Life Sciences Commons^™

First Report Of Goss's Bacterial Wilt And Leaf Blight (Clavibacter Michiganensis Subsp. Nebraskensis) Of Corn In Texas, K. A. Korus, A. D. Timmerman, R. D. French-Monar, T. A. Jackson

Department of Plant Pathology: Faculty Publications

In September 2009, the University of Nebraska–Lincoln Plant and Pest Diagnostic Clinic received leaf samples of hybrid corn (Zea mays L.) displaying long, necrotic lesions with wavy margins. The lesions had discontinuous water-soaked spots that are indicative of Goss's bacterial wilt and leaf blight. The symptomatic leaves were submitted from Dallam County, located in the Texas Panhandle (northwest Texas). According to the USDA Farm Service Agency and the National Agricultural Statistics Service, in 2009 Dallam County had 54,025 ha planted to corn. This is approximately 19% of the total corn planted in the 26 counties in the Texas Panhandle …

Effects Of Integrating Cultivar Resistance And Fungicide Application On Fusarium Head Blight And Deoxynivalenol In Winter Wheat, Stephen N. Wegulo, William W. Bockus, John Hernandez Nopsa, Erick D. De Wolf, Kent M. Eskridge, Kamaranga H. S. Peiris, Floyd E. Dowell

Department of Plant Pathology: Faculty Publications

Fusarium head blight (FHB) or scab, incited by Fusarium graminearum, can cause significant economic losses in small grain production. Five field experiments were conducted from 2007 to 2009 to determine the effects on FHB and the associated mycotoxin deoxynivalenol (DON) of integrating winter wheat cultivar resistance and fungicide application. Other variables measured were yield and the percentage of Fusarium-damaged kernels (FDK). The fungicides prothioconazole + tebuconazole (formulated as Prosaro 421 SC) were applied at the rate of 0.475 liters/ha, or not applied, to three cultivars (experiments 1 to 3) or six cultivars (experiments 4 and 5) differing in their …

Characterization Of Three-Dimensional Spatial Aggregation And Association Patterns Of Brown Rot Symptoms Within Intensively Mapped Sour Cherry Trees, Sydney E. Everhart, Ashley Askew, Lynne Seymour, Imre J. Holb, Harald Scherm

Department of Plant Pathology: Faculty Publications

Characterization of spatial patterns of plant disease can provide insights into important epidemiological processes such as sources of inoculum, mechanisms of dissemination, and reproductive strategies of the pathogen population. While two-dimensional patterns of disease (among plants within fields) have been studied extensively, there is limited information on three-dimensional patterns within individual plant canopies. Reported here are the detailed mapping of different symptom types of brown rot (caused by Monilinia laxa) in individual sour cherry tree (Prunus cerasus) canopies, and the application of spatial statistics to the resulting data points to de-termine patterns of symptom aggregation and association. …

Comparison Of Plant‐Adapted Rhabdovirus Protein Localization And Interactions, Kathleen Marie Martin

University of Kentucky Doctoral Dissertations

Sonchus yellow net virus (SYNV), Potato yellow dwarf virus (PYDV) and Lettuce Necrotic yellows virus (LNYV) are members of the Rhabdoviridae family that infect plants. SYNV and PYDV are Nucleorhabdoviruses that replicate in the nuclei of infected cells and LNYV is a Cytorhabdovirus that replicates in the cytoplasm. LNYV and SYNV share a similar genome organization with a gene order of Nucleoprotein (N), Phosphoprotein (P), putative movement protein (Mv), Matrix protein (M), Glycoprotein (G) and Polymerase protein (L). PYDV contains an additional predicted gene between N and P, denoted as X, that has an unknown function. In order to gain …

Molecular, Genetic And Biochemical Characterization Of Resistance Protein-Mediated Signaling Against Turnip Crinkle Virus, Rae-Dong Jeong

University of Kentucky Doctoral Dissertations

Infection of the resistant Arabidopsis ecotype Di-17 with Turnip Crinkle Virus (TCV) elicits hypersensitive response (HR), accompanied by increased expression of defense genes. HR to TCV is conferred by HRT, which encodes a coiled-coil (CC)-nucleotide-binding site (NBS)-leucine-rich repeat (LRR) class of resistance (R) protein. In contrast to HR, resistance requires HRT and a recessive locus designated rrt. Unlike most CC-NBS-LRR R proteins, HRT-mediated resistance is dependent on EDS1 and independent of NDR1. Resistance is also dependent on salicylic acid (SA) pathway and light. A dark treatment, immediately following TCV inoculation, suppresses HR, resistance and activation of a majority of the …

