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New insights into plant glycoside hydrolase family 32 in Agave species.

Avila de Dios E, Gomez Vargas AD, Damián Santos ML, Simpson J - Front Plant Sci (2015)

Bottom Line: Notably however neither 1-FFT nor 6-SFT encoding cDNAs were identified.In silico analysis revealed that distinct isoforms for certain enzymes found in a single species, showed different levels and tissue specific patterns of expression whereas in other cases expression patterns were conserved both within the species and between different species.Thin layer chromatography confirmed the presence of fructans with degree of polymerization (DP) greater than DP three in both immature buds and fully opened flowers also obtained from A. tequilana.

View Article: PubMed Central - PubMed

Affiliation: Department of Genetic Engineering, Centro de Investigación y Estudios Avanzados Irapuato, Mexico.

ABSTRACT
In order to optimize the use of agaves for commercial applications, an understanding of fructan metabolism in these species at the molecular and genetic level is essential. Based on transcriptome data, this report describes the identification and molecular characterization of cDNAs and deduced amino acid sequences for genes encoding fructosyltransferases, invertases and fructan exohydrolases (FEH) (enzymes belonging to plant glycoside hydrolase family 32) from four different agave species (A. tequilana, A. deserti, A. victoriae-reginae, and A. striata). Conserved amino acid sequences and a hypervariable domain allowed classification of distinct isoforms for each enzyme type. Notably however neither 1-FFT nor 6-SFT encoding cDNAs were identified. In silico analysis revealed that distinct isoforms for certain enzymes found in a single species, showed different levels and tissue specific patterns of expression whereas in other cases expression patterns were conserved both within the species and between different species. Relatively high levels of in silico expression for specific isoforms of both invertases and fructosyltransferases were observed in floral tissues in comparison to vegetative tissues such as leaves and stems and this pattern was confirmed by Quantitative Real Time PCR using RNA obtained from floral and leaf tissue of A. tequilana. Thin layer chromatography confirmed the presence of fructans with degree of polymerization (DP) greater than DP three in both immature buds and fully opened flowers also obtained from A. tequilana.

No MeSH data available.


Comparison of conserved amino acid motifs specific to PGHF32. Motifs are indicated above each column. Dark green squares indicate residues conserved in all sequences.
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Figure 2: Comparison of conserved amino acid motifs specific to PGHF32. Motifs are indicated above each column. Dark green squares indicate residues conserved in all sequences.

Mentions: The conserved motifs close to the active sight that characterize PGHF32 are shown in Figure 2. The FRDP motif is not displayed since this motif was perfectly conserved in all sequences analyzed. In general the WMNDNPG, WSGSAT, ILYTGG, WECPD (WECVD), and GWAS motifs are well conserved within all PGHF32 members from the four Agave species analyzed. Differences observed previously for the WMNDNPG and WSGSAT motifs in fructosyltransferases of A. tequilana were confirmed and also shown to be present in other agave species. The undefined invertase group (clade c in Figure 1) shows no specific pattern of amino acid conservation for these motifs.


New insights into plant glycoside hydrolase family 32 in Agave species.

Avila de Dios E, Gomez Vargas AD, Damián Santos ML, Simpson J - Front Plant Sci (2015)

Comparison of conserved amino acid motifs specific to PGHF32. Motifs are indicated above each column. Dark green squares indicate residues conserved in all sequences.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4524927&req=5

Figure 2: Comparison of conserved amino acid motifs specific to PGHF32. Motifs are indicated above each column. Dark green squares indicate residues conserved in all sequences.
Mentions: The conserved motifs close to the active sight that characterize PGHF32 are shown in Figure 2. The FRDP motif is not displayed since this motif was perfectly conserved in all sequences analyzed. In general the WMNDNPG, WSGSAT, ILYTGG, WECPD (WECVD), and GWAS motifs are well conserved within all PGHF32 members from the four Agave species analyzed. Differences observed previously for the WMNDNPG and WSGSAT motifs in fructosyltransferases of A. tequilana were confirmed and also shown to be present in other agave species. The undefined invertase group (clade c in Figure 1) shows no specific pattern of amino acid conservation for these motifs.

Bottom Line: Notably however neither 1-FFT nor 6-SFT encoding cDNAs were identified.In silico analysis revealed that distinct isoforms for certain enzymes found in a single species, showed different levels and tissue specific patterns of expression whereas in other cases expression patterns were conserved both within the species and between different species.Thin layer chromatography confirmed the presence of fructans with degree of polymerization (DP) greater than DP three in both immature buds and fully opened flowers also obtained from A. tequilana.

View Article: PubMed Central - PubMed

Affiliation: Department of Genetic Engineering, Centro de Investigación y Estudios Avanzados Irapuato, Mexico.

ABSTRACT
In order to optimize the use of agaves for commercial applications, an understanding of fructan metabolism in these species at the molecular and genetic level is essential. Based on transcriptome data, this report describes the identification and molecular characterization of cDNAs and deduced amino acid sequences for genes encoding fructosyltransferases, invertases and fructan exohydrolases (FEH) (enzymes belonging to plant glycoside hydrolase family 32) from four different agave species (A. tequilana, A. deserti, A. victoriae-reginae, and A. striata). Conserved amino acid sequences and a hypervariable domain allowed classification of distinct isoforms for each enzyme type. Notably however neither 1-FFT nor 6-SFT encoding cDNAs were identified. In silico analysis revealed that distinct isoforms for certain enzymes found in a single species, showed different levels and tissue specific patterns of expression whereas in other cases expression patterns were conserved both within the species and between different species. Relatively high levels of in silico expression for specific isoforms of both invertases and fructosyltransferases were observed in floral tissues in comparison to vegetative tissues such as leaves and stems and this pattern was confirmed by Quantitative Real Time PCR using RNA obtained from floral and leaf tissue of A. tequilana. Thin layer chromatography confirmed the presence of fructans with degree of polymerization (DP) greater than DP three in both immature buds and fully opened flowers also obtained from A. tequilana.

No MeSH data available.