Analysis Of ITS Sequences Research Articles

Tubaria asiatica (Tubariaceae), a new species from Khyber Pakhtunkhwa, Pakistan

A new Tubaria (Tubariaceae) species, Tubaria asiatica is described and illustrated from district Swabi, Khyber Pakhtunkhwa Pakistan, based on morphological and phylogenetic analysis of ITS and 28S sequences. The most closely related species is Tubaria conspersa, but it differs from it due to its pale orange to orange‐tinged dark brown pileus adnate lamellae, dark brown–orange, cylindrical to subcylindrical stipe, oblong, ellipsoid, lacrymoid, rhomboid basidiospores, clavate cheilocystidia and basidia with long sterigmata. Morphological descriptions, colored photographs, microscopic structures and a comparative combined ITS‐28S‐based phylogenetic analysis is presented with reference to the newly described species.

Three new species of Talaromyces sect. Talaromyces discovered in China.

Talaromyces species play an important role in the nutrient cycle in natural ecosystems, degradation of vegetal biomass in industries and the implications in medicine. However, the species diversity of this genus is still far from fully understood. The polyphasic taxonomic approach integrating morphological comparisons and molecular phylogenetic analyses based on BenA, CaM, Rpb2 and ITS sequences was used to propose three new Talaromyces species. Three new species of sect. Talaromyces isolated from soil are proposed, namely, T. disparis (ex-type AS3.26221), T. funiformis (ex-type AS3.26220) and T. jianfengicus (ex-type AS3.26253). T. disparis is unique in low growth rate, velvety texture, limited to moderate sporulation, biverticillate, monoverticillate and irregular penicilli bearing a portion of abnormally large globose conidia, it has no close relatives in phylogeny. Being a member of T. pinophilus complex, T. funiformis produces mycelial funicles on Czapek yeast autolysate agar (CYA), 5% malt extract agar (MEA) and yeast extract (YES), sparse sporulation on Czapek agar (Cz), CYA, MEA and YES while abundant on oatmeal agar (OA), bearing appressed biverticillate penicilli and globose to pyriform conida with smooth to finely rough walls. T. jianfengicus belongs to T. verruculosus complex, is characterized by velvety colony texture with moderate to abundant elm-green conidia en masse, producing biverticillate penicilli, globose conidia with verrucose walls. It is now a common practice in establishing new species of Aspergillus, Penicillium and Talaromyces based on morphological characters and phylogenetic analyses of BenA, CaM, Rpb2 and ITS sequences. The proposal of the three novelties of Talaromyces in this article is not only supported by their morphological distinctiveness, but also confirmed by the phylogenetic analyses of the concatenated BenA-CaM-Rpb2 and BenA-CaM-ITS, as well as the individual BenA, CaM, Rpb2 and ITS sequence matrices.

Environmental Factors Shaping the Culturable Freshwater Fungi Diversity of Four Lakes in Yunnan Province, China

Our study focused on freshwater fungal diversity, an important aspect in assessing the ecology of aquatic ecosystems. We carefully explored the diversity and influencing factors of culturable fungi across Dianchi Lake, Fuxian Lake, Xingyun Lake, and Yangzonghai Lake in Yunnan Province, China. Through fungi culture, morphological characterization, and ITS sequence analysis, we identified a total of 565 isolates belonging to 405 species across 133 genera. The diversity indices viz. H′, D, and J were evaluated for fungal diversity across the lakes. Interestingly, although diversity indices were highest during summer and at Yangzonghai Lake, no significant differences in fungal community diversity were observed between seasons and regions. Water variables were analyzed and indicated that changes with rapid fluctuations in temperature, pH, and dissolved oxygen likely influence fungal diversity. These findings significantly contribute to our understanding of fungal communities within plateau lake ecosystems, thereby aiding in managing and conserving vital aquatic resources.

