Plant Death Research Articles

Living and decaying roots as regulators of soil aggregation and organic matter formation—from the rhizosphere to the detritusphere

In dryland ecosystems, typically characterized by sparse vegetation and nutrient scarcity, pioneer plants exert a critical role in the build-up of soil carbon (C). Continuous root-derived C inputs, including rhizodeposition and structural root litter, create hotspots of increased microbial activity and nutrient availability where biogeochemical processes, such as soil aggregation and the accumulation and stabilization of organic matter (OM), are promoted. Our study aims to disentangle the effects of root C inputs on soil aggregate formation, microbial community structures, and on the fate of OM—both before and after plant death, i.e., during the transition from rhizosphere to detritusphere. This was realized in a two-phase incubation approach, tracing the natural and undisturbed transition from growth to subsequent decomposition of a pioneer plant-root system (Helenium aromaticum) in a semi-arid topsoil and subsoil. We quantified water-stable aggregates, investigated the fate and composition of OM separated into particulate and mineral-associated OM fractions (POM and MAOM), and observed successional changes in the root-associated microbiome. Our results underscore the significance of roots as vectors for macroaggregation within the rhizosphere in both topsoil and subsoil, associated with a particularly strong increase in fungal abundance in the subsoil. In topsoil, we identified root legacy effects in the detritusphere, as root-induced macroaggregation persisted after plant death, a phenomenon not observed in subsoil. These root legacy effects were accompanied by a clear succession towards gram + bacteria, which appeared to outcompete fungi during root decomposition. The increased availability of decaying litter surfaces further facilitated the protection of particulate OM via the occlusion into aggregates. Overall, to gain a holistic understanding of plant-microbe-soil interactions, we emphasize the need for more studies that span over the full temporal dimension from living to dying plants in intact soil systems.

First Report of Exserohilum rostratum Causing Brown Spot on Spartina alterniflora in China.

Spartina alterniflora Loisel, a perennial grass, has become an invasive species in China's coastal wetlands (Zhang et al. 2018). In July 2021, brown spot symptoms were observed on S. alterniflora in a coastal wetland (21°45'48″N, 108°44'00″E) in Beihai City, Guangxi Province, China. The disease affected approximately 50% of the plants in the surveyed area (0.2 ha) and was also observed in other regions of Beihai. It caused brown lesions with a gray or whitish center on the leaves and stems of S. alterniflora. As the disease developed, it ultimately led to leaf shedding and plant death. To isolate the causal agent, 18 fragments (~ 5 mm) from six symptomatic plants (3 leaf pieces per plant) were surface sterilized with 1% NaOCl solution for 2 min and rinsed three times with sterilized water. Subsequently, the tissues were placed on potato dextrose agar (PDA) medium supplemented with chloramphenicol (0.1 g/liter) and incubated at 28°C for three days. The hyphal tips were transferred onto fresh PDA to obtain pure cultures. A total of 25 isolates were obtained, 20 of which shared similar morphologies, while the remaining five exhibited distinct morphological characteristics and were non-pathogenic to S. alterniflora. Three isolates (MC16.1.3, MC16.6.2, and MC16.8.3) were randomly selected from the 20 for further investigation. The colonies on PDA were flat with dense aerial mycelia. The colony margins were entire, light brown in the centre, white to grey at the margin; reverse dark brown in the centre, gray at the margin. Conidia were straight to slightly curved, light olive-brown to dark olive-brown, septate, measured 33.5 to 79.1 μm × 10.4 to 18.7 μm (average 52.9 × 14.4 μm, n = 100), with a distinctly protruding hilum swelled from the basal cell. For molecular identification, the genomic DNA was extracted from mycelium on PDA using the CTAB method (Guo et al. 2000). The internal transcribed spacer (ITS), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and translation elongation factor 1 alpha (TEF1-α) genes were amplified and sequenced with the primer pairs ITS1/ITS4 (White et al. 1990), GPD1/GPD2 (Berbee et al. 1999), and EF1-983/EF1-2218 (Rehner et al. 2005), respectively. A BLAST analysis revealed that the ITS (OR516787-9), GAPDH (OR523686-8), and TEF-α (OR523683-5) had 99.1 to 99.7% identity with those of E. rostratum strains BRIP 11417 (LT837836, LT882553, and LT896656) and CBS 128061 (KT265240, LT715900, and LT896658) (Hernández-Restrepo et al. 2018). Based on the concatenated sequences, a phylogenetic tree generated by PhyloSuite software (Zhang et al., 2020) through Bayesian inference (BI) and Maximum Likelihood (ML) methods placed the isolates within E. rostratum. These morphological characteristics and molecular analyses confirmed the pathogen as E. rostratum (Hernández-Restrepo et al. 2018; Kaboré et al. 2022). To confirm pathogenicity, a conidial water suspension (~ 1 × 106 conidia/ml) of each of the three strains was inoculated on nine healthy S. alterniflora plants that had been grown for six months. Control plants were treated with sterile water. All plants were then enclosed in plastic bags and incubated in a greenhouse at 28°C. Six days after inoculation, the plants exhibited symptoms similar to those observed in nature. The control plants developed no symptoms. These experiments were replicated three times with similar results. To fulfill Koch's postulates, E. rostratum was consistently re-isolated from symptomatic tissue and confirmed by morphology and sequencing, whereas no fungus was isolated from the control plants. In recent years, S. alterniflora has posed a serious threat to the indigenous biodiversity of wetland ecosystems (Zhang et al. 2018). To our knowledge, this is the first report of E. rostratum causing brown spot on S. alterniflora worldwide.

