Non-inoculated Plants Research Articles

First Report of Anthracnose Caused by Colletotrichum spaethianum on Hosta longipes in Korea.

Hosta longipes (Franch. & Sav.) Matsum. (Asparagaceae) is a perennial, herbaceous plant, native to Japan and Korea (Lee et al. 2021). In Korea, the plant is used as an edible vegetable and ornamental (Kang and Ju 2015). During 2021-2022, anthracnose symptoms were observed on leaves of H. longipes with over 70% disease incidence in Wanju-gun (35°38'47''N; 127°31'16''E) and Jangsu-gun (35°35'31''N; 127°30'03''E) in Jeollabuk-do, Korea. The disease initially appeared on old leaves, gradually spreading to young ones. The symptoms were characterized as yellow to white discoloration on the upper leaf surface with black necrotic tissue in the center of the lesion. Three H. longipes samples with anthracnose symptoms were collected. From each, a monoconidial isolate was obtained and then deposited in the Korea Agricultural Culture Collection (accession Nos. KACC 410038, 410391, and 410443). The dried specimens were housed at the herbarium of Jeonbuk National University (JBNU0129, 0137) and Korea University (KUS-F33379). Conidiomata was acervular, 65 to 80 × 56 to 70 µm in diam. Setae were dark brown, 2 to 4-septate, 63 to 161 µm long, being formed on a pale brown cushion. Conidia were hyaline, smooth-walled, aseptate, slightly curved, base truncate, 3.9 to 5.1 × 17 to 23 µm. The appressoria were solitary, olivaceous-brown, ovoid or irregularly shaped. Two-week-old colonies grown on PDA at 25 ℃ were 20-25 mm in diameter, initially white, then turned gray with age, with cottony aerial mycelium. The morphological and cultural characteristics of the fungus were consistent with those of Colletotrichum spaethianum (Allesch.) Damm, P.F. Cannon & Crous (Damm et al. 2012). To confirm morphology-based identification, the nucleotide sequences of the internal transcribed spacer (ITS) rDNA region, glyceraldehyde 3-phosphate dehydrogenase (GAPDH), actin (actA), chitin synthase (CHS1), histone (HIS3) and tubulin (TUB2) genes were determined for KACC410443, as outlined by Cannon et al. (2012) and Damm et al. (2009). The resulting sequences were submitted into GenBank (PP000829 for ITS, PP133094 for GAPDH, PP083418 for actA, PP133091 for CHS1, PP133097 for HIS3, and PP133099 for TUB2) and compared with reference sequences in GenBank using BLASTn search tool. The results showed a 100% match with C. spaethianum (MT611068), C. incanum (MN880260) and C. truncatum (EF016303) for ITS, and 100% with C. spaethianum for GAPDH (MH370513), actA (MH045677), CHS1 (MH370520), HIS3 (MH985161), and TUB (MH456884). Pathogenicity was tested by inoculating conidial suspension (1 ×104 cfu/ml) of three-week-old fungal colonies of the isolate KACC410443 onto leaves of three healthy potted plants. Prior to inoculation, leaves were deliberately wounded by pinpricking with a sterilized needle. Two wounded but non-inoculated plants served as controls. Plants were maintained in a greenhouse at 25 to 30 °C. Inoculated plants developed anthracnose symptoms after eight days, while the control plants remained symptomless. The fungus isolated from the inoculated plants was morphologically identical to that observed initially, fulfilling Koch's postulates. To our knowledge, there is no previous record of C. spaethianum on H. longipes, although C. spaethianum has been reported to infect another species, H. plantaginea (Cheon and Jeon 2016). This is the first report of this fungus on H. longipes in Korea (KSPP 2024) and globally (Farr and Rossman 2024). The anthracnose on this ornamental plant can be considered a new severe threat to planting strategies in gardens.

First Report of Powdery Mildew Caused by Podosphaera xanthii on Acalypha indica in China.

Acalypha indica L. is an annual erect herb of the Euphorbiaceae family. This plant is found widely in the tropics and parts of Africa and Asia (Chakraborty et al. 2023). In China, A. indica is a vegetable and also used as a folk medicine due to its antipyretic and hemostatic, antibacterial and anti-inflammatory properties. In February 2022 and 2023, powdery mildew symptoms were observed on 70% of A. indica plants on the Hainan Medical University campus (19° 58' 53″ N; 110° 19' 47″ E) in Haikou, Hainan Province, China. Powdery mildew colonies covered the leaf surfaces and stems of affected plants, causing discoloration and defoliation. Mycelia were superficial and hyphal appressoria were nipple-shaped. Conidiophores (n =30) were unbranched, cylindrical, 66 to 150 × 10 to 15 µm, and produced three to five immature conidia in chains with a crenate outline. Foot cells (n =30) were cylindrical, straight or sometimes curved at the base, and 31 to 59 µm long. Conidia (n =100) were ellipsoid-ovoid to doliiform, 20 to 33 ×12 to 20 µm (length/width ratio = 1.3 to 2.4), with well-developed fibrosin bodies, and produced germ tubes from the lateral position. Based on these morphological characteristics, the pathogen was provisionally identified as Podosphaera xanthii (Braun and Cook 2012). The teleomorph was not observed. A specimen was deposited in the Hainan Medical University Plant Pathology Herbarium as HMAI-23. To confirm the genus identification and ascertain a putative species, genomic DNA was extracted from mycelium, conidiophores, and conidia using a fungal DNA kit (Omega Bio-Tek, USA). The rDNA internal transcribed spacer (ITS) region was amplified with primers ITS1/ITS4 (White et al. 1990) and sequenced directly. The resulting 575-bp sequence was deposited in GenBank (accession no. OR775733). A BLASTn search in GenBank of this sequence showed 99% similarity with the ITS sequences of P. xanthii on plants of Fabaceae, Malvaceae and Cucurbitaceae family from China (MH143485, MT242593, MK439611 and MH143483), Thailand (LC270779 and LC270778), Korea (MG754404), Vietnam (KM260704), and Puerto Rico (OP882310). Additionally, the 28S rDNA region was amplified using the primer pairs NL1 and NL4 (O´Donnell 1993; accession no. OR784547). This region shared 99% similarity with P. xanthii isolates (LC371333, LC270780, AB936277, and OP765401) as well. To confirm pathogenicity, five healthy potted plants of A. indica were inoculated by gently pressing a powdery mildew-infected leaf onto 15 young leaves. Five non-inoculated plants served as controls. All plants were maintained in a greenhouse at 24 to 30°C, 70% relative humidity, with a 16-h photoperiod. After 7 days, inoculated leaves showed powdery mildew symptoms whereas no symptoms were observed on control plants. The fungal colonies observed on inoculated plants were morphologically identical to those found on the originally infected leaves collected from Hainan Province. Based on the morphological characteristics and molecular identification, the fungus was identified as P. xanthii. In different countries and regions, P. xanthii has been previously reported on A. indica from Sudan and India (Amano 1986). To our knowledge, this is the first record of P. xanthii infecting A. indica in China. We are concerned that the pathogen could become a threat to the widespread planting of A. indica in the future.

