Changes in the Density and Composition of Rhizosphere Pathogenic Fusarium and Beneficial Trichoderma Contributing to Reduced Root Rot of Intercropped Soybean.

The dynamic of soil-borne disease is closely related to the rhizosphere microbial communities. Maize-soybean relay strip intercropping has been shown to significantly control the type of soybean root rot that tends to occur in monoculture. However, it is still unknown whether the rhizosphere microbial community participates in the regulation of intercropped soybean root rot. In this study, rhizosphere Fusarium and Trichoderma communities were compared in either healthy or root-rotted rhizosphere soil from monocultured and intercropped soybean, and our results showed the abundance of rhizosphere Fusarium in intercropping was remarkably different from monoculture. Of four species identified, F. oxysporum was the most aggressive and more frequently isolated in diseased soil of monoculture. In contrast, Trichoderma was largely accumulated in healthy rhizosphere soil of intercropping rather than monoculture. T. harzianum dramatically increased in the rhizosphere of intercropping, while T. virens and T. afroharzianum also exhibited distinct isolation frequency. For the antagonism test in vitro, Trichoderma strains had antagonistic effects on F. oxysporum with the percentage of mycelial inhibition ranging from 50.59-92.94%, and they displayed good mycoparasitic abilities against F. oxysporum through coiling around and entering into the hyphae, expanding along the cell-cell lumen and even dissolving cell walls of the target fungus. These results indicate maize-soybean relay strip intercropping significantly increases the density and composition proportion of beneficial Trichoderma to antagonize the pathogenic Fusarium species in rhizosphere, thus potentially contributing to the suppression of soybean root rot under the intercropping.

Development of a multiplex RT-PCR assay for the detection of soybean mosaic virus, bean common mosaic virus and cucumber mosaic virus in field samples of soybean.

Soybean is susceptible to viral diseases which are often present as mixed infections. The individual simplex RT-PCR methods used for the identification of multiple viruses are more tedious and time-consuming than the corresponding multiplex RT-PCR. This study used soybean mosaic virus (SMV), bean common mosaic virus (BCMV) and cucumber mosaic virus (CMV)-infected leaf samples from southern China as the test materials to evaluate a multiplex RT-PCR assay developed for the simultaneous detection of these viruses. The parameters optimised included the annealing temperature, extension time, number of cycles, and primer type and concentration. The specific fragments sizes obtained by the multiplex RT-PCR were 550 bp (SMV), 288 bp (BCMV) and 99 bp (CMV). The assay was tested using infected soybean samples obtained from farmers' fields in Sichuan Province, China. The multiplex RT-PCR assay had high sensitivity, was rapid and simple, and could be used for the diagnosis of soybean infected with various combinations of these viruses in the field.

Transcriptional Responses of Fusarium graminearum Interacted with Soybean to Cause Root Rot.

Fusarium graminearum is the most devastating pathogen of Fusarium head blight of cereals, stalk and ear of maize, and it has recently become a potential threat for soybean as maize-soybean strip relay intercropping is widely practiced in China. To elucidate the pathogenesis mechanism of F. graminearum on intercropped soybean which causes root rot, transcriptional profiling of F. graminearum at 12, 24, and 48 h post-inoculation (hpi) on soybean hypocotyl tissues was conducted. In total, 2313 differentially expressed genes (DEGs) of F. graminearum were annotated by both KEGG pathway and Gene Ontology (GO) analysis. Among them, 128 DEGs were commonly expressed at three inoculation time points while the maximum DEGs were induced at 24 hpi. In addition, DEGs were also rich in carbon metabolism, ribosome and peroxisome pathways which might contribute to carbon source utilization, sexual reproduction, virulence and survival of F. graminearum when infected on soybean. Hence, this study will provide some basis for the deep understanding the pathogenesis mechanism of F. graminearum on different hosts and its effective control in maize-soybean strip relay intercropping systems.

Diversity of the Seedborne Fungi and Pathogenicity of Fusarium Species Associated with Intercropped Soybean.

