Phenotypic and Genotypic Variation of Cultivated Panax quinquefolius.

American ginseng (Panax quinquefolius) is widely used due to its medicinal properties. Ontario is a major producer of cultivated American ginseng, where seeds were originally collected from the wild without any subsequent scientific selection, and thus the crop is potentially very diverse. A collection of 162 American ginseng plants was harvested from a small area in a commercial garden and phenotyped for morphological traits, such as root grade, stem length, and fresh and dry weights of roots, leaves, stems, and seeds. All of the traits showed a range of values, and correlations were observed between root and stem weights, root dry weight and leaf dry weight, as well as root and leaf fresh weights. The plants were also genotyped using single nucleotide polymorphisms (SNPs) at the PW16 locus. SNP analysis revealed 22 groups based on sequence relatedness with some groups showing no SNPs and others being more diverse. The SNP groups correlated with significant differences in some traits, such as stem length and leaf weight. This study provides insights into the genetic and phenotypic diversity of cultivated American ginseng grown under similar environmental conditions, and the relationship between different phenotypes, as well as genotype and phenotype, will aid in future selection programs to develop American ginseng cultivars with desirable agronomic traits.

Role of a LORELEI- like gene from Phaseolus vulgaris during a mutualistic interaction with Rhizobium tropici.

Reactive oxygen species (ROS), produced by NADPH oxidases known as RBOHs in plants, play a key role in plant development, biotic and abiotic stress responses, hormone signaling, and reproduction. Among the subfamily of receptor-like kinases referred to as CrRLK, there is FERONIA (FER), a regulator of RBOHs, and FER requires a GPI-modified membrane protein produced by LORELEI (LRE) or LORELEI-like proteins (LLG) to reach the plasma membrane and generate ROS. In Arabidopsis, AtLLG1 is involved in interactions with microbes as AtLLG1 interacts with the flagellin receptor (FLS2) to trigger the innate immune response, but the role of LLGs in mutualistic interactions has not been examined. In this study, two Phaseolus vulgaris LLG genes were identified, PvLLG2 that was expressed in floral tissue and PvLLG1 that was expressed in vegetative tissue. Transcripts of PvLLG1 increased during rhizobial nodule formation peaking during the early period of well-developed nodules. Also, P. vulgaris roots expressing pPvLLG1:GFP-GUS showed that this promoter was highly active during rhizobium infections, and very similar to the subcellular localization using a construct pLLG1::PvLLG1-Neon. Compared to control plants, PvLLG1 silenced plants had less superoxide (O2-) at the root tip and elongation zone, spotty hydrogen peroxide (H2O2) in the elongation root zone, and significantly reduced root hair length, nodule number and nitrogen fixation. Unlike control plants, PvLLG1 overexpressing plants showed superoxide beyond the nodule meristem, and significantly increased nodule number and nodule diameter. PvLLG1 appears to play a key role during this mutualistic interaction, possibly due to the regulation of the production and distribution of ROS in roots.

Genetic variability of the honey bee mite, Varroa destructor, from a humid continental climatic region of Canada, and temperate and tropical climatic regions of Mexico.

Varroa destructor is a damaging mite of Western honey bees (Apis mellifera). Genetic variability of the mite in different regions of the world could be related to the movement of infested bees or other factors, such as climate. In this study, V. destructor samples were collected from tropical and temperate climate regions of Mexico, and a humid continental climate region of Canada. COX-1 AFLPs showed that all the mites were the Korean haplotype. Four microsatellites revealed nine haplogroups from the continental climate region of Canada, compared to three haplogroups from the tropical and temperate climate regions of Mexico. CytII-ATP sequences showed seven haplogroups from the humid continental climate region vs. two haplogroups from the temperate region and one haplogroup from the tropical region. CytB sequences revealed seven haplogroups from Canada vs. three from Mexico. A comparison of the cytB sequences of the samples from Canada and Mexico to those from a worldwide collection showed that one sequence, designated the cytB1 type, predominated, comprising 57% of the 86 sequences; it clustered with similar sequences that comprised 80% of the sequences, designated family A. CytB1 was predominant in Mexico, but not in Canada. The other 20% of sequences were in families B and C, and all those samples originated from East and Southeast Asia. The microsatellite, cytII-ATP, and cytB markers, all showed higher variability in mites collected in Canada than in Mexico, which could be related to the cooler climate or an earlier invasion and/or multiple mite invasions in Canada.

