Plant Vascular Tissues Research Articles

Genome-Wide Analysis of the Serine Carboxypeptidase-like (SCPL) Protein Family of Bitter Gourd and Functional Validation of McSCPL22 in Fusarium oxysporum f. sp. Momordicae (FOM) Resistance

Bitter gourd is increasingly being recognized for its value as a vegetable and medicinal use, but the molecular mechanisms of pathogen resistance remain relatively poorly understood. The serine carboxypeptidase-like (SCPL) protein family plays a key role in plant growth, pathogen defense, and so on. However, a comprehensive identification and functional characterization of the SCPL gene family has yet to be conducted in bitter melon. In this study, 32 SCPL genes were identified in bitter gourd and divided into three classes. The number of SCPL genes contained in the three clusters was 7, 7, and 18, respectively. Most SCPL gene promoters contain cis-acting elements with light, hormone, and stress responses. The RNA sequencing data showed that the expression of several SCPL genes changed significantly after pathogen infection. In particular, expression of the McSCPL4, 10, 17, 22, and 25 genes increased substantially in the resistant varieties after infection, and their expression levels were higher than those in the susceptible varieties. These results suggested that genes such as McSCPL4, 10, 17, 22, and 25 may play a significant role in conferring resistance to fungal infections. Moreover, the expression levels of the McSCPL10, 17, 22, 23, and 25 genes were likewise significantly changed after being induced by salicylic acid (SA) and jasmonic acid (JA). In situ hybridization showed that McSCPL22 was expressed in the vascular tissues of infected plants, which largely overlapped with the location of Fusarium oxysporum f. sp. Momordicae (FOM) infection and the site of hydrogen peroxide production. Our results showed that McSCPL22 may be involved in the regulation of the SA and JA pathways and enhance resistance to FOM in bitter gourd plants. This is the first study to perform SCPL gene family analysis in bitter gourd. McSCPL22 may have the potential to enhance FOM resistance in bitter gourd, and further investigation into its function is warranted. The results of this study may enhance the yield and molecular breeding of bitter gourd.

A gain-of-function mutation in BnaIAA13 disrupts vascular tissue and lateral root development in Brassica napus.

Rapeseed (Brassica napus) is an important oilseed crop worldwide. Plant vascular tissues are responsible for long-distance transport of water and nutrients and for providing mechanical support. The lateral roots absorb water and nutrients. The genetic basis of vascular tissue and lateral root development in rapeseed remains unknown. This study characterized an ethyl methanesulfonate-mutagenized rapeseed mutant, T16, which showed dwarf stature, reduced lateral roots, and leaf wilting. SEM observations showed that the internode cells were shortened. Observations of tissue sections revealed defects in vascular bundle development in the stems and petioles. Genetic analysis revealed that the phenotypes of T16 were controlled by a single semi-dominant nuclear gene. Map-based cloning and genetic complementarity identified BnaA03.IAA13 as the functional gene; a G-to-A mutation in the second exon changed glycine at position 79 to glutamic acid, disrupting the conserved degron motif VGWPP. Transcriptome analysis in roots and stems showed that auxin and cytokinin signaling pathways were disordered in T16. Evolutionary analysis showed that AUXIN/INDOLE-3-ACETIC ACID is conserved during plant evolution. The heterozygote of T16 showed significantly reduced plant height while maintaining other agronomic traits. Our findings provide novel insights into the regulatory mechanisms of vascular tissue and lateral root development, and offer a new germplasm resource for rapeseed breeding.

Restorer of fertility like 30, encoding a mitochondrion-localized pentatricopeptide repeat protein, regulates wood formation in poplar.

