Microalgae cultivation in wastewater is an emerging approach to remove its contaminants and generate microalgal biomass. This study aimed to screen and isolate potential strains in a cassava biogas effluent wastewater (CBEW) treatment system and produce algal biomass. Chlorella sorokiniana strains P21 and WB1DG were isolated from CBEW and found to grow by utilizing various carbon sources. Experiments conducted in a batch reactor using an unsterilized substrate were done to evaluate the nutrient removal and growth of isolated strains from CBEW. The results showed that C. sorokiniana P21 and WB1DG could achieve biomass accumulation of more than 2564 and 1301 mg L−1, respectively. The removal efficiencies of chemical oxygen demand (COD), total phosphorous (TP), and total inorganic nitrogen (TIN) were found up to be 63.42, 91.68, and 70.66%, respectively, in a WB1DG culture and 73.78, 92.11, and 67.33%, respectively, in a P21 culture. Harvestability of the P21 strain was examined using several coagulant–flocculants. FeCl3 was found to remove more than 90% of the cells. Nutrient removal and growth rates resulting from these indigenous strains with application of untreated CBEW support the possibility of this strain being a promising candidate to couple a CBEW treatment and algal biomass generation with minimal process adjustment.
Various technologies, such as light-emitting diodes (LEDs) and beneficial plant micro-organisms, have been applied to enhance plant growth and development. We aimed to develop appropriate technology by incorporating the benefits of LED light, plant growth promoting rhizobacteria (PGPR), and arbuscular mycorrhiza fungi (AMF) into sweet girl cherry tomato (Solanum lycopersicum L.) seedling production. Our results demonstrated that incorporating red (R) and blue (B) LED lights, PGPR, and AMF positively affected tomato seedling growth. The optimal lighting conditions for tomato seedling growth were LEDs at 200 mol/m2/s with a ratio of R60:B40 and 20 h/d exposure. The optimum LED-illuminated tomato seedlings significantly upregulated photosynthesis-related genes, including psbA, psbB, fdx, atpB, and rbcL. Plants inoculated with PGPR Bradyrhizobium sp. SUTN9-2, Bacillus velezensis SD10 and B. megaterium A20 had a high health index after inoculation. Furthermore, the optimized LED-illuminated tomato seedlings inoculated with SD10 had the highest health index. In addition, the optimum LED-illuminated tomato seedlings inoculated with SD10 and AMF had the highest biomass. Our experiment demonstrated that tomato seedlings produced under optimized LED lights inoculated with SD10 and AMF increased yield by about 16% under field conditions. Therefore, these results provided the optimum conditions for a high-quality tomato seedling production system.
A substrate-based production system is a simple and low-cost method for arbuscular mycorrhizal (AM) fungal inoculum production. However, it is time-consuming and typically yields low numbers of AM fungal spores due to several factors affecting plant growth efficiency. Our study investigated the use of light-emitting diode (LED) technology to expedite AM fungal spore production in planta. We performed experiments with Rhizophagus irregularis inoculated in maize (Zea mays L.), contrasting LED light with greenhouse (GH) conditions. Our results exhibited a significant improvement in AM fungal colonization and spore production, as well as a reduction in the production period from 120 to 90 days under the LED light condition. This was achieved using a red-and-blue light ratio of 60:40 with a total light intensity of 300 µmol m−2 s−1. The LED light treatments improved maize growth by increasing nitrogen (N) and phosphorus (P) concentrations in shoots and roots, respectively. Our gene expression analyses revealed that in AMF-inoculated plants, genes related to photosynthesis were significantly upregulated under LED light compared to the GH condition. Moreover, LED increased the expression of marker genes linked to the AM fungi-related cell cycle, indicating enhanced AM fungal growth during symbiosis. These findings advance our comprehension of LED applications in agriculture, offering promising prospects for acceleration of AM fungal spore production.
