Intercropping can enable more efficient resource use and increase yield. Most current studies focus on the correlation between soil nutrients and crop yield under intercropping conditions. However, the mechanisms related to root exudates and soil nutrients remain unclear. Therefore, this study explored the correlation between rhizosphere soil nutrients and root exudates in sugarcane/peanut intercropping. Root extracts, root exudates, rhizosphere soil enzyme activities, and soil nutrients were analyzed and compared in monocultured and intercropped peanut and sugarcane at different growth stages. The root metabolites were annotated using the Kyoto Encyclopedia of Genes and Genomes pathways to further identify the connection between soil nutrients and root exudates. The effects of intercropping differed in peanut and sugarcane at different growth stages, and the difference between podding and pod-filling stages was significant. Intercropping generally had a great effect on peanut; it not only significantly increased the organic acid, soluble sugars, and phenolic acids in root exudates and extracts from peanuts, but also significantly increased rhizosphere soil enzyme activities and soil nutrient levels. Intercropping peanuts promoted fumaric acid secretion from roots and significantly affected the metabolic pathways of alanine, aspartate, and glutamate. Sugarcane/peanut intercropping can increase root exudates and effectively improve soil nutrients. The changes in soil nutrients are closely related to the effects of fumaric acid on alanine, aspartate, and glutamate metabolism.

Cytochrome P450s (CYPs) constitute extensive enzyme superfamilies in the plants, playing pivotal roles in a multitude of biosynthetic and detoxification pathways essential for growth and development, such as the flavonoid biosynthesis pathway. However, CYPs have not yet been systematically studied in the cultivated peanuts (Arachis hypogaea L.), a globally significant cash crop. This study addresses this knowledge deficit through a comprehensive genome-wide analysis, leading to the identification of 589 AhCYP genes in peanuts. Through phylogenetic analysis, all AhCYPs were systematically classified into 9 clans, 43 gene families. The variability in the number of gene family members suggests specialization in biological functions. Intriguingly, both tandem duplication and fragment duplication events have emerged as pivotal drivers in the evolutionary expansion of the AhCYP superfamily. Ka/Ks analysis underscored the substantial influence of strong purifying selection on the evolution of AhCYPs. Furthermore, we selected 21 genes encoding 8 enzymes associated with the flavonoid pathway. The results of quantitative real-time PCR (qRT-PCR) experiments unveiled stage-specific expression patterns during the development of peanut testa, with discernible variations between pink and red testa. Importantly, we identified a direct correlation between gene expression levels and the accumulation of metabolites. These findings offer valuable insights into elucidating the comprehensive functions of AhCYPs and the underlying mechanisms governing the divergent accumulation of flavonoids in testa of different colors.

Aspergillus flavus is a fungal pathogen that infects maize and produces aflatoxins. Host-Induced Gene Silencing (HIGS) has been shown to reduce host infection by various fungal pathogens. Here, the A. flavus alkaline protease (alk) gene was targeted for silencing through HIGS. An RNAi vector carrying a portion of the alk gene was incorporated into the B104 maize genome. Four out of eight transformation events containing the alk gene, Alk-3, Alk-4, Alk-7 and Alk-9, were self-pollinated to T4/T6 generations. At T3, the Alk-transgenic lines showed up to 87% reduction in aflatoxin accumulation under laboratory conditions. T4 transgenic Alk-3 and Alk-7 lines, and T5 and T6 Alk-4 and Alk-9 showed an average of 84% reduction in aflatoxin accumulation compared to their null controls under field inoculations (p < 0.05). F₁ hybrids of three elite maize inbred lines and the transgenic lines also showed significant improvement in aflatoxin resistance (p < 0.006 to p < 0.045). Reduced A. flavus growth and levels of fungal ß-tubulin DNA were observed in transgenic kernels during in vitro inoculation. Alk-4 transgenic leaf and immature kernel tissues also contained about 1000-fold higher levels of alk-specific small RNAs compared to null controls, indicating that the enhanced aflatoxin resistance in the transgenic maize kernels is due to suppression of A. flavus infection through HIGS of alk gene.

The intercropping of peanuts and sugarcane is a sustainable planting model that deserves in-depth research. For this study, two variables, i.e., intercropping status (peanut monocropping or sugarcane/peanut intercropping) and the level of nitrogen fertilization (low, medium, or high), were evaluated to analyze the effects of intercropping and nitrogen application on soil fertility and microbial communities in peanut rhizosphere soil. These analyses revealed that higher nitrogen application led to increased total nitrogen (TN), available nitrogen (AN), and soil organic matter (OM) levels in rhizosphere soil for both monocropped and intercropped peanuts, with a decrease in pH. Monocropped peanuts had higher TN, total phosphorus (TP), and total potassium (TK) levels compared to intercropped peanuts at the same nitrogen level but lower AN content and pH levels. The diversity of microbial communities in the rhizosphere soil of intercropped peanuts was significantly higher than that of monocropped peanuts under high levels of nitrogen fertilizer application. Higher levels of Gemmatimonadetes abundance were observed in intercropping rhizosphere soil, compared to that associated with peanut monocropping under low, middle, and high levels of nitrogen fertilizer application, whereas the opposite trend was observed for Chloroflexi abundance. Nitrospira abundance levels rose gradually in the monocropping treatment group, whereas the opposite trend was evident under intercropping conditions. Further analyses of nitrogen cycle-related genes demonstrated higher levels of nitrogen conversion cycle activity in intercropping peanut rhizosphere soil under low nitrogen levels, whereas nitrogen transformation cycle activity levels were higher in monocropping peanut rhizosphere soil under high levels of nitrogen amendment. It can be concluded that intercropping and nitrogen fertilizer application change the physical and chemical properties of soil, thus affecting the diversity and function of soil microbial communities in the peanut rhizosphere. These results offer a theoretical foundation for more efficient sugarcane/peanut intercropping systems.

Maize is one of the major crops that are susceptible to Aspergillus flavus infection and subsequent aflatoxin contamination, which poses a serious health threat to humans and domestic animals. Here, an RNA interference (RNAi) approach called Host-Induced Gene Silencing (HIGS) was employed to suppress the O-methyl transferase gene (omtA, also called aflP), a key gene involved in aflatoxin biosynthesis. An RNAi vector carrying part of the omtA gene was introduced into the B104 maize line. Among the six transformation events that were positive for containing the omtA transgene, OmtA-6 and OmtA-10 were self-pollinated from T1 to T4, and OmtA-7 and OmtA-12 to the T6 generation. These four lines showed at least an 81.3% reduction in aflatoxin accumulation at the T3 generation under laboratory conditions. When screened under field conditions with artificial inoculation, OmtA-7 at T5 and T6 generations and OmtA-10 at T4 generation showed a reduction in aflatoxin contamination between 60% and 91% (p < 0.02 to p < 0.002). In order to develop commercial maize lines with enhanced aflatoxin resistance, the omtA transgene in OmtA-7 was introduced into three elite inbred lines through crossing, and the resulting crosses also exhibited significantly lower aflatoxin accumulation compared to crosses with non-transgenic controls (p < 0.04). In addition, high levels of omtA-specific small RNAs were only detected in the transgenic kernel and leaf tissues. These results demonstrate that suppression of omtA through HIGS can enhance maize resistance to aflatoxin contamination, and this resistance can be transferred to elite backgrounds, providing a viable and practical approach to reduce aflatoxin contamination in maize.