Soybean Root Rot Research Articles

Bacillus subtilis strain BS06 protects soybean roots from Fusarium oxysporum infection.

Soybean, as a major oil crop, is one of the most widely planted crops in the world. Fusarium oxysporum causes soybean root rot, leading to great economic losses to soybean planting every year globally. Chemical fungicide for controlling soybean F. oxysporum diseases may cause environmental problems and has human health risks. Biological control methods avoid these shortcomings; however, few studies have focused on biocontrol of soybean diseases caused by F. oxysporum. Aiming at this problem, we obtained biocontrol bacteria against soybean F. oxysporum by plate confrontation method. The type of the strain with the highest biocontrol activity was identified by molecular biological methods, and then its biocontrol effects were verified through greenhouse experiments. One of our isolated strain named BS06 strain had the highest activity, which was identified as Bacillus subtilis. Our study showed that BS06 strain could effectively control soybean F. oxysporum disease and significantly reduce F. oxysporum to infect soybean roots. Compared with control and carbendazim treatments, BS06 treatment had higher root biomass, plant height, leaf chlorophyll content, stem base diameter and control efficiency. Our results indicated that BS06 could effectively protect soybean root (BS06 strain might produce substances to inhibitF. oxysporum), which was potentially useful for soybean planting.

Lauric Acid Is a Potent Biological Control Agent That Damages the Cell Membrane of Phytophthora sojae

Sustainable management of plant pathogens is becoming more challenging, and novel solutions are needed. Plant biologically active secondary metabolites are important sources of novel crop protection chemistry. Effective individual compounds of these natural products have the potential to be successful new agrochemicals. In this study, we identified lauric acid (LA) from soybean defense leaf volatiles. LA inhibited the growth of Phytophthora sojae, the causal agent of soybean root rot. It influenced mycelial development, sporangium formation, and zoospore generation and germination by damaging the P. sojae cell membrane. Additionally, we showed that LA and several of its derivatives, such as glycerol monolaurate (GML), had similar biological activities. Both LA and GML were safe to soybean plants when used at less than 0.3 g a.i./plant and could promote soybean growth, implying their potential as eco-friendly biological control agents.

The BTB/POZ domain protein GmBTB/POZ promotes the ubiquitination and degradation of the soybean AP2/ERF-like transcription factor GmAP2 to regulate the defense response to Phytophthora sojae.

Phytophthora root and stem rot in soybean (Glycine max) is a destructive disease worldwide, and hence improving crop resistance to the causal pathogen, P. sojae, is a major target for breeders. However, it remains largely unclear how the pathogen regulates the various affected signaling pathways in the host, which consist of complex networks including key transcription factors and their targets. We have previously demonstrated that GmBTB/POZ enhances soybean resistance to P. sojae and the associated defense response. Here, we demonstrate that GmBTB/POZ interacts with the transcription factor GmAP2 and promotes its ubiquitination. GmAP2-RNAi transgenic soybean hairy roots exhibited enhanced resistance to P. sojae, whereas roots overexpressing GmAP2 showed hypersensitivity. GmWRKY33 was identified as a target of GmAP2, which represses its expression by directly binding to the promoter. GmWRKY33 acts as a positive regulator in the response of soybean to P. sojae. Overexpression of GmBTB/POZ released the GmAP2-regulated suppression of GmWRKY33 in hairy roots overexpressing GmAP2 and increased their resistance to P. sojae. Taken together, our results indicate that GmBTB/POZ-GmAP2 modulation of the P. sojae resistance response forms a novel regulatory mechanism, which putatively regulates the downstream target gene GmWRKY33 in soybean.

