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BcMF26a and BcMF26b Are Duplicated Polygalacturonase Genes with Divergent Expression Patterns and Functions in Pollen Development and Pollen Tube Formation in Brassica campestris.

Lyu M, Yu Y, Jiang J, Song L, Liang Y, Ma Z, Xiong X, Cao J - PLoS ONE (2015)

Bottom Line: We found that these two genes were orthologous genes of At4g33440, and they originated from a chromosomal segmental duplication.Promoter activity analysis showed that, at reproductive development stages, BcMF26b promoter was active in tapetum, pollen grains, and pistils, whereas BcMF26a promoter was only active in pistils.In the subcellular localization experiment, BcMF26a and BcMF26b proteins could be localized to the cell wall.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Cell & Molecular Biology, Institute of Vegetable Science, Zhejiang University, Hangzhou, China; Zhejiang Provincial Key Laboratory of Horticultural Plant Integrative Biology, Zhejiang University, Hangzhou, China.

ABSTRACT
Polygalacturonase (PG) is one of the cell wall hydrolytic enzymes involving in pectin degradation. A comparison of two highly conserved duplicated PG genes, namely, Brassica campestris Male Fertility 26a (BcMF26a) and BcMF26b, revealed the different features of their expression patterns and functions. We found that these two genes were orthologous genes of At4g33440, and they originated from a chromosomal segmental duplication. Although structurally similar, their regulatory and intron sequences largely diverged. QRT-PCR analysis showed that the expression level of BcMF26b was higher than that of BcMF26a in almost all the tested organs and tissues in Brassica campestris. Promoter activity analysis showed that, at reproductive development stages, BcMF26b promoter was active in tapetum, pollen grains, and pistils, whereas BcMF26a promoter was only active in pistils. In the subcellular localization experiment, BcMF26a and BcMF26b proteins could be localized to the cell wall. When the two genes were co-inhibited, pollen intine was formed abnormally and pollen tubes could not grow or stretch. Moreover, the knockout mutants of At4g33440 delayed the growth of pollen tubes. Therefore, BcMF26a/b can participate in the construction of pollen wall by modulating intine information and BcMF26b may play a major role in co-inhibiting transformed plants.

No MeSH data available.


Related in: MedlinePlus

Sequence characterization and phylogenetic tree analysis.(A) The phylogenetic trees of PG genes from Brassica campestris and Arabidopsis thaliana genomes were generated using the neighbor-joining (NJ) method with 1000 bootstrap repeats (part of data displayed). At4g33440, BcMF26a, and BcMF26b are indicated by solid circle and triangles, respectively. (B) BcMF26a, BcMF26b, At4g33440, and their flanking regions representing ~45 kb of chromosomes are drawn to scale. Collinear conserved blocks are identified. The black solid lines indicate noncolinear chromosome fragments. The dotted lines represent the other regions of the chromosomes, which are not drawn to scale. The positions of BcMF26a, BcMF26b, and their orthologous gene At4g33440 in A. thaliana are labeled with blue, red and green, respectively. Segmental chromosomal duplication and rearrangement are shown. (C) Phylogenetic tree constructed based on the amino acid sequence of BcMF26a, BcMF26b, At4g33440, and 35 PG genes from different plant species in S2 Table by NJ method. Confidence values from the bootstrap test (1000 replicates) are indicated by the numbers on the tree. The genes were clustered into six clades (Clade A to Clade F). BcMF26a, BcMF26b, and At4g33440 are grouped in Clade E.
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pone.0131173.g001: Sequence characterization and phylogenetic tree analysis.(A) The phylogenetic trees of PG genes from Brassica campestris and Arabidopsis thaliana genomes were generated using the neighbor-joining (NJ) method with 1000 bootstrap repeats (part of data displayed). At4g33440, BcMF26a, and BcMF26b are indicated by solid circle and triangles, respectively. (B) BcMF26a, BcMF26b, At4g33440, and their flanking regions representing ~45 kb of chromosomes are drawn to scale. Collinear conserved blocks are identified. The black solid lines indicate noncolinear chromosome fragments. The dotted lines represent the other regions of the chromosomes, which are not drawn to scale. The positions of BcMF26a, BcMF26b, and their orthologous gene At4g33440 in A. thaliana are labeled with blue, red and green, respectively. Segmental chromosomal duplication and rearrangement are shown. (C) Phylogenetic tree constructed based on the amino acid sequence of BcMF26a, BcMF26b, At4g33440, and 35 PG genes from different plant species in S2 Table by NJ method. Confidence values from the bootstrap test (1000 replicates) are indicated by the numbers on the tree. The genes were clustered into six clades (Clade A to Clade F). BcMF26a, BcMF26b, and At4g33440 are grouped in Clade E.

Mentions: BcMF26a and BcMF26b, the orthologous genes of At4g33440, were found in BRAD using the basic local alignment search tool service. The phylogenetic tree analysis of PG genes from A. thaliana and B. campestris genomes also proved the high homology between BcMF26a, BcMF26b, and At4g33440 (Fig 1A, part of data displayed). Collinear analysis was performed on BcMF26a, BcMF26b, and At4g33440 (Fig 1B). BcMF26a was localized on Chromosome 1 (chr1) and BcMF26b was localized on Chromosome 3 (chr3) in the B. campestris genome. Meanwhile, BcMF26a and BcMF26b were aligned to the same locus, the PG gene At4g33440, in the A. thaliana genome. The regions immediately surrounding BcMF26a and BcMF26b on chr1 and chr3, respectively, exhibited the reverse orientation. The collinearly duplicated chromosomal blocks of the two genes were interspersed with chromosomal fragments of no apparent similarity. Moreover, the collinear conserved blocks between the two chromosomal regions preserved their orientations with respect to the target genes.


