Počet záznamů: 1
Pollen and Pollen Tube Biology
- 1.
SYSNO ASEP 0541115 Druh ASEP M - Kapitola v monografii Zařazení RIV C - Kapitola v knize Název Isolation of the pistil-stimulated pollen tube secretome Tvůrce(i) Hafidh, Said (UEB-Q) RID, ORCID
Honys, David (UEB-Q) RID, ORCIDCelkový počet autorů 2 Zdroj.dok. Pollen and Pollen Tube Biology, Methods in Molecular Biology. - New York : Humana Press, 2020 / Geitmann A. - ISSN 1064-3745 - ISBN 978-1-0716-0671-1 Rozsah stran s. 41-72 Poč.str. 32 s. Poč.výt. 2400 Poč.str.knihy 332 Forma vydání Tištěná - P Jazyk dok. eng - angličtina Země vyd. US - Spojené státy americké Klíč. slova Cell–cell communication ; Glycosylation ; lc-ms/ms ; Pollen tube guidance ; Protein secretion ; Sexual plant reproduction Vědní obor RIV EB - Genetika a molekulární biologie Obor OECD Biochemical research methods CEP LM2018129 GA MŠMT - Ministerstvo školství, mládeže a tělovýchovy GA17-23203S GA ČR - Grantová agentura ČR GA18-02448S GA ČR - Grantová agentura ČR GA18-07027S GA ČR - Grantová agentura ČR EF16_019/0000738 GA MŠMT - Ministerstvo školství, mládeže a tělovýchovy Institucionální podpora UEB-Q - RVO:61389030 UT WOS 000680070700005 EID SCOPUS 85086424545 DOI 10.1007/978-1-0716-0672-8_4 Anotace Detection of secreted proteins and peptides during pollen tube guidance has been impeded due to lack of techniques to capture the pollen tube secretome without contamination from the female secreted proteins. Here we present a protocol to detect tobacco pollen tube secreted proteins, semi-in vivo pollen tube secretome assay (SIV-PS), following pollen tube crosstalk with the female reproductive tissues. This method combines the advantages of in vivo pollen tube–pistil interaction and filter-aided sample preparation (FASP) techniques to obtain an in-depth proteome coverage. The SIV-PS method is rapid, efficient, inexpensive, does not require specialized equipment or expertise, and provides a snapshot of the ongoing molecular interplay. We show that the secretome obtained is of greater purity (<,1.4% ADH activities) and that pollen tubes are physiologically and cytologically unaffected. A compendium of quality controls is described and a rough guide on downstream bioinformatics analysis is outlined. The SIV-PS method is applicable to all studies of protein secretion using pollen tube as a model and can be easily adapted to other flowering species with modification. The overall duration for this protocol is approximately 8 hours spanning 4 days (an average of 2 h/day per two workers) excluding microscopy and LC-MS/MS analysis. Pracoviště Ústav experimentální botaniky Kontakt David Klier, knihovna@ueb.cas.cz, Tel.: 220 390 469 Rok sběru 2021 Elektronická adresa http://doi.org/10.1007/978-1-0716-0672-8_4
Počet záznamů: 1