Morina, Filis

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orcid::0000-0003-1521-125X
  • Morina, Filis (4)

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Efficient protein extraction for shotgun proteomics from hydrated and desiccated leaves of resurrection Ramonda serbica plants

Vidović, Marija; Franchin, Cinzia; Morina, Filis; Veljović-Jovanović, Sonja; Masi, Antonio; Arrigoni, Giorgio

(2020)

TY  - JOUR
AU  - Vidović, Marija
AU  - Franchin, Cinzia
AU  - Morina, Filis
AU  - Veljović-Jovanović, Sonja
AU  - Masi, Antonio
AU  - Arrigoni, Giorgio
PY  - 2020
UR  - http://www.ncbi.nlm.nih.gov/pubmed/33037906
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/4505
AB  - Resurrection plant Ramonda serbica is a suitable model to investigate vegetative desiccation tolerance. However, the detailed study of these mechanisms at the protein level is hampered by the severe tissue water loss, high amount of phenolics and polysaccharide, and possible protein modifications and aggregations during the extraction and purification steps. When applied to R. serbica leaves, widely used protein extraction protocols containing polyvinylpolypyrrolidone and ascorbate, as well as the phenol/SDS/buffer-based protocol recommended for recalcitrant plant tissues failed to eliminate persistent contamination and ensure high protein quality. Here we compared three protein extraction approaches aiming to establish the optimal one for both hydrated and desiccated R. serbica leaves. To evaluate the efficacy of these protocols by shotgun proteomics, we also created the first R. serbica annotated transcriptome database, available at http://www.biomed.unipd.it/filearrigoni/Trinity_Sample_RT2.fasta . The detergent-free phenol-based extraction combined with dodecyl-β-D-maltoside-assisted extraction enabled high-yield and high-purity protein extracts. The phenol-based protocol improved the protein-band resolution, band number, and intensity upon electrophoresis, and increased the protein yield and the number of identified peptides and protein groups by LC-MS/MS. Additionally, dodecyl-β-D-maltoside enabled solubilisation and identification of more membrane-associated proteins. The presented study paves the way for investigating the desiccation tolerance in R. serbica, and we recommend this protocol for similar recalcitrant plant material.
T2  - Analytical and Bioanalytical Chemistry
T1  - Efficient protein extraction for shotgun proteomics from hydrated and desiccated leaves of resurrection Ramonda serbica plants
VL  - 412
IS  - 30
SP  - 8299
EP  - 8312
DO  - 10.1007/s00216-020-02965-2
ER  - 
@article{
author = "Vidović, Marija and Franchin, Cinzia and Morina, Filis and Veljović-Jovanović, Sonja and Masi, Antonio and Arrigoni, Giorgio",
year = "2020",
abstract = "Resurrection plant Ramonda serbica is a suitable model to investigate vegetative desiccation tolerance. However, the detailed study of these mechanisms at the protein level is hampered by the severe tissue water loss, high amount of phenolics and polysaccharide, and possible protein modifications and aggregations during the extraction and purification steps. When applied to R. serbica leaves, widely used protein extraction protocols containing polyvinylpolypyrrolidone and ascorbate, as well as the phenol/SDS/buffer-based protocol recommended for recalcitrant plant tissues failed to eliminate persistent contamination and ensure high protein quality. Here we compared three protein extraction approaches aiming to establish the optimal one for both hydrated and desiccated R. serbica leaves. To evaluate the efficacy of these protocols by shotgun proteomics, we also created the first R. serbica annotated transcriptome database, available at http://www.biomed.unipd.it/filearrigoni/Trinity_Sample_RT2.fasta . The detergent-free phenol-based extraction combined with dodecyl-β-D-maltoside-assisted extraction enabled high-yield and high-purity protein extracts. The phenol-based protocol improved the protein-band resolution, band number, and intensity upon electrophoresis, and increased the protein yield and the number of identified peptides and protein groups by LC-MS/MS. Additionally, dodecyl-β-D-maltoside enabled solubilisation and identification of more membrane-associated proteins. The presented study paves the way for investigating the desiccation tolerance in R. serbica, and we recommend this protocol for similar recalcitrant plant material.",
journal = "Analytical and Bioanalytical Chemistry",
title = "Efficient protein extraction for shotgun proteomics from hydrated and desiccated leaves of resurrection Ramonda serbica plants",
volume = "412",
number = "30",
pages = "8299-8312",
doi = "10.1007/s00216-020-02965-2"
}
Vidović, M., Franchin, C., Morina, F., Veljović-Jovanović, S., Masi, A.,& Arrigoni, G.. (2020). Efficient protein extraction for shotgun proteomics from hydrated and desiccated leaves of resurrection Ramonda serbica plants. in Analytical and Bioanalytical Chemistry, 412(30), 8299-8312.
https://doi.org/10.1007/s00216-020-02965-2
Vidović M, Franchin C, Morina F, Veljović-Jovanović S, Masi A, Arrigoni G. Efficient protein extraction for shotgun proteomics from hydrated and desiccated leaves of resurrection Ramonda serbica plants. in Analytical and Bioanalytical Chemistry. 2020;412(30):8299-8312.
doi:10.1007/s00216-020-02965-2 .
Vidović, Marija, Franchin, Cinzia, Morina, Filis, Veljović-Jovanović, Sonja, Masi, Antonio, Arrigoni, Giorgio, "Efficient protein extraction for shotgun proteomics from hydrated and desiccated leaves of resurrection Ramonda serbica plants" in Analytical and Bioanalytical Chemistry, 412, no. 30 (2020):8299-8312,
https://doi.org/10.1007/s00216-020-02965-2 . .
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Supplementary data for the article: Vidović, M.; Franchin, C.; Morina, F.; Veljović-Jovanović, S.; Masi, A.; Arrigoni, G. Efficient Protein Extraction for Shotgun Proteomics from Hydrated and Desiccated Leaves of Resurrection Ramonda Serbica Plants. Anal Bioanal Chem 2020, 412 (30), 8299–8312. https://doi.org/10.1007/s00216-020-02965-2.

