An endo-polygalacturonase (PG) of<i>Fusarium moniliforme</i>escaping inhibition by plant polygalacturonase-inhibiting proteins (PGIPs) provides new insights into the PGâPGIP interaction

Sella, Luca; Castiglioni, Carla; Roberti, Serena; D’Ovidio, R.; Favaron, Francesco

doi:10.1016/j.femsle.2004.09.019

Cited by 33 publications

(32 citation statements)

References 29 publications

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“…This observation was confirmed by the absence of the white precipitate around the negative control well. Similar findings have been described of the appearance of the white halo surrounding the well loaded with polygalacturonase of Fusarium moniliforme and no halo was detected with the same sample mixed with polygalacturonaseinhibitor protein of Phaseolus vulgaris (common bean) [44]. The cellulase activities of the cloned celB and celC products were also confirmed by the appearance of a yellow halo against a red background in CMC-based medium.…”

Section: Discussionsupporting

confidence: 84%

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Molecular Cloning and Expression of Cellulase and Polygalacturonase Genes in E. coli as a Promising Application for Biofuel Production

Ibrahim¹,

Jones²,

Hossenya³

2013

J Pet Environ Biotechnol

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Lignocellulosic biomass has potential for bioethanol, a renewable fuel. A limitation is that bioconversion of the complex lignocellulosic material to simple sugars and then to bioethanol is a challenging process. Recent work has focused on the genetic engineering of a biocatalyst that may play a critical role in biofuel production. Escherichia coli have been considered a convenient host for biocatalysts in biofuel production for its fermentation of glucose into a wide range of short-chain alcohols and production of highly deoxygenated hydrocarbon. The bacterium Pectobacterium carotovorum subsp. carotovorum (P. carotovorum) is notorious for its maceration of the plant cell wall causing soft rot. The ability to destroy plants is due to the expression and secretion of a wide range of hydrolytic enzymes that include cellulases and polygalacturonases. P. carotovorum ATCC™ no. 15359 was used as a source of DNA for the amplification of celB, celC and peh. These genes encode 2 cellulases and a polygalacturonase, respectively. Primers were designed based on published gene sequences and used to amplify the open reading frames from the genomic DNA of P. carotovorum. The individual PCR products were cloned into the pTAC-MAT-2 expression vector and transformed into Escherichia coli. The deduced amino acid sequences of the cloned genes have been analyzed for their catalytically active domains. Estimation of the molecular weights of the expressed proteins was performed using SDS-PAGE analysis and celB, celC and peh products were approximately 29.5 kDa, 40 kDa 41.5 kDa, respectively. Qualitative determination of the cellulase and polygalacturonase activities of the cloned genes was carried out using agar diffusion assays.

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Section: Discussionsupporting

confidence: 84%

“…The determination of polygalacturonase activity was done using a modified method described by [44]. Polygalacturonic acid-based substrate agar was prepared by dissolving 1% polygalacturonic acid (Sigma, cat.…”

Section: Detection Of Polygalacturonase Activitymentioning

confidence: 99%

Molecular Cloning and Expression of Cellulase and Polygalacturonase Genes in E. coli as a Promising Application for Biofuel Production

Ibrahim¹,

Jones²,

Hossenya³

2013

J Pet Environ Biotechnol

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“…3) suggesting that this may be a common mechanism for PG-PGIP interaction. Non-competitive inhibition is presumed to result from binding at other PG locations (Sella et al 2004;Sicilia et al 2005), but such interactions are likely to be diverse in nature and have not yet been fully elucidated. In fact, some PGIPs are capable of activating PG activity in certain ionic environments (Kemp et al 2004).…”

Section: Discussionmentioning

confidence: 99%

Brassica napus possesses an expanded set of polygalacturonase inhibitor protein genes that are differentially regulated in response to Sclerotinia sclerotiorum infection, wounding and defense hormone treatment

