Calcium dynamics in tomato pollen tubes using the Yellow Cameleon 3.6 sensor

María Laura Barberini, Lorena Sigaut, Weijie Huang, Silvina Mangano, Silvina Paola Denita Juarez, Eliana Marzol, José Estevez, Mariana Obertello, Lía Pietrasanta, Weihua Tang, Jorge Muschietti

Resultado de la investigación: Article

2 Citas (Scopus)

Resumen

Key message: In vitro tomato pollen tubes show a cytoplasmic calcium gradient that oscillates with the same period as growth. Abstract: Pollen tube growth requires coordination between the tip-focused cytoplasmic calcium concentration ([Ca2+]cyt) gradient and the actin cytoskeleton. This [Ca2+]cyt gradient is necessary for exocytosis of small vesicles, which contributes to the delivery of new membrane and cell wall at the pollen tube tip. The mechanisms that generate and maintain this [Ca2+]cyt gradient are not completely understood. Here, we studied calcium dynamics in tomato (Solanum lycopersicum) pollen tubes using transgenic tomato plants expressing the Yellow Cameleon 3.6 gene under the pollen-specific promoter LAT52. We use tomato as an experimental model because tomato is a Solanaceous plant that is easy to transform, and has an excellent genomic database and genetic stock center, and unlike Arabidopsis, tomato pollen is a good system to do biochemistry. We found that tomato pollen tubes showed an oscillating tip-focused [Ca2+]cyt gradient with the same period as growth. Then, we used a pharmacological approach to disturb the intracellular Ca2+ homeostasis, evaluating how the [Ca2+]cyt gradient, pollen germination and in vitro pollen tube growth were affected. We found that cyclopiazonic acid (CPA), a drug that inhibits plant PIIA-type Ca2+-ATPases, increased [Ca2+]cyt in the subapical zone, leading to the disappearance of the Ca2+ oscillations and inhibition of pollen tube growth. In contrast, 2-aminoethoxydiphenyl borate (2-APB), an inhibitor of Ca2+ released from the endoplasmic reticulum to the cytoplasm in animals cells, completely reduced [Ca2+]cyt at the tip of the tube, blocked the gradient and arrested pollen tube growth. Although both drugs have antagonistic effects on [Ca2+]cyt, both inhibited pollen tube growth triggering the disappearance of the [Ca2+]cyt gradient. When CPA and 2-APB were combined, their individual inhibitory effects on pollen tube growth were partially compensated. Finally, we found that GsMTx-4, a peptide from spider venom that blocks stretch-activated Ca2+ channels, inhibited tomato pollen germination and had a heterogeneous effect on pollen tube growth, suggesting that these channels are also involved in the maintenance of the [Ca2+]cyt gradient. All these results indicate that tomato pollen tube is an excellent model to study calcium dynamics.

Idioma originalEnglish
Páginas (desde-hasta)159-169
Número de páginas11
PublicaciónPlant Reproduction
Volumen31
N.º2
DOI
EstadoPublished - 1 jun 2018

Huella dactilar

Pollen Tube
Lycopersicon esculentum
pollen tubes
tomatoes
Calcium
calcium
Growth
Pollen
Germination
cyclopiazonic acid
YCaM3.6 protein
borates
pollen germination
Spider Venoms
Genetic Databases
Calcium-Transporting ATPases
Genetically Modified Plants
Exocytosis
Matthiola
Actin Cytoskeleton