Mung bean (Vigna radiata L.), a vital legume in Asia with significant nutritional benefits, is highly susceptible to Cercospora leaf spot (CLS) caused by Cercospora canescens, leading to significant yield losses. As an alternative to chemical fungicides, bio-priming with rhizobacteria can enhance plant resistance. This study explores the potential of Bradyrhizobium sp. strain DOA9 to augment resistance in mung bean against CLS via root priming. The results reveal that short (3 days) and double (17 and 3 days) priming with DOA9 before fungal infection considerably reduces lesion size on infected leaves by activating defense-related genes, including Pti1, Pti6, EDS1, NDR1, PR-1, PR-2, Prx, and CHS, or by suppressing the inhibition of PR-5 and enhancing peroxidase (POD) activity in leaves. Interestingly, the Type 3 secretion system (T3SS) of DOA9 may play a role in establishing resistance in V. radiata CN72. These findings suggest that DOA9 primes V. radiata CN72′s defense mechanisms, offering an effective bio-priming strategy to alleviate CLS. Hence, our insights propose the potential use of DOA9 as a bio-priming agent to manage CLS in V. radiata CN72, providing a sustainable alternative to chemical fungicide applications.
The symbiotic interaction between leguminous and Bradyrhizobium sp. SUTN9-2 mainly relies on the nodulation process through Nod factors (NFs), while the type IV secretion system (T4SS) acts as an alternative pathway in this symbiosis. Two copies of T4SS (T4SS1 and T4SS2) are located on the chromosome of SUTN9-2. ΔT4SS1 reduces both nodule number and nitrogenase activity in all SUTN9-2 nodulating legumes. The functions of three selected genes (copG1, traG1, and virD21) within the region of T4SS1 were examined. We generated deleted mutants and tested them in Vigna radiata cv. SUT4. ΔtraG1 and ΔvirD21 exhibited lower invasion efficiency at the early stages of root infection but could be recently restored. In contrast, ΔcopG1 completely hindered nodule organogenesis and nitrogenase activity in all tested legumes. ΔcopG1 showed low expression of the nodulation gene and ttsI but exhibited high expression levels of the T4SS genes, traG1 and trbE1. The secreted proteins from ΔT4SS1 were down-regulated compared to the wild-type. Although ΔcopG1 secreted several proteins after flavonoid induction, T3SS (nopP and nopX) and the C4-dicarboxylate transporter (dct) were not detected. These results confirm the crucial role of the copG1 gene as a novel key regulator in the symbiotic relationship between SUTN9-2 and legumes.
8 August 2025 in Applied and Environmental Microbiology
Symbiosis between Bradyrhizobium strains isolated from Lao People’s Democratic Republic (Lao PDR) and intercropped legumes ( Arachis hypogaea , Vigna radiata , and V. mungo ) was regulated by the type III secretion system (T3SS), which delivers effector proteins (T3Es) into host plant cells to modulate nodulation. To explore this mechanism, we sequenced and analyzed seven Bradyrhizobium genomes, identifying putative T3Es across five T3SS groups (G.1–G.5), which were classified based on the sequence of rhcN , a conserved ATPase gene essential for T3SS function. Phylogenetic analysis of rhcN more closely reflected the evolutionary relationships of nodulation genes than those based on 16S rRNA or whole-genome comparisons, underscoring its symbiotic relevance. Functional assays using rhcN mutants revealed group-specific effects on nodulation; G.1 strains showed neutral effects on A. hypogaea , negative effects on V. radiata , and positive effects on V. mungo . G.2 strains consistently promoted nodulation across all hosts and lacked effectors related to SUMO (small ubiquitin-like modifier) pathways, which have been implicated in host defense regulation. G.3 strains reduced nodulation in A. hypogaea but enhanced it in Vigna species. G.4 strains suppressed nodulation in A. hypogaea , and G.5 strains inhibited nodulation across all tested legumes. These findings highlight the diversity in T3SS organization, effector composition, and symbiotic responses among native Bradyrhizobium strains. The identification of known and uncharacterized effectors suggests roles in host compatibility and specificity. These strains, along with their effector profiles, provide a foundation for future functional studies to better understand T3SS-mediated interactions and support the development of targeted inoculants for legume hosts. IMPORTANCE: This study advances our understanding of legume– Bradyrhizobium symbiosis by examining the genetic organization and evolutionary patterns of T3SS genes. Our findings revealed that T3SS gene evolution does not always align with phylogenies based on 16S rRNA or whole-genome sequences, suggesting that horizontal gene transfer and functional adaptation may shape diversification. The observed variation in T3SS architecture and effector profiles among the five distinct Bradyrhizobium groups was correlated with host-specific nodulation outcomes in A. hypogaea , V. radiata , and V. mungo . We also identified novel candidate genes influencing symbiotic signaling and compatibility. These insights into the diversity and function of T3SS components contribute to a broader understanding of host–microbe communication and may support the development of more targeted and efficient rhizobial inoculants for sustainable legume cultivation and improved biological nitrogen fixation.