Assessment of Kweilingia divina as a potential augmentative biological control agent against the gramineous weed Microstegium vimineum

Microstegium vimineum (Trin.) A. Camus, commonly called stiltgrass, is an exotic and indigenous troublesome weed in the United States and eastern Asia, respectively. Between 2020 and 2022, a damaging disease was observed on M. vimineum plants at the foot of Daluo Mountain in Wenzhou City, China, with an incidence of 90 %. Typical rust disease symptoms were prominently visible during the early monsoon season (June to July), consisting of chlorotic spots with many orange or orange-yellow uredinia on the abaxial surfaces of the M. vimineum leaves. Based on morphological and molecular characteristics, the fungus was identified as Kweilingia divina (Syd.) Buriticá. This article describes the studies carried out on one strain of the fungus (WZ-2) to ascertain its potential as a biocontrol agent of the weed. Host specificity testing on 61 plant species from 15 families proved that 25 major crops and 36 weeds were insensitive to this fungus, with only two other gramineous weed species being slightly susceptible. However, urediniospores were not observed on these slightly infected plants. Further research is required to assess the pathogenicity and safety of various plant species to K. divina, especially bamboos, which have been identified as susceptible. Temperature and dew period experiments have shown that the fungus has a relatively wide tolerance for infection between 15 and 40 °C, and with only 4 h of dew exposure some uredinia appeared on artificially infected M. vimineum plants. A field trial proved that urediniospores from the strain WZ-2 could infect naturally occurring M. vimineum plants, causing severe disease symptoms and plant death. Thus, strain WZ-2 has great potential as an augmentative biological control agent against M. vimineum. K. divina is a heteroecious rust fungus that requires an alternate host to complete its life-cycle. What the alternative hosts are, and the impact of the alternate hosts on outbreaks of the disease requires further study.

Apoptotic chromatin condensation inducer in the nucleus: Genome-wide analysis in plants and expression profile during Cowpea Severe Mosaic Virus infection in Vigna unguiculata [L.] Walp

Apoptotic Chromatin Condensation Inducer in the Nucleus (ACIN1) is a scaffold protein that was first described as a complex component responsible for triggering apoptosis in human cells. In plants, ACIN1 participates in silencing of Flowering Locus C (FLC), involved in vernalization in Arabidopsis thaliana. Contrary to what has been observed for humans, there are no reports on ACIN1 linked to programmed cell death (PCD) in plants. Actually, the function of ACIN1 in plants is still poorly understood. In the present study, a genome-wide analysis of the ACIN1 gene family in plants identified 27 ACIN1 orthologs from 19 species belonging to 12 plant families. The phylogenetic relationships, physicochemical properties, gene structure, conserved motifs, promoter cis-elements, chromosomal localization, syntenic regions, and protein network were investigated. Altogether, these analyzes revealed highly conserved domains in the structure of the ACIN1 proteins, as well as putative metacaspase cleavage sites, which suggest that they play a conserved function probably associated with the programmed cell death in plants. For instance, differential expression pattern and modulation of ACIN1 were noticed after inoculation of cowpea with Cowpea severe mosaic virus (CPSMV). Therefore, this study was conducted to provide, for the first time, information on the evolutionary, structural, and functional characteristics of the ACIN1 gene family as an initial effort towards understanding the role of these proteins in studied plant development and stress responses.

Variation in symptoms and morphology of Fusarium spp. on shallot associated with basal plate rot disease in Brebes District, Central Java Province, Indonesia

Abstract. Marianah L, Nawangsih AA, Munif A, Giyanto, Tondok ET. 2024. Variation in symptoms and morphology of Fusarium spp. on shallot associated with basal plate rot disease in Brebes District, Central Java Province, Indonesia. Biodiversitas 25: 2198-2208. Basal plate rot disease is an important disease of shallots that causes losses in the field and storage. The disease is caused by Fusarium complex species, with different levels of virulence and host susceptibility. The six species of Fusarium spp. reported to cause basal plate rot disease of onion were F. oxysporum f.sp. cepae, F. proliferatum, F. redolens, F. solani, F. acutatum and F. tricinctum. Information about the variation in symptoms and morphology of fungi that cause basal plate rot disease on shallots, especially in Indonesia, is still very limited. Besides, the species that cause basal plate rot disease on shallots has not been molecularly confirmed using specific primers. The objective of this study was to isolate, identify, and study the variation in symptoms of the fungus associated with basal plate rot disease. The fungi were isolated from shallot plants symptomatic of basal plate rot disease collected from Brebes, Central Java Province. Eight isolates of Fusarium with different morphologies were successfully isolated and identified. Five isolates were identified as F. solani (isolate BC1, BC2, BC3, BBS1, and BBS4), two isolates as F. oxysporum f.sp. cepae (isolate BC4 and BBS6) and one isolate F. proliferatum (isolate BBS5). All eight isolates were pathogenic on shallots with different levels of virulence and symptom variations. BC4 isolate had the highest level of virulence, resulting in plant death, and was identified as F. oxysporum f.sp. cepae.