ASJC Scopus subject areas

  • Plant Science
  • Cell Biology

Citar esto

Barberini, M. L., Sigaut, L., Huang, W., Mangano, S., Juarez, S. P. D., Marzol, E., ... Muschietti, J. (2018). Calcium dynamics in tomato pollen tubes using the Yellow Cameleon 3.6 sensor. Plant Reproduction, 31(2), 159-169. https://doi.org/10.1007/s00497-017-0317-y
Barberini, María Laura ; Sigaut, Lorena ; Huang, Weijie ; Mangano, Silvina ; Juarez, Silvina Paola Denita ; Marzol, Eliana ; Estevez, José ; Obertello, Mariana ; Pietrasanta, Lía ; Tang, Weihua ; Muschietti, Jorge. / Calcium dynamics in tomato pollen tubes using the Yellow Cameleon 3.6 sensor. En: Plant Reproduction. 2018 ; Vol. 31, N.º 2. pp. 159-169.
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abstract = "Key message: In vitro tomato pollen tubes show a cytoplasmic calcium gradient that oscillates with the same period as growth. Abstract: Pollen tube growth requires coordination between the tip-focused cytoplasmic calcium concentration ([Ca2+]cyt) gradient and the actin cytoskeleton. This [Ca2+]cyt gradient is necessary for exocytosis of small vesicles, which contributes to the delivery of new membrane and cell wall at the pollen tube tip. The mechanisms that generate and maintain this [Ca2+]cyt gradient are not completely understood. Here, we studied calcium dynamics in tomato (Solanum lycopersicum) pollen tubes using transgenic tomato plants expressing the Yellow Cameleon 3.6 gene under the pollen-specific promoter LAT52. We use tomato as an experimental model because tomato is a Solanaceous plant that is easy to transform, and has an excellent genomic database and genetic stock center, and unlike Arabidopsis, tomato pollen is a good system to do biochemistry. We found that tomato pollen tubes showed an oscillating tip-focused [Ca2+]cyt gradient with the same period as growth. Then, we used a pharmacological approach to disturb the intracellular Ca2+ homeostasis, evaluating how the [Ca2+]cyt gradient, pollen germination and in vitro pollen tube growth were affected. We found that cyclopiazonic acid (CPA), a drug that inhibits plant PIIA-type Ca2+-ATPases, increased [Ca2+]cyt in the subapical zone, leading to the disappearance of the Ca2+ oscillations and inhibition of pollen tube growth. In contrast, 2-aminoethoxydiphenyl borate (2-APB), an inhibitor of Ca2+ released from the endoplasmic reticulum to the cytoplasm in animals cells, completely reduced [Ca2+]cyt at the tip of the tube, blocked the gradient and arrested pollen tube growth. Although both drugs have antagonistic effects on [Ca2+]cyt, both inhibited pollen tube growth triggering the disappearance of the [Ca2+]cyt gradient. When CPA and 2-APB were combined, their individual inhibitory effects on pollen tube growth were partially compensated. Finally, we found that GsMTx-4, a peptide from spider venom that blocks stretch-activated Ca2+ channels, inhibited tomato pollen germination and had a heterogeneous effect on pollen tube growth, suggesting that these channels are also involved in the maintenance of the [Ca2+]cyt gradient. All these results indicate that tomato pollen tube is an excellent model to study calcium dynamics.",
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Barberini, ML, Sigaut, L, Huang, W, Mangano, S, Juarez, SPD, Marzol, E, Estevez, J, Obertello, M, Pietrasanta, L, Tang, W & Muschietti, J 2018, 'Calcium dynamics in tomato pollen tubes using the Yellow Cameleon 3.6 sensor', Plant Reproduction, vol. 31, n.º 2, pp. 159-169. https://doi.org/10.1007/s00497-017-0317-y

Calcium dynamics in tomato pollen tubes using the Yellow Cameleon 3.6 sensor. / Barberini, María Laura; Sigaut, Lorena; Huang, Weijie; Mangano, Silvina; Juarez, Silvina Paola Denita; Marzol, Eliana; Estevez, José; Obertello, Mariana; Pietrasanta, Lía; Tang, Weihua; Muschietti, Jorge.

En: Plant Reproduction, Vol. 31, N.º 2, 01.06.2018, p. 159-169.

Resultado de la investigación: Article

TY - JOUR

T1 - Calcium dynamics in tomato pollen tubes using the Yellow Cameleon 3.6 sensor

