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Comparative transcriptome analysis reveals carbohydrate and lipid metabolism blocks in Brassica napus L. male sterility induced by the chemical hybridization agent monosulfuron ester sodium.

Li Z, Cheng Y, Cui J, Zhang P, Zhao H, Hu S - BMC Genomics (2015)

Bottom Line: Pathway visualization showed that the tightly regulated gene network for metabolism was reprogrammed to respond to MES treatment.The results of cytological observation and transcriptome analysis in the MES-treated rapeseed plants were mirrored by carbohydrate content analysis.MES treatment led to decrease in soluble sugars content in leaves and early stage buds, but increase in soluble sugars content and decrease in starch content in middle stage buds.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Crop Stress Biology in Arid Areas, Northwest A&F University, Yangling, Shaanxi, 712100, P. R. China. lzj8705@163.com.

ABSTRACT

Background: Chemical hybridization agents (CHAs) are often used to induce male sterility for the production of hybrid seeds. We previously discovered that monosulfuron ester sodium (MES), an acetolactate synthase (ALS) inhibitor of the herbicide sulfonylurea family, can induce rapeseed (Brassica napus L.) male sterility at approximately 1% concentration required for its herbicidal activity. To find some clues to the mechanism of MES inducing male sterility, the ultrastructural cytology observations, comparative transcriptome analysis, and physiological analysis on carbohydrate content were carried out in leaves and anthers at different developmental stages between the MES-treated and mock-treated rapeseed plants.

Results: Cytological analysis revealed that the plastid ultrastructure was abnormal in pollen mother cells and tapetal cells in male sterility anthers induced by MES treatment, with less material accumulation in it. However, starch granules were observed in chloroplastids of the epidermis cells in male sterility anthers. Comparative transcriptome analysis identified 1501 differentially expressed transcripts (DETs) in leaves and anthers at different developmental stages, most of these DETs being localized in plastid and mitochondrion. Transcripts involved in metabolism, especially in carbohydrate and lipid metabolism, and cellular transport were differentially expressed. Pathway visualization showed that the tightly regulated gene network for metabolism was reprogrammed to respond to MES treatment. The results of cytological observation and transcriptome analysis in the MES-treated rapeseed plants were mirrored by carbohydrate content analysis. MES treatment led to decrease in soluble sugars content in leaves and early stage buds, but increase in soluble sugars content and decrease in starch content in middle stage buds.

Conclusions: Our integrative results suggested that carbohydrate and lipid metabolism were influenced by CHA-MES treatment during rapeseed anther development, which might responsible for low concentration MES specifically inducing male sterility. A simple action model of CHA-MES inducing male sterility in B. napus was proposed. These results will help us to understand the mechanism of MES inducing male sterility at low concentration, and might provide some potential targets for developing new male sterility inducing CHAs and for genetic manipulation in rapeseed breeding.

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Photographs of the leaves and developmental anthers for microarrays from the mock-treated (A, C, E, G) and MES-treated (B, D, F, H) plants.A-B, young leaves from the main inflorescences (Ls); C-D, small buds (SBs); E-F, anthers from middle buds (An-MBs); G-H, anthers form large buds (An-LBs). Scale bar in leaves was 1 cm, scale bars in SBs, An-MBs, and An-LBs were 1 mm.
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Fig2: Photographs of the leaves and developmental anthers for microarrays from the mock-treated (A, C, E, G) and MES-treated (B, D, F, H) plants.A-B, young leaves from the main inflorescences (Ls); C-D, small buds (SBs); E-F, anthers from middle buds (An-MBs); G-H, anthers form large buds (An-LBs). Scale bar in leaves was 1 cm, scale bars in SBs, An-MBs, and An-LBs were 1 mm.

Mentions: To obtain genome-wide gene expression profiles in the MES-treated and mock-treated plants of B. napus, the Agilent Single Channel Brassica Oligo Microarray (4 × 44 K) was used. Three independent biological replicates of four pairs of tissues (organs) from the MES-treated and mock-treated plants were collected for gene expression analysis, resulting in a dataset of 24 microarrays. The four tissues (organs) (Figure 2) included the leaves from main inflorescences (Ls), the small buds less than 1 mm in length containing microgametocytes before and during pollen mother stage (SBs), the anthers from middle buds with length between 1 mm and 3 mm containing microgametocytes from meiosis to early uninucleate microspore stage (An-MBs), and the anthers from large buds more than 3 mm in length containing microgametocytes from vacuolated stage to mature pollen stage (An-LBs). The data quality was assessed using two measurements: (1) correlation coefficients between biological replicates, which ranged from 0.8495 to 0.9906, with a mean of 0.9447 (Additional file 1), and (2) quantitative real time RT-PCR (qRT-PCR), which was performed on 62 randomly selected genes. The results of qRT-PCT analysis showed a high degree of concordance (R2 = 0.8775) with microarray results (Additional file 2). Taken together, these demonstrated that the microarray results obtained in this study were reliable.Figure 2


Comparative transcriptome analysis reveals carbohydrate and lipid metabolism blocks in Brassica napus L. male sterility induced by the chemical hybridization agent monosulfuron ester sodium.