Nuclear-mitochondrial communication is crucial for plant growth, particularly in the context of cytoplasmic male sterility (CMS) repair mechanisms linked to mitochondrial genome mutations. The restorer of fertility-like (RFL) genes, known for their role in CMS restoration, remain largely unexplored in plant development. In this study, we focused on the evolutionary relationship of RFL family genes in poplar specifically within the dioecious Salicaceae plants. PtoRFL30 was identified to be preferentially expressed in stem vasculature, suggesting a distinct correlation with vascular cambium development. Transgenic poplar plants overexpressing PtoRFL30 exhibited a profound inhibition of vascular cambial activity and xylem development. Conversely, RNA interference-mediated knockdown of PtoRFL30 led to increased wood formation. Importantly, we revealed that PtoRFL30 plays a crucial role in maintaining mitochondrial functional homeostasis. Treatment with mitochondrial activity inhibitors delayed wood development in PtoRFL30-RNAi transgenic plants. Further investigations unveiled significant variations in auxin accumulation levels within vascular tissues of PtoRFL30-transgenic plants. Wood development anomalies resulting from PtoRFL30 overexpression and knockdown were rectified by NAA and NPA treatments, respectively. Our findings underscore the essential role of the PtoRFL30-mediated mitochondrion-auxin signaling module in wood formation, shedding light on the intricate nucleus-organelle communication during secondary vascular development.

First report of Fusarium oxysporum f. sp. lactucae causing wilt on greenhouse-grown lettuce in North Carolina.

In August 2023, butterhead lettuce (Lactuca sativa L.) presented wilting, chlorosis, and about 2 cm of reddish-tan internal discoloration in the crown from a commercial greenhouse in Orange County, North Carolina. Plant collapse beginning with the outer leaves near the soil surface was observed with 25% disease incidence. Symptomatic lettuce plants were submitted to North Carolina State University's Plant Disease and Insect Clinic. Vascular tissue from symptomatic crowns were cut into pieces, and surface-sterilized in 10% NaOCl for two minutes. The tissue was rinsed in sterile deionized water three times, blotted dry, and placed on acidified potato dextrose agar (APDA). Three isolates, each from a different symptomatic plant, were transferred to APDA and Spezieller Nährstoffarmer agar (SNA) with pieces of sterile filter paper on the surface of the SNA media and incubated for 14 days at 23°C in the dark. Each isolate produced micro and macroconidia consistent with the morphological description of Fusarium oxysporum Schlechtendahl emend. Snyder & Hansen (Leslie and Summerell 2006). DNA was extracted from 15-day-old fungal colonies grown on APDA media using the DNeasy Plant Mini Kit (Qiagen, Germantown, MD, U.S.A.). The intergenic spacer locus was amplified using two primer pairs, iNL11/CNSa and iCNS1/NLa, and sequences were aligned together to form a single contig (O'Donnell et al., 2009). Primers EF1/EF2 were used to amplify the elongation factor 1-alpha region (O'Donnell et al., 1998). Each isolate was deposited into GenBank with accession numbers PP216479, PP216480, PP216481, PP235836, PP235837, and PP235838. Individual isolates revealed a 100% query cover and identity match with sequences of F. oxysporum f. sp. lactucae (FOL) and >99% identity with F. oxysporum CBS 144134 type material accessions in GenBank using BLASTn. A comparison with previously described lettuce isolates showed a homologous match with FOL race 1 isolates from California (MH412701), Arizona (DQ837658), and Greece (OQ466116), and race 4 isolates from Italy (MK801787) and Spain (OP903519). Each isolate was verified as FOL using specific primers FLA0001F/FLA0001R for FOL based on sequence tagged site markers designed by Shimazu et al. (2005). To confirm Koch's postulates, fifteen 21-day-old lettuce cv. Red Tide plants were inoculated with FOL (isolate FOLNC_660). During transplanting, lettuce roots were submerged in a 1 × 105 conidia/mL suspension for five minutes, following an inoculation protocol from Schmale and Gordon (2003). The lettuce plants were placed separately in 8.9 × 8.3 cm pots containing potting soil and maintained in a greenhouse with 31°C daytime and 25°C nighttime temperature, relative humidity of 60%, and 12-hour photoperiod. After 15 days, 80% and 86.7% of infected plants exhibited wilting, chlorosis, and vascular discoloration. The fifteen control plants remained symptomless for both experimental runs. FOL was recovered from the vascular tissue of all symptomatic plants. To our knowledge, this constitutes the first report of FOL infecting lettuce in North Carolina. Fusarium wilt of lettuce has been reported in California (Hubbard and Gerik 1993), Arizona (Matheron and Koike 2003), and most recently in Florida (Murray et al., 2020). The presence of FOL in North Carolina may result in significant crop loss for commercial growers. One of the most effective management strategies is to plant lettuce cultivars that are resistant against FOL.

Stem Cells and Differentiation in Vascular Tissues.

