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Ubiquitous urease affects soybean susceptibility to fungi.

ABSTRACT: The soybean ubiquitous urease (encoded by GmEu4) is responsible for recycling metabolically derived urea. Additional biological roles have been demonstrated for plant ureases, notably in toxicity to other organisms. However, urease enzymatic activity is not related to its toxicity. The role of GmEu4 in soybean susceptibility to fungi was investigated in this study. A differential expression pattern of GmEu4 was observed in susceptible and resistant genotypes of soybeans over the course of a Phakopsora pachyrhizi infection, especially 24 h after infection. Twenty-nine adult, transgenic soybean plants, representing six independently transformed lines, were obtained. Although the initial aim of this study was to overexpress GmEu4, the transgenic plants exhibited GmEu4 co-suppression and decreased ureolytic activity. The growth of Rhizoctonia solani, Phomopsis sp., and Penicillium herguei in media containing a crude protein extract from either transgenic or non-transgenic leaves was evaluated. The fungal growth was higher in the protein extracts from transgenic urease-deprived plants than in extracts from non-transgenic controls. When infected by P. pachyrhizi uredospores, detached leaves of urease-deprived plants developed a significantly higher number of lesions, pustules and erupted pustules than leaves of non-transgenic plants containing normal levels of the enzyme. The results of the present work show that the soybean plants were more susceptible to fungi in the absence of urease. It was not possible to overexpress active GmEu4. For future work, overexpression of urease fungitoxic peptides could be attempted as an alternative approach.

SUBMITTER: Wiebke-Strohm B

PROVIDER: S-EPMC3332383 | biostudies-literature | 2012 May

REPOSITORIES: biostudies-literature

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Ubiquitous urease affects soybean susceptibility to fungi.

Wiebke-Strohm Beatriz B Pasquali Giancarlo G Margis-Pinheiro Márcia M Bencke Marta M Bücker-Neto Lauro L Becker-Ritt Arlete B AB Martinelli Anne H S AH Rechenmacher Ciliana C Polacco Joseph C JC Stolf Renata R Marcelino Francismar C FC Abdelnoor Ricardo V RV Homrich Milena S MS Del Ponte Emerson M EM Carlini Celia R CR De Carvalho Mayra C C G MC Bodanese-Zanettini Maria Helena MH

Plant molecular biology 20120301 1-2

The soybean ubiquitous urease (encoded by GmEu4) is responsible for recycling metabolically derived urea. Additional biological roles have been demonstrated for plant ureases, notably in toxicity to other organisms. However, urease enzymatic activity is not related to its toxicity. The role of GmEu4 in soybean susceptibility to fungi was investigated in this study. A differential expression pattern of GmEu4 was observed in susceptible and resistant genotypes of soybeans over the course of a Phak ...[more]

PMID: 22382992

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Project description:Purpose: Soybean aphid (Aphis glycines Matsumura; SBA) is major pest of soybean (Glycine max) in the United States of America. One previous study on soybean, soybean-aphid interactions showed that avirulent (biotype 1) and virulent (biotype 2) biotypes can co-occur and potentially interact on resistant and susceptible soybean resulting induced susceptibility. The main objective of this research was to employ RNA sequencing technique to characterize the induced susceptibility effect in which initial feeding by virulent aphids can increase the suitability of avirulent aphids in both susceptible and resistant cultivars. Methods: The data in this submission come from the green house experiment using two genotypes of soybean: susceptible soybean cultivar was LD12-15838R and the resistant cultivar was LD12-15813Ra (with Rag1 gene) and two aphid populations: biotype 1 (avirulent) and biotype 2 (virulent biotype 2). RNA was extracted from the leave samples from resistant and susceptible cultivars treated with no aphids, biotype 2: biotype1 collected at day 1 and no aphids, biotype 2: biotype1 and no aphids: biotype1 at day 11 using PureLink RNA mini kit (Invitrogen, USA). RNA samples were treated with TURBOTM DNase (Invitrogen, USA) to remove any DNA contamination following the manufacturer’s instructions. Assessment of the isolated RNA integrity was performed by 1% agarose gel electrophoresis, and RNA concentration was measured by Nanodrop 2000 (Thermo Fisher Scientific, USA). Three replicates from these treatments in resistant and susceptible cultivars were pooled in equimolar concentration. RNAseq library construction was prepared using Illumina’s TruSeq Stranded mRNA Kit v1 (San Diego, CA). The libraries were quantified by QuBit dsDNA HS Assay (Life Technologies, Carlsbad, CA) and pooled in equimolar concentrations. The libraries were sequenced on an Illumina NextSeq 500 using a NextSeq 500/550 High Output Reagent Cartridge v2 (San Diego, CA) at 75 cycles. Results: A total of 10 RNA libraries were prepared and sequenced with the sequencing depth ranging from 24,779,816 to 29,72,4913. Total reads of 266,535,654 were subjected to FastQC analysis to determine the data quality using various quality metrics such as mean quality scores, per sequence quality scores, per sequence GC content, and sequence length distribution. The phred quality scores per base for all the samples were higher than 30. The GC content ranged from 45 to 46% and followed the normal distribution. After trimming, more than 99% of the reads were retained as the clean and good quality reads. Upon mapping these reads, we obtained high mapping rate ranging from 90.4% to 92.9%. Among the mapped reads, 85.8% to 91.9% reads were uniquely mapped. Conclusions: The objective of this study is to characterize the mechanism of induced susceptibility in soybean via transcriptional response study of soybean in presence of biotype 1 and biotype 2 soybean aphids using RNA-Seq. The data resulted from this study might provide insights into the interactions between soybean and soybean aphids and identify genes, their regulation and enriched pathways that may be associated with resistance or susceptibility to A. glycines.

Project description:Landscape complexity influences soybean aphid suppression by generalist predators in North America, but the role of adjacent habitats as sources of these predators has not been studied directly. We quantified movement of aphidophagous predators between soybean and five adjacent habitats common in Manitoba using bi-directional Malaise traps. To test the contribution of predators from neighboring habitats to soybean aphid suppression, we performed experimental manipulations in adjacent soybean and alfalfa fields and monitored the movement of sevenspotted lady beetles, Coccinella septempunctata, using mark-release-recapture experiments. The identity of adjacent habitats affected the net movement of predators into soybean. The most abundant predators were hover flies (Diptera: Syrphidae), moving from woodlands to soybean. Similar (but non-significant) trends were found for lady beetles, minute pirate bugs, and green and brown lacewings. There was also a net movement of hover flies and green lacewings from soybean to canola. Lady beetles showed higher bidirectional movement in alfalfa and wheat borders than in woodland and canola borders in a high lady beetle abundance year. Soybean aphid populations in predator exclusion cages were 21- to 122- fold higher than populations exposed to predators, both in alfalfa and soybean fields. Aerial predators provide similar levels of aphid suppression as aerial and epigeal predators combined. Mark-release-recapture experiments showed high dispersal of C. septempunctata between soybean and alfalfa, with a net movement towards alfalfa, probably due to the lack of aphids in soybean. These results demonstrate that predator assemblages from both soybeans and alfalfa can suppress soybean aphids. Our findings indicate that the type of adjacent habitat and predator identity affect the directionality of predator movement into soybean. This study suggests that information on predator movement can be used to design the distribution of crops and natural habitats in agricultural landscapes that maximize pest control services.

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Ubiquitous urease affects soybean susceptibility to fungi.

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Ubiquitous urease affects soybean susceptibility to fungi.

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