Giant Viruses, James L. Van Etten

James Van Etten Publications

The common view of viruses, mostly true, is of tiny burglars that sneak into cells, grab the biosynthetic controls and compel the cell to make huge numbers of progeny that break out of the cell and keep the replication cycle going. Viruses are supposed to be diminutive even compared to cells that are just a micrometer (1,000 nanometers) in diameter. They are supposed to travel light, making do with just a few well-adapted genes.

In 1992, a new microorganism was isolated from a power-plant cooling tower in Bradford, England, where Timothy Robotham, a microbiologist at Leeds Public Health Laboratory, was …

Abundant Degenerate Miniature Inverted-Repeat Transposable Elements In Genomes Of Epichloid Fungal Endophytes Of Grasses, Damien J Fleetwood, Anar K Khan, Richard D Johnson, Carolyn A Young, Shipra Mittal, Ruth E Wrenn, Uljana Hesse, Simon J Foster, Christopher L. Schardl, Barry Scott

Plant Pathology Faculty Publications

Miniature inverted-repeat transposable elements (MITEs) are abundant repeat elements in plant and animal genomes; however, there are few analyses of these elements in fungal genomes. Analysis of the draft genome sequence of the fungal endophyte Epichloë festucae revealed 13 MITE families that make up almost 1% of the E. festucae genome, and relics of putative autonomous parent elements were identified for three families. Sequence and DNA hybridization analyses suggest that at least some of the MITEs identified in the study were active early in the evolution of Epichloë but are not found in closely related genera. Analysis of MITE integration …

Role Of The Sexual Cycle In Development Of Genotypic And Phenotypic Diversity In Gibberella Zeae, Sladana Bec

Theses and Dissertations--Plant Pathology

Gibberella zeae (anamorph Fusarium graminearum) is a homothallic ascomycete pathogen that is responsible for causing Fusarium head blight (FHB) of wheat and small grains. In addition to causing a reduction in yield, harvested grain is frequently contaminated with trichothecene mycotoxins that are harmful for human and animal health. Use of wheat varieties with resistance to FHB is an important strategy to lower its impact. In order to produce varieties with durable resistance, we must understand the origin and degree of genetic diversity present in the pathogen population. In my research, I focused my efforts on an investigation of the …

Control Of Glucosylceramide Production And Morphogenesis By The Bar1 Ceramide Synthase In Fusarium Graminearum, William B. Rittenour, Ming Chen, Edgar B. Cahoon, Steven D. Harris

Department of Plant Pathology: Faculty Publications

The contribution of plasma membrane proteins to the virulence of plant pathogenic fungi is poorly understood. Accordingly, the objective of this study was to characterize the acyl-CoA dependent ceramide synthase Bar1 (previously implicated in plasma membrane organization) in the wheat pathogen Fusarium graminearum. The role of Bar1 in mediating cell membrane organization was confirmed as DBAR1 mutants failed to display a distinct sterol-rich domain at the hyphal tip. The DBAR1 mutants were non-pathogenic when inoculated onto wheat heads, and their in vitro growth also was severely perturbed. DBAR1 mutants were incapable of producing perithecia (sexual fruiting structures) and only produced …

The Sugar Sensor, Trehalose-6-Phosphate Synthase (Tps1), Regulates Primary And Secondary Metabolism During Infection By The Rice Blast Fungus: Will Magnaporthe Oryzae’S “Sweet Tooth” Become Its “Achilles’ Heel”?, Jesse Fernandez, Richard Wilson

Department of Plant Pathology: Faculty Publications

Rice blast disease is considered one of the most serious diseases of cultivated rice and is mediated by the causal agent, Magnaporthe oryzae. During infection, dome-shaped fungal cells, called appressoria, form on the surface of the leaf and generate turgor through the accumulation of glycerol. This enormous pressure is directed down onto a thin penetration hypha emerging from the base of the cell, forcing it through the surface of the rice leaf and allowing fungal colonization of the plant interior. The non-reducing disaccharide, trehalose, is present in conidia of M. oryzae and is mobilized during appressorium formation. The first …