Effect of Soil and Root Extracts on the Innate Immune Response of American Ginseng (Panax quinquefolius) to Root Rot Caused by Ilyonectria mors-panacis.

Panax quinquefolius shows much higher mortality to Ilyonectria mors-panacis root rot when grown in soil previously planted with ginseng than in soil not previously planted with ginseng, which is known as ginseng replant disease. Treatment of ginseng roots with methanol extracts of previous ginseng soils significantly increased root lesion sizes due to I. mors-panacis compared to roots treated with water or methanol extracts of ginseng roots or non-ginseng soils. Inoculation of water-treated roots with I. mors-panacis increased expression of a basic chitinase 1 gene (PqChi-1), neutral pathogenesis-related protein 5 gene (PqPR5) and pathogenesis-related protein 10-2 gene (PqPR10-2), which are related to jasmonic acid (JA), ethylene (ET) or necrotrophic infection, and also increased expression of an acidic ß-1-3-glucanase gene (PqGlu), which is related to salicylic acid (SA). Infection did not affect expression of a cysteine protease inhibitor gene (PqCPI). Following infection, roots treated with ginseng root extract mostly showed similar expression patterns as roots treated with water, but roots treated with previous ginseng soil extract showed reduced expression of PqChi-1, PqPR5, PqPR10-2 and PqCPI, but increased expression of PqGlu. Methanol-soluble compound(s) in soil previously planted with ginseng are able to increase root lesion size, suppress JA/ET-related gene expression and trigger SA-related gene expression in ginseng roots during I. mors-panacis infection, and may be a factor contributing to ginseng replant disease.

Isolation and Genome-Based Characterization of Biocontrol Potential of Bacillus siamensis YB-1631 against Wheat Crown Rot Caused by Fusarium pseudograminearum.

Fusarium crown rot (FCR) caused by Fusarium pseudograminearum is one of the most serious soil-borne diseases of wheat. Among 58 bacterial isolates from the rhizosphere soil of winter wheat seedlings, strain YB-1631 was found to have the highest in vitro antagonism to F. pseudograminearum growth. LB cell-free culture filtrates inhibited mycelial growth and conidia germination of F. pseudograminearum by 84.14% and 92.23%, respectively. The culture filtrate caused distortion and disruption of the cells. Using a face-to-face plate assay, volatile substances produced by YB-1631 inhibited F. pseudograminearum growth by 68.16%. In the greenhouse, YB-1631 reduced the incidence of FCR on wheat seedlings by 84.02% and increased root and shoot fresh weights by 20.94% and 9.63%, respectively. YB-1631 was identified as Bacillus siamensis based on the gyrB sequence and average nucleotide identity of the complete genome. The complete genome was 4,090,312 bp with 4357 genes and 45.92% GC content. In the genome, genes were identified for root colonization, including those for chemotaxis and biofilm production, genes for plant growth promotion, including those for phytohormones and nutrient assimilation, and genes for biocontrol activity, including those for siderophores, extracellular hydrolase, volatiles, nonribosomal peptides, polyketide antibiotics, and elicitors of induced systemic resistance. In vitro production of siderophore, ß-1, 3-glucanase, amylase, protease, cellulase, phosphorus solubilization, and indole acetic acid were detected. Bacillus siamensis YB-1631 appears to have significant potential in promoting wheat growth and controlling wheat FCR caused by F. pseudograminearum.

Effect of natural weed and Siratro cover crop on soil fungal diversity in a banana cropping system in southwestern China.

