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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

Analysis of the BcMF26a/b mRNA levels in the inflorescences of the bcmf26a/b and control lines using qRT-PCR analysis.The expression levels of BcMF26a/b in the inflorescences of bcmf26a/b lines were significantly less than that of the empty vector pCAMBIA1301-transformed plants (control). UBC-10 was used as an internal control. Standard errors for three independent experiments are shown.
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pone.0131173.g006: Analysis of the BcMF26a/b mRNA levels in the inflorescences of the bcmf26a/b and control lines using qRT-PCR analysis.The expression levels of BcMF26a/b in the inflorescences of bcmf26a/b lines were significantly less than that of the empty vector pCAMBIA1301-transformed plants (control). UBC-10 was used as an internal control. Standard errors for three independent experiments are shown.

Mentions: To investigate the biological role, multiple-target amiRNA construct co-inhibiting BcMF26a and BcMF26b was transformed into B. campestris ssp. chinensis var. parachinensis (S7A and S7B Fig, S8A to S8H Fig). Seven putative transgenic lines (bcmf26a/b-1 to bcmf26a/b-7) were obtained after hygromycin screening. QRT-PCR analysis showed that the mRNA levels of BcMF26a/b in the inflorescences of the seven transformed lines were much lower than those in the control plants (Fig 6), indicating that the expression of the two genes was co-inhibited simultaneously and fairly.


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)

Analysis of the BcMF26a/b mRNA levels in the inflorescences of the bcmf26a/b and control lines using qRT-PCR analysis.The expression levels of BcMF26a/b in the inflorescences of bcmf26a/b lines were significantly less than that of the empty vector pCAMBIA1301-transformed plants (control). UBC-10 was used as an internal control. Standard errors for three independent experiments are shown.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4495986&req=5

pone.0131173.g006: Analysis of the BcMF26a/b mRNA levels in the inflorescences of the bcmf26a/b and control lines using qRT-PCR analysis.The expression levels of BcMF26a/b in the inflorescences of bcmf26a/b lines were significantly less than that of the empty vector pCAMBIA1301-transformed plants (control). UBC-10 was used as an internal control. Standard errors for three independent experiments are shown.
Mentions: To investigate the biological role, multiple-target amiRNA construct co-inhibiting BcMF26a and BcMF26b was transformed into B. campestris ssp. chinensis var. parachinensis (S7A and S7B Fig, S8A to S8H Fig). Seven putative transgenic lines (bcmf26a/b-1 to bcmf26a/b-7) were obtained after hygromycin screening. QRT-PCR analysis showed that the mRNA levels of BcMF26a/b in the inflorescences of the seven transformed lines were much lower than those in the control plants (Fig 6), indicating that the expression of the two genes was co-inhibited simultaneously and fairly.

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