Impaired expression of the plastidic ferrochelatase by antisense RNA synthesis leads to a necrotic phenotype of transformed tobacco plants

authored by
Jutta Papenbrock, Sanjay Mishra, Hans Peter Mock, Elisabeth Kruse, Eva Kathrin Schmidt, Astrid Petersmann, Hans Peter Braun, Bernhard Grimm
Abstract

Protoporphyrin IX is the last common intermediate of tetrapyrrole biosynthesis. The chelation of a Mg2+ ion by magnesium chelatase and of a ferrous ion by ferrochelatase directs protoporphyrin IX towards the formation of chlorophyll and heme, respectively. A full length cDNA clone encoding a ferrochelatase was identified from a Nicotiana tabacum cDNA library. The encoded protein consists of 497 amino acid residues with a molecular weight of 55.4 kDa. In vitro import of the protein into chloroplasts and its location in stroma and thylakoids confirm its close relationship to the previously described Arabidopsis thaliana plastid-located ferrochelatase (FeChll). A 1700-bp tobacco FeCh cDNA sequence was expressed in Nicotiana tabacum cv. Samsun NN under the control of the CaMV 35S promoter in antisense orientation allowing investigation into the consequences of selective reduction of the plastidic ferrochelatase activity for protoporphyrin IX channeling in chloroplasts and for interactions between plastidic and mitochondrial heme synthesis. Leaves of several transformants showed a reduced chlorophyll content and, during development, a light intensity-dependent formation of necrotic leaf lesions. In comparison with wild-type plants the total ferrochelatase activity was decreased in transgenic lines leading to an accumulation of photosensitizing protoporphyrin IX. Ferrochelatase activity was reduced only in plastids but not in mitochondria of transgenic plants. By means of the specifically diminished ferrochelatase activity consequences of the selective inhibition of protoheme formation for the intracellular supply of heme can be investigated in the future.

Organisation(s)
Institute of Botany
Institute of Plant Genetics
External Organisation(s)
Leibniz Institute of Plant Genetics and Crop Plant Research (IPK)
Humboldt-Universität zu Berlin
Type
Article
Journal
Plant Journal
Volume
28
Pages
41-50
No. of pages
10
ISSN
0960-7412
Publication date
23.12.2001
Publication status
Published
Peer reviewed
Yes
ASJC Scopus subject areas
Genetics, Plant Science, Cell Biology
Electronic version(s)
https://doi.org/10.1046/j.1365-313X.2001.01126.x (Access: Unknown)