Mechanism of benzoxazinoids affecting the growth and development of Fusarium oxysporum f. sp. fabae.

Continuous cropping of faba bean (Vicia faba L.) has led to a high incidence of wilt disease. The implementation of an intercropping system involving wheat and faba bean can effectively control the propagation of faba bean wilt disease. To investigate the mechanisms of wheat in mitigating faba bean wilt disease in a wheat-faba bean intercropping system. A comprehensive investigation was conducted to assess the temporal variations in Fusarium oxysporum f. sp. fabae (FOF) on the chemotaxis of benzoxazinoids (BXs) and wheat root through indoor culture tests. The effects of BXs on FOF mycelial growth, spore germination, spore production, and electrical conductivity were examined. The influence of BXs on the ultrastructure of FOF was investigated through transmission electron microscopy. Eukaryotic mRNA sequencing was utilized to analyze the differentially expressed genes in FOF upon treatment with BXs. FOF exhibited a significant positive chemotactic effect on BXs in wheat roots and root secretions. BXs possessed the potential to exert significant allelopathic effects on the mycelial growth, spore germination, and sporulation of FOF. In addition, BXs demonstrated a remarkable ability to disrupt the structural integrity and stability of the membrane and cell wall of the FOF mycelia. BXs possessed the capability of posing threats to the integrity and stability of the cell membrane and cell wall. This ultimately resulted in physiological dysfunction, effectively inhibiting the regular growth and developmental processes of the FOF.

Benzoxazinoids secreted by wheat root weaken the pathogenicity of Fusarium oxysporum f. sp. fabae by inhibiting linoleic acid and nucleotide metabolisms.

KEY MESSAGE: The regulatory action of BXs secreted by wheat on the pathogenicity of FOF causing Fusarium wilt in faba bean were analyzed. DIMBOA and MBOA weakened the pathogenicity of FOF. A large number of pathogenic bacteria in continuous cropping soil infect faba bean plants, leading to the occurrence of wilt disease, which restricts their production. Faba bean-wheat intercropping is often used to alleviate this disease. This study investigates the effect of benzoxazinoids (BXs) secreted by wheat root on the pathogenicity of Fusarium oxysporum f. sp. Fabae (FOF) and underlying molecular mechanisms. The effects of DIMBOA(2,4-dihydroxy-7-methoxy-1,4-benzoxazine-4-one) and MBOA(6-methoxybenzoxazolin-2-one) on the activity of cell-wall-degrading enzymes in FOF(cellulase, pectinase, amylase, and protease), FOF Toxin (fusaric acid, FA) content were investigated through indoor culture experiments. The effect of BXs on the metabolic level of FOF was analyzed by metabonomics to explore the ecological function of benzoxazines intercropping control of Fusarium wilt in faba bean. The results show that the Exogenous addition of DIMBOA and MBOA decreased the activity of plant-cell-wall-degrading enzymes and fusaric acid content and significantly weakened the pathogenicity of FOF. DIMBOA and MBOA significantly inhibited the pathogenicity of FOF, and metabolome analysis showed that DIMBOA and MBOA reduced the pathogenicity of FOF by down-regulating related pathways such as nucleotide metabolism and linoleic acid metabolism, thus effectively controlling the occurrence of Fusarium wilt in faba bean.

Allelopathic effect of phenolic acids in various extracts of wheat against Fusarium wilt in faba bean.

Allelopathy is the main reason for disease control in intercropping systems. The effects of different extracts, root secretions and phenolic acids of wheat and faba bean on Fusarium oxysporum f. fabae (FOF) growth were studied to explore the allelopathy mechanism of wheat in disease control of faba bean. Various extracts and root exudate of faba bean were promoted but those of wheat inhibited the growth and reproduction of FOF. High-performance liquid chromatography revealed significant differences in the contents of phenolic acids in the various extracts and root exudate of wheat and faba bean. The total content of syringic acid (SA) was much higher, but that of other five phenolic acids were lower in wheat than in faba bean. The in vitro addition of these phenolic acids revealed that cinnamic acid (CA), p-hydroxybenzoic acid (PHBA), benzoic acid (BA), vanillic acid (VA) and ferulic acid (FA) exhibited significant promoting effects and SA exhibited strong inhibitory effects on the growth of FOF. These results suggest that the inhibitory effect of various extracts and root exudates from wheat on FOF growth may be due to differences in phenolic acid content and high levels of SA.

Nitrogen Fertilization in a Faba Bean-Wheat Intercropping System Can Alleviate the Autotoxic Effects in Faba Bean.

Continuous cultivation of the faba bean will lead to its autotoxicity. Faba bean-wheat intercropping can effectively alleviate the autotoxicity of the faba bean. In order to investigate the autotoxicity of water extracts of various parts of the faba bean, we prepared water extracts of various parts of the faba bean, such as the roots, stems, leaves, and rhizosphere soil. The results showed various parts of the faba bean significantly inhibited the germination of faba bean seeds. The main autotoxins in these parts were analyzed using HPLC. Six autotoxins, namely, p-hydroxybenzoic acid, vanillic acid, salicylic acid, ferulic acid, benzoic acid, and cinnamic acid, were identified. The exogenous addition of these six autotoxins significantly inhibited the germination of faba bean seeds in a concentration-dependent manner. Furthermore, field experiments were conducted to investigate the effects of various levels of nitrogen fertilizer on the autotoxin content and the aboveground dry weight of the faba bean in a faba bean-wheat intercropping system. The application of various levels of nitrogen fertilizer in the faba bean-wheat intercropping system could significantly reduce the content of autotoxins and increase the aboveground dry weight in faba bean, particularly at the N2 level (90 kg/hm2). The above results showed that the water extracts of faba bean roots, stems, leaves, and rhizosphere soil inhibited faba bean seed germination. The autotoxicity in faba bean under continuous cropping could be caused by p-hydroxybenzoic acid, vanillic acid, salicylic acid, ferulic acid, benzoic acid, and cinnamic acid. The autotoxic effects in the faba bean were effectively mitigated by the application of nitrogen fertilizer in a faba bean-wheat intercropping system.

tuberculosis detection via rolling circle amplification.

Hybridization-based assays for DNA detection often use single-stranded DNA (ssDNA) probes to capture ssDNA targets in solution. Unfortunately, these assays are often not able to detect double-stranded DNA (dsDNA). Here, we achieve highly sensitive dsDNA target detection by including short oligonucleotide sequences during denaturing and cooling. After performing an isothermal nucleic acid amplification technique (Rolling Circle Amplification, RCA), these captured dsDNA targets are labeled, allowing single amplified molecules to be imaged and counted. This detection method was first applied to the detection of PCR-generated (polymerase chain reaction) dsDNA targets, yielding a limit of detection of 4.25 fM. As an application of the developed assay, the detection of extracted Mycobacterium tuberculosis (M. tb.) genomic DNA was attempted. A M. tb.-specific target was detected with high specificity compared to similar bacteria, and a detection limit of 10 000 colony forming units (cfu) ml-1 was achieved, close to the sensitivity required for clinical diagnosis.