Identification of Available Nutrients NPK of Oil Palm Soil (Elaeis guineensis Jacq) in the Phase of Plants Not Yet Producing Fruit

Oil palms that experience nutrient deficiencies will affect their vegetative and generative growth and even cause plant death. This study aims to determine the chemical properties of soil in oil palm plantations at the plant phase before producing fruit. This study was conducted at an oil palm plantation owned by PTPN III Kebun Aek Nabara Utara, Labuhanbatu Regency, and analyzed at the Soil Laboratory of the Faculty of Agriculture, University of North Sumatra. The method used is the observation method, sampling in the field, and analysis in the laboratory to obtain quantitative data. Observation parameters include soil pH, C-organic, Total Nitrogen, Available P, Cation Exchange Capacity, and Base Saturation. The results of the study showed that the level of soil fertility at the research location was included in the low category. The main limiting factor that causes low soil fertility is the low organic matter content of the soil and to improve the soil fertility status at the research location so that the availability of nutrients for plants is more sufficient

First report of Diaporthe eres causing leaf spot on Rosa roxburghii Tratt in Guizhou Province, China.

Rosa roxburghii Tratt is a plant from the Rosaceae family whose fruits are rich in vitamins, dietary fiber, flavonoids, phenolic acids, and other active components (Jiang, et al. 2024). In July 2023, about R. roxburghii 500 plants were investigated in a field of 6000 m2 in Guiding County (107°14'E, 26°45'N), Guizhou province, China, and the results showed a leaf spot incidence of s 20 to 30%. . The affected leaves had irregular, black lesions with a clear blackish brown boundary and faint black conidiomata in a brown center. Fifteen symptomatic leaves were collected from 10 plants washed with sterile distilled water, and 5 × 5 mm pieces of the infected tissues were cut. After surface sterilization for30 s with 75% ethanol, 2 min with 3% NaOCl, three washes in sterilized distilled water, the leaf pieces were dried and placed on potato dextrose agar (PDA) and incubated at 25℃ for 5 days. Three isolates (H3-Y-1-1, H3-Y-1-2, H3-Y-1-3) with identical morphology were obtained, and the isolate H3-Y-1-1was selected for further study. The colonies on PDA exhibited irregular growth patterns, with white felty aerial mycelium on the upper surface, and white mycelium on the lower surface. Conidiomata were irregularly distributed over the agar surface. The isolate H3-Y-1-1 produced darkly pigmented pycnidia on PDA after 30 days and oozed milky mucilaginous drops. The fungus produced two types of conidia, α and β. Regular α conidia were 4.74 - 5.96 × 1.52 - 2.24 μm (n = 50), hyaline, elongated, biguttulate and non-septate. Beta conidia were 20.13 - 25.74 × 0.86 - 1.29 μm (n = 50), aseptate, hyaline, smooth, spindle shaped, slightly curved to bent. The morphological features were consistent with the description of Diaporthe eres (Pereira, et al. 2022). The pathogen was confirmed to be D. eres by amplification and sequencing of the internal transcribed spacer region (ITS), the partial β-tubulin (TUB), the partial translation elongation factor 1-alpha (TEF) genes using primers ITS1/ITS4, Bt-2a/Bt-2b, EF1-728F/EF1-986R, respectively. Sequences from PCR amplification were deposited in GenBank with accession numbers PP411998 (ITS), PP502153 (TUB), PP502156 (TEF). BLAST searches of the sequences revealed (96%) (500/523nt), 97% (479/494 nt) and 99% (334/338 nt) homology with those of D. eres CBS 138594 from GenBank (OM698848, OM752196 and OM752197), respectively. Phylogenetic analysis using maximum-likelihood and Bayesian methods placed the isolate H3-Y-1-1 in a well-supported cluster with D. eres CBS 101742. The pathogen was thus identified as D. eres based on the morphological characterization and molecular analyses (Feng, et al. 2013; Tao, et al. 2020). To assess its pathogenicity, healthy R. roxburghii potted plants were inoculated with H3-Y-1-1 spore suspensions. Symptomatic leaves mirroring field symptoms were observed after XX days of incubations at XX°C, while control plants exhibited no symptoms. Diaporthe eres was consistently reisolated from the infected leaves showing brown irregular or round lesions at the initial stage of the disease, expanding and become more irregular over time ultimately causing leaf curling and plant death. To our knowledge, this is the first report of leaf spot on R. roxburghii caused by D. eres in China. The disease may become a serious threat to fruit of R. roxburghii production in China. Therefore, detection of this pathogen is very important to ensure timely disease management.

