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Identification and localization of a caleosin in olive (Olea europaea L.) pollen during in vitro germination.

Zienkiewicz K, Castro AJ, Alché Jde D, Zienkiewicz A, Suárez C, Rodríguez-García MI - J. Exp. Bot. (2010)

Bottom Line: In addition, caleosins were also visualized in the cytoplasm at the subapical zone, as well as in the tonoplast of vacuoles present in the pollen tube cytoplasm.The number of oil bodies decreased 20-fold in the pollen grain during germination, whereas the opposite tendency occurred in the pollen tube, suggesting that oil bodies moved from one to the other.The data suggest that this pollen caleosin might have a role in the mobilization of oil bodies as well as in the reorganization of membrane compartments during pollen in vitro germination.

View Article: PubMed Central - PubMed

Affiliation: Departamento de BioquĂ­mica, BiologĂ­a Celular y Molecular de Plantas, EstaciĂłn Experimental del ZaidĂ­n (CSIC), Profesor Albareda 1, 18008, Granada, Spain.

ABSTRACT
In plant organs and tissues, the neutral storage lipids are confined to discrete spherical organelles called oil bodies. Oil bodies from plant seeds contain 0.6-3% proteins, including oleosins, steroleosins, and caleosins. In this study, a caleosin isoform of approximately 30 kDa was identified in the olive pollen grain. The protein was mainly located at the boundaries of the oil bodies in the cytoplasm of the pollen grain and the pollen tube. In addition, caleosins were also visualized in the cytoplasm at the subapical zone, as well as in the tonoplast of vacuoles present in the pollen tube cytoplasm. The cellular behaviour of lipid bodies in the olive pollen was also monitored during in vitro germination. The number of oil bodies decreased 20-fold in the pollen grain during germination, whereas the opposite tendency occurred in the pollen tube, suggesting that oil bodies moved from one to the other. The data suggest that this pollen caleosin might have a role in the mobilization of oil bodies as well as in the reorganization of membrane compartments during pollen in vitro germination.

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(A) Anti-Clo3 Ab-immunoprecipitated proteins after SDS–PAGE and Coomassie staining. (B) Immunoblot as in A probed with the anti-Clo3 Ab. C1, control 1, anti-caleosin Ab probed with a secondary Ab; C2, control 2, anti-Clo3 Ab-immunoprecipitated fraction from pollen OB-associated proteins probed as in C1; TP, anti-Clo3 Ab-immunoprecipitated fraction from crude protein extracts from pollen; OB, anti-Clo3 Ab-immunoprecipitated fraction from pollen OB-associated proteins; Mi, anti-Clo3 Ab-immunoprecipitated fraction from the pollen microsomal fraction-associated proteins. The arrowhead shows the 30 kDa caleosin band.
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fig2: (A) Anti-Clo3 Ab-immunoprecipitated proteins after SDS–PAGE and Coomassie staining. (B) Immunoblot as in A probed with the anti-Clo3 Ab. C1, control 1, anti-caleosin Ab probed with a secondary Ab; C2, control 2, anti-Clo3 Ab-immunoprecipitated fraction from pollen OB-associated proteins probed as in C1; TP, anti-Clo3 Ab-immunoprecipitated fraction from crude protein extracts from pollen; OB, anti-Clo3 Ab-immunoprecipitated fraction from pollen OB-associated proteins; Mi, anti-Clo3 Ab-immunoprecipitated fraction from the pollen microsomal fraction-associated proteins. The arrowhead shows the 30 kDa caleosin band.

Mentions: Immunoprecipitation experiments showed that, when incubated with total protein extracts from mature pollen, the anti-Clo3 Ab bound to one protein of 30 kDa (Fig. 2). This band was also present after incubating the Clo-3 Ab with total proteins isolated from both OBs and the microsomal fraction. MS analyses of the tryptic peptides resulting from the digestion of the 30 kDa protein band matched two peptides of a caleosin (accession number EF015588) from lily pollen (Table 1).


