Depletion of cellular brassinolide decreases embryo production and disrupts the architecture of the apical meristems in Brassica napus microspore-derived embryos

Exogenous applications of brassinolide (BL) increased the number and quality of microspore-derived embryos (MDEs) whereas treatments with brassinazole (BrZ), a BL biosynthetic inhibitor, had the opposite effect. At the optimal concentration (4x10(-6) M) BrZ decreased both embryo yield and conversion...

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Veröffentlicht in:Journal of experimental botany. - 1985. - 61(2010), 10 vom: 18. Juni, Seite 2779-94
1. Verfasser: Belmonte, Mark (VerfasserIn)
Weitere Verfasser: Elhiti, Mohamed, Waldner, Blaine, Stasolla, Claudio
Format: Online-Aufsatz
Sprache:English
Veröffentlicht: 2010
Zugriff auf das übergeordnete Werk:Journal of experimental botany
Schlagworte:Journal Article Research Support, Non-U.S. Gov't Biomarkers Brassinosteroids Cholestanols Steroids, Heterocyclic Triazoles Glutathione GAN16C9B8O brassinazole mehr... N9XRW3TF90 Ascorbic Acid PQ6CK8PD0R brassinolide Y9IQ1L53OX
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100 1 |a Belmonte, Mark  |e verfasserin  |4 aut 
245 1 0 |a Depletion of cellular brassinolide decreases embryo production and disrupts the architecture of the apical meristems in Brassica napus microspore-derived embryos 
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520 |a Exogenous applications of brassinolide (BL) increased the number and quality of microspore-derived embryos (MDEs) whereas treatments with brassinazole (BrZ), a BL biosynthetic inhibitor, had the opposite effect. At the optimal concentration (4x10(-6) M) BrZ decreased both embryo yield and conversion to less than half the value of control embryos. Metabolic studies revealed that BL levels had profound effects on glutathione and ascorbate metabolism by altering the amounts of their reduced forms (ASC and GSH) and oxidized forms [dehydroascorbate (DHA), ascorbate free radicals (AFRs), and GSSG]. Applications of BL switched the glutathione and ascorbate pools towards the oxidized forms, thereby lowering the ASC/ASC+DHA+AFR and GSH/GSH+GSSG ratios. These changes were ascribed to the ability of BL to increase the activity of ascorbate peroxidase (APX) and decrease that of glutathione reductase (GR). This trend was reversed in a BL-depleted environment, effected by BrZ applications. These metabolic alterations were associated with changes in embryo structure and performance. BL-treated MDEs developed zygotic-like shoot apical meristems (SAMs) whereas embryos treated with BrZ developed abnormal meristems. In the presence of BrZ, embryos either lacked a visible SAM, or formed SAMs in which the meristematic cells showed signs of differentiation, such as vacuolation and storage product accumulation. These abnormalities were accompanied by the lack or misexpression of three meristem marker genes isolated from Brassica napus (denoted as BnSTM, BnCLV1, and BnZLL-1) homologous to the Arabidopsis SHOOTMERISTEMLESS (STM), CLAVATA 1 (CLV1), and ZWILLE (ZLL). The expression of BnSTM and BnCLV1 increased after a few days in cultures in embryos treated with BL whereas an opposite tendency was observed with applications of BrZ. Compared with control embryos where these two genes exhibited abnormal localization patterns, BnSTM and BnCLV1 always localized throughout the subapical domains of BL-treated embryos in a zygotic-like fashion. Expression of both genes was often lost in the SAM of BrZ-treated embryos. The results suggest that maintenance of cellular BL levels is required to modulate the ascorbate and glutathione redox status during embryogenesis to ensure proper development of the embryos and formation of functional apical meristems 
650 4 |a Journal Article 
650 4 |a Research Support, Non-U.S. Gov't 
650 7 |a Biomarkers  |2 NLM 
650 7 |a Brassinosteroids  |2 NLM 
650 7 |a Cholestanols  |2 NLM 
650 7 |a Steroids, Heterocyclic  |2 NLM 
650 7 |a Triazoles  |2 NLM 
650 7 |a Glutathione  |2 NLM 
650 7 |a GAN16C9B8O  |2 NLM 
650 7 |a brassinazole  |2 NLM 
650 7 |a N9XRW3TF90  |2 NLM 
650 7 |a Ascorbic Acid  |2 NLM 
650 7 |a PQ6CK8PD0R  |2 NLM 
650 7 |a brassinolide  |2 NLM 
650 7 |a Y9IQ1L53OX  |2 NLM 
700 1 |a Elhiti, Mohamed  |e verfasserin  |4 aut 
700 1 |a Waldner, Blaine  |e verfasserin  |4 aut 
700 1 |a Stasolla, Claudio  |e verfasserin  |4 aut 
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773 1 8 |g volume:61  |g year:2010  |g number:10  |g day:18  |g month:06  |g pages:2779-94 
856 4 0 |u http://dx.doi.org/10.1093/jxb/erq110  |3 Volltext 
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