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The use of resistance genes in elite soybean cultivars is one of the most widely used methods to manage Phytophthora sojae. This method relies on effector-triggered immunity, where a Resistant to P. sojae (Rps) gene product from the plant recognizes a specific effector from the pathogen, encoded by an avirulence (Avr) gene. Many Avr genes from P. sojae have been identified in the last decade, allowing a better exploitation of this type of resistance. The objective of the present study was to identify the Avr gene triggering immunity derived from the soybean resistance gene Rps8. The analysis of a segregating F

Oomycetes form a deep lineage of eukaryotic organisms that includes a large number of plant pathogens which threaten natural and managed ecosystems. We undertook a survey to query the community for their ranking of plant-pathogenic oomycete species based on scientific and economic importance. In total, we received 263 votes from 62 scientists in 15 countries for a total of 33 species. The Top 10 species and their ranking are: (1) Phytophthora infestans; (2, tied) Hyaloperonospora arabidopsidis; (2, tied) Phytophthora ramorum; (4) Phytophthora sojae; (5) Phytophthora capsici; (6) Plasmopara viticola; (7) Phytophthora cinnamomi; (8, tied) Phytophthora parasitica; (8, tied) Pythium ultimum; and (10) Albugo candida. This article provides an introduction to these 10 taxa and a snapshot of current research. We hope that the list will serve as a benchmark for future trends in oomycete research.

A feature of pathogenic and invasive organisms is their adaptability when confronted with host and environmental challenges. Recent studies have demonstrated that plant pathogens rely on epigenetic processes for this purpose. Epiallelic variation of effector genes that results in evasion of host immunity is one emerging phenomenon. Another is the epigenetically induced reprogramming and diversification of transcriptional patterns by de-repression of transposable elements. These observations indicate that epigenetic control of gene expression provides a versatile means of generating phenotypic diversity that is adaptable and heritable across generations.

Cyclophilins (CYPs) belong to the immunophilin superfamily with peptidyl-prolyl cis-trans isomerase (PPIase) activity. They catalyze the interconversion of the cis- and trans-rotamers of the peptidyl-prolyl amide bond of peptides. A yeast-two-hybrid screening using the isoflavonoid regulator GmMYB176 as bait identified GmCYP1 as one of the interacting proteins in soybean embryos. GmCYP1 localizes both in the nucleus and cytoplasm, and interacts in planta with GmMYB176, in the nucleus, and with SGF14l (a soybean 14-3-3 protein) in the nucleus and the cytoplasm. GmCYP1 contains a single cyclophilin-like domain and displays a high sequence identity with other plant CYPs that are known to have stress-specific function. Tissue-specific expression of GmCYP1 revealed higher expression in developing seeds compared to other vegetative tissues, suggesting their seed-specific role. Furthermore, GmCYP1 transcript level was reduced in response to stress. Since isoflavonoids are involved in plant stress resistance against biotic and abiotic factors, the interaction of GmCYP1 with the isoflavonoid regulators GmMYB176 and 14-3-3 protein suggests its role in defense in soybean.

The process of RNA splicing influences many physiological processes, including plant immunity. However, how plant parasites manipulate host RNA splicing process remains unknown. Here we demonstrate that PsAvr3c, an avirulence effector from oomycete plant pathogen Phytophthora sojae, physically binds to and stabilizes soybean serine/lysine/arginine-rich proteins GmSKRPs. The SKRPs are novel proteins that associate with a complex that contains plant spliceosome components, and are negative regulators of plant immunity. Analysis by RNA-seq data indicates that alternative splicing of pre-mRNAs from 401 soybean genes, including defense-related genes, is altered in GmSKRP1 and PsAvr3c overexpressing lines compared to control plants. Representative splicing events mediated by GmSKRP1 and PsAvr3c are tested by infection assays or by transient expression in soybean plants. Our results show that plant pathogen effectors can reprogram host pre-mRNA splicing to promote disease, and we propose that pathogens evolved such strategies to defeat host immune systems., Various effectors of plant pathogens modulate host cell biology. Here, Huang et al. show PsAvr3c, an avirulence effector from oomycete plant pathogen Phytophthora sojae, can reprogram host pre-mRNA splicing for immune modulation.