English

Soybean root rot is a worldwide soil-borne fungal disease threatening soybean production, causing huge losses in yield and quality of soybean. Fusarium species are well recognized as the important causal agent of Fusarium root rot. To screen the beneficial Bacillus strains with capability of suppressing soybean root rot and evaluate the impact of Bacillus combined with biochar against soybean root rot, a pot experiment was conducted with different treatments. In this study, as potential biological control measures, antagonistic Bacillus isolates and different types of biochar were added to soil separately and excellent antagonistic strains mixed with bamboo biochar were applied to the soil. The results showed that seven Bacillus strains promoted the growth of soybean seedlings and reduced root rot severity by 33 to 61%. Bacillus amylolique faciens NH2 was associated with the lowest incidence of soybean root rot, indicating its bio-control potential. The value of plant height, root length and plant dry weight of soybean in the sterilized soil mixed with biochar separately treatment were superior to those of soybean in the inoculated with pathogen treatment, especially the bamboo biochar treatment reduced soybean root rot caused by Fusarium significantly and which bio-control efficacy was 77.41%. The soybean plants shoot and root dry weights in the biochar mixed B. amylolique faciens NH2 or B. subtilis DBK treatments were increased by17.1, 10.7% and 19.51, 19.64%, respectively, which were significantly higher than those of the inoculated pathogen treatment. Compared to antagonistic strain or biochar individual treatments, the disease control efficiency on soybean root rot was up to 64.86% in NH2 strain mixed with bamboo biochar treatment, which reduced root rot severity significantly and showed a synergistic effect. These results suggest that antagonistic Bacillus strains mixed with biochar can be used as an effective alternative in managing soybean root rot. © 2021 Friends Science Publishers

Bacillus subtilis HSY21 can reduce soybean root rot and inhibit the expression of genes related to the pathogenicity of Fusarium oxysporum

Soybean root rot occurs globally and seriously affects soybean production. To avoid the many disadvantages of chemical fungicides, the addition of Bacillus is gradually becoming an alternative strategy to tackle soybean root rot. However, the molecular mechanism of phytopathogenic fungi in this process by Bacillus inhibition is rarely reported. In this study, we isolated a strain of B. subtilis HSY21 from soybean rhizosphere soil, which had an inhibition rate of 81.30 ± 0.15% (P < 0.05) against Fusarium oxysporum. The control effects of this strain against soybean root rot under greenhouse and field conditions were 63.83% and 57.07% (P < 0.05), respectively. RNA-seq analysis of F. oxysporum after treatment with strain HSY21 revealed 1445 downregulated genes and 1561 upregulated genes. Among them, genes involved in mycelial growth, metabolism regulation, and disease-related enzymes were mostly downregulated. The activities of cellulase, β-glucosidase, α-amylase, and pectin-methyl- galacturonase as well as levels of oxalic acid and ergosterol in F. oxysporum were significantly decreased after HSY21 treatment. These results demonstrated that B. subtilis HSY21 could effectively control F. oxysporum by inhibiting its growth and the expression of pathogenic genes, thus indicating that this strain may be an ideal candidate for the prevention and control of soybean root rot.

Gene-Based Allele Specific Marker for Resistance to Phytophthora sojae in Soybean (Glycine max L.)

Phytophthora sojae is a soil-borne oomycete that causes both pre- and post-emergence damping-off disease in soybean that are present in poorly drained soils. Phytophthora root and stem rot of soybean has become an emerging threat to soybean production in South Korea as soybean cultivation in paddy fields has increased recently. The cultivar Daewon was identified as a genetic source for resistance to P. sojae isolate 2457; moreover, a 573 kb resistance locus was previously mapped on chromosome 3 via linkage analysis using Daepung × Daewon recombinant inbred line (RIL) population. This study aimed to develop a gene-based molecular marker associated with P. sojae resistance using single nucleotide polymorphisms (SNPs) at this locus. Three sets of single nucleotide amplified polymorphism (SNAP) markers were initially designed based on genic SNPs in the identified genomic region. Of these, the marker SNAP-Set2 successfully worked for allele-specific amplification for the respective Daepung and Daewon, as well as 20 RILs derived from crosses of the two cultivars. To validate this marker, 11 soybean germplasms were randomly selected and genotyped, which resulted in reliable allele-specific amplification that agreed with the 180 K Axiom® SoyaSNP array data. Phenotypic evaluation of the 20 RILs and the 11 germplasms subsequently demonstrated that Daepung-type and Daewon-type for the SNAP-Set2 are both associated with susceptibility and resistance to P. sojae isolate 2457. The availability of a molecular marker linked to this resistance locus would expedite the use of this valuable resistance allele in soybean breeding programs for increased resistance to P. sojae.

