First report of the beech leaf disease nematode Litylenchus crenatae mccannii (Nematoda: Anguinidae) in Michigan.

The North American beech leaf disease (BLD) nematode, Litylenchus crenatae mccannii Handoo, Li, Kantor, Bauchan, McCann, Gabriel, Yu, Reed, Koch, Martin and Burke, 2020, is recognized as a newly emergent nematode species that causes BLD in beech trees (Fagus spp.) in North America (Carta et al. 2020; Kantor et al. 2022a). Since the first report of BLD on American beech (Fagus grandifolia Ehrh) within the Lake County, located at the north-eastern corner of the state of Ohio in 2012 (Carta et al. 2020), the disease has rapidly spread to other US states and a province in Canada (Erwing et al. 2018; Carta et al 2020; Marra and LaMondia 2020; Reed et al 2020; Kantor et al. 2022b). Currently, besides Ohio, this nematode has been reported in Pennsylvania, New York, Connecticut, Massachusetts, Maine, Rhode Island, New Jersey, West Virginia, and Virginia, as well as Ontario, Canada. Different life stages of L. crenatae mccannii were isolated from symptomatic American beech leaves from an isolated natural maple-beech stand in rural Saint Clair Cty., Michigan, US; presenting typical symptoms of beech leaf disease, i.e., swelling and darkening of interveinal leaf tissues. Samples were taken to the Forest Pathology Laboratory at Michigan State University where L. crenatae mccannii presence was confirmed in the leaves after which samples were sent to the Mycology and Nematology Genetic Diversity and Biology Laboratory (USDA-ARS) in Beltsville, Maryland for official confirmation. Nematodes were identified based on morphology and sequence analysis of the internal transcribed spacer (ITS), and the D2D3 region of the 28S large subunit ribosomal DNA. To validate the morphological identification two different ribosomal DNA loci were amplified, sequenced and the phylogenetic relationships were generated. The amplification yielded fragments of 784 and 741 bp flanked by the ITS (GenBank accession no. OP689654) and D2D3 (GenBank accession no. OP689710) primers, respectively. The sequences obtained for the specimens collected in Michigan revealed 100% similarity to L. crenatae mccannii sequences obtained from specimens collected from other geographical areas in the US, and therefore validating the morphological analyses as well. The ITS sequence shared a 99.75% similarity with the subspecies L. crenatae (GenBank accession no. LC383724.1), and 90.53% similarity to L. coprosma Zhao, Davies, Alexander and Riley, 2011 (GU727548.1). While the D2D3 sequences of both L. crenatae subspecies revealed a 100% similarity (versus LC383725.1), they revealed 95.35% similarity to L. coprosma (KY679564.1). Since the first confirmed detection of BLD in June 2022 in St. Clair Cty, BLD has been reported in Oakland and Wayne Ctys (7 reports total across the three counties), suggesting BLD spread in the SE of Michigan. BLD confirmation was based on either physical symptoms (leaf banding), and/or the presence of the beech leaf nematode by morphological or molecular confirmation. The presence of the beech leaf nematode in symptomatic leaves follow the results obtained by Carta et al. (2020) after inoculation of beech seedlings with L. crenatae mccannii. Based on both morphological and molecular analyses the specimens collected in the state of Michigan were identified as L. crenatae mccannii. To our knowledge, this is the first report of this species in conjunction with symptomatic F. grandifolia leaves in this state.

Characterization of Diaporthe spp. Associated With Spruce Decline on Colorado Blue Spruce in Michigan.

Since 2006 there has been a decline in Colorado blue spruce (CBS; Picea pungens) planted as landscape trees and for Christmas tree production throughout the Lower Peninsula of Michigan. This decline is characterized by a slow loss of needles in the lower portion of the tree starting at branch tips, followed by entire branch dieback, which progresses upward over several years. This dieback has been linked to shallow branch cankers visible in the phloem when the bark layer is removed. Isolates in the fungal genus Diaporthe have been consistently isolated from lesion margins on symptomatic branches. Before the initial reports of declining CBS in landscape and Christmas trees, Diaporthe was known only as a nursery disease of CBS. To determine the species of Diaporthe linked to the decline of CBS in Michigan, seven gene regions were sequenced from a collection of Diaporthe isolates collected in 2011 through 2018 from CBS and other coniferous hosts. Subsequent phylogenetic analyses indicated that Diaporthe eres and a novel Diaporthe clade were present on symptomatic CBS in Michigan. The new species D. brevicancria nov. is described, and Koch's postulates were confirmed for D. brevicancria nov. and D. eres. D. brevicancria nov. produced the largest cankers in greenhouse pathogenicity trials, and dual inoculations of D. brevicancria nov. and D. eres produced intermediate cankers.

Independent Evolution Has Led to Distinct Genomic Signatures in Dutch Elm Disease-Causing Fungi and Other Vascular Wilts-Causing Fungal Pathogens.

Vascular wilts are important diseases caused by plant pathogenic fungi that result in the rapid death of their plant hosts. This is due to a systemic defense mechanism whereby the plant induces the compartmentalization of the infected vascular system in order to reduce the propagation of the fungus. The ascomycete class Sordariomycetes contains several species that cause vascular wilts in diverse plant hosts, and they can be classified into four taxonomic orders. The genetic mechanisms of pathogenesis have already been investigated in Fusarium and Verticillium species, but they have not yet been compared with other well-known wilt-causing species, especially fungi causing oak wilt or Dutch elm disease (DED). Here we analyzed 20 whole genome assemblies of wilt-causing fungi together with 56 other species using phylogenetic approaches to trace expansions and contractions of orthologous gene families and gene classes related to pathogenicity. We found that the wilt-causing pathogens evolved seven times, experiencing the largest fold changes in different classes of genes almost every time. However, some similarities exist across groups of wilt pathogens, particularly in Microascales and Ophiostomatales, and these include the common gains and losses of genes that make up secondary metabolite clusters (SMC). DED pathogens do not experience large-scale gene expansions, with most of the gene classes, except for some SMC families, reducing in number. We also found that gene family expansions in the most recent common ancestors of wilt pathogen groups are enriched for carbohydrate metabolic processes. Our study shows that wilt-causing species evolve primarily through distinct changes in their repertoires of pathogenicity-related genes and that there is the potential importance of carbohydrate metabolism genes for regulating osmosis in those pathogens that penetrate the plant vascular system.