Cover crop alters rhizosphere sediments to recruit plant growth-promoting microorganisms, enhancing peanut production

Peanuts are typically grown in continuous monoculture in China, leading to continuous cropping obstacles and reduced yields. Cover cropping is emerging as a pivotal strategy for enhancing soil quality and promoting sustainable agriculture. However, the effects of cover crops on the rhizosphere environment, and how they benefit subsequent crops, are not well understood. We hypothesize that planting cover crop ryegrass during the winter fallow period could alter peanut rhizosphere sediments, recruit plant growth-promoting microorganisms, and ultimately enhance peanut production. This study aims to fill this gap by investigating the impact of ryegrass cover cropping on rhizosphere sediments and peanut yield. Rhizosphere soil organic carbon, total nitrogen, pH, and soil enzymes, including sucrase, urease, N-acetyl-β-D-glucosaminidase, and β-glucosidase, were analyzed, along with metabolomics, 16S rRNA, and fungal ITS sequence analyses, under traditional planting management (TP) and long-term cover crop (CC) treatments. The results showed that CC treatment significantly increased rhizosphere pH, enhanced soil organic carbon and total nitrogen levels, and elevated soil enzyme activities compared to TP treatment. Metabolomic analysis revealed that CC treatment upregulated compounds such as D-pinitol, palmitoleic acid, fructose, sorbitol, and sucrose, while downregulated compounds including benzoic acid, dodecanoic acid, and carbamic acid. This alteration in metabolite composition influenced the recruitment and function of the microbial community. Specifically, CC treatment markedly enhanced the abundance of growth-promoting microbes such as Bacillus, Paenibacillus, Pseudomonas, Lysobacter, Bradyrhizobium, and Mortierell etc., which are involved in important functions such as chemoheterotrophy, nitrate reduction, and plant saprotroph. Simultaneously, there was a decrease in pathogenic microorganisms, such as Aspergillus flavus and Fusarium oxysporum. Moreover, CC treatment positively influenced peanut root growth, resulting in longer roots and ultimately increase pod yield by 21.15 % compared to traditional winter fallow practices. These findings underscore the potential of cover cropping with ryegrass to improve rhizosphere microecosystem components, and ultimately enhance peanut productivity, providing valuable insights for sustainable agricultural practices.

The genus Peniophora (Russulales, Basidiomycota) from Patagonia revisited

The identity of Peniophora specimens from Patagonia is examined based on morphological, cultural, and biological features as well as phylogenetic analyses of nuclear ribosomal ITS and 28S sequences. Three new species are described from Patagonia: Peniophora patagonica, P. pitrae, and P. revoluta. In total, five species are known in the area, growing on endemic trees in Southern Argentina. Peniophora cinerea and P. rufomarginata are excluded from the Patagonian records, and the presence of P. incarnata and P. lycii is confirmed by molecular methods. A key to Peniophora species from Patagonia is presented.

Fruiting Body Heterogeneity, Dimorphism and Haustorium-like Structure of Naematelia aurantialba (Jin Er Mushroom).

Mushroom Jin Er has attracted widespread attention in Asia over the past two decades due to its medicinal properties and nutritional values. In the present study, Jin Er basidiocarps were often found to be surrounded by Stereum hirsutum fruiting bodies in their natural habitat and occasionally in artificial cultivation. The observation of two different kinds of mycelia within the hymenium and analyses of ITS sequences confirmed that Jin Er basidiocarps were composed of two fungal species, Naematelia aurantialba and S. hirsutum. This heterogeneity of Jin Er fruiting bodies is indeed distinct from the homogeneous hypha of Tremella fuciformis found in Yin Er mushroom, although its development also requires the presence of another fungus Annulohypoxylon stygium. Basidiospores can germinate on the surface of basidiocarps and produce mycelia. However, basidiospores in PDA medium can only bud into yeast-like conidia. The yeast-like conidia of N. aurantialba can transform into pseudohyphae with a change in temperature from 20 °C to 28 °C or switch into filamentous cells on an induction medium (IDM) at 20 °C, 25 °C and 28 °C. This dimorphic was reported for the first time in N. aurantialba. Haustorium-like structures were abundantly observed both within the hymenium and in the aerial mycelia cultured on the IDM. The developmental process was documented firstly in this study, involving the formation of protuberances with basal clamp connections, elongation at the protuberances, branch production, and eventual maturation. However, further observation is required to determine whether the haustorium-like structures can penetrate S. hirsutum hyphae. These findings are expected to provide valuable insights into the relationship and interaction between these two fungi, thereby advancing the cultivation of fruiting bodies.