GmBTB/POZ promotes the ubiquitination and degradation of LHP1 to regulate the response of soybean to Phytophthora sojae

Phytophthora sojae is a pathogen that causes stem and root rot in soybean (Glycine max [L.] Merr.). We previously demonstrated that GmBTB/POZ, a BTB/POZ domain-containing nuclear protein, enhances resistance to P. sojae in soybean, via a process that depends on salicylic acid (SA). Here, we demonstrate that GmBTB/POZ associates directly with soybean LIKE HETEROCHROMATIN PROTEIN1 (GmLHP1) in vitro and in vivo and promotes its ubiquitination and degradation. Both overexpression and RNA interference analysis of transgenic lines demonstrate that GmLHP1 negatively regulates the response of soybean to P. sojae by reducing SA levels and repressing GmPR1 expression. The WRKY transcription factor gene, GmWRKY40, a SA-induced gene in the SA signaling pathway, is targeted by GmLHP1, which represses its expression via at least two mechanisms (directly binding to its promoter and impairing SA accumulation). Furthermore, the nuclear localization of GmLHP1 is required for the GmLHP1-mediated negative regulation of immunity, SA levels and the suppression of GmWRKY40 expression. Finally, GmBTB/POZ releases GmLHP1-regulated GmWRKY40 suppression and increases resistance to P. sojae in GmLHP1-OE hairy roots. These findings uncover a regulatory mechanism by which GmBTB/POZ-GmLHP1 modulates resistance to P. sojae in soybean, likely by regulating the expression of downstream target gene GmWRKY40.

Molecular mapping of quantitative disease resistance loci for soybean partial resistance to Phytophthora sansomeana.

Two soybean QDRL were identified with additive interaction to P. sansomeana isolate MPS17-22. Further analyses uncovered four interaction patterns between the two QDRL and seven additional P. sansomeana isolates. Phytophthora sansomeana is a recently recognized species that contributes to root rot in soybean. Previous studies indicated that P. sansomeana is widely distributed among soybean growing regions and has a much wider host range than P. sojae, a well-known pathogen of soybean. Unlike P. sojae, no known disease resistance genes have been documented that can effectively control P. sansomeana. Therefore, it is important to identify resistance that can be quickly integrated into future soybean varieties. E13901 is an improved soybean line that confers partial resistance to P. sansomeana. A mapping population of 228 F4:5 families was developed from a cross between E13901 and a susceptible improved soybean variety E13390. Using a composite interval mapping method, two quantitative disease resistance loci (QDRL) were identified on Chromosomes 5 (designated qPsan5.1) and 16 (designated qPsan16.1), respectively. qPsan5.1 was mapped at 54.71cM between Gm05_32565157_T_C and Gm05_32327497_T_C. qPsan5.1 was contributed by E13390 and explained about 6% of the disease resistance variation. qPsan16.1 was located at 39.01cM between Gm16_35700223_G_T and Gm16_35933600/ Gm16_35816475. qPsan16.1 was from E13901 and could explain 5.5% of partial disease resistance. Further analysis indicated an additive interaction of qPsan5.1 and qPsan16.1 against P. sansomeana isolate MPS17-22. Marker assisted resistance spectrum analysis and progeny tests verified the two QDRL and their interaction patterns with other P. sansomeana isolates. Both QDRL can be quickly integrated into soybean varieties using marker assisted selection.

First report of Mycoleptodiscus terrestris causing root rot of soybean in Indiana.