Vidović, Marija; Franchin, Cinzia; Morina, Filis; Veljović-Jovanović, Sonja; Masi, Antonio; Arrigoni, Giorgio

(SpringerLink, 2020)

TY  - DATA
AU  - Vidović, Marija
AU  - Franchin, Cinzia
AU  - Morina, Filis
AU  - Veljović-Jovanović, Sonja
AU  - Masi, Antonio
AU  - Arrigoni, Giorgio
PY  - 2020
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/4504
PB  - SpringerLink
T2  - Analytical and Bioanalytical Chemistry
T1  - Supplementary data for the article: Vidović, M.; Franchin, C.; Morina, F.; Veljović-Jovanović, S.; Masi, A.; Arrigoni, G. Efficient Protein Extraction for Shotgun Proteomics from Hydrated and Desiccated Leaves of Resurrection Ramonda Serbica Plants. Anal Bioanal Chem 2020, 412 (30), 8299–8312. https://doi.org/10.1007/s00216-020-02965-2.
UR  - https://hdl.handle.net/21.15107/rcub_cherry_4504
ER  - 
@misc{
author = "Vidović, Marija and Franchin, Cinzia and Morina, Filis and Veljović-Jovanović, Sonja and Masi, Antonio and Arrigoni, Giorgio",
year = "2020",
publisher = "SpringerLink",
journal = "Analytical and Bioanalytical Chemistry",
title = "Supplementary data for the article: Vidović, M.; Franchin, C.; Morina, F.; Veljović-Jovanović, S.; Masi, A.; Arrigoni, G. Efficient Protein Extraction for Shotgun Proteomics from Hydrated and Desiccated Leaves of Resurrection Ramonda Serbica Plants. Anal Bioanal Chem 2020, 412 (30), 8299–8312. https://doi.org/10.1007/s00216-020-02965-2.",
url = "https://hdl.handle.net/21.15107/rcub_cherry_4504"
}
Vidović, M., Franchin, C., Morina, F., Veljović-Jovanović, S., Masi, A.,& Arrigoni, G.. (2020). Supplementary data for the article: Vidović, M.; Franchin, C.; Morina, F.; Veljović-Jovanović, S.; Masi, A.; Arrigoni, G. Efficient Protein Extraction for Shotgun Proteomics from Hydrated and Desiccated Leaves of Resurrection Ramonda Serbica Plants. Anal Bioanal Chem 2020, 412 (30), 8299–8312. https://doi.org/10.1007/s00216-020-02965-2.. in Analytical and Bioanalytical Chemistry
SpringerLink..
https://hdl.handle.net/21.15107/rcub_cherry_4504
Vidović M, Franchin C, Morina F, Veljović-Jovanović S, Masi A, Arrigoni G. Supplementary data for the article: Vidović, M.; Franchin, C.; Morina, F.; Veljović-Jovanović, S.; Masi, A.; Arrigoni, G. Efficient Protein Extraction for Shotgun Proteomics from Hydrated and Desiccated Leaves of Resurrection Ramonda Serbica Plants. Anal Bioanal Chem 2020, 412 (30), 8299–8312. https://doi.org/10.1007/s00216-020-02965-2.. in Analytical and Bioanalytical Chemistry. 2020;.
https://hdl.handle.net/21.15107/rcub_cherry_4504 .
Vidović, Marija, Franchin, Cinzia, Morina, Filis, Veljović-Jovanović, Sonja, Masi, Antonio, Arrigoni, Giorgio, "Supplementary data for the article: Vidović, M.; Franchin, C.; Morina, F.; Veljović-Jovanović, S.; Masi, A.; Arrigoni, G. Efficient Protein Extraction for Shotgun Proteomics from Hydrated and Desiccated Leaves of Resurrection Ramonda Serbica Plants. Anal Bioanal Chem 2020, 412 (30), 8299–8312. https://doi.org/10.1007/s00216-020-02965-2." in Analytical and Bioanalytical Chemistry (2020),
https://hdl.handle.net/21.15107/rcub_cherry_4504 .