Plants produce tens of thousands of different natural products also referred to as secondary metabolites. These metabolites were once thought to be the result of aberrant metabolism, or a form of transient storage of byproducts and intermediates thereof. Although the true role of such metabolites in plants remains mostly unknown, it is evident that plants invest a great deal of resources in synthesizing, accumulating and sorting such metabolites, often produced through complex and highly regulated biosynthetic pathways operating in multiple cellular and sub-cellular compartments. There is also growing evidence indicating that many biosynthetic pathways leading to the accumulation of plant natural products are not fully active. Thus, occult enzymes exist, sometimes without any apparent endogenous substrate or function, suggesting that plants have a reservoir of metabolic capabilities that normally remains hidden or unused. It is often difficult to accurately guess what are the actual biological roles of such enzymes solely based on bioinformatics, due to promiscuity towards substrates and the relatively ease to change substrate or product specificity by introducing minor changes in sequence of the enzymes. It could be that such orphan enzyme activities are relics of a recent past that have not been fully eliminated through selection and evolution. Additionally, it could be that such occult activities possess unknown biochemical roles and coincidentally are able to accept novel substrates. We have coined the term “silent metabolism” to describe occult metabolic capacities present or induced in plants. A few examples illustrating silent metabolism in the terpenoid and phenylpropanoid pathways, as well as their repercussion in the metabolic engineering of plant secondary metabolism are discussed in this review.

Ranathunge, K., Thomas, R. H., Fang, X., Peterson, C. A., Gijzen, M., and Bernards, M. A. 2008. Soybean root suberin and partial resistance to root rot caused by Phytophthora sojae. Phytopathology 98:1179-1189. Phytophthora sojae is the causal agent of root and stem rot of soybean (Glycine max). Various cultivars with partial resistance to the pathogen have been developed to mitigate this damage. Herein, two contrasting genotypes, the cultivar Conrad (with strong partial resistance) and the line OX760-6 (with weak partial resistance), were compared regarding their amounts of preformed and induced suberin components, and to early events during the P. sojae infection process. To colonize the root, hyphae grew through the suberized middle lamellae between epidermal cells. This took 2 to 3 h longer in Conrad than in OX760-6, giving Conrad plants more time to establish their chemical defenses. Subsequent growth of hyphae through the endodermis was also delayed in Conrad. This cultivar had more preformed aliphatic suberin than the line OX760-6 and was induced to form more aliphatic suberin several days prior to that of OX760-6. However, the induced suberin was formed subsequent to the initial infection process. Eventually, the amount of induced suberin (measured 8 days postinoculation) was the same in both genotypes. Preformed root epidermal suberin provides a target for selection and development of new soybean cultivars with higher levels of expression of partial resistance to P. sojae.

Soybean root fluorescence in ultraviolet light is controlled by the Fr loci. Extracts from fluorescent roots of soybean contain three closely related fluorescent compounds that are greatly reduced in extracts from non-fluorescent roots. Root fluorescence is due to their presence, regardless of the Frlocus that conditions this trait. Key words: Coumarin, genetic locus, glycoside, fluorescence, roots, soybean