Mechanisms Of Contact-Mediated Killing Of Yeast Cells On Dry Metallic Copper Surfaces, Davide Quaranta, Travis Krans, Christophe Espirito Santo, Christian Elowsky, Dylan W. Domaille, Christopher J. Chang, Gregor Grass

Department of Plant Pathology: Faculty Publications

Surfaces made of copper or its alloys have strong antimicrobial properties against a wide variety of microorganisms. However, the molecular mode of action responsible for the antimicrobial efficacy of metallic copper is not known. Here, we show that dry copper surfaces inactivate Candida albicans and Saccharomyces cerevisiae within minutes in a process called contact-mediated killing. Cellular copper ion homeostasis systems influenced the kinetics of contact-mediated killing in both organisms. Deregulated copper ion uptake through a hyperactive S. cerevisiae Ctr1p (ScCtr1p) copper uptake transporter in Saccharomyces resulted in faster inactivation of mutant cells than of wild-type cells. Similarly, lack of the …

Role Of Lipids In Tombusvirus Replication, Monika Sharma

University of Kentucky Doctoral Dissertations

Positive-strand RNA virus group are the most abundant among viruses affecting plants and animals. To successfully achieve replication, these viruses usurp or co-opt host proteins. To facilitate the discovery of host factors involved in Tomato bushy stunt virus (TBSV), yeast has been developed as a surrogate model host. Genome-wide approaches covering 95% of yeast genes, has revealed approximately hundred factors that could affect virus replication. Among the identified host factors, there are fourteen yeast genes, which affect/regulate lipid metabolism of the host.

One of the identified host gene is ERG25, which is an important factor for sterol biosynthesis pathway, affecting …

Regalia-Induced Resistance In Powdery Mildew-Infected Grapevine Plants Correlates With The Induction Of Chalcone Synthase, Thomas Crottogini

Horticulture and Crop Science

No abstract provided.

Analysis Of The Response Of Medicago Truncatula Calcium Oxalate Mutants To Abiotic Stress, Wayra Gabriela Navia-Gine

Graduate Theses and Dissertations

Calcium oxalate crystals are found in most plant species. In Medicago truncatula wild-type A17, crystals accumulate in leaves along the secondary veins and the only role attributed to them so far is defense against chewing insects. Calcium oxalate deficient (cod) mutants were isolated in M. truncatula; the cod mutants include cod5, which completely lack crystals in the leaves and cod6 that accumulates fewer and smaller crystals compared to A17. We analyzed gene expression in the cod mutants and A17 using GeneChip® Medicago Genome Arrays and found important differences in transcriptome between the three genotypes. In particular, we found a gene …

An Nadph-Dependent Genetic Switch Regulates Plant Infection By The Rice Blast Fungus, Richard A. Wilson, Robert P. Gibson, Cristian Quispe, Jennifer A. Littlechild, Nicholas J. Talbot

Fungal Molecular Plant-Microbe Interactions

To cause rice blast disease, the fungus Magnaporthe oryzae breaches the tough outer cuticle of the rice leaf by using specialized infection structures called appressoria. These cells allow the fungus to invade the host plant and proliferate rapidly within leaf tissue. Here, we show that a unique NADPH-dependent genetic switch regulates plant infection in response to the changing nutritional and redox conditions encountered by the pathogen. The biosynthetic enzyme trehalose-6-phosphate synthase (Tps1) integrates control of glucose-6-phosphate metabolism and nitrogen source utilization by regulating the oxidative pentose phosphate pathway, the generation of NADPH, and the activity of nitrate reductase. We report …

Population Variability Of Rotylenchulus Reniformis In Cotton Agroecosystems, Megan Leach

All Dissertations

Rotylenchulus reniformis, reniform nematode, is a highly variable species and an economically important pest in many cotton fields across the southeast. Rotation to resistant or poor host crops is a prescribed method for management of reniform nematode. An increase in the incidence and prevalence of the nematode in the United States has been reported over the last two decades. However it is not clear whether the observed increase is related to the emergence of novel populations that are more aggressive or have a higher fitness or shifts in host availability or susceptibility. The objectives of this research were to determine …

Fusarium Head Blight: Winter Wheat Cultivar Responses And Characterization Of Pathogen Isolates, John Fredy Hernandez Nopsa

Department of Agronomy and Horticulture: Dissertations, Theses, and Student Research

