Optimisation of Immunohistochemical Detection of Cytokinins in
Transgenic Arabidopsis Seedlings Expressing Activated ipt, a
Cytokinin Biosynthesis Gene

D 2002

Optimisation of Immunohistochemical Detection of Cytokinins in Transgenic Arabidopsis Seedlings Expressing Activated ipt, a Cytokinin Biosynthesis Gene

DUBOVÁ, Jaroslava, Julie Olegivna ROTKOVSKÁ, Alena REKOVÁ and Břetislav BRZOBOHATÝ

Basic information

Original name

Optimisation of Immunohistochemical Detection of Cytokinins in Transgenic Arabidopsis Seedlings Expressing Activated ipt, a Cytokinin Biosynthesis Gene

Name in Czech

Optimalizace imunohistochemické detekce cytokininů u transgenních klíčních rostlin Arabidopsis exprimujících ipt, gen pro biosyntézu cytokininů

Authors

DUBOVÁ, Jaroslava (203 Czech Republic), Julie Olegivna ROTKOVSKÁ (804 Ukraine), Alena REKOVÁ (203 Czech Republic) and Břetislav BRZOBOHATÝ (203 Czech Republic, guarantor)

Edition

Brno, Progress in Basic, Applied and Diagnostic Histochemistry, p. 15-15, 2002

Publisher

Masarykova univerzita v Brně

Other information

Language

English

Type of outcome

Stať ve sborníku

Field of Study

10600 1.6 Biological sciences

Country of publisher

Czech Republic

Confidentiality degree

není předmětem státního či obchodního tajemství

RIV identification code

RIV/00216224:14310/02:00006761

Organization unit

Faculty of Science

ISBN

80-210-2998-6

Keywords in English

cytokinins; immunohistochemistry; Arabidopsis

Abstract

V originále

Aseptically grown Arabidopsis seedlings were collected, fixed in PFA - GA mixture. Indirect immunohistochemistry on cryostat sections was used. Primary antibodies against zeatin-riboside and isopentenyladenine-riboside were used respectively. Goat anti-rabbit IgG conjugated with AP was used as a secondary antibody. AP activity was detected by reaction with BCIP and NBT. The best results of immunostaining were obtained by higher dilution of primary antibodies and lowering the concentration of BCIP and NBT. Arabidopsis seedlings with activated ipt showed higher the amount of reaction product in comparison with the wild type plants. Accumulation of cytokinins was established immunohistochemically during culture period in transgenic Arabidopsis seedlings, which is in accordance with the biochemical analyses. Modified immunohistochemical detection of free bases of cytokinins can be used for the semi quantitative evaluation of cytokinin content in Arabidopsis seedlings.

Links

IAA5004001, research and development project

Name: Systém aktivace transkripce pro studium vztahu mezi metabolizmem a účinkem cytokininů u Arabidopsis a tabáku

Investor: Academy of Sciences of the Czech Republic, Transkription activation system for studying the relationship between cytokinin metabolism and action in Arabidopsis and tobacco

Citovat

DUBOVÁ, Jaroslava, Julie Olegivna ROTKOVSKÁ, Alena REKOVÁ and Břetislav BRZOBOHATÝ. Optimisation of Immunohistochemical Detection of Cytokinins in Transgenic Arabidopsis Seedlings Expressing Activated ipt, a Cytokinin Biosynthesis Gene. In Progress in Basic, Applied and Diagnostic Histochemistry. Brno: Masarykova univerzita v Brně, 2002, p. 15. ISBN 80-210-2998-6.

@inproceedings{406823,
   author = {Dubová, Jaroslava and Rotkovská, Julie Olegivna and Reková, Alena and Brzobohatý, Břetislav},
   address = {Brno},
   booktitle = {Progress in Basic, Applied and Diagnostic Histochemistry},
   keywords = {cytokinins; immunohistochemistry; Arabidopsis},
   language = {eng},
   location = {Brno},
   isbn = {80-210-2998-6},
   pages = {15-15},
   publisher = {Masarykova univerzita v Brně},
   title = {Optimisation of Immunohistochemical Detection of Cytokinins in Transgenic Arabidopsis Seedlings Expressing Activated ipt, a Cytokinin Biosynthesis Gene},
   year = {2002}
}

TY  - JOUR
ID  - 406823
AU  - Dubová, Jaroslava - Rotkovská, Julie Olegivna - Reková, Alena - Brzobohatý, Břetislav
PY  - 2002
TI  - Optimisation of Immunohistochemical Detection of Cytokinins in Transgenic Arabidopsis Seedlings Expressing Activated ipt, a Cytokinin Biosynthesis Gene
PB  - Masarykova univerzita v Brně
CY  - Brno
SN  - 8021029986
KW  - cytokinins
KW  - immunohistochemistry
KW  - Arabidopsis
N2  - Aseptically grown Arabidopsis seedlings were collected, fixed in PFA - GA mixture. Indirect immunohistochemistry on cryostat sections was used. Primary antibodies against zeatin-riboside and isopentenyladenine-riboside were used respectively. Goat anti-rabbit IgG conjugated with AP was used as a secondary antibody. AP activity was detected by reaction with BCIP and NBT. The best results of immunostaining were obtained by higher dilution of primary antibodies and lowering the concentration of BCIP and NBT. Arabidopsis seedlings with activated ipt showed higher the amount of reaction product in comparison with the wild type plants. Accumulation of cytokinins was established immunohistochemically during culture period in transgenic Arabidopsis seedlings, which is in accordance with the biochemical analyses. Modified immunohistochemical detection of free bases of cytokinins can be used for the semi quantitative evaluation of cytokinin content in Arabidopsis seedlings.
ER  -

DUBOVÁ, Jaroslava, Julie Olegivna ROTKOVSKÁ, Alena REKOVÁ and Břetislav BRZOBOHATÝ. Optimisation of Immunohistochemical Detection of Cytokinins in Transgenic Arabidopsis Seedlings Expressing Activated ipt, a Cytokinin Biosynthesis Gene. In \textit{Progress in Basic, Applied and Diagnostic Histochemistry}. Brno: Masarykova univerzita v Brně, 2002, p.~15. ISBN~80-210-2998-6.

Detailed Information on Publication Record