Introduction: Natural weed cover and a legume cover crop were examined to determine if they could impact soil fungal diversity as an indicator of soil quality in banana production. Methods: Banana in Yunnan Province, China, was grown under three treatments: conventional tillage (bare soil), natural weed cover (primarily goosegrass (Eleusine indica (L.) Gaerth)), or a cover crop (Siratro (Macroptilium atropurpureum (DC.) Urb.)). Analysis of the soil fungal communities between 2017 and 2020 was done by Illumina Miseq high-throughput sequencing. Results: Most significant effects were in the intercropping area for the treatments, whereas it was rarely observed in the furrow planted with banana. Based on the Shannon and Simpson diversity indices, soil fungal diversity in the intercropping area significantly decreased following planting banana in 2017 with all three treatments. However, both the Shannon and Simpson diversity indices showed that there were significant increases in fungal soil diversity in 2019 and 2020 with natural weed cover or Siratro compared to bare soil. At the end of the experiment, significant increases in fungal genera with Siratro compared to bare soil were observed with Mortierella, Acremonium, Plectophaerella, Metarhizium and Acrocalymma, and significant decreases were observed with Fusicolla, Myrothecium, Exserohilum, Micropsalliota and Nigrospora. Siratro resulted in higher stability of the soil fungal microbiome by increasing the modularity and the proportion of negative co-occurrences compared to bare soil. For fungal guilds, Siratro significantly increased saprotrophs_symbiotrophs in 2019 and 2020 and significantly decreased pathogens_saprotrophs in 2020 compared to bare soil. Discussion: Using Siratro as a cover crop in the intercropping area of banana helped maintain soil fungal diversity, which would be beneficial for soil health with more symbiotrophs and less pathogens in the soil. However, further research is needed to determine the long-term impact of weed or Siratro cover crop on the fungal soil ecosystem and growth of banana.

Selection of reference genes for RT-qPCR analysis of rice with Rhizoctonia solani infection and biocontrol PGPR/KSi application.

BACKGROUND: Rhizoctonia solani (AG1 IA) is an important pathogen of rice (Oryza sativa L.) that causes rice sheath blight (RSB). Since control of RSB by breeding and fungicides have had limited success, novel strategies like biocontrol with plant growth-promoting rhizobacteria (PGPR) can be an effective alternative. METHOD AND RESULTS: Seven commonly used reference genes (RGs), 18SrRNA, ACT1, GAPDH2, UBC5, RPS27, eIF4a and CYP28, were evaluated for their stability in rice-R. solani-PGPR interaction for real-time quantitative PCR (RT-qPCR) analysis. Different algorithms were examined, Delta Ct, geNorm, NormFinder, BestKeeper, and comprehensive ranking by RefFinder, to evaluate RT-qPCR of rice in tissues infected with R. solani and treated with the PGPR strains, Pseudomonas saponiphilia and Pseudomonas protegens, with potassium silicate (KSi) alone or in combination with each PGPR strain. RG stability was affected for each treatment and treatment-specific RG selection was suggested. Validation analysis was done for nonexpressor of PR-1(NPR1) for each treatment. CONCLUSION: Overall, ACT1 was the most stable RG with R. solani infection alone, GAPDH2 with R. solani infection plus KSi, UBC5 with R. solani infection plus P. saponiphilia, and eIF4a with R. solani infection plus P. protegens. Both ACT1 and RPS27 were the most stable with the combination of KSi and P. saponiphilia, while RPS27 was the most stable with the combination of KSi and P. protegens.

Ginsenosides and Biotic Stress Responses of Ginseng.

Ginsenosides are saponins that possess a sugar moiety attached to a hydrophobic aglycone triterpenoid. They have been widely studied for their various medicinal benefits, such as their neuroprotective and anti-cancer activities, but their role in the biology of ginseng plants has been much less widely documented. In the wild, ginsengs are slow-growing perennials with roots that can survive for approximately 30 years; thus, they need to defend themselves against many potential biotic stresses over many decades. Biotic stresses would be a major natural selection pressure and may at least partially explain why ginseng roots expend considerable resources in order to accumulate relatively large amounts of ginsenosides. Ginsenosides may provide ginseng with antimicrobial activity against pathogens, antifeedant activity against insects and other herbivores, and allelopathic activity against other plants. In addition, the interaction of ginseng with pathogenic and non-pathogenic microorganisms and their elicitors may trigger increases in different root ginsenosides and associated gene expression, although some pathogens may be able to suppress this behavior. While not covered in this review, ginsenosides also have roles in ginseng development and abiotic stress tolerance. This review shows that there is considerable evidence supporting ginsenosides as important elements of ginseng's defense against a variety of biotic stresses.

Occurrence and Distribution of Physiological Races of Exserohilum turcicum in Maize-Growing Regions of Mexico.