Fusarium Head Blight (FHB) is a destructive disease of wheat (Triticum aestivum L). FHB reduces yield and grain quality and causes accumulation of the mycotoxin deoxynivalenol (DON) in grain. Cultivar resistance is one of the most effective management strategies for FHB. Experiments were conducted to 1) identify winter wheat cultivars with resistance to FHB and DON accumulation, 2) determine the effect of winter wheat cultivar on the relationship between FHB and DON concentration, and 3) identify the major species of Fusarium causing FHB in Nebraska and characterize its isolates. Differences (P ≤ 0.05) were detected among cultivars in …

Making Of Viral Replication Organelles By Remodeling Interior Membranes, Zsuzsanna Sasvari, Peter D. Nagy

Plant Pathology Faculty Publications

Positive-stranded RNA (+RNA) viruses exploit host cell machinery by subverting host proteins and membranes and altering cellular pathways during infection. To achieve robust replication, some +RNA viruses, such as poliovirus (PV), build special intracellular compartments, called viral replication organelles. A recent work from the Altan-Bonnett laboratory [1] gave new insights into the formation of poliovirus replication organelles, which are unique subcellular structures containing many individual replication complexes as a result of dynamic cellular membrane remodeling.

Translation Elongation Factor 1a Facilitates The Assembly Of The Tombusvirus Replicase And Stimulates Minus-Strand Synthesis, Zhenghe Li, Judit Pogany, Steven Tupman, Anthony M. Esposito, Terri Goss Kinzy, Peter D. Nagy

Plant Pathology Faculty Publications

Replication of plus-strand RNA viruses depends on host factors that are recruited into viral replicase complexes. Previous studies showed that eukaryotic translation elongation factor (eEF1A) is one of the resident host proteins in the highly purified tombusvirus replicase complex. Using a random library of eEF1A mutants, we identified one mutant that decreased and three mutants that increased Tomato bushy stunt virus (TBSV) replication in a yeast model host. Additional in vitro assays with whole cell extracts prepared from yeast strains expressing the eEF1A mutants demonstrated several functions for eEF1A in TBSV replication: facilitating the recruitment of the viral RNA template …

Identification Of Sources Of Rhizoctonia Root Rot Resistance In Common Bean And Mapping A New Source Of Bean Rust Resistance From The Tertiary Gene Pool Of Common Bean, Pamela A. Peña-Perdomo

Department of Agronomy and Horticulture: Dissertations, Theses, and Student Research

An efficient screening method was developed and used to identify bean lines resistant to Rhizoctonia Root Rot caused by Rhizoctonia solani Kuhn. Two sets of 163 and 111 lines previously evaluated for drought tolerance at Mitchell, NE and Isabela, PR were evaluated for Rhizoctonia Root Rot resistance under greenhouse conditions. This root rot data was also correlated with yield under drought stress and non stress conditions. In the first set of lines the rhizoctonia mean score ranged from 1.7 to 3.9; and in the second set the rhizoctonia mean score was between 2.6 and 5.7. There was no significant correlation …

The Combined Effect Of Environmental And Host Factors On The Emergence Of Viral Rna Recombinants, Hannah M. Jaag, Peter D. Nagy

Plant Pathology Faculty Publications

Viruses are masters of evolution due to high frequency mutations and genetic recombination. In spite of the significance of viral RNA recombination that promotes the emergence of drug-resistant virus strains, the role of host and environmental factors in RNA recombination is poorly understood. Here we report that the host Met22p/Hal2p bisphosphate-3'-nucleotidase regulates the frequency of viral RNA recombination and the efficiency of viral replication. Based on Tomato bushy stunt virus (TBSV) and yeast as a model host, we demonstrate that deletion of MET22 in yeast or knockdown of AHL, SAL1 and FRY1 nucleotidases/phosphatases in plants leads to increased TBSV recombination …

Plant Immunity Directly Or Indirectly Restricts The Injection Of Type Iii Effectors By The Pseudomonas Syringae Type Iii Secretion System, Emerson Crabill, Anna Joe, Anna Block, Jennifer M. Van Rooyen, James R. Alfano