Li Z, Cheng Y, Cui J, Zhang P, Zhao H, Hu S - BMC Genomics (2015)

Photographs of the leaves and developmental anthers for microarrays from the mock-treated (A, C, E, G) and MES-treated (B, D, F, H) plants.A-B, young leaves from the main inflorescences (Ls); C-D, small buds (SBs); E-F, anthers from middle buds (An-MBs); G-H, anthers form large buds (An-LBs). Scale bar in leaves was 1 cm, scale bars in SBs, An-MBs, and An-LBs were 1 mm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4376087&req=5

Fig2: Photographs of the leaves and developmental anthers for microarrays from the mock-treated (A, C, E, G) and MES-treated (B, D, F, H) plants.A-B, young leaves from the main inflorescences (Ls); C-D, small buds (SBs); E-F, anthers from middle buds (An-MBs); G-H, anthers form large buds (An-LBs). Scale bar in leaves was 1 cm, scale bars in SBs, An-MBs, and An-LBs were 1 mm.
Mentions: To obtain genome-wide gene expression profiles in the MES-treated and mock-treated plants of B. napus, the Agilent Single Channel Brassica Oligo Microarray (4 × 44 K) was used. Three independent biological replicates of four pairs of tissues (organs) from the MES-treated and mock-treated plants were collected for gene expression analysis, resulting in a dataset of 24 microarrays. The four tissues (organs) (Figure 2) included the leaves from main inflorescences (Ls), the small buds less than 1 mm in length containing microgametocytes before and during pollen mother stage (SBs), the anthers from middle buds with length between 1 mm and 3 mm containing microgametocytes from meiosis to early uninucleate microspore stage (An-MBs), and the anthers from large buds more than 3 mm in length containing microgametocytes from vacuolated stage to mature pollen stage (An-LBs). The data quality was assessed using two measurements: (1) correlation coefficients between biological replicates, which ranged from 0.8495 to 0.9906, with a mean of 0.9447 (Additional file 1), and (2) quantitative real time RT-PCR (qRT-PCR), which was performed on 62 randomly selected genes. The results of qRT-PCT analysis showed a high degree of concordance (R2 = 0.8775) with microarray results (Additional file 2). Taken together, these demonstrated that the microarray results obtained in this study were reliable.Figure 2

Bottom Line: Pathway visualization showed that the tightly regulated gene network for metabolism was reprogrammed to respond to MES treatment.The results of cytological observation and transcriptome analysis in the MES-treated rapeseed plants were mirrored by carbohydrate content analysis.MES treatment led to decrease in soluble sugars content in leaves and early stage buds, but increase in soluble sugars content and decrease in starch content in middle stage buds.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Crop Stress Biology in Arid Areas, Northwest A&F University, Yangling, Shaanxi, 712100, P. R. China. lzj8705@163.com.

ABSTRACT

Background: Chemical hybridization agents (CHAs) are often used to induce male sterility for the production of hybrid seeds. We previously discovered that monosulfuron ester sodium (MES), an acetolactate synthase (ALS) inhibitor of the herbicide sulfonylurea family, can induce rapeseed (Brassica napus L.) male sterility at approximately 1% concentration required for its herbicidal activity. To find some clues to the mechanism of MES inducing male sterility, the ultrastructural cytology observations, comparative transcriptome analysis, and physiological analysis on carbohydrate content were carried out in leaves and anthers at different developmental stages between the MES-treated and mock-treated rapeseed plants.

Results: Cytological analysis revealed that the plastid ultrastructure was abnormal in pollen mother cells and tapetal cells in male sterility anthers induced by MES treatment, with less material accumulation in it. However, starch granules were observed in chloroplastids of the epidermis cells in male sterility anthers. Comparative transcriptome analysis identified 1501 differentially expressed transcripts (DETs) in leaves and anthers at different developmental stages, most of these DETs being localized in plastid and mitochondrion. Transcripts involved in metabolism, especially in carbohydrate and lipid metabolism, and cellular transport were differentially expressed. Pathway visualization showed that the tightly regulated gene network for metabolism was reprogrammed to respond to MES treatment. The results of cytological observation and transcriptome analysis in the MES-treated rapeseed plants were mirrored by carbohydrate content analysis. MES treatment led to decrease in soluble sugars content in leaves and early stage buds, but increase in soluble sugars content and decrease in starch content in middle stage buds.

Conclusions: Our integrative results suggested that carbohydrate and lipid metabolism were influenced by CHA-MES treatment during rapeseed anther development, which might responsible for low concentration MES specifically inducing male sterility. A simple action model of CHA-MES inducing male sterility in B. napus was proposed. These results will help us to understand the mechanism of MES inducing male sterility at low concentration, and might provide some potential targets for developing new male sterility inducing CHAs and for genetic manipulation in rapeseed breeding.

Show MeSH
Related in: MedlinePlus