Plant vascular tissues are crucial for the long-distance transport of water, nutrients, and a multitude of signal molecules throughout the plant body and, therefore, central to plant growth and development. The intricate development of vascular tissues is orchestrated by unique populations of dedicated stem cells integrating endogenous as well as environmental cues. This review summarizes our current understanding of vascular-related stem cell biology and of vascular tissue differentiation. We present an overview of the molecular and cellular mechanisms governing the maintenance and fate determination of vascular stem cells and highlight the interplay between intrinsic and external cues. In this context, we emphasize the role of transcription factors, hormonal signaling, and epigenetic modifications. We also discuss emerging technologies and the large repertoire of cell types associated with vascular tissues, which have the potential to provide unprecedented insights into cellular specialization and anatomical adaptations to distinct ecological niches.

Draft genomes of two phylotype I and II Ralstonia solanacearum species complex (RSSC) isolates causing bacterial wilt in tomato plants from Costa Rica.

Bacteria from RSSC hold agricultural significance as they are the causal agents of bacterial wilt. Here, we report the draft genomes of two bacteria extracted from vascular tissues of infected tomato plants. Isolate RALF-MA was classified as Ralstonia pseudosolanacearum (phylotype I) and RALSA-MA as Ralstonia solanacearum (phylotype II).

Evidence for poplar PtaPLATZ18 in the regulation of plant growth and vascular tissues development.

Plant A/T-rich protein and zinc-binding protein (PLATZ) are plant-specific transcription factors playing a role in plant development and stress response. To assess the role of PLATZs in vascular system development and wood formation in poplar, a functional study for PtaPLATZ18, whose expression was associated with the xylem, was carried out. Poplar dominant repressor lines for PtaPLATZ18 were produced by overexpressing a PtaPLATZ18-SRDX fusion. The phenotype of three independent transgenic lines was evaluated at morphological, biochemical, and molecular levels and compared to the wild type. The PtaPLATZ18-SRDX lines showed increased plant height resulting from higher internode length. Besides, a higher secondary xylem thickness was also evidenced in these dominant repression lines as compared to the wild type suggesting an activation of cambial activity. A higher amount of lignin was evidenced within wood tissue as compared to the wild type, indicating an alteration in cell wall composition within xylem cell types. This latter phenotype was linked to an increased expression of genes involved in lignin biosynthesis and polymerization. The phenotype observed in the PtaPLATZ18-SRDX lines argues that this transcription factor targets key regulators of plant growth and vascular tissues development.

Development of novel LAMP and qPCR assays for rapid and specific identification of Bronze birch borer (Agrilus anxius)

AbstractBuprestids are an emerging threat to broadleaf forests across the world. Bronze birch borer (Agrilus anxius, BBB) poses a serious threat to European birch species if the insect were to be introduced. Due to their cryptic lifestyle feeding on the vascular tissue of their host plants, buprestids and other wood borers can be difficult to observe or detect. Early detection tools are vital to swiftly implement eradication measures and prevent the establishment of introduced species. In this study, we developed novel qPCR and LAMP assays for BBB and investigated the specificity and sensitivity for their use as early detection tools in European forests. Plant chemicals may limit these assays, so we conducted sensitivity testing with extracted foliage and plant vascular tissues to determine potential inhibition effects on DNA amplification. Both assays were specific to the target species when tested against the DNA of 17 other European Agrilus/buprestid species, two Scolytinae, and five Cerambycids (N = 24). Both assays varied in sensitivity with the qPCR assay amplifying at a concentration as low as 20 fg/μL, whereas the LAMP assay amplified as low as 3.2 pg/μL. Plant chemicals in DNA extracts from leaves did not impact the sensitivity of either assay, reaching similar detection levels. In contrast, vascular tissue reduced the sensitivity of the LAMP assay, amplifying as low as 0.04 ng/μL compared with 0.008 ng/μL in the control. These results demonstrate that both assays are highly specific and sensitive tools that can be used to detect frass and identify larvae as well as monitor the spread of A. anxius. qPCR resulted in more sensitive than LAMP overall. Thus, if results are needed quickly to make fast management decisions or as an initial screening of samples, the LAMP method is optimal. However, if fine detection is critical, then qPCR is preferential.

Monitoring Xylem Transport in Arabidopsis thaliana Seedlings Using Fluorescent Dyes.

