Limits...
Expression profile analysis of genes involved in cell wall regeneration during protoplast culture in cotton by suppression subtractive hybridization and macroarray.

Yang X, Tu L, Zhu L, Fu L, Min L, Zhang X - J. Exp. Bot. (2008)

Bottom Line: As confirmed by reverse-transcription PCR (RT-PCR) and real-time quantitative reverse-transcription PCR (QRT-PCR) analysis, the selected genes displayed complex expression patterns during cell wall regeneration from protoplasts and included most previously published cell-wall-associated genes.Our findings also highlighted the function of some transcription factors during cell wall regeneration from protoplasts, including the squamosa promoter binding protein-like 14 (SPL14), NAC, Gbiaa-re, MYB, WRKY, swellmap 1 (SMP1), RAD5, and zinc finger family protein, as well as the enrichment of Ca(2+)-calmodulin signal molecules.On the basis of the gene expression profiles, a model of cell wall regeneration from protoplasts derived from cotton suspension cultures is proposed.

View Article: PubMed Central - PubMed

Affiliation: National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan, Hubei 430070, PR China.

ABSTRACT
The molecular mechanisms underlying cell wall biosynthesis are poorly understood. In this study, microscopic analysis showed that protoplasts generated a new cell wall within 48 h after transfer to a wall-regeneration medium. To identify genes related to cell wall biosynthesis in cotton, suppression subtractive hybridization was used to visualize differential gene expression at seven time points within the first 48 h. In total, 412 differentially expressed sequence tags (ESTs; >3-fold) were identified, and 210 unigenes were sequenced successfully. As confirmed by reverse-transcription PCR (RT-PCR) and real-time quantitative reverse-transcription PCR (QRT-PCR) analysis, the selected genes displayed complex expression patterns during cell wall regeneration from protoplasts and included most previously published cell-wall-associated genes. ESTs similar to cell-wall-protein genes, such as proline-rich protein (PRPL), glycine-rich protein (GRP), extension (EPR1), fasciclin-like arabinogalactan protein (FLA2), and expensing-like protein (EXLA and EXLB), which might participate in primary cell wall or secondary cell wall construction and modification, were up-regulated during cell wall regeneration from protoplasts. Sucrose synthase, an important enzyme in the sugar signalling pathway, played important roles in cellulose biosynthesis. Our findings also highlighted the function of some transcription factors during cell wall regeneration from protoplasts, including the squamosa promoter binding protein-like 14 (SPL14), NAC, Gbiaa-re, MYB, WRKY, swellmap 1 (SMP1), RAD5, and zinc finger family protein, as well as the enrichment of Ca(2+)-calmodulin signal molecules. On the basis of the gene expression profiles, a model of cell wall regeneration from protoplasts derived from cotton suspension cultures is proposed.

Show MeSH

Related in: MedlinePlus

Model of cell wall regeneration from cotton protoplast based on gene expression patterns.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC2561149&req=5

fig7: Model of cell wall regeneration from cotton protoplast based on gene expression patterns.

Mentions: Ideally, a study of the process of cell wall biosynthesis should be carried out in a synchronous system with a clearly defined time point at which cell wall biosynthesis is initiated. Milioni et al. (2001) showed that the protoplast system was a highly synchronized system that allowed the selection of specific time points for transcript profiling. The choice of this system in the present study was based on the expectation that strongly enhanced cell wall regeneration would derive from markedly increased expression levels of the genes involved, and more than 90% of protoplasts regenerated a new cell wall within 48 h of culture. Unlike Oomen et al. (2003), it was possible to isolate many genes related to cell wall biosynthesis. Although cellulose synthase and callose/glucan synthase were not isolated in our library, possibly because primary cell wall synthesis basically takes place in all young growth tissues, a model of cell wall regeneration in protoplasts derived from cotton suspension cultures according to the expression profile has been proposed (Fig. 7).


Expression profile analysis of genes involved in cell wall regeneration during protoplast culture in cotton by suppression subtractive hybridization and macroarray.

Yang X, Tu L, Zhu L, Fu L, Min L, Zhang X - J. Exp. Bot. (2008)

Model of cell wall regeneration from cotton protoplast based on gene expression patterns.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC2561149&req=5

fig7: Model of cell wall regeneration from cotton protoplast based on gene expression patterns.
Mentions: Ideally, a study of the process of cell wall biosynthesis should be carried out in a synchronous system with a clearly defined time point at which cell wall biosynthesis is initiated. Milioni et al. (2001) showed that the protoplast system was a highly synchronized system that allowed the selection of specific time points for transcript profiling. The choice of this system in the present study was based on the expectation that strongly enhanced cell wall regeneration would derive from markedly increased expression levels of the genes involved, and more than 90% of protoplasts regenerated a new cell wall within 48 h of culture. Unlike Oomen et al. (2003), it was possible to isolate many genes related to cell wall biosynthesis. Although cellulose synthase and callose/glucan synthase were not isolated in our library, possibly because primary cell wall synthesis basically takes place in all young growth tissues, a model of cell wall regeneration in protoplasts derived from cotton suspension cultures according to the expression profile has been proposed (Fig. 7).

Bottom Line: As confirmed by reverse-transcription PCR (RT-PCR) and real-time quantitative reverse-transcription PCR (QRT-PCR) analysis, the selected genes displayed complex expression patterns during cell wall regeneration from protoplasts and included most previously published cell-wall-associated genes.Our findings also highlighted the function of some transcription factors during cell wall regeneration from protoplasts, including the squamosa promoter binding protein-like 14 (SPL14), NAC, Gbiaa-re, MYB, WRKY, swellmap 1 (SMP1), RAD5, and zinc finger family protein, as well as the enrichment of Ca(2+)-calmodulin signal molecules.On the basis of the gene expression profiles, a model of cell wall regeneration from protoplasts derived from cotton suspension cultures is proposed.

View Article: PubMed Central - PubMed

Affiliation: National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan, Hubei 430070, PR China.

ABSTRACT
The molecular mechanisms underlying cell wall biosynthesis are poorly understood. In this study, microscopic analysis showed that protoplasts generated a new cell wall within 48 h after transfer to a wall-regeneration medium. To identify genes related to cell wall biosynthesis in cotton, suppression subtractive hybridization was used to visualize differential gene expression at seven time points within the first 48 h. In total, 412 differentially expressed sequence tags (ESTs; >3-fold) were identified, and 210 unigenes were sequenced successfully. As confirmed by reverse-transcription PCR (RT-PCR) and real-time quantitative reverse-transcription PCR (QRT-PCR) analysis, the selected genes displayed complex expression patterns during cell wall regeneration from protoplasts and included most previously published cell-wall-associated genes. ESTs similar to cell-wall-protein genes, such as proline-rich protein (PRPL), glycine-rich protein (GRP), extension (EPR1), fasciclin-like arabinogalactan protein (FLA2), and expensing-like protein (EXLA and EXLB), which might participate in primary cell wall or secondary cell wall construction and modification, were up-regulated during cell wall regeneration from protoplasts. Sucrose synthase, an important enzyme in the sugar signalling pathway, played important roles in cellulose biosynthesis. Our findings also highlighted the function of some transcription factors during cell wall regeneration from protoplasts, including the squamosa promoter binding protein-like 14 (SPL14), NAC, Gbiaa-re, MYB, WRKY, swellmap 1 (SMP1), RAD5, and zinc finger family protein, as well as the enrichment of Ca(2+)-calmodulin signal molecules. On the basis of the gene expression profiles, a model of cell wall regeneration from protoplasts derived from cotton suspension cultures is proposed.

Show MeSH
Related in: MedlinePlus