BcMF26a and BcMF26b Are Duplicated Polygalacturonase Genes with Divergent Expression Patterns and Functions in Pollen Development and Pollen Tube Formation in Brassica campestris.

Lyu M, Yu Y, Jiang J, Song L, Liang Y, Ma Z, Xiong X, Cao J - PLoS ONE (2015)

Sequence characterization and phylogenetic tree analysis.(A) The phylogenetic trees of PG genes from Brassica campestris and Arabidopsis thaliana genomes were generated using the neighbor-joining (NJ) method with 1000 bootstrap repeats (part of data displayed). At4g33440, BcMF26a, and BcMF26b are indicated by solid circle and triangles, respectively. (B) BcMF26a, BcMF26b, At4g33440, and their flanking regions representing ~45 kb of chromosomes are drawn to scale. Collinear conserved blocks are identified. The black solid lines indicate noncolinear chromosome fragments. The dotted lines represent the other regions of the chromosomes, which are not drawn to scale. The positions of BcMF26a, BcMF26b, and their orthologous gene At4g33440 in A. thaliana are labeled with blue, red and green, respectively. Segmental chromosomal duplication and rearrangement are shown. (C) Phylogenetic tree constructed based on the amino acid sequence of BcMF26a, BcMF26b, At4g33440, and 35 PG genes from different plant species in S2 Table by NJ method. Confidence values from the bootstrap test (1000 replicates) are indicated by the numbers on the tree. The genes were clustered into six clades (Clade A to Clade F). BcMF26a, BcMF26b, and At4g33440 are grouped in Clade E.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4495986&req=5

pone.0131173.g001: Sequence characterization and phylogenetic tree analysis.(A) The phylogenetic trees of PG genes from Brassica campestris and Arabidopsis thaliana genomes were generated using the neighbor-joining (NJ) method with 1000 bootstrap repeats (part of data displayed). At4g33440, BcMF26a, and BcMF26b are indicated by solid circle and triangles, respectively. (B) BcMF26a, BcMF26b, At4g33440, and their flanking regions representing ~45 kb of chromosomes are drawn to scale. Collinear conserved blocks are identified. The black solid lines indicate noncolinear chromosome fragments. The dotted lines represent the other regions of the chromosomes, which are not drawn to scale. The positions of BcMF26a, BcMF26b, and their orthologous gene At4g33440 in A. thaliana are labeled with blue, red and green, respectively. Segmental chromosomal duplication and rearrangement are shown. (C) Phylogenetic tree constructed based on the amino acid sequence of BcMF26a, BcMF26b, At4g33440, and 35 PG genes from different plant species in S2 Table by NJ method. Confidence values from the bootstrap test (1000 replicates) are indicated by the numbers on the tree. The genes were clustered into six clades (Clade A to Clade F). BcMF26a, BcMF26b, and At4g33440 are grouped in Clade E.
Mentions: BcMF26a and BcMF26b, the orthologous genes of At4g33440, were found in BRAD using the basic local alignment search tool service. The phylogenetic tree analysis of PG genes from A. thaliana and B. campestris genomes also proved the high homology between BcMF26a, BcMF26b, and At4g33440 (Fig 1A, part of data displayed). Collinear analysis was performed on BcMF26a, BcMF26b, and At4g33440 (Fig 1B). BcMF26a was localized on Chromosome 1 (chr1) and BcMF26b was localized on Chromosome 3 (chr3) in the B. campestris genome. Meanwhile, BcMF26a and BcMF26b were aligned to the same locus, the PG gene At4g33440, in the A. thaliana genome. The regions immediately surrounding BcMF26a and BcMF26b on chr1 and chr3, respectively, exhibited the reverse orientation. The collinearly duplicated chromosomal blocks of the two genes were interspersed with chromosomal fragments of no apparent similarity. Moreover, the collinear conserved blocks between the two chromosomal regions preserved their orientations with respect to the target genes.

Bottom Line: We found that these two genes were orthologous genes of At4g33440, and they originated from a chromosomal segmental duplication.Promoter activity analysis showed that, at reproductive development stages, BcMF26b promoter was active in tapetum, pollen grains, and pistils, whereas BcMF26a promoter was only active in pistils.In the subcellular localization experiment, BcMF26a and BcMF26b proteins could be localized to the cell wall.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Cell & Molecular Biology, Institute of Vegetable Science, Zhejiang University, Hangzhou, China; Zhejiang Provincial Key Laboratory of Horticultural Plant Integrative Biology, Zhejiang University, Hangzhou, China.

ABSTRACT
Polygalacturonase (PG) is one of the cell wall hydrolytic enzymes involving in pectin degradation. A comparison of two highly conserved duplicated PG genes, namely, Brassica campestris Male Fertility 26a (BcMF26a) and BcMF26b, revealed the different features of their expression patterns and functions. We found that these two genes were orthologous genes of At4g33440, and they originated from a chromosomal segmental duplication. Although structurally similar, their regulatory and intron sequences largely diverged. QRT-PCR analysis showed that the expression level of BcMF26b was higher than that of BcMF26a in almost all the tested organs and tissues in Brassica campestris. Promoter activity analysis showed that, at reproductive development stages, BcMF26b promoter was active in tapetum, pollen grains, and pistils, whereas BcMF26a promoter was only active in pistils. In the subcellular localization experiment, BcMF26a and BcMF26b proteins could be localized to the cell wall. When the two genes were co-inhibited, pollen intine was formed abnormally and pollen tubes could not grow or stretch. Moreover, the knockout mutants of At4g33440 delayed the growth of pollen tubes. Therefore, BcMF26a/b can participate in the construction of pollen wall by modulating intine information and BcMF26b may play a major role in co-inhibiting transformed plants.

No MeSH data available.


Related in: MedlinePlus