First Report of Shoot Blight of Grapevine Caused by Sclerotinia sclerotiorum in Illes Balears, Mallorca, Spain.

In 2021, grapevines (Vitis vinifera L.) cv. Callet growing in a commercial vineyard located at Pollença (northeast of the island of Majorca, Spain) showed severe symptoms of shoot blight during spring and early summer, with an incidence of 70%. Symptoms consisted of elongated cankered-like lesions, surrounded by water-soaked darker tissues, that developed at the base or around the middle nodes of the shoot. For fungal isolation, shoot samples with lesions were collected, surface disinfected with 2% NaCl for 90s, rinsed twice with deionized water and placed in Petri plates containing potato dextrose agar (PDA). The plates were incubated at 25°C under 12 h light-darkness for 6 days. Isolations consistently yielded on kind of fungal colonies that produced white mycelium and black spherical to elongated sclerotia (2 to 10 mm in diameter). Morphological characterization was consistent with the description of Sclerotinia sclerotiorum (Lib.) de Bary (Bolton et al. 2006). Three isolates (UIB 118-1, UIB 118-26, and UIB 129-41) were preserved and deposited in the Culture Collection of Microbiology-Faculty of Sciences, University of Balearic Islands, Spain. Genomic DNA was extracted from isolates UIB 118-26 and UIB 129-41 using the EZNA Miniprep Kit (Omega Bio-Tek, Norcross, GA). The internal transcribed spacer (ITS) region of ribosomal DNA, β-tubulin (BTUB) and calmodulin (CAL) gene regions were amplified using ITS1F-ITS4 (Gardes and Bruns, 1996; White et al. 1990), Bt-2a/Bt-2b (Glass and Donaldson 1995) and CAL228F/CAL737R (Carbone and Kohn 1999) primer sets, respectively. Amplicons were sequenced and deposited in GenBank with accession numbers MZ604647 and MZ604648 for ITS, OK634402 and OK634403 for BTUB and OK634404 and OK634405 for CAL. BLASTn search showed that isolates were >99 % (ITS, BTUB and CAL) identical to S. sclerotiorum GenBank accession no. KF859933, CP017815 and KF871381, respectively. Pathogenicity tests were conducted using eight one-year old grapevines cv. Cabernet Sauvignon. Old and new green shoots were inoculated by inserting a 6-mm plug of mycelium taken from actively growing cultures on PDA into cuts made at the base and at the distal part of each shoot with a sterile scalpel with a total of eight inoculation points per plant. Inoculated wounds were sealed with Parafilm tape to avoid rapid dehydration. Inoculated plants and an equal number of wounded but non-inoculated plants (negative controls) were maintained at 25 ± 1°C for 48 h in plastic containers to ensure a high relative humidity (>90%). After 5 days, the infection girdled and rotted the green new shoots, whereas the older partially lignified shoots developed a localized long brown lesion that reached 16 cm in length. Due to the rotting of the basal part of the petiole, leaves turned gray, wilted, and died, easily detaching from the stem. In advanced stages of the disease, 7 days after infection, branches died and fell with the leaves remained attached (Fig 1 A, B). Reisolations from diseased shoots were successfully performed on PDA to fulfill Koch's postulates. S. slerotiorum was previously reported on grapevine causing shoot blight in Chile (Latorre and Guerrero, 2001), Korea (Jong-Han et al. 2009), California-USA (Boland and Hall, 1994) and Australia (Hall et al. 2002). AlsoS. sclerotiorum was reported among the endophytic mycobiota associated with Vitis vinifera in the Iberian Peninsula (Gonzalez and Tello, 2011) but not as a pathogen causing visible symptoms on that crop. So, this is the first report of the occurrence of S. slerotiorum as a pathogen of grapevines in Spain causing symptoms of canker and shoot blight. This finding highlights a potential risk of this fungal disease for the wine industry in the Mediterranean region and specially for Spain, the country with the largest acreage devoted to grapevines. Although chemical and biological are suitable control strategies, disease management is difficult as sclerotia of Sclerotinia can remain in the soil for up to eight years (Adams and Ayears, 1979), and preventive surveys are greatly recommended as an important epidemiological tool to monitor the epidemiology of disease and identify potential outbreaks of this new pathogen on grapevine in Spain.

First Report of Powdery Mildew Caused by Golovinomyces adenophorae on Adenophora triphylla var. japonica in Korea.

Adenophora triphylla var. japonica (Campanulaceae), known as Japanese lady bell, is native to East Asia. It has been used as a medicinal plant but is widely cultivated in Korea as an indigenous vegetable (Park et al. 2011). In the summer of 2020, about 100 plants in an experimental plot at the National Institute of Forest Science, Seoul, Korea, showed powdery mildew symptoms with a 100% disease incidence. Signs first appeared as white colonies, subsequently expanding over the leaves, stems, and inflorescences. Infected young shoots were elongated and became slender. Chasmothecia were found in late October. Voucher specimens were deposited in the Korea University Herbarium (KUS-F). Conidiophores arising from the lateral part of the hyphae were upright, 100 to 220 × 10 to 12 µm, and produced 2 to 5 immature conidia in chains with sinuate edge lines. Basal parts of foot-cells in conidiophores were curved. Conidia were barrel-shaped to ellipsoid, 26 to 40 × 14 to 20 µm, and produced germ tubes on the perihilar position of the conidia. Chasmothecia with short mycelioid appendages were gregarious, 144 to176 µm in diam., and contained 8 to 22 asci. Asci were clavate-saccate with short stalks, 60 to 82 × 28 to 42 µm, and contained two spores. Ascospores were broadly ellipsoid, cytoplasm-dense without vacuoles, colorless, and 22 to 28 × 12 to 18 µm. The structures and measurements were consistent with those of Golovinomyces adenophorae (R.Y. Zheng & G.Q. Chen) Heluta (Braun & Cook, 2012). To confirm the morphology-based identification, two herbarium specimens (KUS-F29252 and F31898) were sequenced for the internal transcribed spacer (ITS) and large subunit (LSU) regions with PM10/ITS4 and PM3/TW14 primers, respectively (Bradshaw and Tobin, 2020). A Blastn search revealed high similarities in the ITS and LSU sequences, with 99.81% (538/539 bp) and 99.86% (697/698 bp) to G. adenophorae sequences (AB077633 and AB077632), respectively. All resulting sequences were deposited in GenBank under accession numbers OR841069-70 for ITS and OR841071 for LSU. A pathogenicity test was performed through inoculation by gently dusting the conidia from a detached symptomatic leaf onto the leaves of five healthy plants. Five non-inoculated plants served as controls. Following inoculation, plants were covered with plastic film and maintained in a greenhouse (24 to 32°C) until symptoms developed. Powdery mildew colonies developed on the inoculated plants after twelve days, whereas the control plants remained symptomless. The inoculated pathogen was confirmed morphologically and molecularly by the sequence comparison aforementioned, fulfilling Koch's postulates. Based on morphological characteristics and the sequencing data, the powdery mildew was identified as G. adenophorae. The association of G. adenophorae and Adenophora spp. has been known in China, Japan, Kazakhstan, and the Far East of Russia (Farr and Rossman, 2023). This is the first report of powdery mildew caused by G. adenophorae on A. triphylla var. japonica in Korea. Since the commercial cultivation of this plant aims to harvest young shoots as one of the most popular vegetables in Korea, appropriate control measures for the powdery mildew should be considered.