Fungal endophytes Fusarium solani SGGF14 and Alternaria tenuissima SGGF21 enhance the glycyrrhizin production by modulating its key biosynthetic genes in licorice (Glycyrrhiza glabra L.).

To identify promising fungal endophytes that are able to produce glycyrrhizin and enhance it in licorice and the mechanisms involved. Fifteen fungal endophytes were isolated from Glycyrrhiza glabra L. rhizomes among which SGGF14 and SGGF21 isolates were found to produce glycyrrhizin by 4.29 and 2.58µg g-1 dry weight in the first generation of their culture. These isolates were identified as Fusarium solani and Alternaria tenuissima, respectively, based on morphological characteristics and sequence analysis of internal transcribed spacer, TEF1, ATPase, and CAL regions. Subsequently, G. glabra plants were inoculated with these fungal isolates to examine their effect on glycyrrhizin production, plant growth parameters and the expression of key genes involved in glycyrrhizin pathway: SQS1, SQS2, bAS, CAS, LUS, CYP88D6, and CYP72A154. Endophytes were able to enhance glycyrrhizin content by 133%-171% in the plants. Natural control (NC) plants, harboring all natural endophytes, had better growth compared to SGGF14- and SGGF21-inoculated and endophyte-free (EF) plants. Expression of SQS1, SQS2, CYP88D6, and CYP72A154 was upregulated by inoculation with endophytes. LUS and CAS were downregulated after endophyte inoculation. Expression of bAS was higher in SGGF21-inoculated plants when compared with NC, EF, and SGGF14-inoculated plants. Two selected fungal endophytes of G. glabra can produce glycyrrhizin and enhance glycyrrhizin content in planta by modulating the expression of key genes in glycyrrhizin biosynthetic pathway.

Quantification using a HPLC-FLD-based Detector and Molecular Identifi-cation of Ochratoxin-A Producing Aspergillus Species Isolated from Stored Grain Samples.

Quantification using an HPLC-FLD based detector and Molecular identification of Ochratoxin-A producing Aspergillus Species isolated from stored grain samples. Fungi are cosmopolitan in origin and are known to grow in any suitable substra-tum. In the present investigation, Aspergillus species isolated from stored grain samples were analyzed for ochratoxin-A production. The objective of this study is the quantification of Ochratoxin and identification of ochratoxigenic fungi. A total of n=34 black Aspergilli and n=1 Ochre Aspergilli were isolated from grain samples of Bihar, India and it was tested for OTA production. The limit of detection (LOD) is found to be 0.33μg/Kg and the limit of quantification (LOQ) is found to be 1μg/Kg for OTA in HPLC-FLD. In the present study, out of all the fungal isolates, only TiB fungal isolate was able to produce the ochratoxin-A above the level of LOQ. The positive isolate TiB obtained from stored sesame seed samples was able to produce 25.54 μg/Kg of OTA. ITS sequence analysis of TiB isolate was able to matche 100% with Aspergillus welwitschiae and Aspergillus foetidus. This is the initial report of ochratoxigenic Aspergillus fungal isolate isolated from stored sesame seed samples of flood-prone areas of Bihar, India.

Neoscytalidium dimidiatum causing stem canker on Hylocereus megalanthus in China.

