Pathogenicity and Mefenoxam Sensitivity of Pythium, Globisporangium, and Fusarium Isolates From Coconut Coir and Rockwool in Marijuana (Cannabis sativa L.) Production

Oomycetes and fungi were recovered from coconut coir and rockwool substrates where marijuana (Cannabis sativa L. cv. Silver and Citron) plants with root rot and wilt symptoms were grown in a commercial growing facility in Connecticut. The objectives of this study were to identify the isolates collected from these substrates, determine the pathogenicity of the isolates on hemp seedlings in vitro and in vivo, and evaluate the pathogens' sensitivity to mefenoxam. Pythium and Globisporangium isolates were identified by sequencing the mitochondrially-encoded cytochrome oxidase genes (COI and COII) and Fusarium sp. with the translation elongation factor (EF-1α) region and internal transcribed spacer region (ITS4 and ITS5) genes. Three isolates were identified as Globisporangium irregulare (formerly Pythium irregulare), 21 isolates were Pythium myriotylum, and one was Fusarium oxysporum. All the isolates tested were pathogenic to hemp plants in vitro and in vivo, with disease incidence between 6.7 and 100%. Inoculated plants were smaller by 32% or more compared with the non-inoculated control. On average, hemp plants infected with Pythium myriotylum produced the lowest biomass and relative greenness values. None of the Pythium and Globisporangium isolates were resistant to mefenoxam—all were sensitive to ≥5 μg·mL−1 mefenoxam. This is the first report of G. irregulare causing root rot on marijuana and hemp plants. The results of this study provide information about the characteristics of pathogens that can be found potentially in soilless substrates in controlled environment agriculture.

Identification of Quantitative Disease Resistance Loci Toward Four Pythium Species in Soybean

In this study, four recombinant inbred line (RIL) soybean populations were screened for their response to infection by Pythium sylvaticum, Pythium irregulare, Pythium oopapillum, and Pythium torulosum. The parents, PI 424237A, PI 424237B, PI 408097, and PI 408029, had higher levels of resistance to these species in a preliminary screening and were crossed with “Williams,” a susceptible cultivar. A modified seed rot assay was used to evaluate RIL populations for their response to specific Pythium species selected for a particular population based on preliminary screenings. Over 2500 single-nucleotide polymorphism (SNP) markers were used to construct chromosomal maps to identify regions associated with resistance to Pythium species. Several minor and large effect quantitative disease resistance loci (QDRL) were identified including one large effect QDRL on chromosome 8 in the population of PI 408097 × Williams. It was identified by two different disease reaction traits in P. sylvaticum, P. irregulare, and P. torulosum. Another large effect QDRL was identified on chromosome 6 in the population of PI 408029 × Williams, and conferred resistance to P. sylvaticum and P. irregulare. These large effect QDRL will contribute toward the development of improved soybean cultivars with higher levels of resistance to these common soil-borne pathogens.

Plant SWEETs: from sugar transport to plant–pathogen interaction and more unexpected physiological roles

Sugars Will Eventually be Exported Transporters (SWEETs) have important roles in numerous physiological mechanisms where sugar efflux is critical, including phloem loading, nectar secretion, seed nutrient filling, among other less expected functions. They mediate low affinity and high capacity transport, and in angiosperms this family is composed by 20 paralogs on average. As SWEETs facilitate the efflux of sugars, they are highly susceptible to hijacking by pathogens, making them central players in plant–pathogen interaction. For instance, several species from the Xanthomonas genus are able to upregulate the transcription of SWEET transporters in rice (Oryza sativa), upon the secretion of transcription-activator-like effectors. Other pathogens, such as Botrytis cinerea or Erysiphe necator, are also capable of increasing SWEET expression. However, the opposite behavior has been observed in some cases, as overexpression of the tonoplast AtSWEET2 during Pythium irregulare infection restricted sugar availability to the pathogen, rendering plants more resistant. Therefore, a clear-cut role for SWEET transporters during plant–pathogen interactions has so far been difficult to define, as the metabolic signatures and their regulatory nodes, which decide the susceptibility or resistance responses, remain poorly understood. This fuels the still ongoing scientific question: what roles can SWEETs play during plant–pathogen interaction? Likewise, the roles of SWEET transporters in response to abiotic stresses are little understood. Here, in addition to their relevance in biotic stress, we also provide a small glimpse of SWEETs importance during plant abiotic stress, and briefly debate their importance in the particular case of grapevine (Vitis vinifera) due to its socioeconomic impact.