Investigating the simultaneous effect of chitosan and arbuscular mycorrhizal fungi on growth, phenolic compounds, PAL enzyme activity and lipid peroxidation in Salvia nemorosa L.

Considering the importance of Salvia nemorosa L. in the pharmaceutical and food industries, and also beneficial approaches of arbuscular mycorrhizal fungi (AMF) symbiosis and the use of bioelicitors such as chitosan to improve secondary metabolites, the aim of this study was to evaluate the performance of chitosan on the symbiosis of AMF and the effect of both on the biochemical and phytochemical performance of this plant and finally introduced the best treatment. Two factors were considered for the factorial experiment: AMF with four levels (non-inoculated plants, Funneliformis mosseae, Rhizophagus intraradices and the combination of both), and chitosan with six levels (0, 50, 100, 200, 400 mg L-1 and 1% acetic acid). Four months after treatments, the aerial part and root length, the levels of lipid peroxidation, H2O2, phenylalanine ammonia lyase (PAL) activity, total phenol and flavonoid contents and the main secondary metabolites (rosmarinic acid and quercetin) in the leaves and roots were determined. The flowering stage was observed in R. intraradices treatments and the highest percentage of colonization (78.87%) was observed in the treatment of F. mosseae×400 mg L-1 chitosan. Furthermore, simultaneous application of chitosan and AMF were more effective than their separate application to induce phenolic compounds accumulation, PAL activity and reduce oxidative compounds. The cluster and principal component analysis based on the measured variables indicated that the treatments could be classified into three clusters. It seems that different treatments in different tissues have different effects. However, in an overview, it can be concluded that 400 mg L-1 chitosan and F. mosseae×R. intraradices showed better results in single and simultaneous applications. The results of this research can be considered in the optimization of this medicinal plant under normal conditions and experiments related to abiotic stresses in the future.

First Report of Powdery Mildew on Berchemia floribunda Caused by Erysiphe berchemiae in China.

The genus Berchemia (family Rhamnaceae), a group of climbing plants, is mainly distributed in Asia, Africa, and South America. Berchemia plants are widely used in traditional medicine in some Asian countries (Inoshiri et al. 1987). For example, in Japan, B. racemosa (synonym B. floribunda) is used for the treatment of gallstones, liver diseases, neuralgia, and stomach cramps, and in China, B. floribunda is used for the treatment of rheumatism and lumbago. In August 2022, typical powdery mildew symptoms were observed on wild B. floribunda plants in Huaxi District, Guiyang, Guizhou Province, China. The incidence was approximately 60% among 100 B. floribunda plants observed outdoors. White colonies almost entirely covered on both adaxial and abaxial surfaces of all leaves on symptomatic plants. Infected leaves appeared curled or chlorotic, infection occasionally leading to defoliation. To describe the pathogen morphologically, fungal samples were collected from two individual B. floribunda plants and microscopically characterized. In these samples, hyphae were flexuous to straight, branched, septate, 3-6 μm wide, with lobed hyphal appressoria. Conidiophores were erect, flexuous to straight, and 50-160 µm long (n = 30). Foot cells were subcylindrical to slightly curved-sinuous at the base, 20-40 µm long (n = 30), followed by 2-3 shorter cells. Conidia formed singly, occasionally 2-3 in a chain. Conidia were ellipsoid to ovoid, 20-42 × 12-18 µm (n = 50), without fibrosin bodies. Chasmothecia were not found. For molecular identification, the ribosomal DNA internal transcribed spacers (ITSs) of the two fungal samples were amplified and sequenced using the ITS1/ITS4 primer pair (White et al. 1990). The obtained 649-bp ITS sequences (GenBank accession nos. OR414364 and OR414365, respectively) shared 100% identity, and they showed 99.52% identity with the ITS sequence (GenBank accession no. LC009934) of Erysiphe berchemiae. Phylogenetic analysis grouped OR414364 and OR414365 in a clade with LC009934. Based on morphological and molecular characteristics, the powdery mildew fungus from B. floribunda was identified as E. berchemiae (Braun and Cook 2012). The voucher specimen (accession no. GZNU-BFEE/0820/2022) were deposited at the School of Life Sciences, Guizhou Normal University. Pathogenicity was assessed by gently pressing naturally diseased leaves of B. floribunda onto leaves of three healthy potted 1-year-old B. floribunda plants. Three non-inoculated healthy plants were used as controls. The plants were incubated in a greenhouse at 25 ± 2°C, 80% relative humidity. Similar powdery mildew symptoms were observed on the inoculated plants 9 days after inoculation, whereas the control plants remained symptomless. The reisolated fungus from inoculated leaves was morphologically identical to that observed on the original diseased leaves, and the ITS sequence of the reisolated fungus shared 100% identity with OR414364 and OR414365, fulfilling Koch's postulates. E. berchemiae has previously been described as a powdery mildew pathogen on B. yunnanensis (Chen et al. 1987) and B. kulingensis (Chen 1993) in China and B. racemosa (synonym B. floribunda) in Japan (Braun and Cook 2012; Takamatsu et al. 2015), but this is the first report of E. berchemiae causing disease of B. floribunda in China. This work suggests that E. berchemiae is an important pathogen of Berchemia plants, at least for some species in the genus Berchemia.

Transcriptome Analysis of Resistant Cotton Germplasm Responding to Reniform Nematodes.