Hylocereus megalanthus (syn. Selenecereus megalanthus), commonly known as Yanwo fruit (bird's nest fruit), is an important tropical fruit, which is popular and widely planted due to its high nutritional and economic value in southern China. In September 2022, a serious stem and fruit canker was observed on Ecuadorian variety of Yanwo fruit plant in a 0.2 ha orchard in Guangdong (N21°19'1.24" E110°7'28.49"). Almost all plants were infected and disease incidence of fruits and stems was about 80% and 90% respectively. Symptoms on the stem and fruits were small, circular or irregular, sunken, orangish brown spots that developed into cankers (Fig 1 A, B and C). Black pycnidia were embedded under the surface of the cankers at the initial stage, subsequently they became erumpent from the surface, and the infected parts rotted. Five symptomatic stems from five plants were collected, 0.2 cm2 tissues adjacent to cankers were surface sterilized and placed on potato dextrose agar (PDA) to incubate at 25 to 28 ℃. Fungal isolates each with similar morphology grew from 100% of the tissues. Colonies covered with aerial mycelium were grayish white, and then gradually turned to grayish black. Septate hyphae were hyaline to brown and constricted into arthroconidial chains. The arthroconidia were variously shaped and colored, orbicular to rectangular, hyaline to dark brown, thick-walled, and zero- to one- septate, averaging 7.7 × 3.6 μm (n>50) (Fig 1 D, E, F and G). To identify the fungus, the internal transcribed spacer region (ITS), translation elongation factor 1-alpha (tef1), beta-tubulin (tub2), histone H3 (his3) and chitin synthase (chs) gene of isolate ACCC 35488 and ACCC 35489 (Agricultural Culture Collection of China) were amplified and sequenced with primer pairs: ITS1/ITS4 (White et al. 1990), EF1-728F/EF2-rd (Carbone & Kohn 1999; O'Donnell et al.1998), TUB2Fd/ TUB4Rd（Aveskamp et al 2009）, CYLH3F/H3-1b (Crous et al. 2004) and CHS-79F/CHS-345R (Carbone & Kohn 1999) (ITS: OQ381102 and PP488350; tef1: OQ408545 and PP510454; tub2: OQ408546 and PP510455; his3: OQ408544 and PP510453; chs: OQ408543 and PP510452). Sequence Blastn results showed above 99% identical with those of Neoscytalidium dimidiatum ex-type strain CPC38666. Phylogenetic tree inferred from Maximum Likelihood analysis of the combined ITS, tub2 and tef1 sequences revealed two isolates clustered with N. dimidiatum (Fig 2). Pathogenicity was tested on healthy one-year-old cuttings and fruits of Ecuadorian variety at room temperature. Six sites were pin-pricked on each stem and fruit. Both wounded stems and fruits were inoculated with spore suspensions (106 spore/ml) and 6-mm fungal plugs respectively. Sterile water and agar were used as control. The test was repeated twice. Stems and fruits were enclosed in plastic boxes with 80% relative humidity. Symptoms described above were observed on inoculated stems and fruits at five days post inoculation (Fig 1 H and I). No symptoms developed on the controls. Neoscytaliudium dimidiatum was reisolated from the cankers with a frequency of 100% via morphological and molecular analysis. This is first report of stem and fruit canker caused by N. dimidiatum on H. megalanthus in China and this disease represents a serious risk of Yanwo fruit yield losses. This fungus is widespread occurring throughout the world causing diseases on a wide variety of plants. The finding will be helpful for its prevention and control.

Characterization of Fungal Species Isolated from Cankered Apple Barks Demonstrates the Alternaria alternata Causing Apple Canker Disease.

Apple canker disease, also named as apple Valsa canker, is one of the most destructive diseases for apples (Malus domestica Borkh.). Cytospora/Valsa spp. are the dominant causal agent of this disease, but many studies have revealed that fungi from some other genus can also cause typical apple canker symptoms. In this study, we performed fungal pathogen isolation from cankered 'Fuji' apple barks. Six representative morphologically different fungi (Strain 1-6) were further subjected to ITS sequencing and evolutionary analysis. Molecular identification results revealed that Strains 1-6 are Cytospora mali, Fusarium cf. solani, Alternaria alternata, C. mali, Diplodia seriata and F. proliferatum, respectively. All these fungi have been reported to be causal agents of apple diseases. By inoculating fungal plugs onto trunks of 'Fuji' apple trees, the pathogenicity of the six fungi were accessed. Only the inoculations of the two C. mali strains (Strain 1 and Strain 4) and the A. alternata strain (Strain 3) resulted in typical apple canker symptoms in trunks. It is worth noting that Strain 1 caused much more severe canker symptoms and higher pathogenicity incidence than the other two fungi. A. alternata has been identified as a pathogen causing diseases on apple fruits and leaves. By further assessing its pathogenicity on apple fruits and leaves, we verified that it can also cause typical fruit rot and leaf spot symptoms. To the best of our knowledge, this is the first report on apple canker disease caused by A. alternata in China. Our present study can provide a theoretical foundation for the prevention and control of apple canker disease.