Composting Spent Mushroom Substrate from Agaricus bisporus and Pleurotus ostreatus Production as a Growing Media Component for Baby Leaf Lettuce Cultivation under Pythium irregulare Biotic Stress

Composts of spent mushrooms substrates can be an alternative for the partial replacement of peat as growing media in horticulture. Three mature composts from Agaricus bisporus (Ag), Pleurotus ostreatus (Pl), and 70% Ag:30% Pl (AgPl) production were used as partial components of peat growing media, used at a 1:4 compost:peat ratio for growing red baby leaf lettuce. They showed higher yields, between 3 and 7 times more than that for peat itself, even under the pressure of the plant pathogen Pythium irregulare. AgPl showed the higher suppressiveness (50%) against Pythium irregulare than Ag- (38%) or Pl- (15%) supplemented media. The combination of these raw materials and a suitable composting process is important for obtaining mature compost for use as a partial component of peat-based growing media.

Pythium spp. associated with root rot and stunting of winter wheat in North Carolina

In eastern North Carolina, mild to severe stunting and root rot have reduced yields of winter wheat, especially during years with abundant rainfall. Causal agents of root rot of wheat in this region were previously identified as Pythium irregulare, P. vanterpoolii, and P. spinosum. To investigate species prevalence, 114 isolates of Pythium were obtained from symptomatic wheat plants collected in 8 counties. Twelve species were recovered, with P. irregulare (32%), P. vanterpoolii (17%), and P. spinosum (16%) the most common. Pathogenicity screens were performed with selected isolates of each species, and slight to severe necrosis of young roots was observed. The aggressiveness of five isolates each of P. irregulare, P. vanterpoolii, and P. spinosum was compared on a single cultivar of wheat at 14°C, and very aggressive isolates were found within all species. In vitro growth of these isolates was measured at 14°C and 20°C, and all isolates grew faster at the warmer temperature. The effects of varying temperatures and rates of nitrogen on root rot caused by Pythium species alone or in combination were investigated. All inoculation treatments caused severe root rot under all conditions tested, and disease was more severe at 12/14°C compared to 18/20°C, but there was no effect of nitrogen application.

Novel Disease Host Resistances in the World Core Collection of Trifolium subterraneum

Glasshouse and field investigations were undertaken into the phenotypic expressions of resistance of a 97-member World Core Collection of subterranean clover (Trifolium subterraneum), collected from its native Mediterranean habitat and representing around 80% of the total genetic diversity within the known 10,000 accessions of the species, against the most important damping-off and root rot (Phytophthora clandestina, and Pythium irregulare) and foliar (Kabatiella caulivora, Uromyces trifolii-repentis and Erysiphe trifoliorum)pathogens. An additional 28 diverse cultivars were also included. Associations were also examined for these genotypes between 18 disease parameters and 17 morphological traits and between these disease parameters and 24 climatic and eco-geographic variables from their collection sites. Many genotypes showed strong phenotypic expression of novel host disease resistance(s) against one or more pathogens, enabling their potential deployment as disease resistant parents into subterranean clover breeding programs. These new sources of resistance enable future ‘pyramiding’ of different resistance genes to improve resistance against these pathogens. Of particular value were genotypes with multiple disease resistances across soilborne and/or foliar diseases, as many of these pathogens co-occur. All diseases had some parameters significantly correlated with one or more morphological traits and with one or more site of origin variables. In particular, there were significant negative correlations between damping-off (i.e., germination) and eight of the 17 morphological characters. The outcome of these studies provides crucial information to subterranean clover breeding programs, enabling them to simultaneously select genotypes with multiple resistances to co-occurring soilborne and foliar diseases, in addition to desirable traits, to offer renewed hope for re-establishing a more productive subterranean clover livestock feedbase despite multiple diseases prevailing widely.