During the summers in 2019 and 2020, 137 soybean (Glycine max (L.) Merr) seedlings (V1-V3 stage) showing stunting, delayed emergence, and/or crown lesions were collected at Purdue's Agronomy Center for Research and Education in West Lafayette, Indiana. Four seedlings were stunted with reddish-brown girdled lesions along the hypocotyl and crown, rotted tap and lateral roots, and brown discoloration of the cortex and vascular tissues. Four fungal isolates (AC4, AC58, AC96, and AC127) were recovered by plating surface-sterilized symptomatic root tissue onto water agar plates and incubating on the benchtop until mycelia emerged. The growing hyphal tips were transferred to the semi-selective medium DCPA (Andrews and Pitt 1986). On potato dextrose agar, the fungal colonies developed olivaceous green mycelia which melanized into a mat of black microsclerotia with time and no conidia were observed. On 1.5% water agar plates amended with twice autoclaved soybean leaf and root tissue collected from flowering soybean plants, conidia were formed in sporodochia in darkness at 28 οC within one week. Conidia were 1-2 septate, cylindrical with two setae on either end, and measured 20.8 to 26.4 x 4 to 5.6 μm (average 23.9 x 4.7 μm, n=20). The morphological characters matched with the description of Mycoleptodiscus terrestris (Gerd.) Ostaz (Gerdemann 1953). Species identification was further confirmed by sequencing the internal transcribed spacers (ITS) region of rDNA amplified by ITS1 and ITS4 primers (White et al. 1990) and the translation elongation factor 1 alpha (TEF1-α) gene using 983F and 1567R primers with annealing temperature at 53 ○C (Rehner and Buckley 2005). The sequences were deposited in GenBank under the following accession numbers: ITS: MW002684, MT998441, MW010258, and MW010260; and TEF1-α: MW015941-MW015944. The GenBank BLAST searches revealed 100% identity in the ITS region (accession NR_145373.1) and 99.75% identity in the TEF1-α region (MK495977.1) to M. terrestris. Pathogenicity test was conducted on soybean seedlings (cv. Williams) at V1 growth stage using a root dip assay. Isolate AC58 was grown in a modified cotton seed meal broth (CSMB) to produce microsclerotia as inoculum (Gray 1978; Shearer and Jackson 2006). Microsclerotia concentration was measured using a hemocytometer and adjusted to 1.5 x 104 per ml. Five soybean seedlings each were dipped into inoculum or sterile CSMB for 30 minutes then planted individually in vermiculite-filled Styrofoam cups placed on flooded trays in 16-hr photoperiod light racks at room temperature. Seven days after inoculation, all inoculated plants were visibly stunted with root and crown symptoms identical to field symptoms while all controls were healthy. M. terrestris was successfully re-isolated from inoculated plants, but not from the controls, and identified by morphology and sequencing as above. M. terrestris has been previously reported causing root rot of soybean in Illinois (Gray 1978) and Wisconsin (Smith et al. 1998). To our knowledge, this is the first report of M. terrestris infecting soybean in Indiana. Increased geographic distribution of this pathogen warrants more attention for its control. M. terrestris has been proposed as a biological control agent against multiple aquatic weeds (Verma and Charudattan 1993; Shearer and Jackson 2006). Introduction of this fungus into soybean production regions should be avoided.

First report of Fusarium brachygibbosum causing root rot on soybean in Northeastern China.

In August 2017, soybean root rot plants exhibiting root rot were observed in Baiquan County (47°60'N, 126°10'E), Heilongjiang province, China. The disease occurred on approximately 65% of soybean (cv. Heihe43) plantsroots in five fields (>10 ha). The disease resulted in yellowing or wilting and smaller sized leaves, absence of lateral roots and black lesions on tap roots. Infected root tissues from 10 individual plants (2 plants/each field) were surface disinfested with 0.5% NaOCl for 2 min, rinsed three times in sterile distilled water, placed on potato dextrose agar PDA, and incubated at 26℃ for 3 days. Eight fungal isolates were obtained by transferring hyphal tips.isolated and subcultured by transferring hyphal tips. Colonies on PDA were initially white to rose, then yellow in color with abundant aerial mycelium. The fungal colonies grew to a size of 7.4 cm in diameter four days after inoculation. Macroconidia were scarce and scattered, measuring 19.7 μm× 3.5 μm (n = 50) on carnation leaf agar. Typical macroconidium had 3-5 septa, slightly sharp apices with a distinct basal foot cell. Microconidia had 0-2 septa, and were slightly curved, measuring 10.7 μm × 3.2 μm (n = 50). Spherical chlamydospores had a mean diameter of 13.7μm (n = 50), were terminal and intercalary on PDA. According to these morphological characteristics, the fungus was identified as F. brachygibbosum (Padwick1945). Genomic DNA of a representative isolate P13-1was extracted. The Ef-1α, RPB1 and RPB2 regions were amplified using primers ef1/ef2, Fa/G2R and 5f2/7cr (O'Donnell et al. 2010).The consensus sequences (accession nos. MH748277, MH748278 and MH748279) showed 98.65%, 98.91% and 99.54% identity to the sequences of F. brachygibbosum strain NRRL 34033(accession no.GQ505418.1, HM347172.1 and GQ505482.1). Isolate P13-1was preserved in Agricultural Culture Collection of China, Stock ID number is ACCC 39715.To confirm pathogenicity of P13-1, soybean (cv. Heihe43) seeds were grown in 15-cm pots containing a commercial potting mix (5seeds per pot,3 pots/ treatment). Sorghum seeds (10 g) fully colonized by F. brachygibbosum (Li et al., 2018)were uniformly distributed in each pot and then covered with a 0.5-cm layer of sterile potting soil. , Sterilized sterilized sorghum seeds(10 g) were added to control pots. , incubated in a growth chamber at 25°C (12h day) / 20°C (12h night). 10 Ten days after inoculation , all inoculated plants showed symptoms consistent with those observed in the fields. The experiment was repeated two times. F. brachygibbosum was reisolated from diseased plants and identified as F. brachygibbosum based on morphological and gene sequences analysismolecular characteristics. No fungal pathogens were isolated from nontreated controls. To our knowledge, this is the first report of F. brachygibbosum on soybean in China. The soybean is the prime oil seed crop and the source of protein cultivated in Northeast China and this disease seriously affects the seedling growth. So, our findings are very important for the establishment of control strategies and breeding for resistance to soybean root rot.