Turcicum leaf blight (TLB) is a common foliar disease of maize in Mexico that is caused by the fungal pathogen Exserohilum turcicum. The most effective management strategy against TLB is monogenic race-specific resistance. Among the 140 E. turcicum isolates from symptomatic leaves collected from maize fields in Mexico, 100 were obtained from tropical (Veracruz) and temperate areas (Estado de México) between 2010 and 2019, and 40 isolates were obtained from tropical (Sinaloa, Tamaulipas, Veracruz, and Chiapas), subtropical (Nayarit, Jalisco, and Guanajuato), and temperate areas (Estado de Mexico, Hidalgo, and Puebla) collected in 2019. All the isolates caused TLB symptoms on the positive control (ht4), showing that they were all pathogenic. Six physiological races of E. turcicum (2, 3, 23, 3N, 23N, and 123N) were identified based on resistant or susceptible responses displayed by five maize differential genotypes (A619Ht1, A619Ht2, A619Ht3, B68HtN, and A619ht4). The most common was race 23, accounting for 68% of the isolates, followed by races 23N, 123N, 3, 2, and 3N at 15, 8, 6, 2, and 1%, respectively. Race 123N was able to infect the greatest number of maize differential genotypes used in the study. Race 123N was detected in Sinaloa and Estado de México. Race 3 was detected in Nayarit and Jalisco. Race 2 was detected in Jalisco, Estado de México, and Veracruz, and race 3N was detected in Tamaulipas. Race 23 was equally dominant in the tropical, subtropical, and temperate regions, while race 123N was more common in the tropical environment, and race 23N was more common in the tropical and temperate environments. There was no evidence for shifts in the races between 2010 and 2019.

Interaction of Ginseng with Ilyonectria Root Rot Pathogens.

The Ilyonectria radicicola species complex (A.A. Hildebr.) A. Cabral and Crous 2011 contains species of soilborne necrotrophic plant pathogens. The most aggressive to ginseng roots is I. mors-panacis, whereas I. robusta, I. crassa, I. panacis and I. radicicola are less aggressive. Infected ginseng roots show orange-red to black-brown lesions that can expand into a severe root rot, known as disappearing root rot, where only epidermal root tissue remains. Leaves become red-brown with wilting, and stems can have vascular discoloration with black-brown lesions at the base. Less aggressive Ilyonectria species trigger jasmonic acid (JA)-related defenses inducing host ginsenosides, pathogenesis-related (PR) proteins, wound periderm, and cell wall thickening. In contrast, I. mors-panacis triggers reactive oxygen species (ROS) and salicylic acid (SA) production but suppresses JA-related defenses and ginsenoside accumulation. It is also able to suppress SA-related PR protein production. Virulence factors include potential effectors that may suppress PAMP (Pathogen Associated Molecular Patterns) triggered immunity (PTI), polyphenoloxidases, Hsp90 inhibitors, siderophores and cell-wall-degrading enzymes, such as pectinases. Overall, I. mors-panacis appears to be more aggressive because it can suppress JA and SA-related PTI allowing for more extensive colonization of ginseng roots. While many possible mechanisms of host resistance and pathogen virulence mechanisms have been examined, there is a need for using genetic approaches, such as RNAi silencing of genes of Panax or Ilyonectria, to determine their importance in the interaction.

Response of Fusarium pseudograminearum to Biocontrol Agent Bacillus velezensis YB-185 by Phenotypic and Transcriptome Analysis.

The use of biological control agents (BCAs) is a promising alternative control measure for Fusarium crown rot (FCR) of wheat caused by Fusarium pseudograminearum. A bacterial strain, YB-185, was isolated from the soil of wheat plants with FCR and identified as Bacillus velezensis. YB-185 exhibited strong inhibition of F. pseudograminearum mycelial growth and conidial germination in culture. Seed treatment with YB-185 in greenhouse and field resulted in reductions in disease by 66.1% and 57.6%, respectively, along with increased grain yield. Microscopy of infected root tissues confirmed that YB-185 reduced root invasion by F. pseudograminearum. RNA-seq of F. pseudograminearum during co-cultivation with B. velezensis YB-185 revealed 5086 differentially expressed genes (DEGs) compared to the control. Down-regulated DEGs included genes for glucan synthesis, fatty acid synthesis, mechanosensitive ion channels, superoxide dismutase, peroxiredoxin, thioredoxin, and plant-cell-wall-degrading enzymes, whereas up-regulated DEGs included genes for chitin synthesis, ergosterol synthesis, glutathione S-transferase, catalase, and ABC transporters. In addition, fungal cell apoptosis increased significantly, as indicated by TUNEL staining, and the scavenging rate of 2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt radical cation (ABTS·+) in the fungus significantly decreased. Thus, F. pseudograminearum may be trying to maintain normal cell functions by increasing cell wall and membrane synthesis, antioxidant and anti-stress responses, detoxification of bacterial antimicrobial compounds, and transportation of damaging compounds from its cells. However, cell death and free radical accumulation still occurred, indicating that the responses were insufficient to prevent cell damage. Bacillus velezensis YB-185 is a promising BCA against FCR that acts by directly damaging F. pseudograminearum, thus reducing its ability to colonize roots and produce symptoms.

