Biologically active Phytophthora mating hormone prepared by catalytic asymmetric total synthesis

A Phytophthora mating hormone with an array of 1,5-stereogenic centers has been synthesized by using our recently developed methodology of catalytic enantioselective conjugate addition of Grignard reagents. We applied this methodology in a diastereo- and enantioselective iterative route and obtained two of the 16 possible stereoisomers of Phytophthora hormone α1. These synthetic stereoisomers induced the formation of sexual spores (oospores) in A2 mating type strains of three heterothallic Phytophthora species, P. infestans, P. capsici, and P. nicotianae but not in A1 mating type strains. The response was concentration-dependent, and the oospores were viable. These results demonstrate that the biological activity of the synthetic hormone resembles that of the natural hormone α1. Mating hormones are essential components in the sexual life cycle of a variety of organisms. For plant pathogens like Phytophthora, sexual reproduction is important as a source of genetic variation. Moreover, the thick-walled oospores are the most durable propagules that can survive harsh environmental conditions. Sexual reproduction can thus greatly affect disease epidemics. The availability of synthetic compounds mimicking the activity of Phytophthora mating hormone will be instrumental for further unravelling sexual reproduction in this important group of plant pathogens.

The aspartic proteinase family of three Phytophthora species

Background - Phytophthora species are oomycete plant pathogens with such major social and economic impact that genome sequences have been determined for Phytophthora infestans, P. sojae and P. ramorum. Pepsin-like aspartic proteinases (APs) are produced in a wide variety of species (from bacteria to humans) and contain conserved motifs and landmark residues. APs fulfil critical roles in infectious organisms and their host cells. Annotation of Phytophthora APs would provide invaluable information for studies into their roles in the physiology of Phytophthora species and interactions with their hosts. Results - Genomes of Phytophthora infestans, P. sojae and P. ramorum contain 11-12 genes encoding APs. Nine of the original gene models in the P. infestans database and several in P. sojae and P. ramorum (three and four, respectively) were erroneous. Gene models were corrected on the basis of EST data, consistent positioning of introns between orthologues and conservation of hallmark motifs. Phylogenetic analysis resolved the Phytophthora APs into 5 clades. Of the 12 sub-families, several contained an unconventional architecture, as they either lacked a signal peptide or a propart region. Remarkably, almost all APs are predicted to be membrane-bound. Conclusions - One of the twelve Phytophthora APs is an unprecedented fusion protein with a putative G-protein coupled receptor as the C-terminal partner. The others appear to be related to well-documented enzymes from other species, including a vacuolar enzyme that is encoded in every fungal genome sequenced to date. Unexpectedly, however, the oomycetes were found to have both active and probably-inactive forms of an AP similar to vertebrate BACE, the enzyme responsible for initiating the processing cascade that generates the Aß peptide central to Alzheimer's Disease. The oomycetes also encode enzymes similar to plasmepsin V, a membrane-bound AP that cleaves effector proteins of the malaria parasite Plasmodium falciparum during their translocation into the host red blood cell. Since the translocation of Phytophthora effector proteins is currently a topic of intense research activity, the identification in Phytophthora of potential functional homologues of plasmepsin V would appear worthy of investigation. Indeed, elucidation of the physiological roles of the APs identified here offers areas for future study. The significant revision of gene models and detailed annotation presented here should significantly facilitate experimental design

Some factors influencing the sudden death syndrome in cut flower plants : a thesis presented in partial fulfilment of the requirements for the degree of Master of Horticultural Science at Massey University

