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Decreased Polysaccharide Feruloylation Compromises Plant Cell Wall Integrity and Increases Susceptibility to Necrotrophic Fungal Pathogens.

Reem NT, Pogorelko G, Lionetti V, Chambers L, Held MA, Bellincampi D, Zabotina OA - Front Plant Sci (2016)

Bottom Line: Transgenic Arabidopsis plants expressing AnFAE showed a significant reduction of monomeric ferulic acid, decreased amounts of wall-associated extensins, and increased susceptibility to Botrytis cinerea, compared with wild type.Transgenic Brachypodium showed reductions in monomeric and dimeric ferulic acids and increased susceptibility to Bipolaris sorokiniana.These results demonstrate a role, in both monocot and dicot plants, of polysaccharide feruloylation in plant CWI, which contributes to plant resistance to necrotrophic pathogens.

View Article: PubMed Central - PubMed

Affiliation: Roy J. Carver Department of Biochemistry, Biophysiscs and Molecular Biology, Iowa State University, Ames, IA USA.

ABSTRACT
The complexity of cell wall composition and structure determines the strength, flexibility, and function of the primary cell wall in plants. However, the contribution of the various components to cell wall integrity (CWI) and function remains unclear. Modifications of cell wall composition can induce plant responses known as CWI control. In this study, we used transgenic expression of the fungal feruloyl esterase AnFAE to examine the effect of post-synthetic modification of Arabidopsis and Brachypodium cell walls. Transgenic Arabidopsis plants expressing AnFAE showed a significant reduction of monomeric ferulic acid, decreased amounts of wall-associated extensins, and increased susceptibility to Botrytis cinerea, compared with wild type. Transgenic Brachypodium showed reductions in monomeric and dimeric ferulic acids and increased susceptibility to Bipolaris sorokiniana. Upon infection, transgenic Arabidopsis and Brachypodium plants also showed increased expression of several defense-related genes compared with wild type. These results demonstrate a role, in both monocot and dicot plants, of polysaccharide feruloylation in plant CWI, which contributes to plant resistance to necrotrophic pathogens.

No MeSH data available.


Related in: MedlinePlus

Real-time qPCR analysis of selected pathogen-inducible defense genes inmock-infected (A-Arabidopsis,B-Brachypodium) and 48 HPI(C-Arabidopsis,D-Brachypodium) plants. Geneexpression levels in transgenic plants normalized to the expression of the samegene detected in wild-type plants (for which gene expression was set to 1),ACTIN2 was used as reference gene forArabidopsis and GAPDH forBrachypodium expression analysis. The comparative thresholdcycle method was used for determining differences between transcript copynumbers in wild-type and transgenic plants. Data represent average obtained forthree independent transgenic lines. Asterisks indicate significant differencesbetween transgenic plants and wild-type plants (Student’st-test, P < 0.05;n = 3).
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Figure 4: Real-time qPCR analysis of selected pathogen-inducible defense genes inmock-infected (A-Arabidopsis,B-Brachypodium) and 48 HPI(C-Arabidopsis,D-Brachypodium) plants. Geneexpression levels in transgenic plants normalized to the expression of the samegene detected in wild-type plants (for which gene expression was set to 1),ACTIN2 was used as reference gene forArabidopsis and GAPDH forBrachypodium expression analysis. The comparative thresholdcycle method was used for determining differences between transcript copynumbers in wild-type and transgenic plants. Data represent average obtained forthree independent transgenic lines. Asterisks indicate significant differencesbetween transgenic plants and wild-type plants (Student’st-test, P < 0.05;n = 3).

Mentions: To investigate whether the expression of fungal AnFAE and theconsequent decrease of FA content in the cell wall affect the pathways involved inplant response to infection, we used RT-qPCR to measure the transcript levels ofseveral known defense genes in infected and un-infected Arabidopsisand Brachypodium transgenic and wild-type plants. In healthy,un-infected transgenic plants, expression of these genes did not differ from theirexpression in wild-type plants (Figures 4A,B). However, several defense-related genes were significantlyup-regulated in transgenic plants in comparison with wild-type plants upon infection(Figures 4C,D). Thus, inArabidopsis transgenic plants, expression ofPGIP1, bG2, and WRKY40 wassignificantly higher than in wild-type plants, whereas the level ofPR1 transcript was significantly lower (Figure 4C). Some increase ofCYP81F2 was also observed in infectedArabidopsis plants but the difference was not statisticallysignificant. In Brachypodium, expression of WRKY40,WR3, and RetOX was higher in comparison withinfected wild-type plants (Figure 4D). A full list of genes assayed, including those notsignificantly different from wild-type and not amplified inBrachypodium, is included in Supplementary Figure S3.