Comparison of Phytophthora sojae Populations in Iowa and Nebraska to Identify Effective Rps Genes for Phytophthora Stem and Root Rot Management

Phytophthora stem and root rot (PSRR) of soybean, caused by the oomycete Phytophthora sojae, is prevalent in Iowa and Nebraska. Reducing losses to PSRR primarily relies on growing cultivars with specific resistance (Rps) genes. Predominant genes used in commercial soybean cultivars include Rps 1a, Rps 1c, Rps 1k, and Rps 3a. Knowing which Rps gene to deploy depends on knowledge of which genes are effective against the pathogen. From 2016 to 2018, 326 isolates of P. sojae from were recovered from fields in Iowa and Nebraska and classified into pathotypes based on their virulence on 15 soybean genotypes. A total of 15 and 10 pathotypes were identified in Iowa and Nebraska, respectively. Almost all isolates were virulent on Rps 1a, and over 70% of isolates were virulent on Rps 1c and Rps 1k. Only 2.3% of isolates from Iowa were virulent on Rps 3a. Among commercial soybean cultivars tested in the Illinois Soybean Variety trials from 2010 to 2020, Rps 1c was always the most frequently reported gene, followed by Rps 1k. In contrast, Rps 1a and Rps 3a were present in less than 10% and less than 5% of the cultivars tested, respectively. Because many of the P. sojae isolates in our study were virulent on Rps 1a, Rps 1c, and Rps 1k, soybean cultivars with these genes are unlikely to provide protection against PSRR unless they have a high level of partial resistance.

Phosphite translocation in soybean and mechanisms of Phytophthora sojae inhibition

Although phosphite (Phi)-based fertilizers are used in large quantities in agriculture, the use of Phi-based fungicides against soybean root rot caused by Phytophthora sojae are limited. While, their low toxicity are of high ecological and economic focus. Limited attention has been paid to Phi translocation efficiency in soybeans and the efficacy of Phi as a fungicide against P. sojae. In this study, we evaluated the efficiency of Phi translocation in the Williams soybean cultivar by determining the Phi concentrations in roots, stems, and leaves using high-performance ion chromatography after the application of Phi to the roots. Phi was translocated from roots to leaves within 1 h and its concentration increased significantly in leaves within 36 h after Phi application. Results of an in vitro growth inhibition assay and an in vivo infection assay showed that Phi inhibited P. sojae. Additionally, we examined the activation of the salicylic acid (SA) and ethylene (ET) defense pathways by Phi. The expression of SA and ET pathway–related genes was upregulated in most soybean tissues after Phi application. Our results provide evidence that Phi translocation suppresses root rot caused by P. sojae in soybean.