Fluorescent dyes are often used to observe transport mechanisms in plant vascular tissues. However, it has been technically challenging to apply fluorescent dyes on roots to monitor xylem transport in vivo. Here, we present a fast, noninvasive, and high-throughput protocol to monitor xylem transport in seedlings. Using the fluorescent dyes 5(6)-carboxyfluorescein diacetate (CFDA) and Rhodamine WT, we were able to observe xylem transport on a cellular level in Arabidopsis thaliana roots. We describe how to apply these dyes on primary roots of young seedlings, how to monitor root-to-shoot xylem transport, and how to measure xylem transport velocity in roots. Moreover, we show that our protocol can also be applied to lateral roots and grafted seedlings to assess xylem (re)connection. Altogether, these techniques are useful for investigating xylem functionality in diverse experimental setups.

An Overview on Paddy Crop Residue Decomposition: A Biochemical Analysis of the Process

The most common crop in India is rice, which is grown on 43 million planted hectares and produces 746 million tones of grain annually. Due to the fact that straw accounts for 50% of the dry weight of the rice plant, a tremendous amount of straw is produced as a byproduct of rice farming each year. We produce 65% of our biomass on land, according to estimates. Lignin is the most prevalent natural polymer of that biomass after cellulose and a significant renewable supply of aromatic carbon on earth. Since lignin, cellulose, and hemicelluloses make up the structural elements of higher land plant vascular tissues, the biodegradation of these elements is a crucial step in the recycling of terrestrial biosynthetic carbon. This study focus on biochemical analysis of decomposition of paddy straw and find out different fungi present in soil after decomposition.

BIOGENIC AND ANTHROPOGENIC MAGNETIC NANOPARTICLES IN THE PHLOEM SIEVE TUBES OF PLANTS

The samples of leaves and roots of Nicotiana tabacum, the stems and tubers of Solanum tuberosum and the stems of pea Pisum sativum were examined by scanning probe microscopy in atomic force and magnetic force modes after cultivation without and with addition of magnetite nanoparticles in soil. Chains of both biogenic and anthropogenic magnetic nanoparticles are detected in the wall of the phloem sieve tubes i.e., the vascular tissue of plants. Such a localization of biogenic magnetic nanoparticles supports the idea that the chains of magnetic nanoparticles in different organs of plants have common metabolic functions. Stray gradient magnetic fields about several thousand Oe, which are created by chains of both biogenic and anthropogenic magnetic nanoparticles, can significantly affect the metabolic processes through the influence on mass transfer of vesicles, granules, organelles and other components. In view of the results of this investigation, both BMNs and anthropogenic magnetite nanoparticles chains can significantly affect the processes of mass transfer of vesicles, organelles, structural elements of the membrane and others components because they create stray magnetic fields and gradient magnetic forces.

Verticillium dahliae Asp1 regulates the transition from vegetative growth to asexual reproduction by modulating microtubule dynamic organization.

Verticillium dahliae is a devastating pathogenic fungus that causes severe vascular wilts in more than 400 dicotyledonous plants. The conidiation of V. dahliae in plant vascular tissues is the key strategy for its adaptation to the nutrient-poor environment and is required for its pathogenicity. However, it remains unclear about the regulatory mechanism of conidium production of V. dahliae in vascular tissues. Here, we found that VdAsp1, encoding an inositol polyphosphate kinase, is indispensable for the pathogenicity of V. dahliae. Loss of VdAsp1 function does not affect the invasion of the host, but it impairs the colonization and proliferation in vascular tissues. The ΔVdAsp1 mutant shows defective initiation of conidiophore formation and reduced expression of genes associated with the central developmental pathway. By live-cell imaging, we observed that some of ΔVdAsp1 mutant hyphae are swollen, and microtubule arrangements at the apical region of these hyphae are disorganized. These results indicate that VdAsp1 regulates the transition from vegetative growth to asexual reproduction by modulating microtubule dynamic organization, which is essential for V. dahliae to colonize and proliferate in vascular tissues. These findings provided a potential new direction in the control of vascular wilt pathogen by targeting conidium production in vascular tissues.

Combined Transcriptomic and Metabolomic Analysis Reveals Insights into Resistance of Arabidopsis bam3 Mutant against the Phytopathogenic Fungus Fusarium oxysporum.