Evaluation and Genome Analysis of Bacillus subtilis YB-04 as a Potential Biocontrol Agent Against Fusarium Wilt and Growth Promotion Agent of Cucumber.

Cucumber wilt caused by Fusarium oxysporum f.sp. cucumerinum (Foc) is a highly destructive disease that leads to reduced yield in cucumbers. In this study, strain YB-04 was isolated from wheat straw and identified as Bacillus subtilis. It displayed strong antagonistic activity against F. oxysporum f.sp. cucumerinum in dual culture and exhibited significant biocontrol of cucumber Fusarium wilt with a higher control effect than those of previously reported Bacillus strains and displayed pronounced growth promotion of cucumber seedlings. B. subtilis YB-04 could secrete extracellular protease, amylase, cellulose, and ß-1,3-glucanase and be able to produce siderophores and indole acetic acid. Inoculation with B. subtilis YB-04 or Foc increased cucumber defense-related enzyme activities for PPO, SOD, CAT, PAL, and LOX. However, the greatest increase was with the combination of B. subtilis YB-04 and Foc. Sequencing the genome of B. subtilis YB-04 showed that it had genes for the biosynthesis of various secondary metabolites, carbohydrate-active enzymes, and assimilation of nitrogen, phosphorous, and potassium. B. subtilis YB-04 appears to be a promising biological control agent against the Fusarium wilt of cucumber and promotes cucumber growth by genomic, physiological, and phenotypic analysis.

The Rhizosphere Microbiome of Ginseng.

The rhizosphere of ginseng contains a wide range of microorganisms that can have beneficial or harmful effects on the plant. Root exudates of ginseng, particularly ginsenosides and phenolic acids, appear to select for particular microbial populations through their stimulatory and inhibitory activities, which may account for the similarities between the rhizosphere microbiomes of different cultivated species of Panax. Many practices of cultivation attempt to mimic the natural conditions of ginseng as an understory plant in hilly forested areas. However, these practices are often disruptive to soil, and thus the soil microbiome differs between wild and cultivated ginseng. Changes in the microbiome during cultivation can be harmful as they have been associated with negative changes of the soil physiochemistry as well as the promotion of plant diseases. However, isolation of a number of beneficial microbes from the ginseng rhizosphere indicates that many have the potential to improve ginseng production. The application of high-throughput sequencing to study the rhizosphere microbiome of ginseng grown under a variety of conditions continues to greatly expand our knowledge of the diversity and abundance of those organisms as well as their impacts of cultivation. While there is much more to be learnt, many aspects of the ginseng rhizosphere microbiome have already been revealed.

First insights into the honey bee (Apis mellifera) brain lipidome and its neonicotinoid-induced alterations associated with reduced self-grooming behavior.

Introduction: Honey bees (Apis mellifera) play key roles in food production performing complex behaviors, like self-grooming to remove parasites. However, the lipids of their central nervous system have not been examined, even though they likely play a crucial role in the performance of cognitive process to perform intricate behaviors. Lipidomics has greatly advanced our understanding of neuropathologies in mammals and could provide the same for honey bees. Objectives: The objectives of this study were to characterize the brain lipidome of adult honey bees and to assess the effect of clothianidin (a neurotoxic insecticide) on the brain lipid composition, gene expression, and performance of self-grooming behavior under controlled conditions (cage experiments). Methods: After seven days of exposure to oral sublethal doses of clothianidin, the bees were assessed for self-grooming behavior; their brains were dissected to analyze the lipidome using an untargeted lipidomics approach and to perform a high throughput RNAseq analysis. Results: Compared to all other organisms, healthy bee brain lipidomes contain unusually high levels of alkyl-ether linked (plasmanyl) phospholipids (51.42%) and low levels of plasmalogens (plasmenyl phospholipids; 3.46%). This could make it more susceptible to the effects of toxins in the environment. A positive correlation between CL 18:3/18:1/14:0/22:6, TG 6:0/11:2/18:1, LPE 18:0e and intense self-grooming was found. Sublethal doses of a neonicotinoid altered PC 20:3e/15:0, PC 16:0/18:3, PA 18:0/24:1, and TG 18:1/18:1/18/1 levels, and affected gene expression linked to GPI-anchor biosynthesis pathway and energy metabolism that may be partially responsible for the altered lipid composition. Conclusion: This study showed that lipidomics can reveal honey bee neuropathologies associated with reduced grooming behavior due to sublethal neonicotinoid exposure. The ease of use, unusual brain lipidome as well as characterized behaviors that are affected by the environment make honey bees a promising model organism for studying the neurolipidome and associations with neurobehavioral disorders.