First report of Fusarium sporotrichioides causing root rot of soybean in Canada and detection of the pathogen in host tissues by PCR

Soybean plants showing symptoms of root rot were collected from fields in Manitoba, Canada, in 2017, to determine the causal agent of the disease. Of 240 isolated strains, including some known as root rot pathogens, five were identified as Fusarium sporotrichioides Sherb. based on morphological characteristics. All five isolates were pathogenic when inoculated on soybean under controlled conditions, and resulted in typical root rot symptoms. In addition, shoot length, root length, and fresh weight were significantly reduced. A PCR-based diagnostic test was developed, which specifically amplified the Trichothecene gene cluster in F. sporotrichioides. The primer SPO1 amplified a 541 bp fragment in fungal isolates originally isolated from commercial soybean crops, and from artificially infected roots of soybean. Trichothecene (TRI) gene expression was detected in infected soybean roots using RT-PCR analysis with TRI gene-specific primers. This is the first report of F. sporotrichioides as one of causal agents of root rot in soybean in Canada.

Mining germplasm panels and phenotypic datasets to identify loci for resistance to Phytophthora sojae in soybean.

Phytophthora sojae causes Phytophthora root and stem rot of soybean and has been primarily managed through deployment of qualitative Resistance to P. sojae genes (Rps genes). The effectiveness of each individual or combination of Rps gene(s) depends on the diversity and pathotypes of the P. sojae populations present. Due to the complex nature of P. sojae populations, identification of more novel Rps genes is needed. In this study, phenotypic data from previous studies of 16 panels of plant introductions (PIs) were analyzed. Panels 1 and 2 consisted of 448 Glycine max and 520 G. soja, which had been evaluated for Rps gene response with a combination of P. sojae isolates. Panels 3 and 4 consisted of 429 and 460 G. max PIs, respectively, which had been evaluated using individual P. sojae isolates with complex virulence pathotypes. Finally, Panels 5-16 (376 G. max PIs) consisted of data deposited in the USDA Soybean Germplasm Collection from evaluations with 12 races of P. sojae. Using these panels, genome-wide association (GWA) analyses were carried out by combining phenotypic and SoySNP50K genotypic data. GWA models identified two, two, six, and seven novel Rps loci with Panels 1, 2, 3, and 4, respectively. A total of 58 novel Rps loci were identified using Panels 5-16. Genetic and phenotypic dissection of these loci may lead to the characterization of novel Rps genes that can be effectively deployed in new soybean cultivars against diverse P. sojae populations.

Role of resistance inducing chemicals against soybean dry root rot (R. bataticola) disease

Role of resistance inducing chemicals against soybean dry root rot (R. bataticola) disease

Transcriptome and metabolite profiling reveals the effects of Funneliformis mosseae on the roots of continuously cropped soybeans

BackgroundArbuscular mycorrhizal fungi are the most widely distributed mycorrhizal fungi, which can form mycorrhizal symbionts with plant roots and enhance plant stress resistance by regulating host metabolic activities. In this paper, the RNA sequencing and ultra-performance liquid chromatography (UPLC) coupled with tandem mass spectrometry (MS/MS) technologies were used to study the transcriptome and metabolite profiles of the roots of continuously cropped soybeans that were infected with F. mosseae and F. oxysporum. The objective was to explore the effects of F. mosseae treatment on soybean root rot infected with F. oxysporum.ResultsAccording to the transcriptome profiles, 24,285 differentially expressed genes (DEGs) were identified, and the expression of genes encoding phenylalanine ammonia lyase (PAL), trans-cinnamate monooxygenase (CYP73A), cinnamyl-CoA reductase (CCR), chalcone isomerase (CHI) and coffee-coenzyme o-methyltransferase were upregulated after being infected with F. oxysporum; these changes were key to the induction of the soybean’s defence response. The metabolite results showed that daidzein and 7,4-dihydroxy, 6-methoxy isoflavone (glycine), which are involved in the isoflavone metabolic pathway, were upregulated after the roots were inoculated with F. mosseae. In addition, a substantial alteration in the abundance of amino acids, phenolic and terpene metabolites all led to the synthesis of defence compounds. An integrated analysis of the metabolic and transcriptomic data revealed that substantial alterations in the abundance of most of the intermediate metabolites and enzymes changed substantially under pathogen infection. These changes included the isoflavonoid biosynthesis pathway, which suggests that isoflavonoid biosynthesis plays an important role in the soybean root response.ConclusionThe results showed that F. mosseae could alleviate the root rot caused by continuous cropping. The increased activity of some disease-resistant genes and disease-resistant metabolites may partly account for the ability of the plants to resist diseases. This study provides new insights into the molecular mechanism by which AMF alleviates soybean root rot, which is important in agriculture.