The Phytophthora sojae avirulence gene Avr3a encodes an effector that is capable of triggering immunity on soybean plants carrying the resistance gene Rps3a. P. sojae strains that express Avr3a are avirulent to Rps3a plants, while strains that do not are virulent. To study the inheritance of Avr3a expression and virulence towards Rps3a, genetic crosses and self-fertilizations were performed. A cross between P. sojae strains ACR10 X P7076 causes transgenerational gene silencing of Avr3a allele, and this effect is meiotically stable up to the F5 generation. However, test-crosses of F1 progeny (ACR10 X P7076) with strain P6497 result in the release of silencing of Avr3a. Expression of Avr3a in the progeny is variable and correlates with the phenotypic penetrance of the avirulence trait. The F1 progeny from a direct cross of P6497 X ACR10 segregate for inheritance for Avr3a expression, a result that could not be explained by parental imprinting or heterozygosity. Analysis of small RNA arising from the Avr3a gene sequence in the parental strains and hybrid progeny suggests that the presence of small RNA is necessary but not sufficient for gene silencing. Overall, we conclude that inheritance of the Avr3a gene silenced phenotype relies on factors that are variable among P. sojae strains.

Hydrophobic protein from soybean (HPS) is present in soybean dust and is an allergen (Gly m 1) that causes asthma in allergic individuals. Past studies have shown that HPS occurs on the seed surface. To determine the microscopic localization of HPS during seed development, monoclonal antibodies to HPS were used to visualize the protein by fluorescence and transmission electron microscopy. Seed coat and endocarp sections were also examined for pectin, cellulose, callose, starch and protein by histochemical staining. HPS is present in the endocarp epidermal cells at 18 to 28 days post anthesis. At later stages of seed development, HPS occurs in extracellular secretions that accumulate unevenly on the endocarp epidermis and seed surface. HPS is synthesized by the endocarp epidermis and deposited on the seed surface as part of a heterogeneous matrix.

We have used cDNA microarray analysis to examine changes in gene expression during embryo development in soybean (Glycine max) and to compare gene expression profiles of two soybean cultivars that differ in seed isoflavonoid content. The analysis identified 5,910 genes that were differentially expressed in both soybean cultivars grown at two different locations for two consecutive years in one of the five different stages of embryo development. An ANOVA analysis with P value < 0.05 and < 0.01 indicated that gene expression changes due to environmental factors are greater than those due to cultivar differences. Most changes in gene expression occurred at the stages when the embryos were at 30 or 70 d after pollination. A significantly larger fraction of genes (48.5%) was expressed throughout the development and showed little or no change in expression. Transcript accumulation for genes related to the biosynthesis of storage components in soybean embryos showed several unique temporal expressions. Expression patterns of several genes involved in isoflavonoid biosynthesis, such as Phenylalanine Ammonia-Lyase, Chalcone Synthase (CHS) 7, CHS8, and Isoflavone Synthase2, were higher at 70 d after pollination in both the cultivars. Thus, expression of these genes coincides with the onset of accumulation of isoflavonoids in the embryos. A comparative analysis of genes involved in isoflavonoid biosynthesis in RCAT Angora (high seed isoflavonoid cultivar) and Harovinton (low seed isoflavonoid cultivar) revealed that CHS7 and CHS8 were expressed at significantly greater level in RCAT Angora than in Harovinton. Our study provides a detailed transcriptome profiling of soybean embryos during development and indicates that differences in the level of seed isoflavonoids between these two cultivars could be as a result of differential expression of CHS7 and CHS8 during late stages of seed development.

A new genetic locus mediating avirulence in the potato late blight pathogen Phytophthora infestans has been discovered. The Avr3b-Avr10-Avr11 locus is recognized by three different potato resistance genes, and is different from other Avr loci that have been identified thus far. This locus encodes a large protein with a WD40 domain and sequence similarities to transcription factors. Multiple, truncated copies of this gene have arisen by gene amplification and are characteristic of avirulent strains of P. infestans. Here, we describe the new avirulence locus and discuss the importance of this finding.

An emerging group of proteins found in many plant pathogens are related to their ability to cause plant cell death. These proteins may be identified by the presence of a common NPP1 (necrosis-inducing Phytophthora protein) domain, and have collectively been named NLPs (Nep1-like proteins). The NLPs are distinguished by their wide distribution across taxa and their broad spectrum of activity against dicotyledonous plants. The function of NLPs is not known but there is strong evidence that they may act as positive virulence factors, accelerating disease and pathogen growth in plant hosts. Interest in NLPs is gaining momentum as more members of this protein family are discovered in more species of plant pathogens.