Comparative analysis of phenolic profiles of ovipositional fluid of Rhinusa pilosa (Mecinini, Curculionidae) and its host plant Linaria vulgaris (Plantaginaceae)

Sedlarević, Ana; Morina, Filis; Toševski, Ivo; Gašić, Uroš M.; Natić, Maja; Jović, Jelena; Krstić, Oliver; Veljović-Jovanović, Sonja

(Springer, Dordrecht, 2016)

TY  - JOUR
AU  - Sedlarević, Ana
AU  - Morina, Filis
AU  - Toševski, Ivo
AU  - Gašić, Uroš M.
AU  - Natić, Maja
AU  - Jović, Jelena
AU  - Krstić, Oliver
AU  - Veljović-Jovanović, Sonja
PY  - 2016
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/2279
AB  - Rhinusa pilosa (Gyllenhal) is a highly specific weevil that induces stem galls on the common toadflax Linaria vulgaris Mill. females oviposit the eggs near the apex of a growing shoot. The act of oviposition is accompanied by secretion of an ovipositional fluid, which is considered to be cecidogen, directly involved in gall induction. The remains of cecidogenic fluid were collected from the surface of the oviposition point on the stem. We performed a comparative analysis of the phenolics extracted from cecidogen, the stem and galls of L. vulgaris and adult and larva of R. pilosa by HPLC-DAD. One compound with A (max) at 273, 332 nm (R (t) 30.65 min) was exclusively found in the methanol extract of cecidogen. To further characterize the cecidogen and stem phenolic profiles, we used UHPLC coupled with an OrbiTrap mass analyzer. Among 49 phenolic compounds extracted from both the ovipositional fluid and the plant, protocatechuic acid and two phenolic glycosides were exclusively found in cecidogen: diosmetin-O-acetylrutinoside and an unidentified compound. The unknown compound produced an MS2 base peak at 387 and 327 and 267 m/z base peaks at MS3 and MS4 fragmentation, respectively, and had the molecular formula C32H31O18. The plausible role of phenolic compounds in the induction of gall formation on L. vulgaris is discussed.
PB  - Springer, Dordrecht
T2  - Arthropod-Plant Interactions
T1  - Comparative analysis of phenolic profiles of ovipositional fluid of Rhinusa pilosa (Mecinini, Curculionidae) and its host plant Linaria vulgaris (Plantaginaceae)
VL  - 10
IS  - 4
SP  - 311
EP  - 322
DO  - 10.1007/s11829-016-9435-y
UR  - Kon_3095
ER  - 
@article{
author = "Sedlarević, Ana and Morina, Filis and Toševski, Ivo and Gašić, Uroš M. and Natić, Maja and Jović, Jelena and Krstić, Oliver and Veljović-Jovanović, Sonja",
year = "2016",
abstract = "Rhinusa pilosa (Gyllenhal) is a highly specific weevil that induces stem galls on the common toadflax Linaria vulgaris Mill. females oviposit the eggs near the apex of a growing shoot. The act of oviposition is accompanied by secretion of an ovipositional fluid, which is considered to be cecidogen, directly involved in gall induction. The remains of cecidogenic fluid were collected from the surface of the oviposition point on the stem. We performed a comparative analysis of the phenolics extracted from cecidogen, the stem and galls of L. vulgaris and adult and larva of R. pilosa by HPLC-DAD. One compound with A (max) at 273, 332 nm (R (t) 30.65 min) was exclusively found in the methanol extract of cecidogen. To further characterize the cecidogen and stem phenolic profiles, we used UHPLC coupled with an OrbiTrap mass analyzer. Among 49 phenolic compounds extracted from both the ovipositional fluid and the plant, protocatechuic acid and two phenolic glycosides were exclusively found in cecidogen: diosmetin-O-acetylrutinoside and an unidentified compound. The unknown compound produced an MS2 base peak at 387 and 327 and 267 m/z base peaks at MS3 and MS4 fragmentation, respectively, and had the molecular formula C32H31O18. The plausible role of phenolic compounds in the induction of gall formation on L. vulgaris is discussed.",
publisher = "Springer, Dordrecht",
journal = "Arthropod-Plant Interactions",
title = "Comparative analysis of phenolic profiles of ovipositional fluid of Rhinusa pilosa (Mecinini, Curculionidae) and its host plant Linaria vulgaris (Plantaginaceae)",
volume = "10",
number = "4",
pages = "311-322",
doi = "10.1007/s11829-016-9435-y",
url = "Kon_3095"
}
Sedlarević, A., Morina, F., Toševski, I., Gašić, U. M., Natić, M., Jović, J., Krstić, O.,& Veljović-Jovanović, S.. (2016). Comparative analysis of phenolic profiles of ovipositional fluid of Rhinusa pilosa (Mecinini, Curculionidae) and its host plant Linaria vulgaris (Plantaginaceae). in Arthropod-Plant Interactions
Springer, Dordrecht., 10(4), 311-322.
https://doi.org/10.1007/s11829-016-9435-y
Kon_3095
Sedlarević A, Morina F, Toševski I, Gašić UM, Natić M, Jović J, Krstić O, Veljović-Jovanović S. Comparative analysis of phenolic profiles of ovipositional fluid of Rhinusa pilosa (Mecinini, Curculionidae) and its host plant Linaria vulgaris (Plantaginaceae). in Arthropod-Plant Interactions. 2016;10(4):311-322.
doi:10.1007/s11829-016-9435-y
Kon_3095 .
Sedlarević, Ana, Morina, Filis, Toševski, Ivo, Gašić, Uroš M., Natić, Maja, Jović, Jelena, Krstić, Oliver, Veljović-Jovanović, Sonja, "Comparative analysis of phenolic profiles of ovipositional fluid of Rhinusa pilosa (Mecinini, Curculionidae) and its host plant Linaria vulgaris (Plantaginaceae)" in Arthropod-Plant Interactions, 10, no. 4 (2016):311-322,
https://doi.org/10.1007/s11829-016-9435-y .,
Kon_3095 .
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2
2