Hegedus

Buchwaldt

et al. 2008

Planta

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Most plants encode a limited set of polygalacturonase inhibitor (PGIP) genes that may be involved in aspects of plant development, but more importantly in the inactivation of polygalacturonases (PG) secreted by pathogens. Previously, we characterized two Brassica napus PGIP genes, BnPgip1 and BnPgip2, which were differentially expressed in response to pathogen infection and wounding. Here we report that the B. napus genome encodes a set of at least 16 PGIP genes that are similar to BnPgip1 or BnPgip2. This is the largest Pgip gene family reported to date. Comparison of the BnPGIPs revealed several sites within the xxLxLxx region of leucine rich repeats that form beta-sheets along the interacting face of the PGIP that are hypervariable and represent good candidates for generating PGIP diversity. Characterization of the regulatory regions and RT-PCR studies with gene-specific primers revealed that individual genes were differentially responsive to pathogen infection, mechanical wounding and signaling molecules. Many of the BnPgip genes responded to infection by the necrotic pathogen, Sclerotinia sclerotiorum; however, these genes were also induced either by jasmonic acid, wounding and salicylic acid or some combination thereof. The large number of PGIPs and the differential manner in which they are regulated likely ensures that B. napus can respond to attack from a broad spectrum of pathogens and pests.

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“…In rice it has also been shown that OsPGIP1 is less effective in inhibiting fungal PGs from A. niger and B. cinera in comparison to PGIPs from dicots such as soy bean GmPGIP3 (Sella et al, 2004). In dicotyledons, soy bean (GmPGIP) and Arabidopsis (AtPGIP1 and AtPGIP2) the genes are up regulated in response to Botrytis cinerea infection.…”

Section: Discussionmentioning

confidence: 99%

Identification of a partial oil palm polygalacturonase-inhibiting protein (EgPGIP) gene and its expression during basal stem rot infection caused by Ganoderma boninense

Al‐Obaidi¹,

Mohd-Yusuf²,

ChinChong³

et al. 2010

Afr. J. Biotechnol.

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Basal stem rot disease (BSR) is a common and serious fungal disease of the oil palm caused byGanoderma boninense. This fungal disease infects thousands of hectares of plantings in Southeast Asia every year causing not only yield but also tree losses. A natural plant self defence mechanism against fungal infection is the production of fungal resistance protein. A fungal resistance gene that has been reported previously in other monocotyledonous plants such as rice and barley is polygalacturonase-inhibiting protein (PGIP) gene, a plant defence cell wall glycoprotein that has been shown to inhibit the activity of fungal endopolygalacturonase (endo-PGs) and modulate their activity and has the potential to be developed as a disease or resistance biomarker for the oil palm. The identification and isolation of this gene in oil palm allowed for the study of its differential expression during the fungal infection. The oil palm PGIP gene (EgPGIP) has between 60-100% similarities with the database sequence of PGIP from other monocotyledons. Interestingly, we found that the expression of EgPGIP gene measured using Real-Time PCR showed that the expression level of EgPGIP in infected oil palm was temporally down regulated. The results suggest that, down regulation of the EgPGIP is related to the establishment of infection by G. boninense.

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An endo-polygalacturonase (PG) ofFusarium moniliformeescaping inhibition by plant polygalacturonase-inhibiting proteins (PGIPs) provides new insights into the PGâPGIP interaction

Cited by 33 publications

References 29 publications

Molecular Cloning and Expression of Cellulase and Polygalacturonase Genes in E. coli as a Promising Application for Biofuel Production

Molecular Cloning and Expression of Cellulase and Polygalacturonase Genes in E. coli as a Promising Application for Biofuel Production

Brassica napus possesses an expanded set of polygalacturonase inhibitor protein genes that are differentially regulated in response to Sclerotinia sclerotiorum infection, wounding and defense hormone treatment

Identification of a partial oil palm polygalacturonase-inhibiting protein (EgPGIP) gene and its expression during basal stem rot infection caused by Ganoderma boninense

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An endo-polygalacturonase (PG) ofFusarium moniliformeescaping inhibition by plant polygalacturonase-inhibiting proteins (PGIPs) provides new insights into the PGâPGIP interaction

Cited by 33 publications

References 29 publications

Molecular Cloning and Expression of Cellulase and Polygalacturonase Genes in E. coli as a Promising Application for Biofuel Production

Molecular Cloning and Expression of Cellulase and Polygalacturonase Genes in E. coli as a Promising Application for Biofuel Production

Brassica napus possesses an expanded set of polygalacturonase inhibitor protein genes that are differentially regulated in response to Sclerotinia sclerotiorum infection, wounding and defense hormone treatment

Identification of a partial oil palm polygalacturonase-inhibiting protein (EgPGIP) gene and its expression during basal stem rot infection caused by Ganoderma boninense

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An endo-polygalacturonase (PG) ofFusarium moniliformeescaping inhibition by plant polygalacturonase-inhibiting proteins (PGIPs) provides new insights into the PGâPGIP interaction