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Isolation and characterization of CaMF3, an anther-specific gene in Capsicum annuum L.

Hao X, Chen C, Chen G, Cao B, Chen Q, Lei J - Genet. Mol. Biol. (2012)

Bottom Line: The predicted protein of CaMF3 shared sequence similarity with members of the isoamyl acetate-hydrolyzing esterase (IAH1) protein family.Fine expression analysis revealed that CaMF3 was expressed specifically in anthers of the fertile line.These results suggest that CaMF3 is an anther-specific gene that may be essential for anther or pollen development in C. annuum.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, Taiyuan Normal University, Taiyuan, Shanxi Province, China. ; College of Horticulture, South China Agricultural University, Guangzhou, Guangdong Province, China.

ABSTRACT
Previous work on gene expression analysis based on RNA sequencing identified a variety of differentially expressed cDNA fragments in the genic male sterile-fertile line 114AB of Capsicum annuum L. In this work, we examined the accumulation of one of the transcript-derived fragments (TDFs), CaMF3 (male fertile 3), in the flower buds of a fertile line. The full genomic DNA sequence of CaMF3 was 1,951 bp long and contained 6 exons and 5 introns, with the complete sequence encoding a putative 25.89 kDa protein of 234 amino acids. The predicted protein of CaMF3 shared sequence similarity with members of the isoamyl acetate-hydrolyzing esterase (IAH1) protein family. CaMF3 expression was detected only in flower buds at stages 7 and 8 and in open flowers of a male fertile line; no expression was observed in any organs of a male sterile line. Fine expression analysis revealed that CaMF3 was expressed specifically in anthers of the fertile line. These results suggest that CaMF3 is an anther-specific gene that may be essential for anther or pollen development in C. annuum.

No MeSH data available.


Related in: MedlinePlus

The full-length cDNA and deduced amino acid sequence of CaMF3 (accession number JN975046). The start codon (ATG) and stop codon (TGA) are shown in bold. The nucleotide sequences of primers for the full-length cDNA and DNA amplification are underlined. Boxed amino acids indicate the Ser12/Asp189/His192 sequence in its catalytic triad. The triangles indicate three oxyanion holes (specific amino acids: Ser12, Gly44 and Asn78).
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f1-gmb-35-810: The full-length cDNA and deduced amino acid sequence of CaMF3 (accession number JN975046). The start codon (ATG) and stop codon (TGA) are shown in bold. The nucleotide sequences of primers for the full-length cDNA and DNA amplification are underlined. Boxed amino acids indicate the Ser12/Asp189/His192 sequence in its catalytic triad. The triangles indicate three oxyanion holes (specific amino acids: Ser12, Gly44 and Asn78).

Mentions: The full-size cDNA of CaMF3 was 974 bp long and contained a 705-bp open reading frame (Figure 1). The genomic DNA of CaMF3 was isolated using primer pairs that were used to clone the cDNA sequence. Sequencing results showed that the DNA sequence was 1,951 bp long and contained five introns (96, 541, 133, 94 and 113 bp long) (Figure 2A). The complete coding sequence encoded a putative 25.89 kDa protein of 234 amino acids with a theoretical pI of 6.62. The SignalP3.0 program was used to detect signal peptide sequences. However, no significant signal peptide was found in the amino acid sequence of CaMF3. Hydrophobic character prediction based on the ProtParam tool indicated that CaMF3 was hydrophilic with a grand average of hydropathicity (GRAVY) of −0.145. The instability index (II) was calculated to be 37.53, which classified the protein as stable. The deduced protein contained an isoamyl acetate hydrolase-like domain (amino acids 3–212) (cd01838, NCBI Conserved Domain Database) (Wei et al., 1995; Marchler-Bauer et al., 2011), indicating that CaMF3 belongs to this protein family (Wei et al., 1995). The protein also contained three conserved features/sites: the active site (Ser12, Gly44, Asn78, Asp189 and His192), the typical catalytic triad (Ser12, Asp189 and His192) and three oxyanion holes (amino acids Ser12, Gly44 and Asn78) (Figure 2B).