Evaluating the efficacy of probiotic bacterial strain Lactobacillus plantarum for inhibition of fungal strains associated with historical manuscript deterioration: An experimental study

The aim of this study is to develop safe biological methods for controlling fungal deterioration of historical manuscripts. Therefore, fifteen fungal isolates were obtained from paper sheets and leather skins of a deteriorated historical manuscript (dated back to the 13th century). Those isolates were identified using both traditional methods and ITS-sequencing analysis. Aspergillus niger accounted for seven strains, Penicillium citrinum for one strain, Aspergillus flavus for three, Aspergillus fumigatus for one, Aspergillus nidulans for one, and Penicillium chrysogenum for two of the fungal strains that were obtained. The ability of fungal strains for the secretion of cellulase, amylase, gelatinase, and pectinase as hydrolytic enzymes was evaluated. The capability of the probiotic-bacterial strain Lactobacillus plantarum DSM 20174 for inhibition of fungal strains that cause severe deterioration was studied using ethyl acetate-extract. The metabolic profile of the ethyl acetate-extract showed the presence of both high- and low-molecular-weight active compounds as revealed by GC-MS analysis. The safe dose to prevent fungal growth was determined by testing the ethyl acetate extract's biocompatibility against Wi38 and HFB4 as normal cell lines. The extract was found to have a concentration-dependent cytotoxic impact on Wi38 and HFB4, with IC50 values of 416 ± 4.5 and 349.7 ± 5.9 μg mL−1, respectively. It was suggested that 100 μg mL−1 as a safe concentration could be used for paper preservation. Whatman filter paper treated with ethyl acetate extract was used to cultivate the fungal strain Penicillium citrinum AX2. According to data analysis, fungal inhibition measurement, SEM, ATR-FT-IR, XRD, color change measurement, and mechanical property assessment, the recommended concentration of ethyl acetate extract was adequate to protect paper inoculated with the highest enzymatic producer fungi, P. citrinum AX2.

Genotypic Identification of Trees Using DNA Barcodes and Microbiome Analysis of Rhizosphere Microbial Communities.

DNA barcodes can provide accurate identification of plants. We used previously reported DNA primers targeting the internal transcribed spacer (ITS1) region of the nuclear ribosomal cistron, internal transcribed spacer (ITS2), and chloroplast trnL (UAA) intron to identify four trees at Bergen Community College. Two of the four trees were identified as Acer rubrum and Fagus sylvatica. However, Quercus was only identified at the genus level, and the fourth tree did not show similar identification between barcodes. Next-generation sequencing of 16S rRNA genes showed that the predominant bacterial communities in the rhizosphere mainly consisted of the Pseudomonadota, Actinomycetota, Bacteroidota, and Acidobacteriota. A. rubrum showed the most diverse bacterial community while F. sylvatica was less diverse. The genus Rhodoplanes showed the highest relative bacterial abundance in all trees. Fungal ITS sequence analysis demonstrated that the communities predominantly consisted of the Ascomycota and Basidiomycota. Quercus showed the highest fungi diversity while F. sylvatica showed the lowest. Russula showed the highest abundance of fungi genera. Average similarity values in the rhizosphere for fungi communities at the phylum level were higher than for bacteria. However, at the genus level, bacterial communities showed higher similarities than fungi. Similarity values decreased at lower taxonomical levels for both bacteria and fungi, indicating each tree has selected for specific bacterial and fungal communities. This study confirmed the distinctiveness of the microbial communities in the rhizosphere of each tree and their importance in sustaining and supporting viability and growth but also demonstrating the limitations of DNA barcoding with the primers used in this study to identify genus and species for some of the trees. The optimization of DNA barcoding will require additional DNA sequences to enhance the resolution and identification of trees at the study site.