Soil/root mixes from plants with the Sudden Collapse Syndrome of cut flower plants were tested for Phytophthora infection using a lupin (Lupinus angustifolius) baiting technique. Boronia heterophylla and Leucadendron 'Wilsons Wonder' root samples both caused the lupin seedlings to exhibit symptoms of Phytophthora infection. The efficacy of phosphorous acid (Foschek® 500 at 1000 ppm and 2000 ppm) and a combination of phosphorous acid and an additional product (Foschek® 500 and C408 at 1000/200 ppm and 2000/400 ppm) in controlling Phytophthora cinnamomi root infections of L. 'Wilsons Wonder', B.heterophylla and B megastigma rooted cuttings was compared with fosetyl- Al (Aliette® 80 SP at 1000 ppm and 2000 ppm) under conditions of high disease pressure The fungicides were applied as a root drench 7 days prior to the roots being inoculated by a split wheat technique and the effect of the fungicides and their concentrations on the rate of plant mortality was measured The results were species dependent. The treatments delaying plant mortality most effectively were fosetyl-AI at 2000 ppm on L 'Wilsons Wonder', phosphorous acid at 2000 ppm on B. heterophylla and both fosetyl-AI at 1000 or 2000 ppm and phosphorous acid at 2000 ppm on B. megastigma. The allelopathic activity of the root bark of Protea cynaroides, L. 'Wilsons Wonder'. Macadamia 'Beaumont' and Knightia excelsa was evaluated as a growth inhibitor for Phytophthora cinnamomi. The results indicate that by day 4 the root bark of M. 'Beaumont' reduced the growth rate of Phytophthora cinnamomi by 76.8% while that of Protea cynaroides inhibited the growth totally. The root bark of L. 'Wilsons Wonder' had no effect on the growth rate but that of K. excelsa enhanced the growth rate by 128% by day 4. The root bark of Protea cynaroides plants previously infected with an unnamed, indigenous Phytophthora species provided greater resistance to the growth rate of Phytophthora cinnamomi than the root bark of uninfected plants On the corn meal agar the leachate of the infected Protea cynaroides root bark exhibited a 'zone of inhibition' which prevented the growth of Phytophthora cinnamomi. Possible reasons for this are discussed

Unraveling the senses of Phytophthora; leads to novel control strategies?

Oomycetes cause devastating diseases on plants and animals. They cause major yield losses in many crop plants and their control heavily depends on agrochemicals. This is certainly true for the potato late blight pathogen Phytophthora infestans. Strong concerns about adverse effects of agrochemicals on food safety and environment are incentives for the development of novel, environmental friendly control strategies preferably based on natural products. Cyclic lipopeptides (CLPs) were recently discovered as a new class of natural compounds with strong activities against oomycetes including Phytophthora. CLPs lyse zoospores, inhibit mycelial growth and effectively reduce late blight disease. In order to unravel how Phytophthora senses CLPs and other environmental signals we follow two approaches. On the one hand, we monitor genome wide changes in gene expression induced by CLPs with the aim to identify the cellular pathways targeted by CLPs. On the other hand, we analyse components of ubiquitous signal transduction pathways with the aim to identify features that are unique for Phytophthora or oomycetes and, hence, could be suitable targets for novel anti-oomycete agents. Mining and comparing whole genome sequences have revealed that Phytophthora harbours many novel phospholipid modifying enzymes, unique for oomycetes. They have aberrant combinations of catalytic and regulatory domains occasionally combined with transmembrane domains. The latter resemble receptors that might be activated by extracellular ligands. Phospholipids, the substrates of these enzymes, are structural membrane components that also function in signalling. Together these findings open new avenues of research aimed at target-discovery in oomycetes

Investigation on the Biocontrol of Phytophthora diseases on strawberry based on antagonism

After screening of several rhizosphere bacteria against the soilborne pathogens of red core and crown rot disease of strawberry Phytophthora fragariae var. fragariae and Phytophthora cactorum under in vitro conditions, three of the most active isolates which produced up to 63% of reduction in mycelium growth, such as Raoultella terrigena (G-584), Bacillus amyloliquefaciens (G-V1) and Pseudomonas fluorescens (2R1-7) were selected for further studies under in vivo conditions. In a greenhouse and field experiments, mentioned above three isolates were tested against both Phytophthora diseases under artificial infested soil conditions. Root dip treatment with these bacterial antagonists produced a control effect on both fungal diseases between 27 to 55 % and were in some cases comparable with the chemical fungicide Aliette

A survey in natural forest ecosystems of Vietnam reveals high diversity of both new and described Phytophthora taxa including P. ramorum