The seed coat serves as a multifunctional organ with a role in protection and for the supply of nutrients to the embryo sac during development. The composition of the legume seed coat differs from other seed tissues in many ways including its protein composition. An abundant 24 kDa protein (SC24) has been purified and identified from soybean (Glycine max [L.] Merr) seed hulls. The corresponding cDNA and genomic DNA clones for SC24 were isolated and characterized, and expression patterns were determined. The deduced protein sequence of 219 amino acids included an N-terminal signal peptide. Transcripts encoding SC24 were present in the seed coat from 30 days after pollination (DAP) until maturity, but the protein was not detected until the final stages of seed maturation. In mature seeds, most of the SC24 protein was localized to the parenchyma and aleurone layers of the seed coat. The expression of SC24 was also induced in vegetative tissues by pathogen infection and by wounding. The SC24 protein bound to an affinity column containing an isophthalic acid ligand, and was eluted with 7 mM citrate. Polyclonal antibodies raised against recombinant SC24 cross-reacted with the seed coat peroxidase enzyme, suggesting that these two proteins may share an antigenic determinant. Overall, the results indicate that SC24 belongs to a novel class of plant defence proteins with carboxylate-binding activity.

This study presents the application of soybean seed hulls for the decontamination and/or detoxification of phenol and chlorophenol polluted soils. The effect of soil was examined on both catalytic activity of soybean seed hull peroxidase (SBP) and enzymatic transformation of phenol, 2-chlorophenol, and 2,4-dichlorophenol through the polymerization reaction. The sorption of the enzyme to soil organic matter was found to account for the loss of partial catalytic activity of soybean seed hulls in a soil slurry environment. The organic matter present in loamy soils, rather than other soluble soil constituents and soil micro-organisms, is a factor in SBP inhibition by soil and the corresponding decline in treatment efficiency of phenol and chlorophenols. Under improved conditions, however, soybean seed hulls demonstrated a satisfactory ability to catalyze the polymerization reaction. Over 96% of total phenols were removed using soybean seed hulls in a soil slurry bioreactor, which demonstrates a great potential in use of soybean seed hulls, a readily available and inexpensive source of SBP, for bioremediation of soils contaminated with phenolic compounds.

A soybean (Glycine max (L.) Merr.) cotyledon has a concave region on its abaxial center called a pit, and the seed coat has a corresponding convex region on its inner surface called an antipit. While it is clear that the pit is lined with large epidermal cells, the anatomical identity of the antipit has been a subject of confusion. The inner surface of the seed coat consists of a compressed endosperm tissue that is subtended by an aleurone layer. In the antipit region of the seed coat, additional endosperm cells are situated between the aleurone layer and the compressed endosperm tissue. These endosperm cells, called cone cells, are similar to ordinary aleurone cells in basic structure. In the present study, it is demonstrated that the surface ornamentations of the antipit are a print of the pit on the compressed endosperm tissue. The functional aspects of the antipit–pit complex are not known. It was previously postulated that the antipit–pit region has an enhanced capability of nutrient translocation to the growing embryo, but there is little evidence to support this hypothesis. Alternatively, the antipit–pit association may provide a structural feature that anchors the embryo within the seed coat while contributing to an overall spherical shape of the seed.Key words: antipit, endosperm, Glycine max, pit, seed coat, soybean.

