Despite the similarity of the effect of priming on the tomato methylome to responses seen following various other biotic stresses, we did not detect a widespread correspondence between differential methylation and differential gene expression. There was no enrichment of CG or CHG DMRs in genes differentially regulated by pathogen infection. However, consistent with studies discussed above, there was a significant enrichment in genes overlapping CHH hypoDMRs in our DEG lists ( Table 2). Nevertheless, the overall enrichment was relatively low (less than twofold) and the total proportion of DEGs containing CHH DMRs in their promoters was only around 15% (compared with 8% across the entire genome). The majority of pathogen-responsive genes therefore do not contain DMRs (as defined under the parameters applied in our analysis). Furthermore, enrichment of genes with DMRs was similar for DEG lists identified from both control and BABA-treated plants. This suggests that either targetting of DMRs to stress responsive genes is not causally linked with transcriptional priming, or alternatively that our criteria for defining a primed gene are ineffective. For example, genes in the “control only” and “control and BABA” categories that we considered not to be primed, might exhibit enhanced expression in BABA treated leaves at time points other than those sampled here. Regardless, we did not detect differential methylation in the majority of genes exhibiting primed expression. It is likely, therefore, that the changes in DNA methylation associated with BABA treatment are predominantly linked indirectly to the transcriptional response of primed plants challenged with the pathogen. Nevertheless, the set of genes overlapping CHH DMRs was enriched for several GO terms linked with stress responses, especially in relation to oxidative stress ( Table 1). Since oxidative stress is common to many biotic and abiotic stresses, the preferential CHH hypomethylation of genes linked with oxidative stress may be related to the ability of BABA to confer resistance or tolerance against a wide range of different stressors ( Baccelli and Mauch-Mani, 2016; Cohen et al., 2016). Indeed, improved oxidative stress tolerance is commonly reported as a mechanism for BABA priming (e.g., Dubreuil-Maurizi et al., 2010; Pastor et al., 2013; Roylawar and Kamble, 2017). In addition, GO terms linked to general regulation were also over-represented amongst genes with CHH DMRs, which may be one mechanism for the in trans regulation of responses to infection by (de)methylation identified by López Sánchez et al. (2016) and Stassen et al. (2018). IR responses are triggered in the presence of biotic stress and are recognised by the subsequent increase in resistance both to the current challenge, and in many cases, to future challenges too ( de Kesel et al., 2021). An area of significant interest in longer-lasting IR is the process known as priming. As well as immediate activation of inducible defences following stress, plants can also enhance future defences without long-term production of costly defensive metabolites and proteins. The establishment of priming creates a heightened state of alert in which defence responses can be triggered more rapidly and/or to a greater degree in response to a second stress encounter ( Conrath et al., 2015; Mauch-Mani et al., 2017; Wilkinson et al., 2019). Because it does not involve constitutively elevated defence, priming optimises the trade-off between the costs and benefits of defence ( van Hulten et al., 2006). In many cases, priming can be maintained for long periods, extending for the lifetime of the plant, and even in some instances, into the next generation ( Luna et al., 2012; Rasmann et al., 2012; Slaughter et al., 2012). As well as natural infection by pests and diseases, the application of various chemical agents can also trigger immediate IR and longer-term defence priming responses. Such chemicals include endogenous plant hormones and other signalling molecules such as jasmonic acid (JA; Worrall et al., 2012), pipecolic acid ( Bernsdorff et al., 2016) and C 6 green leaf volatiles ( Scala et al., 2013), biological molecules from non-plant sources, such as plant growth-promoting rhizobacteria (PGPR; Pieterse et al., 2014), and synthetic mimetics of biological signalling molecules, such as ( R)-β-homoserine ( Buswell et al., 2018). I saw a patient once who bought a bottle of roots (tonic) at a school gate, drank it and went mad because the drink had marijuana in it. He was never the same again," the doctor claims. Jonsson M, Wratten SD, Landis DA, Gurr GM (2008) Recent advances in conservation biocontrol of arthropods by arthropods. Biol Control 45:172–175