Reniform nematode (Rotylenchulus reniformis) is an important microparasite for Upland cotton (Gossypium hirsutum L.) production. Growing resistant cultivars is the most economical management method, but only a few G. barbadense genotypes and some diploid Gossypium species confer high levels of resistance. This study conducted a transcriptome analysis of resistant genotypes to identify genes involved in host plant defense. Seedlings of G. arboreum accessions PI 529728 (A2-100) and PI 615699 (A2-190), and G. barbadense genotypes PI 608139 (GB 713) and PI 163608 (TX 110), were inoculated with the reniform nematode population MSRR04 and root samples were collected on the fifth (D5) and ninth (D9) day after inoculation. Differentially expressed genes (DEGs) were identified by comparing root transcriptomes from inoculated plants with those from non-inoculated plants. Accessions A2-100 and A2-190 showed 52 and 29 DEGs on D5, respectively, with 14 DEGs in common, and 18 DEGs for A2-100 and 11 DEGs for A2-190 on chromosome 5. On D9, four DEGs were found in A2-100 and two DEGs in A2-190. For GB 713, 52 and 43 DEGs were found, and for TX 110, 29 and 117 DEGs were observed on D5 and D9, respectively. Six DEGs were common at the two sampling times for these genotypes. Some DEGs were identified as Meloidogyne-induced cotton (MIC) 3 and 4, resistance gene analogs, or receptor-like proteins. Other DEGs have potential roles in plant defense, such as peroxidases, programmed cell death, pathogenesis related proteins, and systemic acquired resistance. Further research on these DEGs will aid in understanding the mechanisms of resistance to explore new applications for the development of resistant cultivars.

Simultaneous Impact of Rhizobacteria Inoculation and Leaf-Chewing Insect Herbivory on Essential Oil Production and VOC Emissions in Ocimum basilicum.

Inoculation with rhizobacteria and feeding by herbivores, two types of abiotic stress, have been shown to increase the production of secondary metabolites in plants as part of the defense response. This study explored the simultaneous effects of inoculation with Bacillus amyloliquefaciens GB03 (a PGPR species) and herbivory by third-instar Spodoptera frugiperda larvae on essential oil (EO) yield and volatile organic compound (VOC) emissions in Ocimum basilicum plants. The density of glandular trichomes was also examined, given that they are linked to EO production and VOC emission. Herbivory increased EO content, but inoculation on its own did not. When combined, however, the two treatments led to a 10-fold rise in EO content with respect to non-inoculated plants. VOC emissions did not significantly differ between inoculated and non-inoculated plants, but they doubled in plants chewed by the larvae with respect to their undamaged counterparts. Interestingly, no changes were observed in VOC emissions when the treatments were tested together. In short, the two biotic stressors elicited differing plant defense responses, mainly when EO was concerned. PGPR did not stimulate EO production, while herbivory significantly enhanced it and increased VOC emissions. The combined treatment acted synergistically, and in this case, PGPR inoculation may have had a priming effect that amplified plant response to herbivory. Peltate trichome density was higher in inoculated plants, those damaged by larvae, and those subjected to the combination of both treatments. The findings highlight the intricate nature of plant defense mechanisms against various stressors and hint at a potential strategy to produce essential oil through the combined application of the two stressors tested here.

First Report of Rust Disease on Alcea rosea caused by Puccinia modiolae in China.

Alcea rosea, belonging to the Alcea genus in the Malvaceae family, originated from China, but it is now grown worldwide. A. rosea has been widely used in traditional Chinese medicine to alleviate constipation, pain, swelling, and sores. In February 2023, typical symptoms of fungal infection were observed on A. rosea at Guizhou Normal University in Guiyang, Guizhou Province, China. The disease incidence was over 90% (n = 100) for the surveyed A. rosea plants, and the disease severity range from 30% to 90%. The initial symptoms of A. rosea rust were the appearance of chlorotic spots on the leaves. Subsequently, numerous reddish to dark-brown erumpent pustules (telia) were observed. Gradually, the entire plant was covered by rust and the center of each lesion turned brown, necrotic, and ruptured over times, eventually causing defoliation. Voucher specimens of infected A. rosea leaves as representative samples have been deposited at Guizhou Normal University (GNU2023LS008). Telia are round in shape, mostly aggregated in mass, with a diameter of 0.28-0.78 mm (0.46 mm, n = 20). They range in color from reddish-brown to dark brown, and are mainly hypophyllous but occasionally formed on the adaxial leaf surface. The teliospores are fusoid with dimensions of 31.3-93.8 × 10.9-21.3 μm (57.5 × 15.1 µm average, n = 50), hyaline or yellowish to light-brown in color, mostly two-celled, with a smooth wall (1.5-3.0 µm) and a thickened apex (3.0-9.0 µm). However, teliospores which are one-, three-, or four-celled with a notch at the apex, are rarely observed. The morphological characteristics of host symptoms and teliospores were similar to those of Puccinia modiolae (Aime and Abbasi 2018; Albu et al. 2019). For phylogenetic analysis, genomic DNA was extracted from the teliospores of infected leaves. To confirm the species-level identification, PCR was performed on the extracted DNA to amplify the ribosomal DNA internal transcribed spacer (ITS) and large subunit (LSU) regions using primer pairs ITS1/ITS4 (Schoch et al. 2012) and NL1/NL4 (Ziemiecki et al. 1990), respectively. The resulting ITS DNA sequence (GenBank accession no. OR607960) showed 100% identity with P. modiolae sequences (OP369291.1), when the query coverage was 100%. The LSU DNA sequence obtained (OR607961.2) shared 99.85% similarity with P. modiolae (MK458702.1). A phylogenetic tree was constructed using MEGA7.0 and the maximum likelihood method based on the ITS and LSU sequences. The fungal isolates collected in this study and several reference sequences of P. modiolae were grouped within a clade that included the isolates reported on A. rosea in Korea (Ryu et al. 2023), with 100% bootstrap support. Pathogenicity testing was conducted by gently pressing spore powder of naturally diseased leaves onto young leaves of three healthy A. rosea plants, with three noninoculated healthy plants serving as controls. The inoculated and noninoculated plants were kept in a growth chamber at the 26°C with a 12 hour light/dark cycle and 80% humidity. After 2 weeks, all inoculated A. rosea plants showed characteristic disease symptoms of rust infection and telia of P. modiolae, while control plants remained symptomless. The pathogen was identical to that observed on the original diseased leaves. The study results indicate that the causal fungus responsible for the disease is P. modiolae, which has been previously reported on Malvaceae plants (Farr and Rossman 2022). To the best of our knowledge, this is the first report of P. modiolae on A. rosea in China. This study will contribute to an increased understanding of the host range of Puccinia modiolae.

