Dataset Information

Soybean endo-?-mannanase GmMAN1 is not associated with leaf abscission, but might be involved in the response to wounding.

ABSTRACT: The objective of this work is to investigate the relationship between endo-?-mannanase and leaf abscission, and response to wounding in soybean (Glycine max). An endo-?-mannanase gene GmMAN1 was cloned from the abscission zone in petiole explants, and was heterologously expressed in E. coli. Polyclonal antibodies were raised against the fusion protein. The increases in activity, isoform numbers, and amounts of transcripts and proteins of GmMAN1 were found not only in the abscission zone but also in the non-abscission zone during petiole abscission in the explants, but not in these two tissues during leaf abscission artificially induced by ethephon treatment in the intact plants. The changes in endo-?-mannanase expression patterns in these two tissues were probably induced by the inherent mechanical wounding during the preparation of explants. When soybean plants were wounded by removing half of the leaf blade of the first pair of true leaves, the transcripts and proteins of GmMAN1 were induced in the leaves and stem, leading to the increases in enzyme activity and isoform numbers in them. It is concluded that the soybean endo-?-mannanase GmMAN1 is not associated with leaf abscission, but might be involved in the response to wounding.

SUBMITTER: Yan M

PROVIDER: S-EPMC3500276 | biostudies-literature | 2012

REPOSITORIES: biostudies-literature

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Soybean endo-β-mannanase GmMAN1 is not associated with leaf abscission, but might be involved in the response to wounding.

Yan Min M Zhang Yifan Y Guo Wenjuan W Wang Xiaofeng X

PloS one 20121116 11

The objective of this work is to investigate the relationship between endo-β-mannanase and leaf abscission, and response to wounding in soybean (Glycine max). An endo-β-mannanase gene GmMAN1 was cloned from the abscission zone in petiole explants, and was heterologously expressed in E. coli. Polyclonal antibodies were raised against the fusion protein. The increases in activity, isoform numbers, and amounts of transcripts and proteins of GmMAN1 were found not only in the abscission zone but also ...[more]

PMID: 23173047

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Project description:Background and aimsThe stimulatory and inhibitory role of ethylene and auxin, respectively, in leaf abscission (leaf drop) is well documented. More recently, IDA (INFLORESCENCE DEFICIENT IN ABSCISSION) peptides and their putative interacting receptor-like-kinase partners, HAESA and HAESA-like2, were shown to be essential components in Arabidopsis floral organ abscission. Prior to research on IDA, it was reported that bean (Phaseolus vulgaris) leaf abscission required a diffusible signal that emanated from the vascular tissue. We were interested in determining whether the IDA signalling path might regulate abscission in plants other than Arabidopsis and whether IDA might act as a diffusible signal in abscission.MethodologyQuantitative polymerase chain reaction was used to monitor gene expression and a GUS reporter gene construct used to determine the need for a diffusible signal in tomato.Principal resultsWe identified 12 IDA-like and 11 HAESA-like genes in soybean (Glycine max) and monitored their gene expression in abscission in relation to the expression of several cell-wall-modifying proteins and aminocyclopropane-1-carboxylic acid synthases. Ethylene evoked the expression of several IDA-like genes in abscission zones (AZ), but also to a lesser degree in the adjacent petiole tissue. Surprisingly, IDA-like gene expression was very high in senescent soybean leaves. We identified five IDA-like genes in tomato (Solanum lycopersicum). Only one IDA-like gene was expressed in the tomato AZ and its expression was approximately equal in the AZ and petioles, but no IDA-like gene showed significant expression in leaves at up to 96 h of exposure to ethylene.ConclusionsIDA-like gene expression is up-regulated during soybean and tomato abscission but up-regulation was not limited to the AZ. Cell separation in the AZ cortex of tomato does not require a diffusible signal emanating from the stele. A role for IDA in soybean and tomato leaf abscission is discussed.

Project description:BackgroundThe study of coffee polysaccharides-degrading enzymes from the coffee berry borer Hypothenemus hampei, has become an important alternative in the identification for enzymatic inhibitors that can be used as an alternative control of this dangerous insect. We report the cloning, expression and biochemical characterization of a mannanase gene that was identified in the midgut of the coffee berry borer and is responsible for the degradation of the most abundant polysaccharide in the coffee bean.MethodsThe amino acid sequence of HhMan was analyzed by multiple sequence alignment comparisons with BLAST (Basic Local Alignment Search Tool) and CLUSTALW. A Pichia pastoris expression system was used to express the recombinant form of the enzyme. The mannanase activity was quantified by the 3,5-dinitrosalicylic (DNS) and the hydrolitic properties were detected by TLC.ResultsAn endo-1,4-β-mannanase from the digestive tract of the insect Hypothenemus hampei was cloned and expressed as a recombinant protein in the Pichia pastoris system. This enzyme is 56% identical to the sequence of an endo-β-mannanase from Bacillus circulans that belongs to the glycosyl hydrolase 5 (GH5) family. The purified recombinant protein (rHhMan) exhibited a single band (35.5 kDa) by SDS-PAGE, and its activity was confirmed by zymography. rHhMan displays optimal activity levels at pH 5.5 and 30°C and can hydrolyze galactomannans of varying mannose:galactose ratios, suggesting that the enzymatic activity is independent of the presence of side chains such as galactose residues. The enzyme cannot hydrolyze manno-oligosaccharides such as mannobiose and mannotriose; however, it can degrade mannotetraose, likely through a transglycosylation reaction. The K(m) and k(cat) values of this enzyme on guar gum were 2.074 mg ml(-1) and 50.87 s(-1), respectively, which is similar to other mannanases.ConclusionThis work is the first study of an endo-1,4-β-mannanase from an insect using this expression system. Due to this enzyme's importance in the digestive processes of the coffee berry borer, this study may enable the design of inhibitors against endo-1,4-β-mannanase to decrease the economic losses stemming from this insect.