Occurrence and molecular characterization of Diaporthe eres causing stem canker on sweet cherry trees (Prunus avium) in northern China

AbstractSweet cherry (Prunus avium) is a commercially important species in China, experiencing a rapid increase in production. In June 2022, a severe infection, affecting more than 8% of sweet cherry trees, was observed on the Tetian cultivar in a 2–3‐year‐old orchard in Dalian City, northern China. Initially, spindle‐shaped brown disease spots formed on the surfaces of the branches. These spots continued to spread and merge, and the middle portion of the cankers sunk inward and gradually dried. Small black particles were found on the surface of the stems. The disease spread was more prevalent during the rainy season, leading to the withering of numerous branches and the death of the whole plant. Seven isolates, named lncy1‐1, lncy4‐1, lncy5‐1, lncy6‐1, lncy7‐1, lncy8‐1 and lncy9‐1, were obtained from 24 disease samples. Phylogenetic analysis based on three loci (rDNA ITS, TEF1 and TUB2) coupled with morphological identification confirmed that these seven isolates belong to Diaporthe eres. A representative isolate, lncy1‐1, was inoculated onto sweet cherry branches in a controlled environment, and showed that the isolate lncy1‐1 was pathogenic. The fungus isolated from diseased tissues was identified as D. eres based on morphological and molecular criteria. To the best of our knowledge, this is the first report of stem canker in sweet cherry caused by D. eres in China, which will promote disease management and expand the known host range of D. eres.

Control biológico de Ralstonia solanacearum y su efecto en el crecimiento vegetativo de banano orgánico

The banana (Musa AAA) is one of the main economically important crops worldwide. Currently, it faces a serious problem of plant death caused by the bacterium Ralstonia solanacearum (Smith) variety 2. The objective of this research was to evaluate the effect of biological control of Moko disease and its relationship with the vegetative growth of banana seedlings under field conditions. Four treatments were employed: absolute control (T0), Trichoderma spp. (T2), Bacillus spp. (Bio-remedy) (T3), and ADMF® (T4) with three replications in a completely randomized block design. The incubation period, the incidence percentage, and the area under the disease progress curve (AUDPC) were evaluated, along with plant survival and height, pseudostem diameter, and leaf emission rate. At 113 days after transplanting (dat), treatments T1, T2, and T3 showed higher seedling survival. Plant height (111-145 cm), pseudostem diameter (7.43-11.28 cm), and leaf emission rate (11.74-13.15 leaves) exhibited significant differences among treatments. Treated plants showed the lowest AUDPC (between 576.1 and 1435.4 units) compared to untreated plants (3156.55 units). Trichoderma, Bacillus, and ADMF® were demonstrated to reduce disease incidence and promote banana vegetative growth, proving to be viable sustainable alternatives for this crop. Keywords: Musa AAA, Trichoderma, Bacillus, ADMF®, Ralstonia solanacearum, AUDPC.

Antagonism and Synergism Characterize the Interactions between Four North American Potato Virus Y Strains

Potato virus Y (PVY) is one of the most important constraints to potato production worldwide. There is an increasing occurrence of recombinant PVY strains PVYNTN and PVYN-Wi and a decline in the incidence of the nonrecombinant PVYO. We hypothesized that this may be due to the ability of these recombinant strains to antagonize and/or outcompete PVYO in mixed infections. To determine this, we investigated interactions between PVYO and three recombinant PVY strains common in North America: PVYNTN, PVYN-Wi, and PVYN:O. Overall, our study showed that these interactions are tissue-dependent. Specifically, PVYNTN, the main causal agent of potato tuber necrotic ringspot disease (PTNRD), was found to be more adaptable than PVYO, especially in potato leaves due, at least in part, to the Ny gene that confers hypersensitive resistance (HR) to PVYO. Furthermore, PVYN-Wi was found to repress PVYO in potato tubers but act synergistically in potato leaves. The PVYO-induced foliage necrosis in cultivar ‘Ranger Russet’ was observed to be more severe in plants co-infected by PVYN-Wi and PVYN:O, respectively, resulting in plant death. Strikingly, this PVYO -induced necrosis was suppressed by PVYNTN in doubly infected plants. These interactions may, at least partially, explain the decreasing incidence of PVYO in United States potato production regions, especially given that many cultivars contain the Ny gene, which likely limits PVYO enabling PVYNTN and PVYN-Wi to outcompete. We also found that replication and cell-to-cell movement of these PVY strains in tubers at 4 °C was similar to levels at ambient temperature.