Center for Plant Science Innovation: Faculty Publications

Plants perceive microorganisms by recognizing microbial molecules known as pathogen-associated molecular patterns (PAMPs) inducing PAMP-triggered immunity (PTI) or by recognizing pathogen effectors inducing effector-triggered immunity (ETI). The hypersensitive response (HR), a programmed cell death response associated with ETI, is known to be inhibited by PTI. Here, we show that PTI-induced HR inhibition is due to direct or indirect restriction of the type III protein secretion system’s (T3SS) ability to inject type III effectors (T3Es). We found that the Pseudomonas syringae T3SS was restricted in its ability to inject a T3E-adenylate cyclase (CyaA) injection reporter into PTI-induced tobacco (Nicotiana tabacum) cells. …

The Chlorella Variabilis Nc64a Genome Reveals Adaptation To Photosymbiosis, Coevolution With Viruses, And Cryptic Sex, Guillaume Blanc, Garry Duncan, Irina Agarkova, Mark Borodovsky, James Gurnon, Alan Kuo, Erika Lindquist, Susan Lucas, Jasmyn Pangilinan, Juergen Polle, Asaf Salamov, Astrid Terry, Takashi Yamada, David D. Dunigan, Igor V. Grigoriev, Jean-Michel Claverie, James L. Van Etten

Department of Plant Pathology: Faculty Publications

Chlorella variabilis NC64A, a unicellular photosynthetic green alga (Trebouxiophyceae), is an intracellular photobiont of Paramecium bursaria and a model system for studying virus/algal interactions. We sequenced its 46-Mb nuclear genome, revealing an expansion of protein families that could have participated in adaptation to symbiosis. NC64A exhibits variations in GC content across its genome that correlate with global expression level, average intron size, and codon usage bias. Although Chlorella species have been assumed to be asexual and nonmotile, the NC64A genome encodes all the known meiosis-specific proteins and a subset of proteins found in flagella. We hypothesize that Chlorella might have …

Increased Plant Uptake Of Nitrogen From 15N-Depleted Fertilizer Using Plant Growth-Promoting Rhizobacteria, A. O. Adesemoye, H. A. Torbert, J. W. Kloepper

Department of Plant Pathology: Faculty Publications

Harmful environmental effects resulting from fertilizer use have spurred research into integrated nutrient management strategies which can include the use of specific microorganisms to enhance nutrient use efficiency by plants. Some strains of plant growth-promoting rhizobacteria (PGPR) have been reported to enhance nutrient uptake by plants, but no studies with PGPR have used ¹⁵N isotope techniques to prove that the increased N in plant tissues came from the N applied as fertilizer. The current study was conducted to demonstrate that a model PGPR system can enhance plant uptake of fertilizer N applied to the soil using different rates of …

Unique Functionality Of 22 Nt Mirnas In Triggering Rdr6-Dependent Sirna Biogenesis From Target Transcripts In Arabidopsis, Josh T. Cuperus, Alberto Carbonell, Noah Fahlgren, Hernan Garcia-Ruiz, Russell T. Burke, Atsushi Takeda, Christopher M. Sullivan, Sunny D. Gilbert, Taiowa A. Montgomery, James C. Carrington

Department of Plant Pathology: Faculty Publications

RNA interference pathways may involve amplification of secondary siRNAs by RNA-dependent RNA polymerases. In plants, RDR6-dependent secondary siRNAs arise from transcripts targeted by some microRNA (miRNA). Here, Arabidopsis thaliana secondary siRNA from mRNA, and trans-acting siRNA, are shown to be triggered through initial targeting by 22 nt miRNA that associate with AGO1. In contrast to canonical 21 nt miRNA, 22 nt miRNA primarily arise from foldback precursors containing asymmetric bulges. Using artificial miRNA constructs, conversion of asymmetric foldbacks to symmetric foldbacks resulted in production of 21 nt forms of miR173, miR472 and miR828. Both 21 and 22 nt forms associated …

Heterokaryotic Nuclear Conditions And A Heterogeneous Nuclear Population Are Observed By Flow Cytometry In Phytophthora Infestans, Mursel Catal, Louis King, Pavani Tumbalam, Prissana Wiriyajitsomboon, William W. Kirk, Gerard C. Adams

Department of Plant Pathology: Faculty Publications

A simple and reliable method for preparation of whole nuclei of a common oomycete, Phytophthora infestans, is described for laser flow cytometry. The ease of preparation, the absence of detectable debris and aggregates, and the precision in determinations of DNA content per nucleus improve interpretation and understanding of the genetics of the organism. Phytophthora infestans is the pathogen that causes potato and tomato late blight. The genetic flexibility of P. infestans and other oomycete pathogens has complicated understanding of the mechanisms of variation contributing to shifts in race structure and virulence profiles on important agricultural crops. Significant phenotypic and …