Occurrence of Pratylenchus coffeae Causing Root Rot of Soybean in Shandong Province of China.

Soybean (Glycine max L.) is a very important commercial crop in China (Li et al. 2019). Pratylenchus coffeae (Zimmermann, 1898) Filipjev & Schuurmans Stekhoven, 1941, is one of the most important root-lesion nematodes that invade the roots of many crops. In August 2018, five root and soil samples were collected in a soybean field, near Xipan village in Linshu county of Linyi City, Shandong Province, China (Fig. S1), to investigate the occurrence of root-lesion nematodes. The collected plants (cv. Lindou No.10) were growing poorly and the roots showed distinct brown lesions (Fig. S2). Pratylenchus spp. were extracted using the modified Baermann funnel method for 2 days (Hooper et al. 2005). On average, 395 root-lesion nematodes per kg of soil and 36 root-lesion nematodes per gram of fresh roots were extracted. The extracted root-lesion nematodes were disinfected with 0.3% streptomycin sulfate and cultured on carrot disks for propagation at 25°C. The species identification was based on morphological and molecular criteria. Key morphological features were determined for females and males. Measurements of females (n = 16) included body length = 561.0 μm ± 37.6 (standard deviation) (520.5 to 654.0 μm), tail length = 30.0 μm ± 1.9 (27.0 to 33.5 μm), stylet = 16.0 μm ± 0.6 (15.0 to 17.5 μm), a = 28.2 ± 2.3 (23.7 to 31.5), b = 6.4 ± 0.5 (5.7 to 7.3), c = 18.7 ± 1.8 (15.7 to 23.8), and V = 80.8% ± 2.1 (76.5 to 83.8%). Measurements of males (n = 16): body length = 511.0 μm ± 28.1 (range= 475.5 to 566.0 μm), tail length = 26.0 μm ± 1.3 (23.5 to 28.5 μm), stylet = 15.0 μm ± 0.5 (14.5 to 16.0 μm), spicule length = 17.0 μm ± 0.9 (16.0 to 18.5 μm), a = 30.8 ± 1.5 (28.0 to 33.2), b = 6.1 ±0.4 (5.6 to 6.9), and c = 19.8 ± 1.3 (18.1 to 22.2) (Fig. S3). All the morphological features of this population matched the description of P. coffeae (Castillo and Vovlas, 2007). DNA was extracted from an individual female as described previously (Wang et al. 2011). The rDNA-internal transcribed spacer (ITS) region and the D2/D3 region of the 28S rRNA gene were amplified by primers 18S/26S (Vrain et al. 1992) and D2A/D3B (De Ley et al. 1999), respectively. The PCR products were purified and sequenced. The obtained sequences of the ITS region (1,253 bp) and the D2/D3 region of 28S rRNA (781 bp) were deposited in GenBank. The ITS sequences of the root-lesion nematode obtained in this study (GenBank Accession no. MT879294) exhibited 99% identity with several P. coffeae sequences available in the GenBank (e.g., KR106219, MT586756, KY424205, and MN749379), and the obtained D2/D3 region sequence (MT879295) exhibited 100% identity with several P. coffeae sequences (e.g., MT586754, MN750755, MK829009, and MH730447). Both morphological and molecular data confirmed the presence of P. coffeae. To confirm reproduction on soybean, the obtained root-lesion nematode population was used in a greenhouse (25°C) assay to fulfill modified Koch's postulates. About 20 days after sowing, eight pots, each with one soybean plant (Lindou No.10) were inoculated with 1000 P. coffeae. The inoculated plants were kept in 1.5 L pots containing 1.2 L sterilized soil. Eight pots of uninoculated soybeans were used as the control. Ten weeks later, the inoculated roots were washed and brown lesions were observed. The number of nematodes/pot was approximately 7360 in soil and 796 in roots, and the reproduction factor was 8.16. Root-lesion nematodes and symptoms were not observed in control groups. P. coffeae has only been reported on soybean in Zhejiang (Wei et al. 2013) and Henan Province (Li et al. 2019) of China. To our knowledge, this is the first report of P. coffeae infecting soybean in Shandong Province, China. Since the root-lesion nematode can cause considerable damage to soybean, care should be taken to prevent the spread of P. coffeae to other regions in China.