Maize/soybean relay strip intercropping has been widely practiced in Southwest China due to its high productivity and effective application of agricultural resources; however, several seedborne diseases such as seedling blight, pod and seed decay are frequently observed causing severe yield loss and low seed quality. So far, the population and pathogenicity of the seedborne fungi associated with intercropped soybean remain unexplored. In this study, seeds of 12 soybean cultivars screened for intercropping were collected from three growing regions in Sichuan Province of Southwest China, and the seedborne fungi were isolated from the surface-sterilized seeds. Based on sequence analysis of ribosomal DNA internal transcribed spacer (rDNA ITS), 148 isolates were identified into 13 fungal genera, among which Fusarium covered 55.0% as the biggest population followed by Colletotrichum. Furthermore, Fusarium isolates were classified into five distinct species comprising F. fujikuroi, F. proliferatum, F. verticillioides, F. asiaticum and F. incarnatum through sequence analysis of translation elongation factor 1 alpha (EF-1α) and DNA-directed RNA ploymerase II second largest subunit (RPB2). Among them, F. fujikuroi accounted for 51.22% (42/82) and was isolated from 91.7% (11/12) soybean varieties. Pathogenicity assay showed that five Fusarium species were able to infect the seeds of soybean cultivar "Nandou12" and caused water-soaked or rot symptoms, while F. fujikuroi and F. asiaticum had much higher aggressiveness than other species with significant reductions of seed fresh weight and germination percentage. Accordingly, this study indicates that Fusarium species are the dominant seedborne fungi in the intercropped soybean in Sichuan, China, and this provides some useful references for the effective management of seedborne fungal diseases as well as soybean resistance breeding in maize/soybean relay strip intercropping.

Maize/Soybean Relay Strip Intercropping Reduces the Occurrence of Fusarium Root Rot and Changes the Diversity of the Pathogenic Fusarium Species.

Fusarium species are the most detrimental pathogens of soybean root rot worldwide, causing large loss in soybean production. Maize/soybean relay strip intercropping has significant advantages on the increase of crop yields and efficient use of agricultural resources, but its effects on the occurrence and pathogen population of soybean root rot are rarely known. In this study, root rot was investigated in the fields of the continuous maize/soybean strip relay intercropping and soybean monoculture. Fusarium species were isolated from diseased soybean roots and identified based on sequence analysis of translation elongation factor 1 (EF-1α) and RNA polymerase II second largest subunit (RPB2), and the diversity and pathogenicity of these species were also analyzed. Our results showed that intercropping significantly decreased soybean root rot over monoculture. A more diverse Fusarium population including Fusarium solani species complex (FSSC), F. incarnatum-equiseti species complex (FIESC), F. oxysporum, F. fujikuroi, F. proliferatum and F. verticillioides, F. graminearum and F. asiaticum was identified from intercropping while FSSC, FIESC, F. oxysporum, F. commune, F. asiaticum and F. meridionale were found from monoculture. All Fusarium species caused soybean root infection but exhibited distinct aggressiveness. The most aggressive F. oxysporum was more frequently isolated in monoculture than intercropping. FSSC and FIESC were the dominant species complex and differed in their aggressiveness. Additionally, F. fujikuroi, F. proliferatum and F. verticillioides were specifically identified from intercropping with weak or middle aggressiveness. Except for F. graminearum, F. meridionale and F. asiaticum were firstly reported to cause soybean root rot in China. This study indicates maize/soybean relay strip intercropping can reduce soybean root rot, change the diversity and aggressiveness of Fusarium species, which provides an important reference for effective management of this disease.

Effects of contrasting shade treatments on the carbon production and antioxidant activities of soybean plants.

In China, maize-soybean relay-intercropping system follow the two main planting-patterns: (i) traditional relay-intercropping; maize-soybean equal row planting, where soybean experience severe maize shading on both sides of plants, and (ii) modern relay-intercropping; narrow-wide row planting, in this new planting pattern only one side of soybean leaves suffer from maize shading. Therefore, in this study, changes in morphological traits, cytochrome content, photosynthetic characteristics, carbon status, and the activities of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT) and ascorbate peroxidase (APX) were investigated at 30 days after treatment (DAT) in shade-tolerant soybean variety Nandou-12 subjected to three different types of shading conditions; normal light (NL, all trifoliate-leaves of soybean plants were under normal light); unilateral shade (US, all right-side trifoliate-leaves of soybean plants from top to bottom were under shade while all the left-side of trifoliate-leaves from top to bottom were in normal light); bilateral shade (BS, all trifoliate-leaves of soybean plants were under complete shade). Compared with BS, US conditions decreased plant height and increased stem diameter, leaf area, and biomass at 30 DAT. Biomass distribution rates to stem, petiole and leaves, and photosynthetic characteristics were markedly improved by the US at all sampling stages, which proved to be a better growing condition than BS with respect to shade tolerance. The enhanced net photosynthesis and transpiration rates in the left-side leaves (LS) of soybean plants in US, when compared with the LS in BS, allowed them to produce higher total soluble sugar (by 70%) and total soluble protein (by 17%) at 30 DAT which reduce the adverse effects of shading at right-side leaves (RS) of the soybean plants. Similarly, soybean leaves under US accumulated higher proline content in US than the leaves of BS plants. Soybean leaves grown in shading conditions (LS and RS of BS and RS of US) developed antioxidative defence-mechanisms, including the accelerated activities of SOD, POD, APX, and CAT. Comparatively, soybean leaves in US displayed lower activity levels of the antioxidative enzymes than the leaves of BS plants, showing that soybean plants experienced less shade stress in US as compared with BS treatment. Overall, these results indicate that the association of improved photosynthetic characteristics, sugar and protein accumulation and optimum antioxidative defences could be an effective approach for growing soybean in intercropping environments.

