The use of biofertilizers is important for sustainable agriculture, and the use of nodule bacteria and endophytic actinomycetes is an attractive way to enhance plant growth and yield. This study tested the effects of a biofertilizer produced from Bradyrhizobium strains and Streptomyces griseoflavus on leguminous, cereal, and vegetable crops. Nitrogen fixation was measured using the acetylene reduction assay. Under N-limited or N-supplemented conditions, the biofertilizer significantly promoted the shoot and root growth of mung bean, cowpea, and soybean compared with the control. Therefore, the biofertilizer used in this study was effective in mung bean, cowpea, and soybean regardless of N application. In this study, significant increments in plant growth, nodulation, nitrogen fixation, nitrogen, phosphorus, and potassium (NPK) uptake, and seed yield were found in mung beans and soybeans. Therefore, Bradyrhizobium japonicum SAY3-7 plus Bradyrhizobium elkanii BLY3-8 and Streptomyces griseoflavus are effective bacteria that can be used together as biofertilizer for the production of economically important leguminous crops, especially soybean and mung bean. The biofertilizer produced from Bradyrhizobium and S. griseoflavus P4 will be useful for both soybean and mung bean production.

This study was conducted to evaluate the effects of Bradyrhizobium japonicum SAY3-7, Bradyrhizobium elkanii BLY3-8, and Streptomyces griseoflavus P4 on the symbiotic effectiveness of soybeans before biofertilizer production, to produce biofertilizer containing the studied three strains (SAY3-7, BLY3-8, and P4), to test the effectiveness of the biofertilizer on soybean varieties, and to assess the varietal effects and interaction effects between variety and biofertilizer on plant growth, nodulation, nitrogen fixation, nutrient absorption, and seed yield. Nitrogen fixation was measured using the acetylene reduction assay and ureide methods. Contents of nutrients (N, P, K, Ca, and Mg) were also measured to calculate their uptakes. In this study, synergistic effects of nitrogen fixation were induced by combined inoculation with SAY3-7, BLY3-8 and P4 in all tested soybean varieties. Therefore, we assumed that an effective biofertilizer could be produced using these effective bacteria (SAY3-7, BLY3-8, and P4). After making biofertilizer using these effective bacteria, packages were stored at 30 °C. The populations of the bacteria in the biofertilizer were maintained at a density of 1 × 10⁸ colony forming units (cfu) g⁻¹ for P4 and 7 × 10⁹ cells g⁻¹ for Bradyrhizobium. Diluting biofertilizer by 10⁻³ proved more effective for nodulation and nitrogen fixation than other dilution treatments. Moreover, this biofertilizer significantly promoted plant growth, nodulation, nitrogen fixation, nutrient uptakes, and seed yield in Yezin-3 and Yezin-6 soybean varieties. Yezin-6 is a more efficient variety than Yezin-3 for improved plant growth, nodulation, nitrogen fixation, nutrient absorption, and seed yield. Taken together, the application of an effective biofertilizer and the use of an efficient soybean variety can play important roles in promoting plant growth, nodulation, nitrogen fixation, and higher seed yield.

Aegle marmelos (L.) Correa is an economically and therapeutically valuable tree. It is cultivated as a fruit plant in southeast Asian countries. In this research, we investigated the allelopathy and possible allelochemicals in the leaves of A. marmelos. Aqueous methanol extracts of A. marmelos exhibited significant inhibitory effects against the growth of Lepidium sativum, Lactuca sativa, Medicago sativa, Echinochloa crusgalli, Lolium multiflorum, and Phleum pratense. Bioassay-directed chromatographic purification of the A. marmelos extracts resulted in identifying five active compounds: umbelliferone (1), trans-ferulic acid (2), (E)-4-hydroxycinnamic acid methyl ester (3), trans-cinnamic acid (4), and methyl (E)-3’-hydroxyl-4’-methoxycinnamate (5). The hypocotyl and root growth of L. sativum were considerably suppressed by these compounds. Methyl (E)-3’-hydroxyl-4’-methoxycinnamate also suppressed the coleoptile and root growth of E. crusgalli. The concentrations of these compounds, causing 50% growth reduction (I₅₀) of L. sativum, were in the range of 74.19–785.4 μM. The findings suggest that these isolated compounds might function in the allelopathy of A. marmelos.