Effects of Soil Conditions on Root Rot of Soybean Caused by Fusarium graminearum.

Fusarium graminearum is an important soybean pathogen that causes seedling disease, root rot, and pre- and postemergence damping-off. However, effects of soil conditions on the disease are not well understood. The objective of this greenhouse study was to determine the impacts of soil texture, pH, and soil moisture on seedling root rot symptoms and detrimental effects on seedling development caused by F. graminearum. F. graminearum-infested millet was added (10%, vol/vol) to soil with four different textures (sand, loamy sand, sandy loam, and loam). Soil moisture was maintained at saturation, field capacity or permanent wilting point at soil pH levels of 6 or 8. Seedlings were evaluated 4 weeks after planting for root rot, root length, root and shoot dry weights, leaf area, and F. graminearum colonization (by qPCR). There was a significant interaction between soil moisture and soil texture for root rot assessed visually (P < 0.0001). Highest severity (67%) and amount of F. graminearum DNA were observed at pH 6 and permanent wilting point in sandy loam soils. Pot saturation resulted in the lowest levels of disease in sandy loam and loam soils (11.6 and 10.8%, respectively). Reductions in seedling growth parameters, including root length, foliar area, shoot and root dry weights, and root tips, relative to the noninfested control, were significantly greater in sandy loam soils. In contrast, there were no significant growth reductions in sand. This study showed that levels of root rot increased under moisture-limiting conditions, producing detrimental effects on plant development.

Bacterial strains for biological control of Fusarium root rot of soybean in Siberia

Two strains of bacteria isolated from agricultural soil in Yenisey Siberia were tested for their ability to suppress Fusarium root rot of soybean (Glycine max (L.) Merr.) in vitro. The strain RSA-1 statistically significantly (p<0.001) reduced the disease incidence by 25 percentage points in case of naturally infected soybean seeds and by 29 percentage points in case of artificially infected seeds. The strain RSA-13 statistically significantly (p<0.001) reduced the disease incidence by 32 percentage points in case of naturally infected soybean seeds but did not affect the disease incidence in case of artificially infected seeds. Both strains statistically significantly stimulated growth of infected plants. RSA-1 is Gram-positive, motile, aerobic, rod-shaped spore forming bacterium belonging to the Bacillaceae family. RSA-13 is Gram-positive, aerobic, filamentous actinobacterium belonging to the Actinomycetaceae family. Both strains can be recommended for biological control of Fusarium root rot of soybean in Siberia.

Transcriptomic analyses revealed the effect of Funneliformis mosseae on genes expression in Fusarium oxysporum.

Soybean root rot is a typical soil-borne disease that severely affects the yield of soybean. Funneliformis mosseae is one of the arbuscular mycorrhizal fungi(AMF) dominant strains in soybean continuous cropping soil. The aim of this study was to providing an experimental basis for the study of the molecular mechanism underlying the alleviation of the obstacles associated with the continuous cropping of soybean by AMF. In this study, F. mosseae was inoculated in soil planted with soybean infected with Fusarium oxysporum. The results showed that the incidence of soybean root rot was significantly reduced after inoculation with F. mosseae. In F. mosseae-treated samples, the significantly upregulated genes encoded transmembrane protein in fungal cell membrane. The significantly downregulated genes encoded some proteins, which took part in composition of essential component of fungal cell wall; hydrolyse cellulose and hemicellulose. The DEGs in each treatment were enriched in antigen processing and presentation, carbon fixation in photosynthetic organisms, glycolysis/gluconeogenesis, the MAPK signalling pathway, protein processing in the endoplasmic reticulum and RNA degradation. Inoculation with F. mosseae could in a variety of ways to promote the growth, development of soybean and improve disease resistance. Such as help fungal build barriers to the disease resistance of host plant and enhance their pathogenicity; damaging the structure of the pathogen; protect plant tissues and so on. This study provides an experimental basis for further research on the molecular mechanism underlying the alleviation of challenges associated with the continuous cropping of soybean by AMF.