Isolation and characterization of CaMF3, an anther-specific gene in Capsicum annuum L.

Hao X, Chen C, Chen G, Cao B, Chen Q, Lei J - Genet. Mol. Biol. (2012)

The full-length cDNA and deduced amino acid sequence of CaMF3 (accession number JN975046). The start codon (ATG) and stop codon (TGA) are shown in bold. The nucleotide sequences of primers for the full-length cDNA and DNA amplification are underlined. Boxed amino acids indicate the Ser12/Asp189/His192 sequence in its catalytic triad. The triangles indicate three oxyanion holes (specific amino acids: Ser12, Gly44 and Asn78).
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3526090&req=5

f1-gmb-35-810: The full-length cDNA and deduced amino acid sequence of CaMF3 (accession number JN975046). The start codon (ATG) and stop codon (TGA) are shown in bold. The nucleotide sequences of primers for the full-length cDNA and DNA amplification are underlined. Boxed amino acids indicate the Ser12/Asp189/His192 sequence in its catalytic triad. The triangles indicate three oxyanion holes (specific amino acids: Ser12, Gly44 and Asn78).
Mentions: The full-size cDNA of CaMF3 was 974 bp long and contained a 705-bp open reading frame (Figure 1). The genomic DNA of CaMF3 was isolated using primer pairs that were used to clone the cDNA sequence. Sequencing results showed that the DNA sequence was 1,951 bp long and contained five introns (96, 541, 133, 94 and 113 bp long) (Figure 2A). The complete coding sequence encoded a putative 25.89 kDa protein of 234 amino acids with a theoretical pI of 6.62. The SignalP3.0 program was used to detect signal peptide sequences. However, no significant signal peptide was found in the amino acid sequence of CaMF3. Hydrophobic character prediction based on the ProtParam tool indicated that CaMF3 was hydrophilic with a grand average of hydropathicity (GRAVY) of −0.145. The instability index (II) was calculated to be 37.53, which classified the protein as stable. The deduced protein contained an isoamyl acetate hydrolase-like domain (amino acids 3–212) (cd01838, NCBI Conserved Domain Database) (Wei et al., 1995; Marchler-Bauer et al., 2011), indicating that CaMF3 belongs to this protein family (Wei et al., 1995). The protein also contained three conserved features/sites: the active site (Ser12, Gly44, Asn78, Asp189 and His192), the typical catalytic triad (Ser12, Asp189 and His192) and three oxyanion holes (amino acids Ser12, Gly44 and Asn78) (Figure 2B).

Bottom Line: The predicted protein of CaMF3 shared sequence similarity with members of the isoamyl acetate-hydrolyzing esterase (IAH1) protein family.Fine expression analysis revealed that CaMF3 was expressed specifically in anthers of the fertile line.These results suggest that CaMF3 is an anther-specific gene that may be essential for anther or pollen development in C. annuum.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, Taiyuan Normal University, Taiyuan, Shanxi Province, China. ; College of Horticulture, South China Agricultural University, Guangzhou, Guangdong Province, China.

ABSTRACT
Previous work on gene expression analysis based on RNA sequencing identified a variety of differentially expressed cDNA fragments in the genic male sterile-fertile line 114AB of Capsicum annuum L. In this work, we examined the accumulation of one of the transcript-derived fragments (TDFs), CaMF3 (male fertile 3), in the flower buds of a fertile line. The full genomic DNA sequence of CaMF3 was 1,951 bp long and contained 6 exons and 5 introns, with the complete sequence encoding a putative 25.89 kDa protein of 234 amino acids. The predicted protein of CaMF3 shared sequence similarity with members of the isoamyl acetate-hydrolyzing esterase (IAH1) protein family. CaMF3 expression was detected only in flower buds at stages 7 and 8 and in open flowers of a male fertile line; no expression was observed in any organs of a male sterile line. Fine expression analysis revealed that CaMF3 was expressed specifically in anthers of the fertile line. These results suggest that CaMF3 is an anther-specific gene that may be essential for anther or pollen development in C. annuum.

No MeSH data available.


Related in: MedlinePlus