Soybean peroxidase (SBP), (EC 1.11.1.7) can be readily extracted from soybean seed hulls. This study reports on the direct use of soybean seed-hull extracts for the bioremediation of phenolic wastes. The crude SBP extract from the hulls, like pure soybean peroxidase, is catalytically active in a broad range of pH and temperatures. As SBP is gradually released into the aqueous solution from seed hulls, the direct use of soybean seed hulls can reduce SBP inactivation by H 2 O 2 and enhance the utilization efficiency of SBP through the slow release of the enzyme from the seed hulls. However, large doses of soybean seed hulls were found to be ineffective in phenol removal. Gradual additions of H 2 O 2 in combination with the SBP released from the hulls were applied to optimize the bioremediation. Since the crude extract contains a mixture of multiple soybean proteins, soybean seed hull slurry required a higher concentration of H 2 O 2 to remove the phenolic substrates than did the purified enzyme. Under the experimental conditions, 80 % of phenol (10.6 mM), 96 % of 2-chlorophenol (3.9 mM), 95 % of 2,4-dichlorophenol (3.1 mM), and 94 % of mixed phenol and chlorophenols were removed using soybean seed hulls in a single batch reactor. These results demonstrate that soybean seed hulls, compared to purified SBP, may be a more cost-effective alternative in the enzymatic removal of phenolic compounds through polymerization reactions.

Isoflavonoids are biologically active natural products that accumulate in soybean seeds during development. The amount of isoflavonoids present in soybean seed is variable, depending on genetic and environmental factors that are not fully understood. Experiments were conducted to determine whether isoflavonoids are synthesized within seed tissues during development, or made in other plant organs and transported to the seeds where they accumulate. An analysis of isoflavonoids by HPLC detected the compounds in all organs of soybean plant, but the amount of isoflavonoids present varied depending on the tissue and developmental stage. The greatest concentrations were found in mature seeds and leaves. The 2-hydroxyisoflavanone synthase genes IFS1 and IFS2 were studied to determine their pattern of expression in different tissues and developmental stages. The highest level of expression of IFS1 was observed in the root and seed coat, while IFS2 was most highly expressed in embryos and pods, and in elicitor-treated or pathogen-challenged tissues. Incorporation of radiolabel into isoflavonoids was observed when developing embryos and other plant organs were fed with [(14)C]phenylalanine. Embryos excised from developing soybean seeds also accumulated isoflavonoids from a synthetic medium. A maternal effect on seed isoflavonoid content was noted in reciprocal crosses between soybean cultivars that differ in seed isoflavonoids. From these results, we propose that developing soybean embryos have an ability to synthesize isoflavonoids de novo, but that transport from maternal tissues may in part contribute to the accumulation of these natural products in the seed.

SUMMARY Transcripts encoding elicitin-like protein domains were identified from similarity searches of Phytophthora sojae expressed sequence tags and were characterized with regard to molecular structure and elicitor activity. The P. sojae elicitin family consists of at least nine genes with products similar to previously described elicitins (SOJA-2, SOJB, SOJ2, SOJ3, SOJ5, SOJ6 and SOJ7) or highly diverged from known sequences (SOJX and SOJY). The predicted structural features of seven (SOJA-2, SOJB, SOJ2, SOJ3, SOJ6, SOJX and SOJY) of the elicitin preproteins were compared. All of the predicted elicitins possess a leader signal sequence and a core elicitin domain. Five (SOJ2, SOJ3, SOJ6, SOJX and SOJY) of the characterized elicitins also contain a variable C-terminal region. In addition, SOJX and SOJY contain a C-terminal hydrophobic membrane-spanning domain. An analysis of expression patterns of the elicitin transcripts showed that SOJA-2, SOJB, SOJ2, SOJ3 and SOJ6 were expressed in axenically grown mycelia and during infection, but not in zoospores. In contrast, SOJX and SOJY were predominantly and specifically expressed in zoospores. Selected elicitin domains were also tested for the induction of the hypersensitive response (HR) in Nicotiana spp. All of the elicitin protein domains tested induced the HR, except for SOJX and SOJY. Overall, the results show that the P. sojae elicitin gene family is large and diverse, with varying patterns of expression and HR-inducing activity.