Medicinal plants are rich sources of bioactive substances that can be used to develop environmentally friendly weed control alternatives. Marsdenia tenacissima (Roxb.) Moon is a traditional medicinal plant well known for its pharmacological activities and several bioactive compounds. However, its allelopathy and related substances have not been reported. Hence, the present study was conducted to explore the allelopathic potential and substances from M. tenacissima leaves. Aqueous methanol extracts of M. tenacissima showed significant inhibitory activities against the growth of cress (Lepidium sativum L.) and Italian ryegrass (Lolium multiflorum Lam.). The extracts were purified through various chromatography steps, and two allelopathic substances were isolated and determined by spectral data to be steroidal glycoside 1 (5,6-dihydrogen-11α-O-acetyl-12β-O-tigloyl-17β-marsdenin), a novel compound, and steroidal glycoside 2 (5,6-dihydrogen-11α,12β-di-O-tigloyl-17β-marsdenin). Both compounds significantly inhibited the growth of cress seedlings. Steroidal glycoside 1 showed 1.6- and 4-times greater growth inhibitory potential against the cress shoots and roots than steroidal glycoside 2. The concentrations needed for 50% growth inhibition of the cress seedling shoots and roots were 0.46 and 0.03 mM for steroidal glycoside 1, respectively, and 0.74 and 0.12 mM for steroidal glycoside 2, respectively. Therefore, these results suggest that steroidal glycosides 1 and 2 may be responsible for the allelopathy of the M. tenacissima leaves.

Plant parts and extracts that are rich in bioactive substances with allelopathic potential can be explored as a possible alternative to herbicides for natural weed control in sustainable agriculture. In the present study, we investigated the allelopathic potential of Marsdenia tenacissima leaves and its active substances. Aqueous methanol extracts of M. tenacissima showed significant inhibitory activities against the growth of lettuce (Lactuca sativa L.), alfalfa (Medicago sativa L.), timothy (Phleum pratense L.), and barnyard grass (Echinochloa crusgalli (L.) Beauv.). The extracts were purified through various chromatography steps, and one active substance was isolated and determined by spectral data to be a novel compound, assigned as steroidal glycoside 3 (8-dehydroxy-11β-O-acetyl-12β-O-tigloyl-17β-marsdenin). Steroidal glycoside 3 significantly inhibited the seedling growth of cress at a concentration of 0.03 mM. The concentrations needed for 50% growth inhibition of the cress shoots and roots were 0.25 and 0.03 mM, respectively. These results suggest that steroidal glycoside 3 may be responsible for the allelopathy of M. tenacissima leaves.

Investigation of allelopathic substances from herbal plants may lead to the development of allelochemical-based natural herbicides. Croton oblongifolius (Roxb.) is a well-known herbal plant with a long history of being used for traditional medicines and for being the source of a diverse range of bioactive compounds. This plant has been reported to have allelopathic potential; however, its allelopathic-related substances have not yet been described. Therefore, we conducted this investigation to explore the allelopathic substances from the leaves of C. oblongifolius. Aqueous methanol extracts of C. oblongifolius leaves exhibited significant growth inhibitory potential against four test plants (monocot barnyard grass and timothy, and dicot cress and lettuce). The leaf extracts were purified in various chromatographic steps and yielded four active compounds identified as (3R,6R,7E)-3-hydroxy-4-7-megastigmadien-9-one (I), 2-hydroxy alpinolide (a novel compound) (II), alpinolide (III), and epialpinolide (IV) via an analysis of the spectral data. These identified compounds significantly restricted the seedling growth of cress. The concentration necessary for 50% growth reduction of the cress seedlings varied from 0.15 to 0.24 mM for (3R,6R,7E)-3-hydroxy-4-7-megastigmadien-9-one, 0.04 to 0.11 mM for 2-hydroxy alpinolide, 0.07 to 0.12 mM for alpinolide, and 0.09 to 0.16 mM for epialpinolide. Therefore, the leaf extracts of C. oblongifolius and the characterized compounds have the potential to be used as weed-suppressive resources for natural weed control.