Decreased Polysaccharide Feruloylation Compromises Plant Cell Wall Integrity and Increases Susceptibility to Necrotrophic Fungal Pathogens.

Reem NT, Pogorelko G, Lionetti V, Chambers L, Held MA, Bellincampi D, Zabotina OA - Front Plant Sci (2016)

Real-time qPCR analysis of selected pathogen-inducible defense genes inmock-infected (A-Arabidopsis,B-Brachypodium) and 48 HPI(C-Arabidopsis,D-Brachypodium) plants. Geneexpression levels in transgenic plants normalized to the expression of the samegene detected in wild-type plants (for which gene expression was set to 1),ACTIN2 was used as reference gene forArabidopsis and GAPDH forBrachypodium expression analysis. The comparative thresholdcycle method was used for determining differences between transcript copynumbers in wild-type and transgenic plants. Data represent average obtained forthree independent transgenic lines. Asterisks indicate significant differencesbetween transgenic plants and wild-type plants (Student’st-test, P < 0.05;n = 3).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4862258&req=5

Figure 4: Real-time qPCR analysis of selected pathogen-inducible defense genes inmock-infected (A-Arabidopsis,B-Brachypodium) and 48 HPI(C-Arabidopsis,D-Brachypodium) plants. Geneexpression levels in transgenic plants normalized to the expression of the samegene detected in wild-type plants (for which gene expression was set to 1),ACTIN2 was used as reference gene forArabidopsis and GAPDH forBrachypodium expression analysis. The comparative thresholdcycle method was used for determining differences between transcript copynumbers in wild-type and transgenic plants. Data represent average obtained forthree independent transgenic lines. Asterisks indicate significant differencesbetween transgenic plants and wild-type plants (Student’st-test, P < 0.05;n = 3).
Mentions: To investigate whether the expression of fungal AnFAE and theconsequent decrease of FA content in the cell wall affect the pathways involved inplant response to infection, we used RT-qPCR to measure the transcript levels ofseveral known defense genes in infected and un-infected Arabidopsisand Brachypodium transgenic and wild-type plants. In healthy,un-infected transgenic plants, expression of these genes did not differ from theirexpression in wild-type plants (Figures 4A,B). However, several defense-related genes were significantlyup-regulated in transgenic plants in comparison with wild-type plants upon infection(Figures 4C,D). Thus, inArabidopsis transgenic plants, expression ofPGIP1, bG2, and WRKY40 wassignificantly higher than in wild-type plants, whereas the level ofPR1 transcript was significantly lower (Figure 4C). Some increase ofCYP81F2 was also observed in infectedArabidopsis plants but the difference was not statisticallysignificant. In Brachypodium, expression of WRKY40,WR3, and RetOX was higher in comparison withinfected wild-type plants (Figure 4D). A full list of genes assayed, including those notsignificantly different from wild-type and not amplified inBrachypodium, is included in Supplementary Figure S3.

Bottom Line: Transgenic Arabidopsis plants expressing AnFAE showed a significant reduction of monomeric ferulic acid, decreased amounts of wall-associated extensins, and increased susceptibility to Botrytis cinerea, compared with wild type.Transgenic Brachypodium showed reductions in monomeric and dimeric ferulic acids and increased susceptibility to Bipolaris sorokiniana.These results demonstrate a role, in both monocot and dicot plants, of polysaccharide feruloylation in plant CWI, which contributes to plant resistance to necrotrophic pathogens.

View Article: PubMed Central - PubMed

Affiliation: Roy J. Carver Department of Biochemistry, Biophysiscs and Molecular Biology, Iowa State University, Ames, IA USA.

ABSTRACT
The complexity of cell wall composition and structure determines the strength, flexibility, and function of the primary cell wall in plants. However, the contribution of the various components to cell wall integrity (CWI) and function remains unclear. Modifications of cell wall composition can induce plant responses known as CWI control. In this study, we used transgenic expression of the fungal feruloyl esterase AnFAE to examine the effect of post-synthetic modification of Arabidopsis and Brachypodium cell walls. Transgenic Arabidopsis plants expressing AnFAE showed a significant reduction of monomeric ferulic acid, decreased amounts of wall-associated extensins, and increased susceptibility to Botrytis cinerea, compared with wild type. Transgenic Brachypodium showed reductions in monomeric and dimeric ferulic acids and increased susceptibility to Bipolaris sorokiniana. Upon infection, transgenic Arabidopsis and Brachypodium plants also showed increased expression of several defense-related genes compared with wild type. These results demonstrate a role, in both monocot and dicot plants, of polysaccharide feruloylation in plant CWI, which contributes to plant resistance to necrotrophic pathogens.

No MeSH data available.


Related in: MedlinePlus