The 90-kD molecular chaperone hsp90 is the key component of a multiprotein chaperone complex that facilitates folding, stabilization, and functional modulation of a number of signaling proteins. The components of the animal chaperone complex include hsp90, hsp70, hsp40, Hop, and p23. The animal Hop functions to link hsp90 and hsp70, and it can also inhibit the ATPase activity of hsp90. We have demonstrated the presence of an hsp90 chaperone complex in plant cells, but not all components of the complex have been identified. Here, we report the isolation and characterization of soybean (Glycine max) GmHop-1, a soybean homolog of mammalian Hop. An analysis of soybean expressed sequence tags, combined with preexisting data in literature, suggested the presence of at least three related genes encoding Hop-like proteins in soybean. Transcripts corresponding to Hop-like proteins in soybean were detected under normal growth conditions, and their levels increased further in response to stress. A recombinant GmHop-1 bound hsp90 and its binding to hsp90 could be blocked by the tetratricopeptide repeat (TPR) domain of rat (Rattus norvegicus) protein phosphatase 5. Deletion of amino acids 325 to 395, adjacent to the TPR2A domain in GmHop-1, resulted in loss of hsp90 binding. In a minimal assembly system, GmHop-1 was able to stimulate mammalian steroid receptor folding. These data show that plant and animal Hop homologs are conserved in their general characteristics, and suggest that a Hop-like protein in plants is an important cochaperone of plant hsp90.

Plant pathogens display impressive versatility in adapting to host immune systems. Pathogen effector proteins facilitate disease but can become avirulence (Avr) factors when the host acquires discrete recognition capabilities that trigger immunity. The mechanisms that lead to changes to pathogen Avr factors that enable escape from host immunity are diverse, and include epigenetic switches that allow for reuse or recycling of effectors. This perspective outlines possibilities of how epigenetic control of Avr effector gene expression may have arisen and persisted in filamentous plant pathogens, and how it presents special problems for diagnosis and detection of specific pathogen strains or pathotypes.

Soybean seed hull peroxidase (SBP) is an inexpensive oxidoreductive enzyme and could potentially be used to oxidise/polymerise various organic pollutants present in the industrial and petrochemical wastes. SBP is able to retain its catalytic properties under wide ranges of pH and at elevated temperatures. In this study, a systematic evaluation of the biocatalytic properties of SBP was carried out. The optimal pH for SBP activity is pH 6.0 and significant activity was observed between 2.2 and 8.0. SBP also showed three times higher activity at an elevated temperature of 80°C and at pH 6.0 when compared to the activity at room temperature. The pH and temperature of the reaction mixture were found to significantly influence the SBP activity. SBP is fairly active in organic solvents. The enzyme exhibited highest activity in the presence of 16.67% (w/v) ethanol followed by acetone, methanol and acetonitrile. The enzyme activity was reduced with an increase in concentration of the organic solvent. SBP also showed maximum activity at different concentrations of acetone using a phosphate buffer, pH 6.0 than with the other pH buffers. Benzene/acetone mixture seems to be another better solvent system for SBP where it showed about 65% of its activity at 16.67% (w/v) concentration.

Phytophthora sojae (Kaufmann and Gerdemann) is an oomycete that causes stem and root rot on soybean (Glycine max L. Merr) plants. We have constructed three cDNA libraries using mRNA isolated from axenically grown mycelium and zoospores and from tissue isolated from plant hypocotyls 48 h after inoculation with zoospores. A total of 3,035 expressed sequence tags (ESTs) were generated from the three cDNA libraries, representing an estimated 2,189 cDNA transcripts. The ESTs were classified according to putative function based on similarity to known proteins, and were analyzed for redundancy within and among the three source libraries. Distinct expression patterns were observed for each library. By analysis of the percentage G+C content of the ESTs, we estimate that two-thirds of the ESTs from the infected plant library are derived from P. sojae cDNA transcripts. The ESTs originating from this study were also compared with a collection of Phytophthora infestans ESTs and with all other non-human ESTs to assess the similarity of the P. sojae sequences to existing EST data. This collection of cDNA libraries, ESTs, and accompanying annotation will provide a new resource for studies on oomycetes and on soybean responses to pathogen challenge.