Rhizobia-legume symbiosis mediates direct and indirect interactions between plants, herbivores and their parasitoids

Microorganisms associated with plant roots significantly impact the quality and quantity of plant defences. However, the bottom-up effects of soil microbes on the aboveground multitrophic interactions remain largely under studied. To address this gap, we investigated the chemically-mediated effects of nitrogen-fixing rhizobia on legume-herbivore-parasitoid multitrophic interactions. To address this, we initially examined the cascading effects of the rhizobia bean association on herbivore caterpillars, their parasitoids, and subsequently investigated how rhizobia influence on plant volatiles and extrafloral nectar. Our goal was to understand how these plant-mediated effects can affect parasitoids. Lima bean plants (Phaseoulus lunatus) inoculated with rhizobia exhibited better growth, and the number of root nodules positively correlated with defensive cyanogenic compounds. Despite increase of these chemical defences, Spodoptera latifascia caterpillars preferred to feed and grew faster on rhizobia-inoculated plants. Moreover, the emission of plant volatiles after leaf damage showed distinct patterns between inoculation treatments, with inoculated plants producing more sesquiterpenes and benzyl nitrile than non-inoculated plants. Despite these differences, Euplectrus platyhypenae parasitoid wasps were similarly attracted to rhizobia- or no rhizobia-treated plants. Yet, the oviposition and offspring development of E. platyhypenae was better on caterpillars fed with rhizobia-inoculated plants. We additionally show that rhizobia-inoculated common bean plants (Phaseolus vulgaris) produced more extrafloral nectar, with higher hydrocarbon concentration, than non-inoculated plants. Consequently, parasitoids performed better when fed with extrafloral nectar from rhizobia-inoculated plants. While the overall effects of bean-rhizobia symbiosis on caterpillars were positive, rhizobia also indirectly benefited parasitoids through the caterpillar host, and directly through the improved production of high quality extrafloral nectar. This study underscores the importance of exploring diverse facets and chemical mechanisms that influence the dynamics between herbivores and predators. This knowledge is crucial for gaining a comprehensive understanding of the ecological implications of rhizobia symbiosis on these interactions.

Mitigation of Salt Stress in Rice by the Halotolerant Plant Growth-Promoting Bacterium Enterobacter asburiae D2.

Salinity is a major abiotic stress that seriously affects crop growth worldwide. In this work, we aimed to isolate potential halotolerant plant growth-promoting rhizobacteria (PGPR) to mitigate the adverse impacts of salt stress in rice. An isolate, D2, with multiple plant growth-promoting (PGP) characteristics was identified as Enterobacter asburiae D2. Strain D2 could produce indole-3-acetic acid and siderophore. It also exhibited phosphate solubilization and 1-aminocyclopropane-1-carboxylic deaminase activity. Genome analysis further provided insights into the molecular mechanism of its PGP abilities. Strain D2 inoculation efficiently stimulated rice growth under both normal and saline conditions. Compared with the non-inoculated plants, a significant increase in plant height (18.1-34.7%), root length (25.9-57.1%), root dry weight (57.1-150%), and shoot dry weight (17.3-50.4%) was recorded in inoculated rice seedlings. Meanwhile, rice seedlings inoculated with strain D2 showed improvement in chlorophyll and proline content, while the oxidant damage was reduced in these plants in comparison with the control group. Moreover, the K+/Na+ ratio of the inoculated rice seedlings exposed to NaCl and Na2CO3 was higher than that of the uninoculated groups. These results imply that Enterobacter asburiae D2 is a potential PGPR that can be used for alleviation of salt stress in rice.

Marine Plant Growth Promoting Bacteria (PGPB) inoculation technology: Testing the effectiveness of different application methods to improve tomato plants tolerance against acute heat wave stress

Regenerative agriculture aims to boost native biodiversity and ensure sustainable production. Plant Growth Promoting Bacteria (PGPB) enhance crop yield through improved nutrient use efficiency or stress tolerance. Utilizing synthetic bacterial communities (SynCom) tailored to specific challenges holds promise for enhancing plant resilience. However, challenges include designing microbial consortia and selecting SynCom application techniques. This study tested four PGPB SynCom application methods on tomato plants to evaluate their effectiveness in promoting plant resistance and recovery from short extreme heat waves. Non-invasive phenotyping techniques were used to assess plant responses to three delivery methods (watering, foliar spray, and alginate spheres immobilization) under heat stress. Plants treated with the marine PGPB SynCom through methods like leaf spraying or encapsulation in alginate spheres exhibited improved resilience in terms of Kautsky curve intensity and shape, indicating better photochemical apparatus function. These techniques also facilitated post-stress recovery. Deep photochemical analysis revealed SynCom-induced improvements in the plant's photochemical apparatus, particularly the PS II donor side. Moreover, it was also possible to observe that upon stress relief these two techniques, alongside the plants inoculated throughout watering were able to recover their photochemical profiles. Analysing the Fv/Fm values, plants inoculated through watering, alginate beads and leaf spray can be considered heat-tolerant, a characteristic conserved throughout the three sampling moments. Heat-wave exposure led to an increase in the energy absorbed by the non-inoculated plants as a counteractive measure to overcome the low energy use efficiency observed (low trapping and transported energy fluxes). The reduction in stress impact was attributed to SynCom's ACC-deaminase production, which lowered ethylene accumulation, promoting growth and photosynthetic efficiency. The study also noted reduced energy dissipation under stress relief, indicating efficient heat dissipation mechanisms due to SynCom application. Statistical models generated with the attained photochemical data allowed depicting specific fluorescence signatures of each thermal and inoculation treatment with a very high degree of accuracy, reinforcing that these different inoculation treatments have in fact different impacts on plant photochemistry but also provide a tool for future phenotyping assessments using the proposed inoculation techniques. Alginate bead based PGPB release emerged as an efficient, scalable technique with continuous benefits for plant growth and stress response.

First report of Powdery Mildew Caused by Podosphaera xanthii on Siraitia grosvenorii in Guizhou, China.