Genomic and Phenotypic Insights into the Potential of Bacillus subtilis YB-15 Isolated from Rhizosphere to Biocontrol against Crown Rot and Promote Growth of Wheat.

Fusarium crown rot caused by Fusarium pseudograminearum is one of the most devastating diseases of wheat worldwide causing major yield and economic losses. In this study, strain YB-15 was isolated from soil of wheat rhizosphere and classified as Bacillus subtilis by average nucleotide identity analysis. It significantly reduced Fusarium crown rot with a control efficacy of 81.50% and significantly improved the growth of wheat seedlings by increasing root and shoot fresh weight by 11.4% and 4.2%, respectively. Reduced Fusarium crown rot may have been due to direct antagonism by the production of ß-1, 3-glucanase, amylase, protease and cellulase, or by the ability of B. subtilis YB-15 to induce defense-related enzyme activities of wheat seedlings, both alone and in seedlings infected with F. pseudograminearum. Improved plant growth may be related to the ability of B. subtilis YB-15 to secrete indole acetic acid and siderophores, as well as to solubilize phosphorus. In addition, the genome of strain YB-15 was determined, resulting in a complete assembled circular genome of 4,233,040 bp with GC content of 43.52% consisting of 4207 protein-encoding genes. Sequencing the B. subtilis YB-15 genome further revealed genes for encoding carbohydrate-active enzymes, biosynthesis of various secondary metabolites, nutrient acquisition, phytohormone production, chemotaxis and motility, which could explain the potential of strain YB-15 to be plant growth-promoting bacteria and biological control agent. B. subtilis YB-15 appears to be a promising biocontrol agent against Fusarium crown rot as well as for wheat growth promotion.

The Endosphere Microbiome of Ginseng.

The endosphere of ginseng contains a variety of fungal, bacterial, archaeal and viral endophytes. Bacterial endophytes are primarily members of the Proteobacteria, Actinobacteria, Firmicutes and Bacteroidetes, and fungal endophytes are primarily members of the Ascomycota, Zygomycota and Basidiomycota. Although archaea and viruses have been detected in symptomless ginseng plants, little is known about them. Many but not all studies have shown roots having the highest abundance and diversity of bacterial and fungal endophytes, with some endophytes showing specificity to above or belowground tissues. Abundance often increases with root age, although diversity can decrease, possibly related to increases in potential latent fungal pathogen infections. The descriptions of many endophytes that can metabolize ginsenosides indicate an adaptation of the microbes to the unique combination of secondary metabolites found in ginseng tissues. Most research on the benefits provided by bacterial and fungal endophytes has concentrated on improved plant nutrition, growth promotion and increased disease resistance, but little on their ability to increase abiotic stress resistance. Some other areas where more research is needed is field trials with endophyte-treated plants grown in various environments, genomic/metagenomic analysis of endophytes, and the effects of endophytes on induced disease resistance and abiotic stress tolerance.

Differences in Gene Expression of Pear Selections Showing Leaf Curling or Leaf Reddening Symptoms Due to Pear Decline Phytoplasma.