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Project description:BACKGROUND:Glutathione S-transferases (GSTs) have been reported to regulate the plant tolerance to environmental stresses. Many plant GSTs exhibited the roles on promoting tolerance to drought stress, oxidative stress and plant hormones. The biological function of GSTs has been well characterized in Arabidopsis thaliana in response to exogenous environmental stresses. However, their regulation function under exogenous environmental stresses regulating leaf abscission in cassava (Manihot esculenta Crantz) remained unknown. RESULTS:Here, 83 GSTs were identified from tropical plant cassava. The amino acid motifs and phylogenetic analyses indicated that MeGSTs were divided into 9 classes. The global expression analyses were carried out to analyze the gene expression patterns of MeGST in cassava abscission zones by comparing the MeGST genes expression patterns in both ethylene and drought induced cassava leaf abscission. Totally, 34 GSTs were detected to express in both ethylene and drought induced leaf abscission in cassava abscission zones. Comparison of GST expression profiling between ethylene and drought induced leaf abscission suggested that Tau GST genes showed with the similar expression in both treatments induced leaf abscission in cassava abscission zone. GO annotation indicated that all 17 Tau GST genes participated in the pathway of toxin catabolism (GO: 0009407). The expression levels of 17 Tau MeGST genes were analyzed in two cassava cultivars, 'SC124' and 'Arg7', the two cultivars exhibit different levels of leaf abscission when suffered from the same environmental stress. Higher expression levels of Tau MeGSTs were detected in the precocious abscission Arg7 cultivar, while lower expression levels in delayed abscission SC124 cultivar. All the results indicated that Tau MeGSTs have the function in regulation the cassava leaf abscission under environmental stresses. CONCLUSION:Analysis of the expression patterns of GSTs in various abscission-promoting treatments in cassava abscission zones helps us to understand the possible roles of GSTs in cassava leaf abscission.

Project description:BackgroundSugarcane (Saccharum officinarum L.) is an economically important crop, mainly due to the production of sugar and biofuel (Azevedo RA, Carvalho RF, Cia MC, & Gratão PL, Trop Plant Biol 4:42-51, 2011). Grown mainly in tropical and subtropical countries, sugarcane is a highly polyploid plant with up to ten copies of each chromosome, which increases the difficulties of genome assembly and genetic, physiologic and biochemical analyses. The increasing demands of sugar and the increasing cost of sugarcane harvest require sugarcane varieties which can shed their leaves during the maturity time, so it is important to study the mechanism of leaf abscission in sugarcane.ResultsTo improve the understanding of miRNA roles in sugarcane leaf abscission, we reported the genome-wide characterization of miRNAs and their putative targets in sugarcane using deep sequencing for six small RNA libraries. In total, 93 conserved miRNAs and 454 novel miRNAs were identified in sugarcane using previously reported transcriptome as reference. Among them, 25 up-regulated and 13 down-regulated miRNAs were identified in leaf abscission sugarcane plants (LASP) compared to leaf packaging sugarcane plants (LPSP). Target prediction revealed several miRNA-mRNA modules including miR156-SPL, miR319-TPR2, miR396-GRF and miR408-LAC3 might be involved in the sugarcane leaf abscission. KEGG pathway enrichment analysis showed differentially expressed miRNAs may regulate pathways like "plant hormone signal transduction" and "plant-pathogen interaction", which is consistent with previous transcriptome study. In addition, we identified 96 variant miRNAs with 135 single nucleotide polymorphisms (SNPs). The expression of sugarcane miRNAs and variant miRNAs were confirmed by qRT-PCR. We identified a possible poaceae specific miRNA called miR5384 for the first time in sugarcane.ConclusionsWe not only reported miR5384, a possible poaceae specific miRNA, for the first time in sugarcane but also presented some miRNA-mRNA modules including miR156-SPL, miR319-TPR2, miR396-GRF and miR408-LAC in sugarcane. These modules might be involved in the regulation of sugarcane leaf abscission during the maturity time. All of these findings may lay ground work for future application of sugarcane breeding program and benefit research studies of sugarcane miRNAs.

Dataset Information

Soybean endo-?-mannanase GmMAN1 is not associated with leaf abscission, but might be involved in the response to wounding.

Publications

Soybean endo-β-mannanase GmMAN1 is not associated with leaf abscission, but might be involved in the response to wounding.

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