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Analysis of the tip-to-base gradient of CaM in pollen tube pulsant growth using in vivo CaM–GFP system

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Abstract

Ca2+-CaM signaling is involved in pollen tube development. However, the distribution and function of CaM and the downstream components of Ca2+-CaM signal in pollen tube development still need more exploration. Here we obtained the CaM–GFP fusion protein transgenic line of Nicotiana tobacum SRI, which allowed us to monitor CaM distribution pattern in vivo and provided a useful tool to observe CaM response to various exogenous stimulations and afforded solid evidences of the essential functions of CaM in pollen tube growth. CaM–GFP fusion gene was constructed under the control of Lat52-7 pollen-specific promoter and transformed into Nicotiana tobacum SRI. High level of CaM–GFP fluorescence was detected at the germinal pores and the tip-to-base gradient of fluorescence was observed in developing pollen tubes. The distribution of CaM at apical dome had close relationship with the pulsant growth mode of pollen tubes: when CaM aggregated at the apical dome, pollen tubes stepped into growth state; When CaM showed non-polarized distribution, pollen tubes stopped growing. In addition, after affording exogenous Ca2+, calmidazolium (antagonism of CaM) or Brefeldin A (an inhibitor of membrane trafficking), CaM turned to a uniform distribution at the apical dome and pollen tube growth was held back. Taken together, our results showed that CaM played a vital role in pollen tube elongation and growth rate, and the oscillation of tip-to-base gradient of CaM was required for the normal pulsant growth of pollen tube.

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Abbreviations

BFA:

Brefeldin A

BK:

Brewbaker and Kwack

CaM:

Calmodulin

CDPK:

Calcium dependent protein kinases

GFP:

Green fluorescent protein

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Acknowledgments

This work is supported by National Natural Science Foundation of China (Grant #30821064).

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Authors and Affiliations

  1. Key Lab of MOE for Plant Developmental Biology, Wuhan University, 430072, Wuhan, People’s Republic of China

    Ya-Ya Shi, Wen-Jing Tao, Shu-Ping Liang, Yingtang Lü & Lei Zhang

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  1. Ya-Ya Shi
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  2. Wen-Jing Tao
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  3. Shu-Ping Liang
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  4. Yingtang Lü
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  5. Lei Zhang
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Correspondence to Lei Zhang.

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Communicated by Y. Lü.

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Shi, YY., Tao, WJ., Liang, SP. et al. Analysis of the tip-to-base gradient of CaM in pollen tube pulsant growth using in vivo CaM–GFP system. Plant Cell Rep 28, 1253–1264 (2009). https://doi.org/10.1007/s00299-009-0725-z

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  • DOI: https://doi.org/10.1007/s00299-009-0725-z

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