterns, total RNA was isolated from needles, stems, and roots, as well as from germinating P. wilsonii pollen mixtures incubated at 6 h intervals, and transcript accumulation was examined by semi-quantitative RT-PCR using gene-specific primers (Table 1). The RW primer was designed based on the 3′-UTR downstream from the stop codon, as this region was hypervariable. The results showed that PwTUA1 was expressed specifically in germinating pollen but was not detected in needles, stems, and roots (Fig. 3A). RT-PCR was next used to examine the expression of PwTUA1 during pollen development. As shown in Fig. 3B, the expression of PwTUA1 in pollen was first detected at 6 h after incubation (just germinated); it was abundant during the middle developmental stages (12, 18, and 24 h after incubation) and then continued weakly at later stages (30 h and 36 h after incubation).


Overexpression of PwTUA1, a pollen-specific tubulin gene, increases pollen tube elongation by altering the distribution of alpha-tubulin and promoting vesicle transport.

Yu Y, Li Y, Li L, Lin J, Zheng C, Zhang L - J. Exp. Bot. (2009)

Expression of PwTUA1 in different tissues of Picea wilsonii. (A) Tissue-specific expression of PwTUA1 in P. wilsonii. Total RNA was isolated from needles (N), stems (S), roots (R), and pollen (P) (incubated for 0, 6, 12, 18, and 24 h). (B) Expression of PwTUA1 in pollen at different growth stages. (C) PwTUA1 gene expression induced by 0.1% (w/v) Ca2+ and 0.1% (w/v) H3BO3 at different time points.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC2692020&req=5

fig3: Expression of PwTUA1 in different tissues of Picea wilsonii. (A) Tissue-specific expression of PwTUA1 in P. wilsonii. Total RNA was isolated from needles (N), stems (S), roots (R), and pollen (P) (incubated for 0, 6, 12, 18, and 24 h). (B) Expression of PwTUA1 in pollen at different growth stages. (C) PwTUA1 gene expression induced by 0.1% (w/v) Ca2+ and 0.1% (w/v) H3BO3 at different time points.
Mentions: To obtain a profile of PwTUA1 expression patterns, total RNA was isolated from needles, stems, and roots, as well as from germinating P. wilsonii pollen mixtures incubated at 6 h intervals, and transcript accumulation was examined by semi-quantitative RT-PCR using gene-specific primers (Table 1). The RW primer was designed based on the 3′-UTR downstream from the stop codon, as this region was hypervariable. The results showed that PwTUA1 was expressed specifically in germinating pollen but was not detected in needles, stems, and roots (Fig. 3A). RT-PCR was next used to examine the expression of PwTUA1 during pollen development. As shown in Fig. 3B, the expression of PwTUA1 in pollen was first detected at 6 h after incubation (just germinated); it was abundant during the middle developmental stages (12, 18, and 24 h after incubation) and then continued weakly at later stages (30 h and 36 h after incubation).

Bottom Line: Here, a TUA1 gene from Picea wilsonii, which is specifically expressed in pollen, was isolated.Transient expression analysis in P. wilsonii pollen indicated that PwTUA1 improved pollen germination and pollen tube growth.The possible role of PwTUA1 in microtubule dynamics and organization was discussed.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Crop Biology, College of Life Sciences, Shandong Agricultural University, Taian, Shandong 271018, PR China.

ABSTRACT
Tubulin genes are intimately associated with cell division and cell elongation, which are central to plant secondary cell wall development. However, their roles in pollen tube polar growth remain elusive. Here, a TUA1 gene from Picea wilsonii, which is specifically expressed in pollen, was isolated. Semi-quantitative RT-PCR analysis showed that the amount of PwTUA1 transcript varied at each stage of growth of the pollen tube and was induced by calcium ions and boron. Transient expression analysis in P. wilsonii pollen indicated that PwTUA1 improved pollen germination and pollen tube growth. The pollen of transgenic Arabidopsis overexpressing PwTUA1 also showed a higher percentage of germination and faster growth than wild-type plants not only in optimal germination medium, but also in medium supplemented with elevated levels of exogenous calcium ions or boron. Immunofluorescence and electron microscopy showed alpha-tubulin to be enriched and more vesicles accumulated in the apex region in germinating transgenic Arabidopsis pollen compared with wild-type plants. These results demonstrate that PwTUA1 up-regulated by calcium ions and boron contributes to pollen tube elongation by altering the distribution of alpha-tubulin and regulating the deposition of pollen cell wall components during the process of tube growth. The possible role of PwTUA1 in microtubule dynamics and organization was discussed.

Show MeSH
Related in: MedlinePlus