Distinction of chia varieties in vivo and in vitro based on the flow cytometry and rosmarinic acid production

Flow cytometry has made a significant contribution to the study of several complex fundamental mechanisms in plant cytogenetics, becoming a useful analytical tool to understand several mechanisms and processes underlying plant growth, development, and function. In this study, the genome size, DNA ploidy level, and A-T/G-C ratio were measured for the first time for two genotypes of chia, Salvia hispanica, an herbaceous plant commonly used in phytotherapy and nutrition. This study also evaluated, for the first time by flow cytometry, the capacity to produce organic acids of tissues stained with LysoTracker Deep Red after elicitation with either yeast extract or cadmium chloride. Rosmarinic acid content differed between the two chia varieties treated with different elicitor concentrations, compared with non-elicited plant material. Elicited tissues of both varieties contained a higher content of rosmarinic acid compared with non-elicited cultures, and cadmium chloride at 500 μM was much better than that at 1000 μM, which led to plant death. For both genotypes, a dose-response was observed with yeast extract, as the higher the concentration of elicitor used, the higher rosmarinic acid content, resulting also in better results and a higher content of rosmarinic acid compared with cadmium chloride. This study demonstrates that flow cytometry may be used as a taxonomy tool, to distinguish among very close genotypses of a given species and, for the first time in plants, that this approach can also be put to profit for a characterization of the cytoplasmic acid phase and the concomitant production of secondary metabolites of interest in vitro, with or without elicitation.Key points• Genome size, ploidy level, A-T/G-C ratio, and cytoplasm acid phase of S. hispanica• Cytometry study of cytoplasm acid phase of LysoTracker Deep Red-stained plant cells• Yeast extract or cadmium chloride elicited rosmarinic acid production of chia tissuesGraphical

Effector Cs02526 from Ciboria shiraiana induces cell death and modulates plant immunity.

Sclerotinia disease is one of the most devastating fungal diseases worldwide, as it reduces the yields of many economically important crops. Pathogen-secreted effectors play crucial roles in infection processes. However, key effectors of Ciboria shiraiana, the pathogen primarily responsible for sclerotinia disease in mulberry (Morus spp.), remain poorly understood. In this study, we identified and functionally characterized the effector Cs02526 in C. shiraiana and found that Cs02526 could induce cell death in a variety of plants. Moreover, Cs02526-induced cell death was mediated by the central immune regulator brassinosteroid insensitive 1-associated receptor kinase 1, dependent on a 67-amino acid fragment. Notably, Cs02526 homologs were widely distributed in hemibiotrophic and necrotrophic phytopathogenic fungi, but the homologs failed to induce cell death in plants. Pretreatment of plants with recombinant Cs02526 protein enhanced resistance against both C. shiraiana and Sclerotinia sclerotiorum. Furthermore, the pathogenicity of C. shiraiana was diminished upon spraying plants with synthetic dsRNA-Cs02526. In conclusion, our findings highlight the cell death-inducing effector Cs02526 as a potential target for future biological control strategies against plant diseases.

Disease prevalence, incidence, morphological and molecular characterisation of Lasiodiplodia pseudotheobromae causing collar rot disease on peanut plants in Turkey

AbstractPeanut (Arachis hypogaea L.) holds significant commercial and dietary importance as a major source of edible oil and protein in Turkey. Stem, collar or root rot, caused by several fungal disease agent, are serious soil-borne diseases of peanut. Accurate and precise identification of the disease agent provides fundamental and precise information for integrated plant management. During the period from June to September 2021, symptoms consistent with collar rot disease, including dark-brown stem rot, chlorotic leaves, wilting, and eventual whole plant death, were observed on peanut plants cultivated in the different districts of Osmaniye Province of Turkey. The disease incidence ranged from 8.0 to 45.0% in the inspected fields with an average of 3.4% overall. Twenty-four single-spore representative isolates were obtained from surface-disinfected symptomatic tissues. Morphological characteristics of fungal mycelium, conidial and pycnidial structures on potato sucrose agar (PSA) and water agar (WA) closely resembled those described for Lasiodiplodia spp. All isolates caused typical collar rot symptoms upon artificial inoculation of peanut seedlings. Morphological identification of Lasiodiplodia spp. isolates was corroborated by MALDI-TOF and molecular analyses utilizing sequences from the internal transcribed spacer (ITS), β-tubulin 2 (tub2) and translation elongation factor-1 alpha (TEF1-α) loci. Phylogenetic analysis confirmed that the representative fungal isolates (MKUBK-B1 and MKUBK-K22) belong to Lasiodiplodia pseudotheobromae. To the best of our knowledge, this is the first report of L. pseudotheobromae infecting peanut plants in Turkey. This work is expected to contribute to previously limited knowledge regarding the host range, incidence and prevalence of L. pseudotheobromae as a soilborne pathogen of peanuts. Due to the potential destructiveness and broad host range of this pathogen, it is essential to develop new strategies to establish more reliable, environmentally sustainable, and cost-effective management approaches for this disease.

Identification of the Coexisting Virus-Derived siRNA in Maize and Rice Infected by Rice Black-Streaked Dwarf Virus.