A seed coat-specific gene, SCS1 (Seed Coat Subtilisin 1), from soybean, Glycine max [L.] Merill, has been identified and studied. The gene belongs to a small family of genes with sequence similarity to the subtilisins, which are serine proteases. Northern blot analysis showed that SCS1 RNA accumulates to maximal levels in seed coats at 12 days post anthesis, preceding the final stages of seed coat differentiation. The SCS1 RNA was not found in other tissues including embryos, seed pods, flowers, stems, roots or leaves. In-situ hybridization studies confirmed the temporal pattern of expression observed by Northern blot analysis and further revealed a restricted pattern of RNA accumulation in thick-walled parenchyma cells of the seed coats. These cells are important in the apoplastic translocation of nutrients en route to the embryo from the vascular tissues. The tissue-specific subtilisin-like gene may be required for regulating the differentiation of the thick-walled parenchyma cells.

Stevia rebaudiana Bertoni leaves accumulate a mixture of at least eight different glycosides derived from the tetracyclic diterpene steviol. These natural products taste intensely sweet and have similar biosynthetic origins to those of gibberellic acid (GA). The initial steps leading to the formation of GA result from the two-step cyclization of geranylgeranyl diphosphate (GGDP) to (-)-kaurene via the action of two terpene cyclases (-)-copalyl diphosphate synthase (CPS) and (-)-kaurene synthase (KS). Steviol biosynthesis probably uses the same mechanism although the genes and enzymes from S. rebaudiana that are involved in the cyclization of GGDP have not been characterized. We have isolated both the CPS and KS genes from S. rebaudiana and found that recombinant CPS and KS were catalytically active, suggesting that the CPS and KS genes participate in steviol biosynthesis. The genes coding for CPS and KS are usually present in single copies in most plant species and their expression is normally low and limited to rapidly growing tissues. The KS gene has been duplicated in the S. rebaudiana genome and both the KS and CPS genes are highly expressed in mature leaves, a pattern opposite to that found with GA biosynthesis. This pattern may, at least in part, lead to temporal and spatial separation of GA and steviol biosynthesis and probably helps to prevent over-expression from interfering with normal GA metabolism. Our results show that CPS and KS are part of the steviol glycoside biosynthetic pathway and that Stevia rebaudiana has recruited two genes to secondary metabolism from a highly regulated pathway involved in hormone biosynthesis.

In this study, the removal of phenol and 2-chlorophenol using soybean seed-hulls in the presence of hydrogen peroxide is demonstrated. The performance of a stirred membrane reactor containing soluble purified SBP was compared with a batch stirred reactor containing raw soybean seed-hulls. The purified enzyme reactor proved to be ineffective while excellent results were obtained with the crude seed-hulls for the removal of phenol and 2-chlorophenol. Four sequential batch reactors containing raw seed-hulls achieved greater than 96% removal of phenol with a retention time of 20min in each reactor. A single batch reactor containing raw seed-hulls was effective in removing greater than 98.5% of 2-chlorophenol (initially at 1000 ppm) in less than 15 min. The performance of these reactors is comparable to existing HRP-based technology. The stability of the soybean peroxidase (SBP) enzyme was also examined in the presence of detergents (SDS, Tween 20 and Triton X-100). Low concentrations of the detergents significantly increased the enzyme activity and higher concentrations of detergents (up to 20% w/v) did not inactivate the SBP enzyme. These results demonstrate that SBP has good potential for the treatment of phenol contaminated solutions.