Carbon allocation from source to sink leaf tissue in relation to flavonoid biosynthesis in variegated Pelargonium zonale under UV-B radiation and high PAR intensity

Vidović, Marija; Morina, Filis; Milic, Sonja; Albert, Andreas; Zechmann, Bernd; Tosti, Tomislav; Winkler, Jana Barbro; Veljović-Jovanović, Sonja

(Elsevier France-Editions Scientifiques Medicales Elsevier, Paris, 2015)

TY  - JOUR
AU  - Vidović, Marija
AU  - Morina, Filis
AU  - Milic, Sonja
AU  - Albert, Andreas
AU  - Zechmann, Bernd
AU  - Tosti, Tomislav
AU  - Winkler, Jana Barbro
AU  - Veljović-Jovanović, Sonja
PY  - 2015
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/1727
AB  - We studied the specific effects of high photosynthetically active radiation (PAR, 400-700 nm) and ecologically relevant UV-B radiation (0.90 W m(-2)) on antioxidative and phenolic metabolism by exploiting the green-white leaf variegation of Pelargonium zonale plants. This is a suitable model system for examining "source-sink" interactions within the same leaf. High PAR intensity (1350 mu mol m(-2) s(-1)) and UV-B radiation induced different responses in green and white leaf sectors. High PAR intensity had a greater influence on green tissue, triggering the accumulation of phenylpropanoids and flavonoids with strong antioxidative function. Induced phenolics, together with ascorbate, ascorbate peroxidase (APX, EC 1.11.1.11) and catalase (CAT, EC 1.11.1.6) provided efficient defense against potential oxidative pressure. UV-B-induced up-regulation of non-phenolic H2O2 scavengers in green leaf sectors was greater than high PAR-induced changes, indicating a UV-B role in antioxidative defense under light excess; on the contrary, minimal effects were observed in white tissue. However, UV-B radiation had greater influence on phenolics in white leaf sections compared to green ones, inducing accumulation of phenolic glycosides whose function was UV-B screening rather than antioxidative. By stimulation of starch and sucrose breakdown and carbon allocation in the form of soluble sugars from "source" (green) tissue to "sink" (white) tissue, UV-B radiation compensated the absence of photosynthetic activity and phenylpropanoid and flavonoid biosynthesis in white sectors. (C) 2015 Elsevier Masson SAS. All rights reserved.
PB  - Elsevier France-Editions Scientifiques Medicales Elsevier, Paris
T2  - Plant Physiology and Biochemistry
T1  - Carbon allocation from source to sink leaf tissue in relation to flavonoid biosynthesis in variegated Pelargonium zonale under UV-B radiation and high PAR intensity
VL  - 93
SP  - 44
EP  - 55
DO  - 10.1016/j.plaphy.2015.01.008
UR  - Kon_2873
ER  - 
@article{
author = "Vidović, Marija and Morina, Filis and Milic, Sonja and Albert, Andreas and Zechmann, Bernd and Tosti, Tomislav and Winkler, Jana Barbro and Veljović-Jovanović, Sonja",
year = "2015",