Swingle (Siraitia grosvenorii), a member of the Cucurbitaceae family, stands out as a distinctive plant with both economic and medicinal significance. In October 2023, severe powdery mildew were observed on S. grosvenorii in Guiyang City (26.50°N; 106.66°E), Guizhou Province, China. About 80 % of the plants in the greenhouse showed powdery mildew symptoms. Three infected plant samples were selected for morphological and molecular analysis (GZAAS 23-0801, GZAAS 23-0802 and GZAAS 23-0803). The voucher specimens are deposited in the Key Laboratory of Agricultural Biotechnology of Guizhou Province. The symptoms initially manifested as irregular to nearly circular, small yellow spots, with distinct depressions as well as surfaces covered in white mycelium. Over time, these spots gradually expanded and merged patches. In the final stages, the entire leaves turned into yellow and withered. Microscopic observations showed that fungal hyphae were septate, branched, and flexuous to straight and 5 to 9 µm wide, and appressoria were indistinct to slightly nipple-shaped. Conidia were hyaline and ellipsoid to oval with fibrosin bodies and measured 31 to 43 × 18 to 24 μm (n = 50) with a length/width ratio of 1.3 to 2.3. Conidiophores were unbranched, straight, 120 to 268 × 14 to 22 µm (n = 30), producing two to five immature conidia in chains. Foot cells of conidiophores were cylindrical, 39 to 84 × 8 to 14 μm (n = 30), followed by one to three short cells. Short cells were cylindrical, 12 to 32 × 8 to 15 μm (n = 50). The morphological characteristics were identical with the previous description of Podosphaera xanthii (Braun and Cook, 2012). Total DNA was extracted from conidia and mycelia by the Chelex method (Walsh et al., 1991). The ribosomal DNA internal transcribed spacer (ITS) and nuclear ribosomal large subunit (LSU) were amplified by using the primers ITS1/ITS4 (White et al., 1990) and LSU1/LSU2 (Scholin et al., 1994), respectively. The ITS (OR825802, OR825803 and OR825804, respectively) and LSU (OR825805, OR825806 and OR825807, respectively) sequences of three isolates, were deposited in GenBank. The BLAST results revealed that both the ITS and LSU region sequence were 100% identical to those of P. xanthii (ITS: MF043939, MG754404 and KJ698669; LSU: OQ061319, AB936277and OP218411). Phylogenetic analyses of ITS and LSU sequences showed that our three isolates were clustered with P. xanthii (KX842351, LC270782 and LC270779) with high statistical support (ML/MP/BI: 100%/97%/1.00). Combined with their morphological characteristics, these three isolates were identified as P. xanthii. Pathogenicity tests were performed by gently brushing conidia onto the leaves of five healthy S. grosvenorii plants. Five non-inoculated plants were used as the control. All plants were maintained in a greenhouse at 25 ± 2°C. One week after inoculation, similar symptoms were observed in the inoculated plants, whereas no symptoms occurred on the control plants. By microscopic observation, the fungus on the inoculated plants was morphologically identical to those on originally diseased plants. Powdery mildew caused by P. xanthii has been reported on Vernonia cinerea (Wu et al., 2023), Vigna unguiculata (Zhang et al., 2023), Cucumis melo (Meesam et al., 2023). To our knowledge, this is the first report of powdery mildew caused by P. xanthii on S. grosvenorii in Guizhou, China. The occurrence of powdery mildew on S. grosvenorii may pose a potential threat to its large-scale cultivation. The pathogen could become a threat to other Cucurbitaceae members in the future.

Effects of thiobacillus bacteria on physiology and vegetative growth of pomegranate

The climate change and consequent degradation of soil and water resources have led to difficult conditions for farmers; thus, they should make intelligent decisions to face these statuses. Managing soil nutrient and mineral uptake by plants and its feasibility under harsh environment must be considered. Therefore, the present research was conducted to investigate the response of pomegranate variety Shishe Kab to Thiobacillus bacteria (B) and sulfur (S) fertilization. Treatments included sulfur fertilization (three levels: 0, 500 and 1000 g−1 tree) and inoculation with Thiobacillus bacteria (two levels: inoculated and non-inoculated). Totally, sulfur fertilization and inoculation with bacteria improved physiological and biochemical aspects and fruit production of pomegranate. Supplying S decreased soil pH, which the lowest value (5.2) was recorded in 500 g S tree−1 when accompanied with bacteria. Soil electrolyte leakage (EL) increased, when sulfur incorporated or when bacteria added to the soil. The interactive effects of B × S was significant on fruit diameter and weight, chlorophyll (Chl), carotenoid and potassium content, which were improved, although non-inoculated plants also showed high values of potassium content. Anthocyanin and total carbohydrate improved when bacteria inoculation was done; however, total acidity (TA) and total soluble solids (TSS) showed a significant decrease. The EL variable significantly decreased and tissue water content increased in this experiment, when inoculation or sulfur fertilization was used. The results obtained from present study emphasized on the necessity of using sulfur and Thiobacillus bacteria in increasing growth and yield and also chilling tolerance of pomegranate trees.

First report of Irpex rosettiformis causing white root rot in avocado trees in Michoacán, México.

In México, avocado production is an important economic source. In the last season it generated $ 3. 27 billion USD of foreign currency in the country. Irpex spp. are wood decay fungi. In the period 2019-2022, in the state of Michoacán (19°13' N; 101°55' W), México, basidiomes of Irpex sp. were observed on the base of trunks and crowns of 5-years-old and older avocado (Persea americana) trees. The trees exhibited disease symptoms that included white root rot, leaf yellowing, small leaves, branch diebacks, generalized defoliation, apical flaccidity, abundant but small sun burnt fruits due to the lack of foliage, and after 2-4 years of first disease appearance, the infected trees died. In the place where fungus was established, abundant white and cottony mycelium was formed, which caused trees decay. The incidence of the disease in the sampled orchards was estimated to be 30% per ha with 350 - 400 trees, which was determined through a simple sampling design focused on trees with signs and symptoms of the disease due to the phytopathogen. Samples of infected tissue (roots and stems) and fungal basidiomes were collected from 90 trees (5-6 per orchard). The symptomatic avocado trees studied were randomly selected from 17 orchards. For the fungal macroscopic characterization, the synoptic keys described by Gilbertson and Ryvarden (1986) and by Largent (1973) were used. The samples showed typical structures corresponding to Irpex sp., including rosettes, annual basidiomes, a system of monomitic hyphae, and subglobose basidiospores. In vitro fungal isolation from basidiomes and infected tree tissues was done according to the protocol of Agrios (2004). The fungal strains were maintained on PDA at 28 °C. At 16 days of incubation the colonies were opaque, whitish with fluffy and corky mycelium. Microscopic analysis of the fungus showed typical yellowish spores, with an ellipsoid shape of 3-4 x 4-5.5 µm (50 accounted structures per isolate [N=19]) and basidia of 20-25 x 4.5-5.5 µm (n=20 basidiomes). For molecular characterization, two molecular markers were used, the internal transcribed spacer rDNA-ITS1 5.8 rDNA-ITS2 (ITS; White et al. 1990) and the large ribosomal subunit (LSU; Vilgalys and Hester 1990). The PCR reaction was performed as described by Martínez-González et al. (2017). The consensus sequences were compared with those deposited in the NCBI-GenBank, using the BLASTN 2.2.19 tool (Zhang et al. 2000), the samples showed 99% match with the species, Irpex rosettiformis. GenBank accession numbers of the submitted isolates are summarized in supplementary Table 4. To test Koch's postulates, 3-months old avocado plants grown in greenhouse conditions were inoculated (n = 10 per each isolate [N= 19]) on the roots with 3 g of I. rosettiformis mycelium. The experiment was done twice with 20 non-inoculated plants as control. After 67 days, basidiomes (50 x 70 x 1.5 mm in average) were observed where the disease incidence was >77%, with subsequent tree decline. The pathogen was re-isolated in vitro in PDA and its identity was confirmed by morphological characteristics of mycelium. This work shows that I. rosettiformis is not only a wood decay fungus, but also a phytopathogen, the causative agent of white root rot disease in P. americana var. drymifolia, cultivar 'Hass', which establishes a precedent for monitoring and preventing its proliferation to other regions in the American continent and the world where nursery avocado seedlings are exported.