While host gene expression has been related to symptoms associated with different phytoplasma diseases, it is unknown why some phytoplasmas are associated with different symptoms in genotypes of the same plant species. Pear tree selections showed symptoms of either leaf reddening (selection 8824-1) or leaf curling (selection 9328-1) associated with pear decline (PD) phytoplasma presence. PD populations were similar in leaves and shoots of the two selections, but in the roots, populations were significantly lower in selection 8824-1 than in 9328-1, indicating greater resistance. For host carbohydrate metabolism gene expression in PD-infected tissues, significant up-regulation in selection 8824-1 was observed for a sucrose synthase gene in leaves and an acid invertase gene in leaves and roots. These features have been associated with localized higher sugar levels in phytoplasma-infected tissues, and thus may be related to leaf reddening. For host stress/defense response gene expression in PD-infected tissues, significant up-regulation of a phenylalanine ammonia lyase gene was observed in PD-infected shoots of both selections; however, up-regulation of alcohol dehydrogenase gene in shoots, a chitinase gene in all tissues and a phloem protein 2 gene in roots was only observed for selection 8824-1. These changes indicate greater triggered innate immunity in roots associated with lower PD populations and leaf reddening. Leaf reddening may be related to changes in gene expression associated with increased sugar levels in leaves and stronger immune responses in several tissues, while leaf curling may be due to water stress resulting from dysfunctional root associated with higher PD populations in the roots.

Effect of azoxystrobin on tobacco leaf microbial composition and diversity.

Azoxystrobin, a quinone outside inhibitor fungicide, reduced tobacco target spot caused by Rhizoctonia solani by 62%, but also affected the composition and diversity of other microbes on the surface and interior of treated tobacco leaves. High-throughput sequencing showed that the dominant bacteria prior to azoxystrobin treatment were Methylobacterium on healthy leaves and Pseudomonas on diseased leaves, and the dominant fungi were Thanatephorous (teleomorph of Rhizoctonia) and Symmetrospora on healthy leaves and Thanatephorous on diseased leaves. Both bacterial and fungal diversity significantly increased 1 to 18 days post treatment (dpt) with azoxystrobin for healthy and diseased leaves. For bacteria on healthy leaves, the relative abundance of Pseudomonas, Sphingomonas, Unidentified-Rhizobiaceae and Massilia declined, while Methylobacterium and Aureimonas increased. On diseased leaves, the relative abundance of Sphingomonas and Unidentified-Rhizobiaceae declined, while Methylobacterium, Pseudomonas and Pantoea increased. For fungi on healthy leaves, the relative abundance of Thanatephorous declined, while Symmetrospora, Sampaiozyma, Plectosphaerella, Cladosporium and Cercospora increased. On diseased leaves, the relative abundance of Thanatephorous declined, while Symmetrospora, Sampaiozyma, Plectosphaerella, Cladosporium, Phoma, Pantospora and Fusarium, increased. Compared to healthy leaves, azoxystrobin treatment of diseased leaves resulted in greater reductions in Thanatephorous, Sphingomonas and Unidentified-Rhizobiaceae, a greater increase in Methylobacterium, and similar changes in Phoma, Fusarium, Plectosphaerella and Cladosporium. Azoxystrobin had a semi-selective effect altering the microbial diversity of the tobacco leaf microbiome, which could be due to factors, such as differences among bacterial and fungal species in sensitivity to quinone outside inhibitors, ability to use nutrients and niches as certain microbes are affected, and metabolic responses to azoxystrobin.

Effect of feeding chitosan or peptidoglycan on Nosema ceranae infection and gene expression related to stress and the innate immune response of honey bees (Apis mellifera).

Nosema ceranae is a microsporidian parasite that causes nosema disease, an infection of the honey bee (Apis mellifera) midgut. Two pathogen-associated molecular patterns (PAMPs), chitosan and peptidoglycan, and N. ceranae spores were fed to worker bees in sucrose syrup and compared to non-inoculated and N. ceranae-inoculated bees without PAMPs. Both chitosan and peptidoglycan significantly increased bee survivorship and reduced spore numbers due to N. ceranae infection. To determine if these results were related to changes in health status, expression of the immune-related genes, hymenoptaecin and defensin2, and the stress tolerance-related gene, blue cheese, was compared to that of control bees. Compared to the inoculated control, bees with the dose of chitosan that significantly reduced N. ceranae spore numbers showed lower expression of hymenoptaecin and defensin2 early after infection, higher expression mid-infection of defensin2 and lower expression of all three genes late in infection. In contrast, higher expression of defensin2 early in the infection and all three genes late in the infection was observed with peptidoglycan treatment. Changes late in the parasite multiplication stage when mature spores would be released from ruptured host cells are less likely to have contributed to reduced spore production. Based on these results, it is concluded that feeding bees chitosan or peptidoglycan can reduce N. ceranae infection, which is at least partially related to altering the health of the bee by inducing immune and stress-related gene expression.