The wilt-inducing strains of Fusarium oxysporum are responsible for severe damage to many economically important plant species. The most cost-effective and environmentally safe method for the management of Fusarium wilt is the use of resistant cultivars when they are available. In the present study, the Arabidopsis genotype with disruptions in the β-amylase 3 (BAM3) gene, which encodes the major hydrolytic enzyme that degrades starch to maltose, had significantly lower susceptibility to Fusarium oxysporum f. sp. raphani (For) compared to wild-type (wt) plants. It showed the lowest disease severity and contained reduced quantities of fungal DNA in the plant vascular tissues when analyzed with real-time PCR. Through metabolomic analysis using gas chromatography (GC)-mass spectrometry (MS) and gene-expression analysis by reverse-transcription quantitative PCR (RT-qPCR), we observed that defense responses of Arabidopsis bam3 mutants are associated with starch-degradation enzymes, the corresponding modification of the carbohydrate balance, and alterations in sugar (glucose, sucrose, trehalose, and myo-inositol) and auxin metabolism.

Hierarchically Assembled Counter Electrode for Fiber Solar Cell Showing Record Power Conversion Efficiency

AbstractFiber solar cells have attracted significant interest as a promising wearable power supply solution for their merits of high flexibility, lightweight, and good compatibility with textile configuration and weaving process. However, because of the limited ion diffusion and charge transfer in the fiber counter electrode, the poor photovoltaic performances have long been one obstacle to hinder their real applications. Herein, in mimicking the efficient mass transport and exchange through the vascular tissue of plants like pine needle, a hierarchically assembled carbon nanotube (HCNT) fiber counter electrode is fabricated by a scalable process. The designed hierarchically aligned channels with large sizes of micrometers and small sizes of tens of nanometers in the HCNT fiber offers high‐flux pathways for rapid ion diffusion and abundant active area for charge transport, thus endowing the fiber dye‐sensitized solar cell with a record power conversion efficiency of 11.94%. By weaving such fiber solar cells in a scalable way, a flexible and breathable large photovoltaic textile (17 cm × 22 cm) is made to present a power output of 22.7 mW. These fiber solar cells are further integrated with fiber lithium‐ion batteries to efficiently power wearable electronics.

Effects of anthropogenic nitrogen additions and elevated CO2 on microbial community, carbon and nitrogen content in a replicated wetland.

Anthropogenic deposition of nitrogen (N) and elevated CO2 (eaCO2) are expected to increase continuously and rapidly in the near future and influence global carbon cycling. These parameters affect the ecosystem by regulating the microbial community and contribute to soil organic matter decomposition. The study was performed to understand the effects of N additions (4 and 6mgl-1) and eaCO2 (700ppm) on carbon (C)/nitrogen (N) content in the soil, microbial community, and plant biomass (Alternanthera philoxeroides species). The results showed that when the atmospheric CO2 concentration was raised, the total organic carbon (TOC) in the soil statistically increased (P < 0.05) by 4% and 3% under low and high N additions respectively, while the inorganic carbon content also increased by 1% and 3% (P > 0.05) under the same conditions. The increase in the soil TOC content was a result of the movement of carbon from water to the soil due to the presence of vascular tissues of plants in the water. The redundancy analysis (RDA) results revealed that the presence of plant species was responsible for the carbon content increment in the soil. The plant biomass content increased by 30.96% (P = 0.081) and 31.36%, (P = 0.002) under low and high N addition respectively due to the increment in atmospheric CO2. The nitrogen content in the plant species decreased (p > 0.05) by 8.62% and 6.25% at low and high N addition respectively when atmospheric CO2 was raised. This suggests that soil microbes competed with the plants for inorganic nitrogen in the soil and the microbes used up the inorganic nitrogen before it got to the plants. The gram-positive bacteria and fungi population decreased under high N addition and eaCO2 while gram-negative bacteria increased, suggesting that N additions and eaCO2 affected the microbial function and correlated with the nitrogen reduction in the soil. The results from this study serve as a guide to researchers and stakeholders in making policies with regard to the constant increasing CO2 concentration in the atmosphere.

Light regulates xylem cell differentiation via PIF in Arabidopsis.