Prevalence and Epidemiology of Apple Powdery Mildew in Northwest Himalayan Region of Himachal Pradesh

A survey was conducted to assess the incidence and severity of powdery mildew of apples during April to July of 2020 and 2021 at the different locations of Kinnaur district, Himachal Pradesh, India. Assessment scale of Hofer et al. (2019) with slight modification was used for cultivar screening. Apple seedlings were grown in the pots in the polyhouse of the Department of Plant Pathology in order to test the pathogenicity of Podosphaera leucotricha. Pathogenicity was confirmed through inoculation tests by gently pressing diseased leaves on young leaves of five asymptomatic, potted one-year-old Gale Gala seedlings of apple. Five non-inoculated plants were used for a control treatment. Overall, twenty different apple cultivars were screened against powdery mildew planted under field conditions at Krishi Vigyan Kendra Sharbo, Kinnaur, Himachal Pradesh, India. Out of these cultivars, five cultivars viz. Red Delicious, Royal Delicious, Red Velox, Auvil Early Fuji and Oregon Spur II, were found to be less susceptible to Podosphaera leucotricha, causing powdery mildew of apples, while Granny Smith, Golden Delicious, and Gale Gala were found extremely susceptible. Gala series of cultivars were found to be extremely susceptible to the pathogen. Incidence of the disease ranged from 25.42% to 53.80% and severity from 39.75% to 66.25%, respectively. According to the present study the disease initiated during the first week of May and peaked in the month of July in district Kinnaur. Temperatures between 18–24°C and RH between 50–65% were found to be favourable for the disease development. The conditions in the region were found to favourable for disease development and spread.

First Report of rattail cactus necrosis-associated virus Infecting Prickly pear (Opuntia macrocentra) in the United States.

Black-spined prickly pear (Opuntia macrocentra Engelmann; Cactaceae) is a cactus native to Arizona, New Mexico, Texas, and northwest Mexico. The plant is often grown for ornamental purposes in the United States. In February 2023, virus-like symptoms such as concentric ringspots and chlorotic spots were observed on O. macrocentra plants grown at the vicinity of Maricopa County Cooperative extension, University of Arizona, Phoenix, AZ (33°24'24.6"N, 111°59'15.3"W). Total RNA was extracted from two samples (YPHC-60-A and YPHC-60-B), following the protocol by Tzanetakis et al. (2007). Reverse transcription polymerase chain reaction (RT-PCR) was performed with degenerate tobamovirus, TobamodF/TobamodR (Li et al. 2018) and potexvirus, 1RC, Potex 2RC, and Potex 5 (van der Vlugt and Berendsen 2002) primers. An expected amplicon of ~880 bp was obtained from both samples using TobamodF/TobamodR primers, while no amplification was observed with potexvirus primers. Further, RT-PCR was carried out using species-specific primers to detect cacti related tobamoviruses: cactus mild mottle virus (CMMoV), rattail cactus necrosis-associated virus (RCNaV) (Park et al. 2018) and Opuntia virus 2 (Salgado-Ortiz et al. 2020). Amplicons of ~540 bp were amplified from both samples using RCNaV specific primers, whereas no amplification was obtained using CMMoV and Opuntia virus 2 specific primers. Then, the amplicons from both YPHC-60 (A-B) isolates (~540 bp) were Sanger sequenced and shared 99.22% nucleotide identity to each other. A BLAST search revealed 93% nucleotide identity with RCNaV CP sequences (KY581586.1, JF729471, and MT130378.1). The sequences were submitted in the GenBank (accessions no. OQ914798 and OR828526). Furthermore, complete RCNaV- RNA dependent RNA polymerase (RdRP) gene was amplified using primers 3490-s-5'GTAGGTGGTACCGCATAGCA-3'; 3490as 5'AAACGCAAGTCMRYGACYGA-3' (designed in this study from accession no. JF729471.1, position 3490-3509 and 4905-4925). The expected amplicons of ~1,500 bp were obtained from both YPHC-60 (A-B) samples and sequenced (GenBank: OQ914799 and OR823954) showing 87.5 % identity with RCNaV sequences (JF729471.1 and NC_016442.1). The maximum-likelihood phylogenetic tree clustered YPHC-60 (A-B) isolates in a single clade with other RCNaV isolates. RCNaV virus particles were isolated from YPHC-60 (A-B) and submitted for RNA extraction, testing positive for RCNaV by RT-PCR. Sap extract of YPHC-60 (A-B) prepared in 0.01 M phosphate buffer (pH =7.0) was used to mechanically inoculate 3 indicator plant species (n=10): Phaseolus vulgaris, Medicago sativa, and Cucumis melo. Also, infected tissue was used to graft Opuntia sp. plants. Symptoms such as local lesions were observed on M. sativa and vein thickening on P. vulgaris 14 days post-inoculation, while Opuntia sp. showed chlorosis 30 days after grafting. RCNaV infection in mechanically inoculated P. vulgaris, M. sativa, and Opuntia sp. was also confirmed through RT-PCR. C. melo and non-inoculated control plants did not show any symptoms, nor tested positive through RT-PCR. RCNaV has been reported earlier to infect cactus species in South Korea (Park et al. 2018) and O. albicarpa in Mexico (De La Torre-Almaráz et al. 2016), where it was found in several orchards. To the best of our knowledge, this is the first report of RCNaV infecting O. macrocentra in the United States. This study highlights that RCNaV is easily transmitted mechanically or by grafting, which could impact the nursery industry as most cacti are clonally propagated.