Transferring Peucedanum macilentum (Apioideae, Apiaceae) to Ligusticopsis based on morphological and molecular evidences

Peucedanum macilentum is a less-studied species endemic to China, previously assigned to the genus Kitagawia. To elucidate its taxonomic affinity, morphological and molecular analyses were conducted. Morphologically, Peucedanum macilentum resembles Ligusticopsis species in several diagnostic characters of the stem base clothed in fibrous sheaths, pinnate bracteoles, conspicuous calyx teeth, strongly dorsally compressed fruits, as well as more vittae on the commissure. Phylogenetic analyses of nrDNA ITS and plastome sequences further reveal that P. macilentum is distant from Kitagawia but nested within Ligusticopsis. Therefore, in the basis of the morphological and molecular evidences, Peucedanum macilentum is transferred into Ligusticopsis and a new combination, Ligusticopsis macilenta, is proposed.

Identification of common human infectious and potentially zoonotic novel genotypes of Enterocytozoon bieneusi in cavernicolous bats in Thailand.

Enterocytozoon bieneusi is a common cause of human microsporidiosis and can infect a variety of animal hosts worldwide. In Thailand, previous studies have shown that this parasite is common in domestic animals. However, information on the prevalence and genotypes of this parasite in other synanthropic wildlife, including bats, remains limited. Several pathogens have been previously detected in bats, suggesting that bats may serve as a reservoir for this parasite. In this study, a total of 105 bat guano samples were collected from six different sites throughout Thailand. Of these, 16 from Chonburi (eastern), Ratchaburi (western), and Chiang Rai (northern) provinces tested positive for E. bieneusi, representing an overall prevalence of 15.2%. Based on ITS1 sequence analysis, 12 genotypes were identified, including two known genotypes (D and type IV) frequently detected in humans and ten novel potentially zoonotic genotypes (TBAT01-TBAT10), all belonging to zoonotic group 1. Lyle's flying fox (Pteropus lylei), commonly found in Southeast Asia, was identified as the host in one sample that was also positive for E. bieneusi. Network analysis of E. bieneusi sequences detected in this study and those previously reported in Thailand also revealed intraspecific divergence and recent population expansion, possibly due to adaptive evolution associated with host range expansion. Our data revealed, for the first time, multiple E. bieneusi genotypes of zoonotic significance circulating in Thai bats and demonstrated that bat guano fertilizer may be a vehicle for disease transmission.

First record of Anopheles claviger (Meigen, 1804) (Diptera, Culicidae) in Karelia, Northwestern Russia.

Anopheles claviger (Meigen, 1804) (Diptera, Culicidae) is widespread in the western Palaearctic Region, but it was recorded in Karelia (Russia) for the first time. This record is one of the northernmost ones in the Palaearctic Region and Russia, updates the northern border of the An. claviger range. Mosquitoes were collected from July to September 2023 in the southern Karelia (the village of Gomselga, Kondopoga District, and Petrozavodsk) using Krishtal trap (from human) and Mosquito Magnet® trap (Pioneer design, Octenol as attractant). Seven females of An. claviger were collected in Gomselga; one specimen was sampled from Petrozavodsk City parks. Morphological identification of eight females was verified by COI and ITS2 sequences. Phylogenetic analysis of ITS2 and COI sequences confirmed the collected specimens to An. claviger s. s., clustering in both cases in a strongly supported clade clearly differentiated from the closely related species An. petragnani. The high diversity of An. claviger haplotypes from Karelia is in agreement with data from other geographical regions and shows that the records of this species in Gomselga and Petrozavodsk are not accidental.

First Report of White Mold Caused by Sclerotinia sclerotiorum on Echeveria gigantea in Mexico.