In 2016 and 2017, surveys of Phytophthora diversity were performed in 25 natural and semi-natural forest stands and 16 rivers in temperate and subtropical montane and tropical lowland regions of Vietnam. Using baiting assays from soil samples and rivers and direct isolations from naturally fallen leaves, 13 described species, five informally designated taxa and 21 previously unknown taxa of Phytophthora were isolated from 58 of the 91 soil samples (63.7%) taken from the rhizosphere of 52 of the 64 woody plant species sampled (81.3%) in 20 forest stands (83.7%), and from all rivers: P. capensis, P. citricola VII, VIII, IX, X and XI, P. sp. botryosa-like 2, P. sp. meadii-like 1 and 2, P. sp. tropicalis-like 2 and P. sp. multivesiculata-like 1 from Phytophthora major phylogenetic Clade 2; P. castaneae and P. heveae from Clade 5; P. chlamydospora, P. gregata, P. sp. bitahaiensis-like and P. sp. sylvatica-like 1, 2 and 3 from Clade 6; P. cinnamomi (Pc), P. parvispora, P. attenuata, P. sp. attenuata-like 1, 2 and 3 and P. ×heterohybrida from Clade 7; P. drechsleri, P. pseudocryptogea, P. ramorum (Pr) and P. sp. kelmania from Clade 8, P. macrochlamydospora, P. sp. ×insolita-like, P. sp. ×kunnunara-like, P. sp. ×virginiana-like s.l. and three new taxa, P. sp. quininea-like, P. sp. ×Grenada 3-like and P. sp. ×Peru 4-like, from Clade 9; and P. sp. gallica-like 1 and 2 from Clade 10. The A1 and A2 mating types of both Pc and Pr co-occurred. The A2 mating type of Pc was associated with severe dieback of montane forests in northern Vietnam. Most other Phytophthora species, including Pr, were not associated with obvious disease symptoms. It is concluded that (1) Vietnam is within the center of origin of most Phytophthora taxa found including Pc and Pr, and (2) Phytophthora clades 2, 5, 6, 7, 8, 9, and 10 are native to Indochina.info:eu-repo/semantics/publishedVersio

Management of Phytophthora cinnamomi for biodiversity conservation in Australia: Part 2. National best practice guidelines

Disease in natural ecosystems of Australia, caused by the introduced plant pathogen Phytophthora cinnamomi, is listed as a key threatening process under the Environment Protection and Biodiversity Conservation Act 1999 (EPBC Act). The Act requires the Australian Government to prepare and implement a threat abatement plan for nationally coordinated action to mitigate the harm caused by P. cinnamomi to Australian species, particularly threatened flora, fauna and ecological communities. The .National Threat Abatement Plan for Dieback Caused by the Root-Rot Fungus Phytophthora cinnamomi. (NTAP) was released in 2001 (Environment Australia, 2001). The NTAP is designed to promote a common understanding of the national threat P. cinnamomi poses to biodiversity in Australia. This project, funded by the Australian Government Department of the Environment and Heritage (DEH), is one of the most significant actions to be implemented from the NTAP to date. The project has two major components: * to review current management approaches and identify benchmarks for best practice * the development of risk assessment criteria and a system for prioritising management of assets that are or could be threatened by P. cinnamomi. The project outputs are presented in a four-part document entitled Management of Phytophthora cinnamomi for Biodiversity Conservation in Australia: Part 1 - A Review of Current Management Part 2 - National Best Practice Guidelines Part 3 - Risk Assessment for Threats to Ecosystems, Species and Communities: A Review Part 4 - Risk Assessment Models for Species, Ecological Communities and Areas. A model of best practice was developed which encompasses all the components necessary for an informed and integrated approach to P. cinnamomi management, from strategic through to on-ground management. The current document (Part 1 . A Review of Current Management) thoroughly reviews the approaches to P. cinnamomi management in Australia within the context of the best practice model

Sexual reproduction in Phytophthora infestans

Potato late blight, caused by the oomycete Phytophthora infestans, is one of the most devastating plant diseases worldwide. It is becoming increasingly difficult to control the disease in Sweden, leading to an ever more intensified use of fungicides in the potato production. This unsustainable development could be caused by changes in the pathogen due to the introduction of sexual reproduction. These changes could result in an enhanced capability of the pathogen to adapt to different factors, e. g. weather, cultivar resistance, chemical control or other cultivation measures. This might manifest itself in earlier infections and a late blight that spreads faster in the potato fields requiring increasing efforts to control. Different approaches were taken to clarify the role of sexual reproduction in P. infestans in the epidemiology of potato late blight. The formation of sexually formed oospores and their ability to serve as primary inoculum was studied, both as field observations and by determining within-field genotypic diversity. To further estimate the importance of reproduction on a large spatial scale a population study based on molecular and phenotypic markers was performed on P. infestans isolates from the whole Nordic region. The variation in the aggressiveness of Nordic populations of P. infestans was studied by determining different components of aggressiveness, e.g. sporulation capacity, lesion growth and infection efficacy. The aggressiveness study was combined with an analysis of the phenotypic structure of the Nordic population of P. infestans. The results clearly indicate that the Swedish populations of P. infestans are influenced by sexual reproduction. The facts that both mating types are found all over the country in near 1:1 proportions and that oospores are commonly formed in field crops and serve as inoculum under field conditions support this. Studies of the genetic diversity also indicate that sexual reproduction has an effect on the population structure of P. infestans in Sweden and the Nordic region as a whole