Rice black-streaked dwarf virus is transmitted by small brown planthoppers, which causes maize rough dwarf disease and rice black-streaked dwarf disease. This virus leads to slow growth or death of the host plants. During the coevolutionary arms race between viruses and plants, virus-derived small interfering RNAs (vsiRNAs) challenge the plant's defense response and inhibit host immunity through the RNA silencing system. However, it is currently unknown if rice black-streaked dwarf virus can produce the same siRNAs to mediate the RNA silencing in different infected species. In this study, four small RNA libraries and four degradome libraries were constructed by extracting total RNAs from the leaves of the maize (Zea mays) inbred line B73 and japonica rice (Oryza sativa) variety Nipponbare exposed to feeding by viruliferous and nonviruliferous small brown planthoppers. We analyzed the characteristics of small RNAs and explored virus-derived siRNAs in small RNA libraries through high-throughput sequencing. On analyzing the characteristics of small RNA, we noted that the size distributions of small RNAs were mainly 24 nt (19.74 to 62.00%), whereas those of vsiRNAs were mostly 21 nt (41.06 to 41.87%) and 22 nt (39.72 to 42.26%). The 5'-terminal nucleotides of vsiRNAs tended to be adenine or uracil. Exploring the distribution of vsiRNA hot spots on the viral genome segments revealed that the frequency of hotspots in B73 was higher than those in Nipponbare. Meanwhile, hotspots in the S9 and S10 virus genome segments were distributed similarly in both hosts. In addition, the target genes of small RNA were explored by degradome sequencing. Analyses of the regulatory pathway of these target genes unveiled that viral infection affected the ribosome-related target genes in maize and the target genes in the metabolism and biosynthesis pathways in rice. Here, 562 and 703 vsiRNAs were separately obtained in maize and rice and 73 vsiRNAs named as coexisting vsiRNAs (co-vsiRNAs) were detected in both hosts. Stem-loop PCR and real-time quantitative PCR confirmed that co-vsiRNA 3.1 and co-vsiRNA 3.5, derived from genome segment S3, simultaneously play a role in maize and rice and inhibited host gene expression. The study revealed that rice black-streaked dwarf virus can produce the same siRNAs in different species and provides a new direction for developing new antiviral strategies.

Analysis of Salt Stress on Soil Microbial Community Composition and Its Correlation with Active Components in the Rhizosphere of Acanthopanax senticosus.

Salt stress can adversely affect plant seed germination, growth and development, and eventually lead to slow growth and even death of plants. The purpose of this study was to investigate the effects of different concentrations of NaCl and Na2SO4 stress on the physicochemical properties, enzyme activities, rhizosphere microbial community and seven active components (L-phenylalanine, Protocatechuic acid, Eleutheroside B, Chlorogenic acid, Caffeic acid, Eleutheroside E, Isofraxidin) of Acanthopanax senticosus rhizosphere soil. Statistical analysis was used to explore the correlation between the rhizosphere ecological factors of Acanthopanax senticosus and its active components. Compared with Acanthopanax senticosus under NaCl stress, Na2SO4 generally had a greater effect on Acanthopanax senticosus, which reduced the richness of fungi in rhizosphere soil and adversely affected the content of multiple active components. Pearson analysis showed that pH, organic matter, ammonium nitrogen, available phosphorus, available potassium, catalase and urease were significantly correlated with active components such as Caffeic acid and Isofraxidin. There were 11 known bacterial genera, 12 unknown bacterial genera, 9 known fungal genera and 1 unknown fungal genus significantly associated with the active ingredient. Salt stress had great changes in the physicochemical properties, enzyme activities and microorganisms of the rhizosphere soil of Acanthopanax senticosus. In conclusion, different types and concentrations of salts had different effects on Acanthopanax senticosus, and the active components of Acanthopanax senticosus were regulated by rhizosphere soil ecological factors.

First Report of Strawberry Root Rot Caused by Fusarium falciforme in China.