The soybean Ep gene encodes an anionic peroxidase enzyme that accumulates in large amounts in seed coat tissues. We have isolated a second peroxidase gene, Prx2, that is also highly expressed in developing seed coat tissues. Sequence analysis of Prx2 cDNA indicates that this transcript encodes a cationic peroxidase isozyme that is far removed from Ep in peroxidase phylogeny. To determine the expression patterns for these two peroxidases in developing seeds, the abundance and localization of the Ep and Prx2 transcripts were compared by in situ hybridization. Results show the expression of Ep begins in a small number of cells flanking the vascular bundle in the seed coat, spreads to encircle the seed, and then migrates to the hourglass cells as they develop. Expression of Prx2 occurs throughout development in all cell layers of the seed coat, and is also evident in the pericarp and embryo. Nonetheless, the Ep-encoded enzyme accounts for virtually all of the peroxidase activity detected in mature seed coats. The Prx2 enzyme is either insoluble in a catalytically inactive form, or is subject to degradation during seed maturation.

Stevia rebaudiana is a member of the Compositae, native to Paraguay. It produces a number of high-potency low-calorie sweeteners in its leaf tissue. The sweeteners are diterpene glycosides and range between 30 and 320 times sweeter than sugar. Increasing consumer interest in natural food ingredients means that products like stevia sweeteners will be subject to increasing demand. Such demand will need to be supported by a modern mechanised production system. The purpose of this review is to summarize the existing agricultural, chemical and biochemical literature to provide a baseline for new research. Key words: Stevia, diterpene, steviol glycoside, sweeteners

Phytophthora sojae is a soil-borne microorganism and a plant pathogen that causes stem and root rot of soybean. P. sojae is one of the ~120 species of plant pathogenic eukaryotes within the genus Phytophthora. These organisms are oomycetes, fungus-like organisms in the kingdom Stramenopila. Together with members of related genera such as Pythium, Peronospora, and Albugo, these organisms cause highly destructive diseases of plants of importance to agriculture, horticulture, forestry, and natural ecosystems. The draft genome sequence of P. sojae, together with that of the forest pathogen P. ramorum, was completed in 2004 and published in 2006. These were the first oomycete genomes sequenced, and have provided a resource of information that has revolutionized our understanding of oomycete infection mechanisms, and propelled oomycete research to the forefront of studies of molecular plant-microbe interactions. Highlights of insights that have emerged from the genome sequence of P. sojae and other oomycetes include: a bipartite genome organization whereby rapidly evolving genes involved in infection are located in dynamic, transposon-rich regions while highly conserved housekeeping genes are located in stable gene-rich regions; two classes of effector proteins that can cross into the cytoplasm of host plant cells to suppress immunity; the role of epigenetic variation in oomycete pathogen adaptation; and the role of transkingdom horizontal gene transfer in introducing key virulence factors into the genomes of plant pathogenic oomycetes.

Grapevine (Vitis vinifera) is susceptible to many pathogens which cause significant losses to viticulture worldwide. Chemical control is available, but agro-ecological concerns have raised interest in alternative methods, especially in triggering plant immunity by elicitor treatments. The b-glucan laminarin (Lam) and its sulfated derivative (PS3) have been previously demonstrated to induce resistance in grapevine against downy mildew (Plasmopara viticola). However, if Lam elicits classical grapevine defenses such as oxidative burst, pathogenesis-related (PR)-proteins and phytoalexin production, PS3 triggered grapevine resistance via a poorly understood priming phenomenon. The aim of this study was to identify the molecular mechanisms of the PS3-induced resistance. For this purpose we studied i) the signaling events and transcriptome reprogramming triggered by PS3 treatment on uninfected grapevine, ii) grapevine immune responses primed by PS3 during P. viticola infection. Our results showed that i) PS3 was unable to elicit reactive oxygen species (ROS) production, cytosolic Ca2+ concentration variations, mitogen-activated protein kinase (MAPK) activation but triggered a long lasting plasma membrane depolarization in grapevine cells, ii) PS3 and Lam shared a common stress-responsive transcriptome profile that partly overlapped the salicylate- (SA) and jasmonate-(JA)-dependent ones. After P. viticola inoculation, PS3 specifically primed the SA- and ROS-dependent defense pathways leading to grapevine induced resistance against this biotroph. Interestingly pharmacological approaches suggested that the plasma membrane depolarization and the downstream ROS production are key events of the PS3-induced resistance