Promising Composts as Growing Media for the Production of Baby Leaf Lettuce in a Floating System

The floating system is a successful strategy for producing baby leaf vegetables. Moreover, compost from agricultural and agri-food industry wastes is an alternative to peat that can be used as a component of growing media in this cultivation system. In this study, we experimented with three composts containing tomato (Solanum lycopersicum L.), leek (Allium porrum L.), grape (Vitis vinifera L.), and/or olive (Olea europaea L.) mill cake residues, which were used as the main component (75/25 volume/volume) of three growing media (GM1, GM2 and GM3) to evaluate their effect on the growth and quality of red baby leaf lettuce (Lactuca sativa L.). We used a commercial peat substrate as a control treatment (100% volume) and in mixtures (25% volume) with the composts. The plants were cultivated over two growing cycles, in spring and summer, and harvested twice in each cycle when the plants had four to five leaves. We found that the percentage of seed germination was significantly higher in plants grown in peat than in those grown in compost growing media. The yield was affected by the growing media in the summer cycle, and we obtained the highest value with GM1. Furthermore, the second cut was more productive than the first one for all the growing media in both cycles. The lettuce quality was also affected by the growing media. In general, the total phenolic content and antioxidant capacity in the leaves was higher in plants grown in the compost growing media, particularly in the second cut, but the nitrate content in the leaves was greater in some of the compost treatments compared with the peat treatment. In addition, an in vitro suppressive activity study demonstrated that the interaction between different fungi and bacteria observed through metagenomics analysis could contribute to the effectiveness of the compost in controlling Pythium irregulare. The use of compost as a component of the growing media in the production of baby leaf vegetables in a floating system does not only favor the crop yield and product quality, but also shows suppressive effects against P. irregulare.

Influence of soybean tissue and oomicide seed treatments on oomycete isolation

Soybean seedlings are vulnerable to different oomycete pathogens. Seed treatments containing the two anti-oomycete (oomicide) chemicals, metalaxyl-M (mefenoxam), and ethaboxam are used for the protection against oomycete pathogens. This study aimed to evaluate the influence of these two oomicides on the isolation probability of oomycetes from soybean taproot or lateral root sections. Soybean plants were collected between the first and third trifoliate growth stages from five Midwest field locations in 2016 and four of the same fields in 2017. Oomycetes were isolated from taproot and lateral root. In 2016, 369 isolation attempts were completed resulting in 121 isolates from the taproot and 154 isolates from the lateral root. In 2017, 468 isolation attempts were completed, with 44 isolates from the taproot and 120 isolates from the lateral roots. In three of nine site-years, the probability of isolating an oomycete from a taproot or lateral root section was significantly different. Seed treatments containing a mixture of ethaboxam and metalaxyl significantly reduced the probability of oomycete isolation from lateral roots in Illinois in 2016 and 2017, but not in other locations, which may have been related to the heavy soil type (clay loam). Among the 439 isolates collected from the two years sampled, 24 oomycete species were identified, and community compositions differed depending on location and year. The five most abundant species were Pythium sylvaticum (28.9%), Pythium heterothallicum (14.3%), Pythium ultimum var. ultimum (11.8%), Pythium attrantheridium (7.9%), and Pythium irregulare (6.6%) which accounted for 61.7% of the isolates collected. Oomicide sensitivity to ethaboxam and mefenoxam was assessed for more than 300 isolates. There were large differences in ethaboxam sensitivity among oomycete species with EC50 ranging from < 0.01 to > 100 μg ml-1, and a median of 0.65 μg ml-1. Isolates with insensitivity to ethaboxam (> 12 μg ml-1) belonging to the species Pythium torulosum and Pythium rostratifingens but were sensitive to mefenoxam. Oomicide sensitivity to mefenoxam ranged from < 0.01 to 0.62 μg ml-1 with a median of 0.03 μg ml-1. The mean EC50 of the five most abundant species to ethaboxam ranged from 0.35 to 0.97 μg ml-1 of ethaboxam and from 0.02 to 0.04 μg ml-1 of mefenoxam. No shift in sensitivity to mefenoxam or ethaboxam was observed due to soybean seed treatment or year relative to the non-treated seed controls. In summary, this study contributed to the understanding of the composition of oomycete populations from different soybean root tissues, locations, years, and seed treatments. Finally, the effectiveness of seed treatments containing mefenoxam or metalaxyl plus ethaboxam can be effective in reducing the probability of oomycete isolation from soybean roots.