Made from 100 percent natural ingredients, this Baba Roots Herbal Drink gives you the energy and boost you need to take on your day. Home-made roots drinks/tonics have been a part of the Jamaican tradition for a long time. The formulas for these roots/tonics are usually handed down by older members of the family or community members. This powerful, traditional herbal tonic helps to stimulate your nerves, cells and organs, helps improve circulation, alleviate pain and many other benefits. In addition to the presence of CHH DMRs in the promoters and bodies of tomato genes, CHH DMRs were also highly enriched in Class II TEs, i.e., DNA transposons with TIRs. In contrast, TEs were relatively unaffected by BABA treatment at mCG positions. In tomato, TIR TEs are preferentially localised in the chromosome arms along with protein-coding genes, supporting the conclusion that BABA priming is marked by hypomethylation in the chromosome arms. Activation of TEs resulting from demethylation is a well-known response to environmental stress, and where such TEs exist close to protein-coding genes, the impact of their demethylation can extend to an influence on the expression of those genes ( Negi et al., 2016; Horváth et al., 2017; Zhang et al., 2018). Several previous studies have identified priming and/or elevated transcriptional activity of stress responsive genes as a consequence of their proximity to demethylated TEs, and particularly DNA TEs ( Dowen et al., 2012; Wibowo et al., 2016; Geng et al., 2019; Wang et al., 2021). Based on the patterns of changes in methylation and gene expression identified in our experiments, we suggest that priming of stress responses by BABA likely arises from a combination of direct changes in methylation at loci encoding genes that either have direct defence functions or which play regulatory roles during stress, the influence of hypomethylation at loci containing TEs on adjacent defence genes, and in trans regulatory effects that operate over longer distances. Specificity of Priming Memory Sarfraz M, Dosdall LM, Keddie BA (2009b) Host plant quality affects the performance of the parasitoid Diadegma insulare. Biol Control 51:34–41 The company, which is the brainchild of entrepreneur William Webb, has been investing heavily in community outreach programmes in Darliston. In addition, it has been a source of full-time employment for numerous residents. Sarsaparilla helps to keep the major hormones that play a role in the health and function of the reproductive system balanced.The product is Jamaican, and we intend to keep it Jamaican," said Dennisur. "All the ingredients that are used in the production of the Baba Roots drink are from indigenous Jamaican plants. There are also some other herbs that are good for fertility. These include red clover. This herb is a source of bioidentical antioxidants. Red clover can be used to treat menstrual symptoms and promote regular periods. Another herb that is good for fertility in women is red raspberry. This herb serves as a tonic for the uterus. Zimmerli L, Jakab G, Metraux J-P, Mauch-Mani B (2000) Potentiation of pathogen-specific defense mechanisms in Arabidopsis by β-aminobutyric acid. Proc Natl Acad Sci 97:12920–12925 Improve nerve function, helping with anxiety, insomnia, mental clarity, focus, brain fog, scattered mind, depression and other nervous conditions The herb saw palmetto has been used over many years for boosting sexual health in men and it has also been used to treat impotence which is a major concern in fertility.

David Dennisur, the chief executive officer of Baba Roots, said that the company's objective is to keep the tonic as unique as possible while promoting local farmers' productivity. Cohen Y, Niderman T, Mosinger E, Fluhr R (1994) β-aminobutyric acid induces the accumulation of pathogenesis-related proteins in tomato ( Lycopersicon esculentum L.) plants and resistance to late blight infection caused by Phytophthora infestans. Plant Physiol 104:59–66

Funding

Ton J, Jakab G, Toquin V, Flors V, Lavicoli A, Maeder MN, Metraux J-P, Mauch-Mani B (2005) Dissecting the β-aminobutyric acid-induced priming phenomenon in Arabidopsis. Plant Cell 17:987–999 Guerrieri E, Pennacchio F, Tremblay E (1997) Effect of adult experience on in-flight orientation to plant and plant–host complex volatiles in Aphidius ervi Haliday (Hymenoptera, Braconidae). Biol Control 10:159–165 We are trying to bring the product to a level where it can not only be profitable, but you can see the benefit that can be generated for the community, and it will give farmers the benefit of expanding their production," added Dennisur.