Understanding the mechanisms that give rise to obstacles in the continuous cultivation of C. pilosula is essential for addressing or mitigating these challenges. The findings of this study suggest that repeated cultivation significantly reduced the content of polysaccharide in roots, and significantly increased the dead seedling rate in the field. The vascular bundles of the affected plant were extensively colonized by fungi. Furthermore, the root vascular bundles exhibit significant woodiness and corkiness, accompanied by cellular fractures and structural collapse. It was determined that the pathogenic endophyte is Fusarium oxysporum, and the exacerbated disease manifestation corresponds to an acute wilting type. Additionally, the root-zone soil microorganisms Cladosporium austroafricanum, Fusarium foetens, Fusarium petersiae, and Acaulium retardatum may significantly contribute to the yield-reducing phenomenon associated with continuous cropping. The proliferation of pathogenic bacteria during continuous cultivation initiates a complex interaction mechanism between the host plant and these pathogens. This process is characterized by a rapid increase in calcium ion (Ca2+) concentration, which subsequently leads to an upsurge in reactive oxygen species (ROS), particularly manifested as elevated levels of hydrogen peroxide (H2O2). Additionally, this response triggers thickening of cell walls and other immune mechanisms aimed at inhibiting the invasion of pathogenic bacteria. At the same time, to prevent ROS from inducing oxidative damage and triggering oxidative stress, there is a notable increase in both antioxidant enzyme activity and antioxidant substances content.

Calcium-dependent protein kinase (CDPK) is an important mediator for Ca2 + signal recognition and transduction, playing a crucial role in plant stress response. Previous studies have shown that PcCDPK5 may be involved in the response of patchouli to p-hydroxybenzoic acid (p-HBA) stress. In this study, we further found that the subcellular localization of PcCDPK5 protein is in the cytoplasm, and its gene expression is closely related to continuous cropping (CC) and p-HBA stress. Under p-HBA stress, silencing the PcCDPK5 homologous gene in Nicotiana tabacum leads to decreased antioxidant enzyme activity and increased malondialdehyde (MDA) content, significantly accumulating reactive oxygen species (ROS) and affecting normal plant growth. On the contrary, overexpression of PcCDPK5 can effectively alleviate the damage caused by p-HBA stress to plant bodies. Through this research, the function of PcCDPK5 in response to p-HBA stress has been preliminarily analyzed, laying a theoretical foundation for alleviating CC obstacles in patchouli.

Root-knot nematodes (Meloidogyne spp.) have garnered significant attention from researchers owing to the substantial damage they cause to crops and their worldwide distribution. However, controlling these nematodes is challenging because a limited number of chemical pesticides and biocontrol agents are effective against them. Here, we demonstrate that pepper rotation markedly reduces Meloidogyne incognita infection in cucumber and diminishes the presence of p-hydroxybenzoic acid in the soil, a compound known to exacerbate M. incognita infection. Pepper rotation also restructures the rhizobacterial community, leading to the colonization of the cucumber rhizosphere by two Pseudarthrobacter oxydans strains (RH60 and RH97), facilitated by enrichment of palmitic acid in pepper root exudates. Both strains exhibit high nematocidal activity against M. incognita and have the ability to biosynthesize indoleacetic acid and biodegrade p-hydroxybenzoic acid. RH60 and RH97 also induce systemic resistance in cucumber plants and promote their growth. These data suggest that the pepper root exudate palmitic acid alleviates M. incognita infection by recruiting beneficial P. oxydans to the cucumber rhizosphere. Our analyses identify a novel chemical component in root exudates and reveal its pivotal role in crop rotation for disease control, providing intriguing insights into the keystone function of root exudates in plant protection against root-knot nematode infection.