Echeveria gigantea, native of Mexico (Reyes et al. 2011), holds economic importance as it is marketed as a potted plant and cut flower due to its drought-tolerant capabilities and aesthetic appeal. In September 2023, a field sampling was conducted at the Research Center in Horticulture and Native Plants (18°55'56.6" N, 98°24'01.5" W) of UPAEP University. Echeveria gigantea cv. Quilpalli plants with white mold symptoms were found in an area of 0.5 ha, with an incidence of 40% and severity of 50% on severely affected stems. The symptoms included chlorosis of older foliage, necrosis at the base of the stem, and soft rot with abundant white to gray mycelium and abundant production of irregular sclerotia resulting in wilted plants. The fungus was isolated from 30 symptomatic plants. Sclerotia were collected, sterilized in 3% NaOCl, rinsed with sterile distilled water (SDW), and plated on Potato Dextrose Agar (PDA) with sterile forceps. Subsequently, a dissecting needle was used to place fragments of mycelium directly on PDA. Plates were incubated at 23 °C in darkness. A total of 30 isolates were obtained using the hyphal-tip method, one from each diseased plant (15 isolates from sclerotia and 15 from mycelium). After 6 days, colonies had fast-growing, dense, cottony-white aerial mycelium forming irregular sclerotia of 3.67 ± 1.13 mm (n=100). Each Petri dish produced 32.47 ± 7.5 sclerotia (n=30), after 12 days. The sclerotia were initially white and gradually turned black. The isolates were tentatively identified as Sclerotinia sclerotiorum based on morphological characteristics (Saharan and Mehta 2008). Two isolates were selected for molecular identification. Genomic DNA was extracted using the CTAB protocol. The ITS region and the glyceraldehyde 3-phosphate dehydrogenase (G3PDH) gene were sequenced for two randomly selected isolates (White et al. 1990; Staats et al. 2005). The ITS and G3PDH sequences of the SsEg9 isolate were deposited in GenBank (ITS-OR816006; G3PDH-OR879212). BLAST analysis of the partial ITS (510 bp) and G3PDH (915 bp) sequences showed 100% and 99.78% similarity to S. sclerotiorum isolates (GenBank: MT101751 and MW082601). Pathogenicity was confirmed by inoculating 30 120-day-old E. gigantea cv. Quilpalli plants grown in pots with sterile soil. Ten sclerotia were deposited at the base of the stem, 10 mm below the soil surface. As control treatment, SDW was applied to 10 plants. The plants were placed in a greenhouse at 23 °C and 90% relative humidity. After 16 days, all inoculated plants displayed symptoms similar to those observed in the field. Control plants did not display any symptoms. The fungus was reisolated from the inoculated stems, fulfilling Koch's postulates. The pathogenicity tests were repeated three times. Recently S. sclerotiorum has been reported causing white mold on cabbage in the state of Puebla, Mexico (Terrones-Salgado et al. 2023). To the best of our knowledge, this is the first report of S. sclerotiorum causing white mold on E. gigantea in Mexico. Information about diseases affecting this plant is very limited, so this research is crucial for designing integrated management strategies and preventing spread to other production areas.

Revealing the diversity of Jojoba-associated fungi using amplicon metagenome approach and assessing the in vitro biocontrol activity of its cultivable community.

Jojoba shrubs are wild plants cultivated in arid and semiarid lands and characterized by tolerance to drought, salinity, and high temperatures. Fungi associated with such plants may be attributed to the tolerance of host plants against biotic stress in addition to the promotion of plant growth. Previous studies showed the importance of jojoba as jojoba oil in the agricultural field; however, no prior study discussed the role of jojoba-associated fungi (JAF) in reflecting plant health and the possibility of using JAF in biocontrol. Here, the culture-independent and culture-dependent approaches were performed to study the diversity of the jojoba-associated fungi. Then, the cultivable fungi were evaluated for in-vitro antagonistic activity and in vitro plant growth promotion assays. The metagenome analysis revealed the existence of four fungal phyla: Ascomycota, Aphelidiomycota, Basidiomycota, and Mortierellomycota. The phylum Ascomycota was the most common and had the highest relative abundance in soil, root, branch, and fruit samples (59.7%, 50.7%, 49.8%, and 52.4%, respectively). Alternaria was the most abundant genus in aboveground tissues: branch (43.7%) and fruit (32.1%), while the genus Discosia had the highest abundance in the underground samples: soil (24%) and root (30.7%). For the culture-dependent method, a total of 14 fungi were isolated, identified, and screened for their chitinolytic and antagonist activity against three phytopathogenic fungi (Fusarium oxysporum, Alternaria alternata and Rhizoctonia solani) as well as their in vitro plant growth promotion (PGP) activity. Based on ITS sequence analysis, the selected potent isolates were identified as AspergillusstellatusEJ-JFF3, Aspergillusflavus EJ-JFF4, Stilbocreasp. EJ-JLF1, Fusariumsolani EJ-JRF3, and AmesiaatrobrunneaEJ-JSF4. The endophyte strain A. flavus EJ-JFF4 exhibited the highest chitinolytic activity (9 Enzyme Index) and antagonistic potential against Fusarium oxysporum, Alternaria alternata, and Rhizoctonia solani phytopathogens with inhibitory percentages of 72, 70, and 80 respectively. Also, A. flavus EJ-JFF4 had significant multiple PGP properties, including siderophore production (69.3%), phosphate solubilization (95.4 µg ml-1). The greatest production of Indol-3-Acetic Acid was belonged to A.atrobrunnea EJ-JSF4 (114.5 µg ml-1). The analysis of FUNGuild revealed the abundance of symbiotrophs over other trophic modes, and the guild of endophytes was commonly assigned in all samples. For the first time, this study uncovered fungal diversity associated with jojoba plants using a culture-independent approach and in-vitro assessed the roles of cultivable fungal strains in promoting plant growth and biocontrol. The present study indicated the significance of jojoba shrubs as a potential source of diverse fungi with high biocontrol and PGP activities.