AU - Barberini, María Laura

AU - Sigaut, Lorena

AU - Huang, Weijie

AU - Mangano, Silvina

AU - Juarez, Silvina Paola Denita

AU - Marzol, Eliana

AU - Estevez, José

AU - Obertello, Mariana

AU - Pietrasanta, Lía

AU - Tang, Weihua

AU - Muschietti, Jorge

PY - 2018/6/1

Y1 - 2018/6/1

N2 - Key message: In vitro tomato pollen tubes show a cytoplasmic calcium gradient that oscillates with the same period as growth. Abstract: Pollen tube growth requires coordination between the tip-focused cytoplasmic calcium concentration ([Ca2+]cyt) gradient and the actin cytoskeleton. This [Ca2+]cyt gradient is necessary for exocytosis of small vesicles, which contributes to the delivery of new membrane and cell wall at the pollen tube tip. The mechanisms that generate and maintain this [Ca2+]cyt gradient are not completely understood. Here, we studied calcium dynamics in tomato (Solanum lycopersicum) pollen tubes using transgenic tomato plants expressing the Yellow Cameleon 3.6 gene under the pollen-specific promoter LAT52. We use tomato as an experimental model because tomato is a Solanaceous plant that is easy to transform, and has an excellent genomic database and genetic stock center, and unlike Arabidopsis, tomato pollen is a good system to do biochemistry. We found that tomato pollen tubes showed an oscillating tip-focused [Ca2+]cyt gradient with the same period as growth. Then, we used a pharmacological approach to disturb the intracellular Ca2+ homeostasis, evaluating how the [Ca2+]cyt gradient, pollen germination and in vitro pollen tube growth were affected. We found that cyclopiazonic acid (CPA), a drug that inhibits plant PIIA-type Ca2+-ATPases, increased [Ca2+]cyt in the subapical zone, leading to the disappearance of the Ca2+ oscillations and inhibition of pollen tube growth. In contrast, 2-aminoethoxydiphenyl borate (2-APB), an inhibitor of Ca2+ released from the endoplasmic reticulum to the cytoplasm in animals cells, completely reduced [Ca2+]cyt at the tip of the tube, blocked the gradient and arrested pollen tube growth. Although both drugs have antagonistic effects on [Ca2+]cyt, both inhibited pollen tube growth triggering the disappearance of the [Ca2+]cyt gradient. When CPA and 2-APB were combined, their individual inhibitory effects on pollen tube growth were partially compensated. Finally, we found that GsMTx-4, a peptide from spider venom that blocks stretch-activated Ca2+ channels, inhibited tomato pollen germination and had a heterogeneous effect on pollen tube growth, suggesting that these channels are also involved in the maintenance of the [Ca2+]cyt gradient. All these results indicate that tomato pollen tube is an excellent model to study calcium dynamics.

AB - Key message: In vitro tomato pollen tubes show a cytoplasmic calcium gradient that oscillates with the same period as growth. Abstract: Pollen tube growth requires coordination between the tip-focused cytoplasmic calcium concentration ([Ca2+]cyt) gradient and the actin cytoskeleton. This [Ca2+]cyt gradient is necessary for exocytosis of small vesicles, which contributes to the delivery of new membrane and cell wall at the pollen tube tip. The mechanisms that generate and maintain this [Ca2+]cyt gradient are not completely understood. Here, we studied calcium dynamics in tomato (Solanum lycopersicum) pollen tubes using transgenic tomato plants expressing the Yellow Cameleon 3.6 gene under the pollen-specific promoter LAT52. We use tomato as an experimental model because tomato is a Solanaceous plant that is easy to transform, and has an excellent genomic database and genetic stock center, and unlike Arabidopsis, tomato pollen is a good system to do biochemistry. We found that tomato pollen tubes showed an oscillating tip-focused [Ca2+]cyt gradient with the same period as growth. Then, we used a pharmacological approach to disturb the intracellular Ca2+ homeostasis, evaluating how the [Ca2+]cyt gradient, pollen germination and in vitro pollen tube growth were affected. We found that cyclopiazonic acid (CPA), a drug that inhibits plant PIIA-type Ca2+-ATPases, increased [Ca2+]cyt in the subapical zone, leading to the disappearance of the Ca2+ oscillations and inhibition of pollen tube growth. In contrast, 2-aminoethoxydiphenyl borate (2-APB), an inhibitor of Ca2+ released from the endoplasmic reticulum to the cytoplasm in animals cells, completely reduced [Ca2+]cyt at the tip of the tube, blocked the gradient and arrested pollen tube growth. Although both drugs have antagonistic effects on [Ca2+]cyt, both inhibited pollen tube growth triggering the disappearance of the [Ca2+]cyt gradient. When CPA and 2-APB were combined, their individual inhibitory effects on pollen tube growth were partially compensated. Finally, we found that GsMTx-4, a peptide from spider venom that blocks stretch-activated Ca2+ channels, inhibited tomato pollen germination and had a heterogeneous effect on pollen tube growth, suggesting that these channels are also involved in the maintenance of the [Ca2+]cyt gradient. All these results indicate that tomato pollen tube is an excellent model to study calcium dynamics.

KW - Calcium ATPases

KW - Calcium concentration gradient

KW - Ion channels

KW - Pollen tube growth

KW - Solanum lycopersicum

KW - Yellow Cameleon 3.6

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Barberini ML, Sigaut L, Huang W, Mangano S, Juarez SPD, Marzol E y otros. Calcium dynamics in tomato pollen tubes using the Yellow Cameleon 3.6 sensor. Plant Reproduction. 2018 jun 1;31(2):159-169. https://doi.org/10.1007/s00497-017-0317-y