Root rot caused by Fusarium spp. is a destructive disease affecting agricultural regions worldwide. Strawberries (Fragaria × ananassa Duch.) are an economically important crop in China. In March 2023, root rot was observed in strawberries grown in Jinan, Shandong Province, China. Symptoms included leaf wilt, necrotic roots, and plant death (Figure 1). Four strawberry samples (two symptomatic and two asymptomatic) were collected from ~2-acre fields where the disease incidence rate ranged from 2 to 3%. Tissue pieces (5 mm × 5 mm × 5 mm) from two healthy and two diseased strawberry root tissues were surface-disinfected with 75% ethanol for 3 min, treated with 10% sodium hypochlorite for 5 min, and washed three times with sterile water. These pieces were cultured for 5 days at 28°C on potato dextrose agar (PDA) containing 200 mg/L timentin. Typical Fusarium spp. like growth was observed on plates with the two symptomatic samples. Two representative fungal isolates (CM1 and CM2) with similar morphological characteristics were purified using the single-spore method (Figure 1). CM1 showed an average growth rate of 5 mm/d in PDA and comprised of several white-to-cream aerial mycelia after 5 d. After cultivation in carnation leaf agar medium for 7 d, falciform macroconidia, with blunt apical cells and slightly hooked basal cells comprising 3 to 4 septa of varying sizes (20 to 39)×(3.6 to 6.7 μm) were observed (n=50) (Figure 1). The chlamydospores were spherical, terminal or intercalary, solitary or chain-forming, and 3.1-10.5 μm in diameter (Figure 1). The microconidia on PDA were (5.8 to 13.6)× (2.5 to 3.3) μm in size (n=50). These morphological characteristics are consistent with previous descriptions of the Fusarium solani species complex (FSSC). DNA was extracted using the CTAB method (Stenglein and Balatti 2006). The internal transcribed spacer (ITS), translation elongation factor 1-α gene (tef1), RNA polymerase II largest subunit (rpb1), and RNA polymerase II second largest subunit (rpb2) were amplified and sequenced using specific primers (O'Donnell et al. 2010). The ITS (OR526528, OR526529), tef1 (OR536947, OR536948), rpb1 (OR536949, OR536950), and rpb2 (OR536951, OR536952) sequences of the CM1 and CM2 isolates were uploaded to the NCBI database. BLASTn analysis revealed that the ITS, tef1, rpb1, and rpb2 sequences were 99.1-100% identical to those of the Fusarium falciforme reference strains NRRL 54989 and NRRL 54978. A phylogenetic tree based on the ITS, tef1, rpb1, and rpb2 sequences was generated using MEGA v.11 via the maximum-likelihood method (Tamura et al. 2021). CM1 clustered with the Fusarium falciforme reference strains NRRL 54989 and NRRL 54978 and belonged to the FSSC based on its morphological and molecular characteristics (Figure 2). To test for pathogenicity, the roots of nine 3-month-old healthy strawberry (cv. Akihime) plants were exposed to conidial suspensions (1×108 spores/mL) of the CM1 isolate. Another nine root samples were treated with sterile water and used as controls. All strawberry plants were maintained in a growth chamber under a 12/12 h light/dark cycle at 28°C and 90% relative humidity and the experiment was repeated three times. After one month, the inoculated plants had withered and died, and the pith became dark red (similar to field plants) (Figure 1). The fungi isolated from the experimental plants were confirmed as F. falciforme using morphological and sequence analyses. F. falciforme causes root rot in several species including Nicotiana tabacum (Qiu et al. 2023) and Weigela florida (Shen et al. 2020); however, this study is the first to report root rot caused by F. falciforme in strawberries in China. Overall, F. falciforme infection poses a threat to strawberry production and breeding.

A new canker on Gleditsia sinensis caused by Thyronectria austroamericana in China

Gleditsia sinensis is a medicinal plant that brings an annual economic value of nearly 400 million to Song County, Henan Province, China. From 2021 to 2022, a canker disease was observed on the branches of G. sinensis. The diseased plants often displayed symptoms of wilting, yellowing, early leaf drop and total plant death. The incidence of canker on G. sinensis was estimated at 20%–40% for the years 2021, 2022 and 2023, with 50% of the infected plants ultimately perishing. The putative pathogen was isolated from symptomatic G. sinensis branches. Thyronectria austroamericana was identified as the causal agent by using morphology and molecular phylogeny of multi-loci (alpha-actin (act), internal transcribed spacer (ITS) region, large nuclear ribosomal RNA subunit (LSU), translation elongation factor 1-alpha (tef1), and β-tubulin (tub2) sequence data. In vitro and field inoculation experiments showed that the fungus could produce canker symptoms in G. sinensis which is similar to those observed in the field. The putative pathogen was re-isolated and Koch's postulates were fulfilled. This study provides the first report of T. austroamericana causing G. sinensis canker disease in China. Furthermore, these results will contribute to the knowledge of the G. sinensis canker disease.

First Report of Enterobacter mori Causing Bacterial Wilt on Tomato in China.