Novelty in Arundinella (Arundinelleae, Panicoideae, Poaceae): Morphological and Molecular Evidences for a New Species from Sikkim Himalaya

Novelty in Arundinella (Arundinelleae, Panicoideae, Poaceae): Morphological and Molecular Evidences for a New Species from Sikkim Himalaya

Characterization and pathogenicity of Pleurostoma richardsiae causing decline of mango trees in Southern Italy

Mango trees (Mangifera indica) showing symptoms of twig and branch dieback, internal wood necroses, and decline, were surveyed in an orchard in Palermo province (Eastern Sicily, Italy). A Pleurostoma-like fungus was consistently isolated from symptomatic wood tissues. Based on morphology and phylogenetic analysis of ITS and tub2 sequences, the fungus was identified as Pleurostoma richardsiae. A pathogenicity test was conducted by inoculating stems of 2-year-old mango seedlings with mycelium plugs and conidium suspensions of a representative isolate. Two months after inoculation, necrotic lesions were observed around the inoculation points, and P. richardsiae was reisolated from the necrotic tissues. This is the first report of P. richardsiae causing dieback and decline of mango trees.

Molecular characterization of apple endophytic bacteria and biological effect on Alternaria rot on apple

AbstractApple rot disease, caused by Alternaria alternata, is causing significant losses in apple production, especially in post‐harvest periods. Alternaria alternata, was isolated from rotten apple fruit of cv Amasya. Identification of the strain isolated from rotten apple fruit was made by ITS sequence and morphological analysis. In the present study, 58 different endophytic bacteria strains were isolated from apple fruit for use in apple rot disease control. Among the isolated strains, 26 endophyte strains were determined to inhibit the radial growth of mycelia of A. alternata in binary culture in PDA media. Based on 16S rRNA sequencing analysis, these antagonistic endophytic bacteria strains were identified as Bacillus ssp. Bacillus methylotrophicus (ib17) showed the highest significant inhibitory effect on the mycelial growth of A. alternata at a rate of 79.5% in PDA medium. The results of in vivo experiments showed that Bacillus licheniformis (ib20) achieved 100% biocontrol when endophyte bacterial strains were inoculated 24 h before the inoculation of A. alternata. When the three bacteria species and A. alternata were inoculated simultaneously, Bacillus myloliquefaciens (ib1), Bacillus licheniformis (ib21) and the Endophytic bacterium (ib16) showed the highest biocontrol efficacy against A. alternata at 81.8%. Furthermore, when the bacterial strains were inoculated at 24 h after inoculation of A. alternata, B. licheniformis (ib21) showed the highest biocontrol efficacy of 83.3% against the fungal pathogen compared to the controls. The endophytic bacterial strains produced fungal cell wall hydrolysing enzymes such as chitinase, cellulase, amylase and protease. According to the results of this study, apple endophytic bacteria are potential candidates to control apple decay caused by A. alternata through cell wall hydrolysing enzymes and reduce disease severity. However, the efficacy of the biological control was dependent on the applied time of epiphytic bacterial strains. Further research is needed on the biochemical mechanisms underlying the activity of these bacteria against A. alternata.