Eryngium foetidum Reported as a New Host of Meloidogyne enterolobii in the State of Pará, in the Eastern Amazon Region, Brazil

Amazon chicory (Eryngium foetidum L. [Apiaceae]), also known as culantro, is native to Tropical America and the West Indies. It belongs to the unconventional food plants (UFPs) group, and in addition to be consumed as a spice herb, it possesses a wide range of ethnomedicinal uses (Paul et al. 2011). In 2019, in the eastern Amazon region of Brazil, state of Pará, producers of E. foetidum in the municipality of Castanhal (01°15'363” S 047°10'232” W) reported the occurrence of underdeveloped plants with leaf yellowing and a large number of galls in the root system, which are typical symptoms of root-knotting nematode. Soil and root samples were collected and sent to the Nematology Laboratory (LabNema) located at the Faculty of Agrarian and Veterinary Sciences, UNESP, Jaboticabal, São Paulo, Brazil. A total of 46 second-stage juveniles (J2s) were extracted per 100 cm3 of soil, and a total of 460 eggs and J2s Meloidogyne spp. were found per gram of root. Morphological and molecular techniques were used to identify the species. The analysis of the perineal patter of ten females revealed thin striations in an oval shape with a high and semi-trapezoidal dorsal arch. No striations were observed in the perivulvar region. The labial region of the ten males analyzed exhibited a non-prominent labial disc, fused and slightly recessed submedian lips, with no apparent annulations. The morphological characteristics observed in the adults were consistent with those originally described for Meloidogyne enterolobii (Yang; Eisenback, 1983), confirming the species purity of the recovered population. Three individual nematodes had their 18S rDNA region sequenced (Holterman et al. 2006) which showed an average identity of 99.7% with other sequences of M. enterolobii available in the GenBank database. A Bayesian phylogenetic tree was constructed, providing insights into the specific relationship of M. enterolobii recovered from E. foetidum with other related nematodes. Each of the three sequenced nematodes represented a unique haplotype, resulting in their separation into distinct clades. Moreover, the obtained sequences presented polymorphisms that differed from the M. enterolobii sequences already available in the database, highlighting the genetic diversity of this species in relation to its original host (Silva et al. 2021). The species M. enterolobii was also confirmed using species-specific primers for M. incognita, M. javanica, and M. enterolobii (Zijlstra et al. 2000; Tigano et al. 2010). To confirm the pathogenicity of M. enterolobii on E. foetidum, a modified Koch Postulate was conducted. Six seedlings of E. foetidum were transplanted individually to 10-liter pots containing autoclaved soil. Each pot was then inoculated with 5 mL of a suspension containing 3,000 eggs and J2s from the original population of M. enterolobii obtained from E. foetidum. After 90 days, the inoculated plants exhibited root galls with a plentiful egg mass, in contrast to the healthy non-inoculated plants. The average number of M. enterolobii nematodes recovered from the roots of the inoculated plants was 42,040 eggs and J2s, resulting in a reproduction factor (RF) of 14.0. The importance of reporting the occurrence of M. enterolobii in E. foetidum is due to the fact that this plant species is cultivated in a crop rotation system with other vegetables such as lettuce and coriander, which are also hosts of M. enterolobii. Consequently, different crop rotation strategies and control alternatives need to be considered in areas where E. foetidum is grown. This is the first report of E. foetidum serving as a host for the root-knot nematode M. enterolobii worldwide.

Sulfur-Oxidizing Bacteria Alleviate Salt and Cadmium Stress in Halophyte Tripolium pannonicum (Jacq.) Dobrocz.

The aim of this study was to investigate how introducing halophilic sulfur-oxidizing bacteria (SOB) Halothiobacillus halophilus to the growth substrate affects the physiological and biochemical responses of the halophyte Tripolium pannonicum (also known as sea aster or seashore aster) under salt and cadmium stress conditions. This study assessed the plant's response to these stressors and bacterial inoculation by analyzing various factors including the accumulation of elements such as sodium (Na), chloride (Cl), cadmium (Cd) and sulfur (S); growth parameters; levels of photosynthetic pigments, proline and phenolic compounds; the formation of malondialdehyde (MDA); and the plant's potential to scavenge 2,2-Diphenyl-1-picrylhydrazyl (DPPH). The results revealed that bacterial inoculation was effective in mitigating the deleterious effect of cadmium stress on some growth criteria. For instance, stem length was 2-hold higher, the growth tolerance index was 3-fold higher and there was a 20% increase in the content of photosynthetic pigments compared to non-inoculated plants. Furthermore, the SOB contributed to enhancing cadmium tolerance in Tripolium pannonicum by increasing the availability of sulfur in the plant's leaves, which led to the maintenance of an appropriate, about 2-fold-higher level of phenolic compounds (phenylpropanoids and flavonols), as well as chloride ions. The level of MDA decreased after bacterial application in all experimental variants except when both salt and cadmium stress were present. These findings provide novel insights into how halophytes respond to abiotic stress following inoculation of the growth medium with sulfur-oxidizing bacteria. The data suggest that inoculating the substrate with SOB has a beneficial effect on T. pannonicum's tolerance to cadmium stress.

IPD3, a master regulator of arbuscular mycorrhizal symbiosis, affects genes for immunity and metabolism of non-host Arabidopsis when restored long after its evolutionary loss

Arbuscular mycorrhizal symbiosis (AM) is a beneficial trait originating with the first land plants, which has subsequently been lost by species scattered throughout the radiation of plant diversity to the present day, including the model Arabidopsis thaliana. To explore if elements of this apparently beneficial trait are still present and could be reactivated we generated Arabidopsis plants expressing a constitutively active form of Interacting Protein of DMI3, a key transcription factor that enables AM within the Common Symbiosis Pathway, which was lost from Arabidopsis along with the AM host trait. We characterize the transcriptomic effect of expressing IPD3 in Arabidopsis with and without exposure to the AM fungus (AMF) Rhizophagus irregularis, and compare these results to the AM model Lotus japonicus and its ipd3 knockout mutant cyclops-4. Despite its long history as a non-AM species, restoring IPD3 in the form of its constitutively active DNA-binding domain to Arabidopsis altered expression of specific gene networks. Surprisingly, the effect of expressing IPD3 in Arabidopsis and knocking it out in Lotus was strongest in plants not exposed to AMF, which is revealed to be due to changes in IPD3 genotype causing a transcriptional state, which partially mimics AMF exposure in non-inoculated plants. Our results indicate that molecular connections to symbiosis machinery remain in place in this nonAM species, with implications for both basic science and the prospect of engineering this trait for agriculture.