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Biopesticide Trunk Injection Into Apple Trees: A Proof of Concept for the Systemic Movement of Mint and Cinnamon Essential Oils.

ABSTRACT: The use of conventional pesticides is debated because of their multiple potential adverse effects on non-target organisms, human health, pest resistance development and environmental contaminations. In this setting, this study focused on developing alternatives, such as trunk-injected essential oil (EO)-based biopesticides. We analysed the ecophysiology of apple trees (Malus domestica) following the injection of Cinnamomum cassia and Mentha spicata nanoemulsions in the tree's vascular system. Targeted and untargeted volatile organic compounds (VOCs) analyses were performed on leaf-contained and leaf-emitted VOCs and analysed through dynamic headspace-gas chromatography-mass spectrometry (DHS-GC-MS) and thermal desorption unit (TDU)-GC-MS. Our results showed that carvone, as a major constituent of the M. spicata EO, was contained in the leaves (mean concentrations ranging from 3.39 to 19.7 ng g_DW ^-1) and emitted at a constant rate of approximately 0.2 ng g_DW ^-1 h^-1. Trans-cinnamaldehyde, C. cassia's major component, accumulated in the leaves (mean concentrations of 83.46 and 350.54 ng g_DW ^-1) without being emitted. Furthermore, our results highlighted the increase in various VOCs following EO injection, both in terms of leaf-contained VOCs, such as methyl salicylate, and in terms of leaf-emitted VOCs, such as caryophyllene. Principal component analysis (PCA) highlighted differences in terms of VOC profiles. In addition, an analysis of similarity (ANOSIM) and permutational multivariate analysis of variance (PERMANOVA) revealed that the VOC profiles were significantly impacted by the treatment. Maximum yields of photosystem II (Fv/Fm) were within the range of 0.80-0.85, indicating that the trees remained healthy throughout the experiment. Our targeted analysis demonstrated the systemic translocation of EOs through the plant's vascular system. The untargeted analysis, on the other hand, highlighted the potential systemic acquired resistance (SAR) induction by these EOs. Lastly, C. cassia and M. spicata EOs did not appear phytotoxic to the treated trees, as demonstrated through chlorophyll fluorescence measurements. Hence, this work can be seen as a proof of concept for the use of trunk-injected EOs given the systemic translocation, increased production and release of biogenic VOCs (BVOCs) and absence of phytotoxicity. Further works should focus on the ecological impact of such treatments in orchards, as well as apple quality and production yields.

SUBMITTER: Werrie PY

PROVIDER: S-EPMC8063119 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Two methods were developed for the generation of cisgenic apples. Both have been successfully applied producing trees. The first method avoids the use of any foreign selectable marker genes; only the gene-of-interest is integrated between the T-DNA border sequences. The second method makes use of recombinase-based marker excision. For the first method we used the MdMYB10 gene from a red-fleshed apple coding for a transcription factor involved in regulating anthocyanin biosynthesis. Red plantlets were obtained and presence of the cisgene was confirmed. Plantlets were grafted and grown in a greenhouse. After 3 years, the first flowers appeared, showing red petals. Pollination led to production of red-fleshed cisgenic apples. The second method used the pM(arker)F(ree) vector system, introducing the scab resistance gene Rvi6, derived from apple. Agrobacterium-mediated transformation, followed by selection on kanamycin, produced genetically modified apple lines. Next, leaves from in vitro material were treated to activate the recombinase leading to excision of selection genes. Subsequently, the leaf explants were subjected to negative selection for marker-free plantlets by inducing regeneration on medium containing 5-fluorocytosine. After verification of the marker-free nature, the obtained plants were grafted onto rootstocks. Young trees from four cisgenic lines and one intragenic line, all containing Rvi6, were planted in an orchard. Appropriate controls were incorporated in this trial. We scored scab incidence for three consecutive years on leaves after inoculations with Rvi6-avirulent strains. One cisgenic line and the intragenic line performed as well as the resistant control. In 2014 trees started to overcome their juvenile character and formed flowers and fruits. The first results of scoring scab symptoms on apple fruits were obtained. Apple fruits from susceptible controls showed scab symptoms, while fruits from cisgenic and intragenic lines were free of scab.