The balance between cell proliferation and differentiation in the cambium defines the formation of plant vascular tissues. As cambium cells proliferate, subsets of daughter cells differentiate into xylem or phloem. TDIF-PXY/TDR signaling is central to this process. TDIF, encoded by CLE41 and CLE44, activates PXY/TDR receptors to maintain proliferative cambium. Light and water are necessary for photosynthesis; thus, vascular differentiation must occur upon light perception to facilitate the transport of water and minerals to the photosynthetic tissues. However, the molecular mechanism controlling vascular differentiation in response to light remains elusive. In this study we show that the accumulation of PIF transcription factors in the dark promotes TDIF signaling and inhibits vascular cell differentiation. On the contrary, PIF inactivation by light leads to a decay in TDIF activity, which induces vascular cell differentiation. Our study connects light to vascular differentiation and highlights the importance of this crosstalk to fine-tune water transport.

Ordovician-Devonian lichen canopies before evolution of woody trees

Devonian evolution of woodlands has been envisaged as a protracted increase in size of vascular plants, which can be reconstructed from fossil stumps and trunks. However, Late Silurian and Early Devonian nematophytes such as Prototaxites would have towered over land plants, including vascular plant trees, in the same fossil plant assemblage, until finally overtaken by vascular land plants during the Early Carboniferous. Nematophytes lack tissues of vascular plants, and some have spherical photobionts encircled and indented by hyphae, as in lichens. Nematophytes were not monolithic poles, but branched, and trunk spacing in paleosols is evidence that they formed closed canopies. Depths of root and hyphal bioturbation and pedogenic calcite precipitation in paleosols though time increase with greater height of trees. In addition to large non-vascular trunks and early land plants in Ordovician to Devonian paleosols, there also were extensive, nutrient-gathering, networks of glomeromycotan mycorrhizae. An Ordovician-Silurian “age of lichens”, when nematophytes were the tallest elements of terrestrial vegetation and soils were riddled with mycorrhizae, may have nurtured and sheltered Ordovician-Silurian land plants and then Devonian woody plants. Fungi preceded and facilitated the evolution of early land plants.

First report of Eutypella species associated with dying cotton (Gossypium hirsutum L.)

A rapid increase in the number of Gossypium hirsutum L. plants suddenly wilting and dying in a commercial cotton field in Central Queensland initiated this study. The aim was to characterise fungal species recovered from discoloured vascular tissue of dead plants and determine if they contributed to their death. Isolations were consistently dominated by one fungus based on culture morphology. Identification was established on sequences of the internal transcribed spacer (ITS) region of ribosomal DNA and revealed that the isolates all belong to Diatrypaceae, with high homology to Eutypella scoparia. Further analyses showed that there were two distinct Eutypella species present in the isolates, which are quite different from E. scoparia from the sequence dissimilarity due to the presence of ITS1 and ITS2 insertions. In diseased root samples, community profiling showed two operational taxonomic units (OTUs) related to E. scoparia were the most abundant fungi accounting for 45 to 99% of all sequences. Pathogenicity tests showed that a Eutypella isolate when inoculated into the stem of healthy G. hirsutum using two inoculation methods caused cankerous growth and necrosis of vascular tissue, typical of trunk disease. The fungus caused a red—brown streaking of the vascular tissue like that observed in diseased field plants. This study shows that the fungal isolates, which form a distinct group within the Eutypella, are associated with the root and stem of dying G. hirsutum and were the dominant fungi of diseased roots. This is the first known case of Eutypella affecting cotton worldwide and is considered an expansion of this genus’ host range.

Patterns of Seed-to-Seedling Transmission of Xanthomonas citri pv. malvacearum, the Causal Agent of Cotton Bacterial Blight.