Tomato (Solanum lycopersicum L.) is one of the most important vegetable crops in China. In October 2023, a new bacterial disease was discovered on tomato plants in a 0.3-acre farm's greenhouse (35.514806N, 118.996106E) in Longshan Town, Shandong Province, China. Over 50% of the tomato plants showed symptoms of stem rot, leaf wilt, or plant death. Three diseased tomato plants were collected for pathogen isolation and purification. Two leaf samples, each about 1 cm2, were cut from the junction area of healthy and diseased parts and disinfected with 75% alcohol for 60 s, followed by 0.1% HgCl2 for 90 s, and then washed three times with sterilized H2O. The samples were subsequently ground with 1.0 mL sterilized H2O. The ground samples were diluted to 10-4, 10-5, and 10-6 and then were plated on a potato dextrose agar (PDA) plate, respectively. White mucous bacterial colonies appeared at 28℃ for 24~48 h, no fungal colony was observed. Six bacterial colonies were randomly selected for gram staining and found to be gram-negative. To further determine their species classification, fragments of the 16SrDNA, hsp60, gyrB, and rpoB genes were separately amplified using previously reported PCR conditions and with primer pairs, including 27F/1492R (Wu et al., 2023), HSP60-F /HSP60-R (Gül et al., 2023), gyrB UP-1 / gyrB UP-2r (Yamamoto et al., 1995) and rpoB CM81-F / rpoB CM32b-R (Brady et al., 2008). Sequence analysis showed that the obtained sequences of the 16SrDNA, hsp60, gyrB, and rpoB genes among these six colonies were identical and 100%, 100%, 99.31%, and 99.36% similar to those of Enterobacter mori accessions OP601841 (with a coverage of 100%), MT199160 (83%), OP676246 (100%), and MN594495 (100%), at nucleotide level, respectively. Sequences of the above four genes of 23LSFQ were submitted to GenBank under the accession numbers PP461247, PP474090, PP136037, and PP136038, respectively. We selected one of these six colonies, 23LSFQ, for further analysis. The phylogenetic tree based on the concatenated sequences of the above four genes using the maximum likelihood method with MEGAX software showed that 23LSFQ is grouped with E. mori LMG25706 (NCBI: txid980518). To determine the pathogenicity of 23LSFQ , we sprayed 23LSFQ (OD600=0.8) onto five 30-day-old healthy plants of the tomato cultivars Alisa Craig, Jinpeng NO.1, and Chaobei, respectively. These seedlings were incubated in a chamber at 28°C with a 16 h light/ 8h dark photoperiod and 60% relative humidity. The leaves of the inoculated plants became curled and wilted at two days post inoculation (dpi) and appeared necrotic at 10 dpi. The symptoms were similar to those observed in field-infected tomato plants. No symptoms were observed on the plants inoculated with water. We further sequenced the re-isolated bacteria from the symptomatic inoculated seedlings. Results showed that they belong to E. mori. The experiment was repeated three times. E. mori has been found to cause diseases on peaches (Ahmad et al., 2021), watermelons (Wu et al., 2023), Canna indica, (Zhang et al., 2023), and strawberries (Ji et al., 2023). E. cloacae has been found to cause diseases on tomatoes in Heilongjiang province (Jin et al., 2023). This is the first report of E. mori causing leaf yellowing and wilting on tomatoes in China. These results are significant for the safe production and disease control of greenhouse tomatoes.

Banana bunchy top virus movement protein induces resistance in banana against Fusarium wilt

Banana Fusarium wilt, which is known as Panama disease and caused by Fusarium oxysporum f. sp. cubense (Foc), is a destructive disease that can lead to plant death and complete loss of banana plantations. For obtaining resistant varieties against Fusarium wilt, research is required to elucidate the mechanisms of genetic resistance to Foc. In this research, the banana plants infected with banana bunchy top virus (BBTV) showed resistance to Foc. RNA-seq results showed that the infection with Foc upregulated many resistance-related genes in BBTV-positive plants, especially genes related to ROS production. It was also found that more H2O2 in BBTV-positive plants was induced during Foc infection. The movement protein (MP) of BBTV could increase H2O2 levels by promoting the accumulation of MaSGT1a (suppressor of the G-two allele of SKP1) via direct interaction. Thus, the resistance to Foc induced by BBTV may attribute to MP increasing MaSGT1a protein levels to induce ROS production. MaSGT1a could promote the accumulation of R proteins MaRPM1 and MaRPP8, and increase H2O2 levels in Nicotiana benthamiana. The expression of R proteins in Nicotiana benthamiana caused an increase in H2O2 levels and promoted resistance to Foc. Thus, the MP of BBTV could induce plant defense through interaction with MaSGT1a. We speculate that MaSGT1a is likely involved in R gene-mediated resistance and thereby promotes the resistance of banana against Foc.

Transcription factor CmHSFA4-CmMYBS3 complex enhances salt tolerance in chrysanthemum by repressing CmMYB121 expression.

Excessive soil salinity not only hampers plant growth and development but can also lead to plant death. Previously, we found that heat-shock factor A4 (CmHSFA4) enhances the tolerance of chrysanthemum (Chrysanthemum morifolium) to salt. However, the underlying molecular mechanism remains unclear. In this study, we identified a candidate MYB transcription factor, CmMYB121, which responded to salt stress. We observed that the CmMYB121 transcription is suppressed by CmHSFA4. Moreover, overexpression of CmMYB121 exacerbated chrysanthemum sensitivity to salt stress. CmHSFA4 directly bound to the promoter of CmMYB121 at the heat-shock element. Protein-protein interaction assays identified an interaction between CmHSFA4 and CmMYBS3, a transcriptional repressor, and recruited the corepressor TOPLESS (CmTPL) to inhibit CmMYB121 transcription by impairing the H3 and H4 histone acetylation levels of CmMYB121. Our study demonstrated that a CmHSFA4-CmMYBS3-CmTPL complex modulates CmMYB121 expression, consequently regulating the tolerance of chrysanthemum to salt. The findings shed light on the responses of plants to salt stress.