Requirement of proline synthesis during Arabidopsis reproductive development.
BACKGROUND
Gamete and embryo development are crucial for successful reproduction and seed set in plants, which is often the determining factor for crop yield. Proline accumulation was largely viewed as a specific reaction to overcome stress conditions, while recent studies suggested important functions of proline metabolism also in reproductive development. Both the level of free proline and proline metabolism were proposed to influence the transition to flowering, as well as pollen and embryo development.


RESULTS
In this study, we performed a detailed analysis of the contribution of individual proline biosynthetic enzymes to vegetative development and reproductive success in Arabidopsis. In contrast to previous reports, we found that pyrroline-5-carboxylate (P5C) synthetase 2 (P5CS2) is not essential for sexual reproduction although p5cs2 mutant plants were retarded in vegetative development and displayed reduced fertility under long-day conditions. Single mutant plants devoid of P5CS1 did not show any developmental defects. Simultaneous absence of both P5CS isoforms resulted in pollen sterility, while fertile egg cells could still be produced. Expression of P5C reductase (P5CR) was indispensable for embryo development but surprisingly not needed for pollen or egg cell fertility. The latter observation could be explained by an extreme stability of P5CR activity, which had a half-life time of greater than 3 weeks in vitro. Expression of P5CR-GFP under the control of the endogenous P5CR promoter was able to restore growth of homozygous p5cr mutant embryos. The analysis of P5CR-GFP-fluorescence in planta supported an exclusively cytoplasmatic localisation of P5CR.


CONCLUSIONS
Our results demonstrate that potential alternative pathways for proline synthesis or inter-generation transfer of proline are not sufficient to overcome a defect in proline biosynthesis from glutamate during pollen development. Proline biosynthesis through P5CS2 and P5CR is limiting for vegetative and reproductive development in Arabidopsis, whereas disruption of P5CS1 alone does not affect development of non-stressed plants.
Department of Plant Physiology and Biochemistry Biology Section, University of Konstanz, Universitätsstraße 10, 78464 Konstanz, Germany. dietmar.funck@uni-konstanz.de
Dietmar
D
Funck
Gudrun
G
Winter
Lukas
L
Baumgarten
Giuseppe
G
Forlani
D017360:Arabidopsis / Q000201:enzymology / Q000235:genetics / Q000254:growth & development*; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D002734:Chlorophyll / Q000378:metabolism; D003593:Cytoplasm / Q000235:genetics / Q000378:metabolism; D004789:Enzyme Activation; D057075:Enzyme Assays; D005298:Fertility; D005453:Fluorescence; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006207:Half-Life; D018613:Microscopy, Confocal; D051479:Plant Infertility; D011058:Pollen / Q000201:enzymology / Q000235:genetics / Q000254:growth & development; D011392:Proline / Q000096:biosynthesis* / Q000235:genetics; D011401:Promoter Regions, Genetic; D055550:Protein Stability; D011762:Pyrroline Carboxylate Reductases / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012639:Seeds / Q000201:enzymology / Q000235:genetics / Q000254:growth & development; D000097502:delta-1-Pyrroline-5-Carboxylate Reductase
D029681:Arabidopsis Proteins; D002734:Chlorophyll; D049452:Green Fluorescent Proteins; D011392:Proline; D011762:Pyrroline Carboxylate Reductases; D011993:Recombinant Fusion Proteins; D000097502:delta-1-Pyrroline-5-Carboxylate Reductase