Alteration In Lignin Biosynthesis Restricts Growth Of Fusarium Spp. In Brown Midrib Sorghum, Deanna L. Funnell-Harris, Jeffrey F. Pedersen, Scott E. Sattler

Department of Plant Pathology: Faculty Publications

To improve sorghum for bioenergy and forage uses, brown midrib (bmr)6 and -12 near-isogenic genotypes were developed in different sorghum backgrounds. The bmr6 and bmr12 grain had significantly reduced colonization by members of the Gibberella fujikuroi species complex compared with the wild type, as detected on two semiselective media. Fusarium spp. were identified using sequence analysis of a portion of the translation elongation factor (TEF) 1-α gene. The pathogens Fusarium thapsinum, F. proliferatum, and F. verticillioides, G. fujikuroi members, were commonly recovered. Other frequently isolated Fusarium spp. likely colonize sorghum asymptomatically. The χ2 analyses showed that the …

A Functional Calcium-Transporting Atpase Encoded By Chlorella Viruses, Maria Cristina Bonza, Holger Martin, Ming Kang, Gentry L. Lewis, Timo Greiner, Sonia Giacometti, James L. Van Etten, Maria Ida De Michelis, Gerhard Thiel, Anna Moroni

James Van Etten Publications

Calcium-transporting ATPases (Ca²⁺ pumps) are major players in maintaining calcium homeostasis in the cell and have been detected in all cellular organisms. Here, we report the identification of two putative Ca²⁺ pumps, M535L and C785L, encoded by chlorella viruses MT325 and AR158, respectively, and the functional characterization of M535L. Phylogenetic and sequence analyses place the viral proteins in group IIB of P-type ATPases even though they lack a typical feature of this class, a calmodulin-binding domain. A Ca²⁺ pump gene is present in 45 of 47 viruses tested and is transcribed during virus infection. Complementation analysis of …

Identification Of An L-Rhamnose Synthetic Pathway In Two Nucleocytoplasmic Large Dna Viruses, Madhu Parakkottil Chothi, Garry A. Duncan, Andrea Armirotti, Chantal Abergel, James R. Gurnon, James L. Van Etten, Cinzia Bernardi, Gianluca Damonte, Michela Tonetti

James Van Etten Publications

Nucleocytoplasmic large DNA viruses (NCLDVs) are characterized by large genomes that often encode proteins not commonly found in viruses. Two species in this group are Acanthocystis turfacea chlorella virus 1 (ATCV-1) (family Phycodnaviridae, genus Chlorovirus) and Acanthamoeba polyphaga mimivirus (family Mimiviridae), commonly known as mimivirus. ATCV-1 and other chlorovirus members encode enzymes involved in the synthesis and glycosylation of their structural proteins. In this study, we identified and characterized three enzymes responsible for the synthesis of the sugar L-rhamnose: two UDP-D-glucose 4,6-dehydratases (UGDs) encoded by ATCV-1 and mimivirus and a bifunctional UDP-4-keto-6-deoxy-D-glucose epimerase/reductase (UGER) from mimivirus. Phylogenetic …

Overall Growth Of Tomato (Lycopersicon Esculentum L. Cv. Glacier) Inoculated With Species Of Glomus And Trichoderma Growing Under Greenhouse Conditions, Brian J. Adkins

Horticulture and Crop Science

The overall plant growth of tomato (Lycopersicon esculentum L. cv. Glacier) cultivated in greenhouse conditions along with inoculation treatments of beneficial fungi species were observed. This study included 2 brand name products containing species of fungi which include: 1)Botanicare Guardian TR containing Trichoderma harzianum 10,000,000 CFU/g, T. hamatum 10,000,000 CFU/g, T. koningii 10,000,000 CFU/g; and 2)Botanicare Guardian MY containing Glomus intraradices 57 propagules/g;G. fasiculatum 57 propagules/g; G. etunicatum 57 propagules/g; G. clarum 57 propagules/g. Treatments include: Treatments: 1) 2g Botanicare Guardian TR; 2) 2g Botanicare Guardian MY; 3) 1g Botanicare Guardian TR + 1g Botanicare Guardian MY; and …