Characterization and Pathogenicity of Fusarium Species Associated with Soybean Pods in Maize/Soybean Strip Intercropping.

Intercropping has been considered as a kind of a sustainable agricultural cropping system. In southwest China, maize/soybean strip intercropping has commonly been practised under local limited agricultural land resources. However, heavy rainfall in combination with high humidity and low temperatures cause severe pod and seed deterioration in the maturity and pre-harvesting stages of intercropped soybean. Numerous Fusarium species have been reported as the dominant pathogens of soybean root rot, seedling blight, as well as pod field mold in this area. However, the diversity and pathogenicity of Fusarium species on soybean pods remain unclear. In the current study, diseased soybean pods were collected during the cropping season of 2018 from five different intercropped soybean producing areas. A total of 83 Fusarium isolates were isolated and identified as F. fujikuroi, F. graminearum, F. proliferatum, and F. incarnatum-equiseti species complex based on morphological characteristics and phylogenetic analysis of the nucleotide sequence of EF1-α and RPB2 genes. Pathogenicity tests demonstrated that all Fusarium species were pathogenic to seeds of the intercropped soybean cultivar Nandou12. Fusarium fujikuroi had the maximum disease severity, with a significant reduction of seed germination rate, root length, and seed weight, followed by F. equiseti, F. graminearum, F. proliferatum, and F. incarnatum. Additionally, the diversity of Fusarium species on soybean pods was also considerably distinct according to the geographical origin and soybean varieties. Thus, the findings of the current study will be helpful for the management and resistance breeding of soybean pod decay in the maize/soybean intercropping system.

Growth and development of soybean under changing light environments in relay intercropping system.

BACKGROUND: Maize-soybean relay-intercropping (MSR) is a famous system of crop production in developing countries. However, maize shading under this system directly affects the light quality and intensity of soybean canopy. This is a challenging scenario in which to implement the MSR system, in terms of varieties selection, planting pattern, and crop management since the duration of crop resource utilization clearly differs. METHODS: Therefore, this experiment aimed to elucidate the effect of leaf excising treatments from maize top to fully clarify the needs and balance of light quality and intensity of intercrop-soybean under MSR in field conditions. The effects of different leaf excising treatments (T0, no removal of leaves; T2, removal of two topmost leaves; T4, removal of four topmost leaves; T6, removal of six topmost leaves from maize plants were applied at first-trifoliate stage (V1) of soybean) on photosynthetically active radiation transmittance (PART), red to far-red ratio (R:FR), morphological and photosynthetic characteristics and total biomass production at second-trifoliate stage (V2), fifth-trifoliate stage (V5), and flowering-stage (R1) of soybean were investigated through field experiments for 2-years under MSR. RESULTS: As compared to T0, treatment T6 increased the PART and R:FR ratio at soybean canopy by 77% and 37% (V2), 70% and 34% (V5), and 41% and 36% (R1), respectively. This improved light environment in T6 considerably enhanced the leaf area index, SPAD values and photosynthetic rate of soybean plants by 66%, 25% and 49% at R1, respectively than T0. Similarly, relative to control, T6 also increased the stem diameter (by 29%) but decreased the plant height (by 23%) which in turn significantly increased stem breaking strength (by 87%) by reducing the lodging rate (by 59%) of soybean plants. Overall, under T6, relay-cropped soybean produced 78% of sole soybean seed-yield, and relay-cropped maize produced 81% of sole maize seed-yield. Our findings implied that by maintaining the optimum level of PART (from 60% to 80%) and R:FR ratio (0.9 to 1.1), we can improve morphological and photosynthetic characteristics of soybean plants in MSR. Therefore, more attention should be paid to the light environment when considering the sustainability of MSR via appropriate planting pattern selection.