Project description:Virginia mountain mint (Pycnanthemum virginianum) is a peppermint-flavored aromatic herb of the Lamiaceae and is mainly used for culinary, medicinal, aromatic, and ornamental purposes. North Alabama's climate is conducive to growing mint for essential oils used in culinary, confectionery, and medicinal purposes. There is, however, a need for varieties of P. virginianum that can be adapted and easily grown for production in North Alabama. Towards this end, four field-grown varieties with three harvesting times (M1H1, M1H2, M1H3; M2H1, M2H2, M2H3; M3H1, M3H2, M3H3, M4H1, M4H2, M4H3) were evaluated for relative differences in essential oil yield and composition. Thirty-day-old greenhouse-grown plants of the four varieties were transplanted on raised beds in the field at the Alabama A & M University Research Station in North Alabama. The plots were arranged in a randomized complete block with three replications. The study's objective was to compare the four varieties for essential oil yield and their composition at three harvest times, 135, 155, and 170 days after planting (DAP). Essential oils were obtained by hydrodistillation with continuous extraction with dichloromethane using a Likens-Nickerson apparatus and analyzed by gas chromatographic techniques. At the first harvest, the essential oil yield of the four varieties showed that M1H1 had a yield of 1.15%, higher than M2H1, M3H1, and M4H1 with 0.91, 0.76, and 1.03%, respectively. The isomenthone concentrations increased dramatically through the season in M1 (M1H1, M1H2, M1H3) by 19.93, 54.7, and 69.31%, and M3 (M3H1, M3H2, M3H3) by 1.81, 48.02, and 65.83%, respectively. However, it increased only slightly in M2 and M4. The thymol concentration decreased slightly but not significantly in all four varieties; the thymol in M2 and M4 was very high compared with M1 and M3. The study showed that mountain mint offers potential for production in North Alabama. Two varieties, M1 and M3, merit further studies to determine yield stability, essential oil yield, composition, and cultivation development practices.

Project description:Infant sociability is generally conceived in terms of dyadic capacities and behaviors. Recently, quantitative evidence has been published to support arguments that infants achieve a criterion for groupness: the capacity to interact simultaneously with two others. Such studies equate this capacity with alternating dyadic acts to the two other members of an interacting trio. Here we propose a stricter threefold criterion for infant groupness, of which the crux is whether the social behavior of an infant at time B is shown to be influenced by what two or more group-members were previously doing at time A. We test the viability of this conceptualization: (a) through its justification of the novel laboratory procedure of studying infant sociability in infant-peer quartets (rather than trios); and, (b) in an analysis of a pilot study of gaze-behavior recorded in 5-min interactions among two quartets of infants aged 6-9 months. We call this a 'proof of concept' because our aim is to show that infants are capable of groupness, when groupness is conceptualized in a supra-dyadic way-not that all infants will manifest it, nor that all conditions will produce it, nor that it is commonplace in infants' everyday lives. We found that both quartets did achieve the minimum criterion of groupness that we propose: mutual gaze predicting coordinated gaze (where two babies, A and B, are looking at each other, and B is then looked at by C, and sometimes D) more strongly than the reverse. There was a significant absence of 'parallel mutual gaze,' where the four babies pair off. We conclude that, under specific conditions, preverbal infants can manifest supra-dyadic groupness. Infants' capacities to exhibit groupness by 9 months of age, and the paucity of parallel mutual gaze in our data, run counter to the assumption that infant sociability, when in groups, is always generated by a dyadic program. Our conceptualization and demonstration of groupness in 8-month-olds thus opens a host of empirical, theoretical, and practical questions about the sociability and care of young babies.

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Biopesticide Trunk Injection Into Apple Trees: A Proof of Concept for the Systemic Movement of Mint and Cinnamon Essential Oils.

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