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Developmental and heat stress-regulated expression of HsfA2 and small heat shock proteins in tomato anthers.

Giorno F, Wolters-Arts M, Grillo S, Scharf KD, Vriezen WH, Mariani C - J. Exp. Bot. (2009)

Bottom Line: A transcriptional analysis at different developmental anther/pollen stages was performed using semi-quantitative and real-time PCR.The messengers were localized using in situ RNA hybridization, and protein accumulation was monitored using immunoblot analysis.These data suggest that HsfA2 may be directly involved in the activation of protection mechanisms in the tomato anther during heat stress and, thereby, may contribute to tomato fruit set under adverse temperatures.

View Article: PubMed Central - PubMed

Affiliation: Department of Plant Cell Biology, IWWR Institute, Radboud University Nijmegen, Heyendaalseweg 135, Nijmegen, The Netherlands.

ABSTRACT
The high sensitivity of male reproductive cells to high temperatures may be due to an inadequate heat stress response. The results of a comprehensive expression analysis of HsfA2 and Hsp17-CII, two important members of the heat stress system, in the developing anthers of a heat-tolerant tomato genotype are reported here. A transcriptional analysis at different developmental anther/pollen stages was performed using semi-quantitative and real-time PCR. The messengers were localized using in situ RNA hybridization, and protein accumulation was monitored using immunoblot analysis. Based on the analysis of the gene and protein expression profiles, HsfA2 and Hsp17-CII are finely regulated during anther development and are further induced under both short and prolonged heat stress conditions. These data suggest that HsfA2 may be directly involved in the activation of protection mechanisms in the tomato anther during heat stress and, thereby, may contribute to tomato fruit set under adverse temperatures.

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Related in: MedlinePlus

Expression analyses of HsfA2 and Hsp17-CII in 2 mm anthers after HS treatments. (A) The pictogram shows the time course of HS treatments. Arrows indicate the time points when the 2 mm anthers were harvested (CT-I). (B–D) qRT-PCR of HsfA2 (B), Hsp17.4-CII (C), and Hsp17.6-CII (D). Expression data were normalized using LeEF1 and 18S rRNA as housekeeping genes. The mRNA levels of the target genes are relative to that of the sample CT (value 1). (E) Immunoblotting analyses showing protein levels of HsfA2 and Hsp17-CII in anthers at the same stages as in B, C, and D. Ponceau staining of total protein was used as control for equal loading.
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fig4: Expression analyses of HsfA2 and Hsp17-CII in 2 mm anthers after HS treatments. (A) The pictogram shows the time course of HS treatments. Arrows indicate the time points when the 2 mm anthers were harvested (CT-I). (B–D) qRT-PCR of HsfA2 (B), Hsp17.4-CII (C), and Hsp17.6-CII (D). Expression data were normalized using LeEF1 and 18S rRNA as housekeeping genes. The mRNA levels of the target genes are relative to that of the sample CT (value 1). (E) Immunoblotting analyses showing protein levels of HsfA2 and Hsp17-CII in anthers at the same stages as in B, C, and D. Ponceau staining of total protein was used as control for equal loading.

Mentions: Heat stress (HS) was applied to plants in the form of high temperature (36 °C). Anthers at the 2 mm stage were collected after being subjected to short HS treatments at 36 °C for 1, 2, and 6 h, respectively (Fig. 4A, samples A, B, and C), after recovery at 26 °C and on the second day of the HS treatments (Fig. 4A, samples D, E, and F). Anthers at the 2 mm stage were harvested from the same treated plants (Fig. 4A, samples G, H and I) at day 7 after daily repeated cycles of mild HS and recovery (36 °C / 26 °C day/night). Recovery samples D and G (Fig. 4) were also harvested at 26 °C after 30 min of light acclimatization to avoid interference from the circadian rhythm.


Developmental and heat stress-regulated expression of HsfA2 and small heat shock proteins in tomato anthers.

Giorno F, Wolters-Arts M, Grillo S, Scharf KD, Vriezen WH, Mariani C - J. Exp. Bot. (2009)

Expression analyses of HsfA2 and Hsp17-CII in 2 mm anthers after HS treatments. (A) The pictogram shows the time course of HS treatments. Arrows indicate the time points when the 2 mm anthers were harvested (CT-I). (B–D) qRT-PCR of HsfA2 (B), Hsp17.4-CII (C), and Hsp17.6-CII (D). Expression data were normalized using LeEF1 and 18S rRNA as housekeeping genes. The mRNA levels of the target genes are relative to that of the sample CT (value 1). (E) Immunoblotting analyses showing protein levels of HsfA2 and Hsp17-CII in anthers at the same stages as in B, C, and D. Ponceau staining of total protein was used as control for equal loading.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC2803211&req=5

fig4: Expression analyses of HsfA2 and Hsp17-CII in 2 mm anthers after HS treatments. (A) The pictogram shows the time course of HS treatments. Arrows indicate the time points when the 2 mm anthers were harvested (CT-I). (B–D) qRT-PCR of HsfA2 (B), Hsp17.4-CII (C), and Hsp17.6-CII (D). Expression data were normalized using LeEF1 and 18S rRNA as housekeeping genes. The mRNA levels of the target genes are relative to that of the sample CT (value 1). (E) Immunoblotting analyses showing protein levels of HsfA2 and Hsp17-CII in anthers at the same stages as in B, C, and D. Ponceau staining of total protein was used as control for equal loading.
Mentions: Heat stress (HS) was applied to plants in the form of high temperature (36 °C). Anthers at the 2 mm stage were collected after being subjected to short HS treatments at 36 °C for 1, 2, and 6 h, respectively (Fig. 4A, samples A, B, and C), after recovery at 26 °C and on the second day of the HS treatments (Fig. 4A, samples D, E, and F). Anthers at the 2 mm stage were harvested from the same treated plants (Fig. 4A, samples G, H and I) at day 7 after daily repeated cycles of mild HS and recovery (36 °C / 26 °C day/night). Recovery samples D and G (Fig. 4) were also harvested at 26 °C after 30 min of light acclimatization to avoid interference from the circadian rhythm.

Bottom Line: A transcriptional analysis at different developmental anther/pollen stages was performed using semi-quantitative and real-time PCR.The messengers were localized using in situ RNA hybridization, and protein accumulation was monitored using immunoblot analysis.These data suggest that HsfA2 may be directly involved in the activation of protection mechanisms in the tomato anther during heat stress and, thereby, may contribute to tomato fruit set under adverse temperatures.

View Article: PubMed Central - PubMed

Affiliation: Department of Plant Cell Biology, IWWR Institute, Radboud University Nijmegen, Heyendaalseweg 135, Nijmegen, The Netherlands.

ABSTRACT
The high sensitivity of male reproductive cells to high temperatures may be due to an inadequate heat stress response. The results of a comprehensive expression analysis of HsfA2 and Hsp17-CII, two important members of the heat stress system, in the developing anthers of a heat-tolerant tomato genotype are reported here. A transcriptional analysis at different developmental anther/pollen stages was performed using semi-quantitative and real-time PCR. The messengers were localized using in situ RNA hybridization, and protein accumulation was monitored using immunoblot analysis. Based on the analysis of the gene and protein expression profiles, HsfA2 and Hsp17-CII are finely regulated during anther development and are further induced under both short and prolonged heat stress conditions. These data suggest that HsfA2 may be directly involved in the activation of protection mechanisms in the tomato anther during heat stress and, thereby, may contribute to tomato fruit set under adverse temperatures.

Show MeSH
Related in: MedlinePlus