Identification and localization of a caleosin in olive (Olea europaea L.) pollen during in vitro germination.

Zienkiewicz K, Castro AJ, Alché Jde D, Zienkiewicz A, Suárez C, Rodríguez-García MI - J. Exp. Bot. (2010)

(A) Anti-Clo3 Ab-immunoprecipitated proteins after SDS–PAGE and Coomassie staining. (B) Immunoblot as in A probed with the anti-Clo3 Ab. C1, control 1, anti-caleosin Ab probed with a secondary Ab; C2, control 2, anti-Clo3 Ab-immunoprecipitated fraction from pollen OB-associated proteins probed as in C1; TP, anti-Clo3 Ab-immunoprecipitated fraction from crude protein extracts from pollen; OB, anti-Clo3 Ab-immunoprecipitated fraction from pollen OB-associated proteins; Mi, anti-Clo3 Ab-immunoprecipitated fraction from the pollen microsomal fraction-associated proteins. The arrowhead shows the 30 kDa caleosin band.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC2837266&req=5

fig2: (A) Anti-Clo3 Ab-immunoprecipitated proteins after SDS–PAGE and Coomassie staining. (B) Immunoblot as in A probed with the anti-Clo3 Ab. C1, control 1, anti-caleosin Ab probed with a secondary Ab; C2, control 2, anti-Clo3 Ab-immunoprecipitated fraction from pollen OB-associated proteins probed as in C1; TP, anti-Clo3 Ab-immunoprecipitated fraction from crude protein extracts from pollen; OB, anti-Clo3 Ab-immunoprecipitated fraction from pollen OB-associated proteins; Mi, anti-Clo3 Ab-immunoprecipitated fraction from the pollen microsomal fraction-associated proteins. The arrowhead shows the 30 kDa caleosin band.
Mentions: Immunoprecipitation experiments showed that, when incubated with total protein extracts from mature pollen, the anti-Clo3 Ab bound to one protein of 30 kDa (Fig. 2). This band was also present after incubating the Clo-3 Ab with total proteins isolated from both OBs and the microsomal fraction. MS analyses of the tryptic peptides resulting from the digestion of the 30 kDa protein band matched two peptides of a caleosin (accession number EF015588) from lily pollen (Table 1).

Bottom Line: In addition, caleosins were also visualized in the cytoplasm at the subapical zone, as well as in the tonoplast of vacuoles present in the pollen tube cytoplasm.The number of oil bodies decreased 20-fold in the pollen grain during germination, whereas the opposite tendency occurred in the pollen tube, suggesting that oil bodies moved from one to the other.The data suggest that this pollen caleosin might have a role in the mobilization of oil bodies as well as in the reorganization of membrane compartments during pollen in vitro germination.

View Article: PubMed Central - PubMed

Affiliation: Departamento de BioquĂ­mica, BiologĂ­a Celular y Molecular de Plantas, EstaciĂłn Experimental del ZaidĂ­n (CSIC), Profesor Albareda 1, 18008, Granada, Spain.

ABSTRACT
In plant organs and tissues, the neutral storage lipids are confined to discrete spherical organelles called oil bodies. Oil bodies from plant seeds contain 0.6-3% proteins, including oleosins, steroleosins, and caleosins. In this study, a caleosin isoform of approximately 30 kDa was identified in the olive pollen grain. The protein was mainly located at the boundaries of the oil bodies in the cytoplasm of the pollen grain and the pollen tube. In addition, caleosins were also visualized in the cytoplasm at the subapical zone, as well as in the tonoplast of vacuoles present in the pollen tube cytoplasm. The cellular behaviour of lipid bodies in the olive pollen was also monitored during in vitro germination. The number of oil bodies decreased 20-fold in the pollen grain during germination, whereas the opposite tendency occurred in the pollen tube, suggesting that oil bodies moved from one to the other. The data suggest that this pollen caleosin might have a role in the mobilization of oil bodies as well as in the reorganization of membrane compartments during pollen in vitro germination.

Show MeSH