Cotton bacterial blight (CBB), caused by Xanthomonas citri pv. malvacearum, was a major disease of cotton in the United States in the early part of the twentieth century. The reemergence of CBB revealed many gaps in our understanding of this important disease. In this study, we employed a wild-type (WT) field isolate of X. citri pv. malvacearum from Georgia (U.S.A.) to generate a nonpathogenic hrcV mutant lacking a functional type-III secretion system (T3SS-). We tagged the WT and T3SS- strains with an auto-bioluminescent Tn7 reporter and compared colonization patterns of CBB-susceptible and CBB-resistant cotton seedlings using macroscopic image analysis and bacterial load enumeration. WT and T3SS- X. citri pv. malvacearum strains colonized cotton cotyledons of CBB-resistant and CBB-susceptible cotton cultivars. However, X. citri pv. malvacearum populations were significantly higher in CBB-susceptible seedlings inoculated with the WT strain. Additionally, WT and T3SS- X. citri pv. malvacearum strains systemically colonized true leaves, although at different rates. Finally, we observed that seed-to-seedling transmission of X. citri pv. malvacearum may involve systemic spread through the vascular tissue of cotton plants. These findings yield novel insights into potential X. citri pv. malvacearum reservoirs for CBB outbreaks.

First Report of Wilt of European hornbeam (Carpinus betulus L.) Caused by Fusarium oxysporum in China.

European hornbeam (Carpinus betulus L.) has been used as an important ornamental species for urban landscaping since the Italian Renaissance (Rocchi et al. 2010). In May 2019, 15% of 3000 C. betulus trees with wilted leaves and root rot were observed in a field (about 26 ha) in Pizhou, Jiangsu Province, China. Internal discoloration of the stem began with brown to black discoloration of the vascular system and gradually spread to inward areas. Roots and stems from symptomatic plants were washed free of soil, surface sterilized with 0.8% NaOCl, rinsed three times in sterile H2O, and blotted dry with a paper towel. Small segments (0.5-cm-long) were cut from the discolored vascular tissues, and then put on potato dextrose agar (PDA) at 25°C in darkness. After 4 days, fungal colonies were observed on the PDA. Pure cultures were obtained by monosporic isolation, and 9 morphologically similar fungal isolates (EJ-1 to EJ-9) were obtained. All purified cultures were incubated on PDA at 25°C in darkness as the initial isolation. Colonies of the 9 isolates on PDA displayed entire margins and showed abundant pink aerial mycelia initially and turned to light violet with age. Microconidia were elliptical or oval in shape, 0 septate, (5.2-)8.7(-12.5) × (3.5-)3.6(-5.5) µm. Macroconidia were falciform, 0-4 septate, and straight to slightly curved with a notched foot cell, (17.1-)20.5(-28.4) × (3.8-)4.1(-4.6) µm. These morphological characteristics resemble Fusarium oxysporum (Leslie and Summerell 2006). Genomic DNA of each isolate was extracted from mycelia using a CTAB method (Mo¨ller et al. 1992). The RPB2, TEF1 and cmdA genes were amplified and sequenced with the primers 5f2/7c (Liu et al. 2000), EF-1Ha/EF-2Tb (Carbone and Kohn 1999) and Cal228F/CAL2Rd (Groenewald et al. 2013), respectively. The sequences were deposited in GenBank (Table 1). A maximum likelihood phylogenetic analysis based on RPB2, TEF1 and cmdA sequences using MEGA7 revealed that the isolates were placed in the F. oxysporum species complex with 98% bootstrap support. Based on the morphological and molecular characters, all 9 isolates were identified as F. oxysporum. A pathogenicity experiment was conducted using 30 2-year-old C. betulus seedlings potted in sterile peat, 27 for inoculation (3 replicate plants per isolate) and 3 for a negative control. The treated plants were planted in the peat mixed with 50 ml of a conidial suspension of each isolate respectively. The negative control was inoculated with sterilized water. Conidia were harvested from colonized plates of PDA using sterilized water and adjusted to a concentration of 1×107 conidia/ml. All 30 seedlings were incubated in a greenhouse at 25°C with a relative humidity of 80% and a 12-h photoperiod. The inoculated seedlings displayed wilt symptoms within 30 to 40 days, and eventually died within 75 to 85 days after inoculation. Control plants remained symptomless. F. oxysporum was successfully reisolated from the vascular tissues of symptomatic plants, and sequences of RPB2, TEF1 and cmdA of re-isolates matched those of the original isolates. No pathogen was isolated from the tissues of control plants. The experiment was repeat twice with the similar results, fulfilling Koch's postulates. F. oxysporum is an important soil-borne pathogen and can cause disease in many economic plants, such as yellowwood (Graney et al. 2016), hickory (Zhang et al. 2015) and larch (Rolim et al. 2020). To our knowledge, this is the first report of wilt on C. betulus caused by F. oxysporum in China.