abstract = "We studied the specific effects of high photosynthetically active radiation (PAR, 400-700 nm) and ecologically relevant UV-B radiation (0.90 W m(-2)) on antioxidative and phenolic metabolism by exploiting the green-white leaf variegation of Pelargonium zonale plants. This is a suitable model system for examining "source-sink" interactions within the same leaf. High PAR intensity (1350 mu mol m(-2) s(-1)) and UV-B radiation induced different responses in green and white leaf sectors. High PAR intensity had a greater influence on green tissue, triggering the accumulation of phenylpropanoids and flavonoids with strong antioxidative function. Induced phenolics, together with ascorbate, ascorbate peroxidase (APX, EC 1.11.1.11) and catalase (CAT, EC 1.11.1.6) provided efficient defense against potential oxidative pressure. UV-B-induced up-regulation of non-phenolic H2O2 scavengers in green leaf sectors was greater than high PAR-induced changes, indicating a UV-B role in antioxidative defense under light excess; on the contrary, minimal effects were observed in white tissue. However, UV-B radiation had greater influence on phenolics in white leaf sections compared to green ones, inducing accumulation of phenolic glycosides whose function was UV-B screening rather than antioxidative. By stimulation of starch and sucrose breakdown and carbon allocation in the form of soluble sugars from "source" (green) tissue to "sink" (white) tissue, UV-B radiation compensated the absence of photosynthetic activity and phenylpropanoid and flavonoid biosynthesis in white sectors. (C) 2015 Elsevier Masson SAS. All rights reserved.",
publisher = "Elsevier France-Editions Scientifiques Medicales Elsevier, Paris",
journal = "Plant Physiology and Biochemistry",
title = "Carbon allocation from source to sink leaf tissue in relation to flavonoid biosynthesis in variegated Pelargonium zonale under UV-B radiation and high PAR intensity",
volume = "93",
pages = "44-55",
doi = "10.1016/j.plaphy.2015.01.008",
url = "Kon_2873"
}
Vidović, M., Morina, F., Milic, S., Albert, A., Zechmann, B., Tosti, T., Winkler, J. B.,& Veljović-Jovanović, S.. (2015). Carbon allocation from source to sink leaf tissue in relation to flavonoid biosynthesis in variegated Pelargonium zonale under UV-B radiation and high PAR intensity. in Plant Physiology and Biochemistry
Elsevier France-Editions Scientifiques Medicales Elsevier, Paris., 93, 44-55.
https://doi.org/10.1016/j.plaphy.2015.01.008
Kon_2873
Vidović M, Morina F, Milic S, Albert A, Zechmann B, Tosti T, Winkler JB, Veljović-Jovanović S. Carbon allocation from source to sink leaf tissue in relation to flavonoid biosynthesis in variegated Pelargonium zonale under UV-B radiation and high PAR intensity. in Plant Physiology and Biochemistry. 2015;93:44-55.
doi:10.1016/j.plaphy.2015.01.008
Kon_2873 .
Vidović, Marija, Morina, Filis, Milic, Sonja, Albert, Andreas, Zechmann, Bernd, Tosti, Tomislav, Winkler, Jana Barbro, Veljović-Jovanović, Sonja, "Carbon allocation from source to sink leaf tissue in relation to flavonoid biosynthesis in variegated Pelargonium zonale under UV-B radiation and high PAR intensity" in Plant Physiology and Biochemistry, 93 (2015):44-55,
https://doi.org/10.1016/j.plaphy.2015.01.008 .,
Kon_2873 .
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