The Effect of Alfalfa Residue Incorporation on Soil Bacterial Communities and the Quantity of <Emphasis Type="Italic">Verticillium dahliae</Emphasis> Microsclerotia in Potato Fields in the Columbia Basin of Washington State,USA

Verticillium wilt, caused by the soil-borne fungus Verticillium dahliae, is one of the most important diseases of potato in North America. Soil incorporation of alfalfa residues prior to planting potato could be a nonchemical Verticillium wilt management tactic by reducing the number of viable microsclerotia in field soil. Verticillium dahliae microsclerotia were quantified in field soils where organic material from alfalfa was incorporated, and numbers of microsclerotia were compared to fields where alfalfa residue was not incorporated. In addition, bacterial metagenomics was utilized to characterize soils where organic material from alfalfa was or was not incorporated to determine if alfalfa residue incorporation facilitates the formation of soils that suppress or kill V. dahliae microsclerotia. The number of V. dahliae microsclerotia in soil was greater (P = 0.0003) in fields where crop residue was incorporated than fields without incorporation when chloropicrin was used as a fumigant. Conversely, the number of V. dahliae microsclerotia observed in potato plants did not differ (P = 0.4020) between fields where residues were or were not incorporated if chloropicrin was used. Alfalfa residue incorporation did not significantly alter the soil bacterial metagenome compared to fields not subject to residue incorporation in both years of study. Despite these conclusions, the method can be employed to analyze the effect of grower practices with the intent of linking a field practice to increasing soil bacterial diversity and decreasing Verticillium wilt severity on potato.     

Maturity-Adjusted Resistance of Potato (<Emphasis Type="Italic">Solanum tuberosum</Emphasis> L.) Cultivars to Verticillium Wilt Caused by <Emphasis Type="Italic">Verticillium dahliae</Emphasis>

Verticillium wilt is a fungal disease of potato caused by two species of Verticillium, V. dahliae and V. albo atrum. The pathogen infects the vascular tissue of potato plants through roots, interfering with the transport of water and nutrition, and reducing both the yield and quality of tubers. We have evaluated the reaction of 283 potato clones (274 cultivars and nine breeding selections) to inoculation with V. dahliae under greenhouse conditions. A significant linear correlation (r = 0.4, p < 0.0001) was detected between plant maturity and partial resistance to the pathogen, with late maturing clones being generally more resistant. Maturity-adjusted resistance, that takes into consideration both plant maturity and resistance, was calculated from residuals of the linear regression between the two traits. Even after adjusting for maturity, the difference in the resistance of clones was still highly significant, indicating that a substantial part of resistance cannot be explained by the effect of maturity. The highest maturity-adjusted resistance was found in the cv. Navajo, while the most susceptible clone was the cv. Pungo. We hope that the present abundance of data about the resistance and maturity of 283 clones will help potato breeders to develop cultivars with improved resistance to V. dahliae.     

Rooting Characteristics of <Emphasis Type="Italic">Solanum chacoense</Emphasis> and <Emphasis Type="Italic">Solanum tuberosum in Vitro</Emphasis>

The precise level of environmental control in vitro may aid in identifying genetically superior plant germplasm for rooting characteristics (RC) linked to increased foraging for plant nitrogen (N). The objectives of this research were to determine the phenotypic variation in root morphological responses of 49 Solanum chacoense (chc), 30 Solanum tuberosum Group Phureja – Solanum tuberosum Group Stenotomum (phu-stn), and three Solanum tuberosum (tbr) genotypes to 1.0 and 0.5 N rate in vitro for 28 d, and identify genotypes with superior RC. The 0.5 N significantly increased density of root length, surface area, and tips. All RC were significantly greater in chc than in either phu-stn or tbr. Based upon clustering on root length, surface area, and volume, the cluster with the greatest rooting values consisted of eight chc genotypes that may be utilized to initiate a breeding program to improve RC in potato.     

Resistance to <Emphasis Type="Italic">Meloidogyne chitwoodi</Emphasis> Identified in Wild Potato Species

Meloidogyne chitwoodi (Columbia root-knot nematode, CRKN) can cause serious damage in potato production systems, decreasing tuber value in the fresh market and processing industries. Genetic resistance to CRKN was first identified from the wild diploid potato species Solanum bulbocastanum accession SB22 and was successfully introgressed into tetraploid potato breeding material. To expand the base of genetic resistance, 40 plant accessions representing nine wild potato species were screened for their resistance to M. chitwoodi. Greenhouse screening identified fifteen clones from S. hougasii, one clone from S. bulbocastanum, and one clone from S. stenophyllidium with moderate to high levels of resistance against three isolates of M. chitwoodi. Geographical mapping showed that the resistance sources identified in this and previous studies primarily originated in the states of Jalisco and Michoacán in west-central Mexico. These new sources of resistance will be introgressed into elite potato populations to facilitate the development of potato cultivars with durable resistance to M. chitwoodi.     

Enhanced <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> f. sp. <Emphasis Type="Italic">tuberosi</Emphasis> Resistance in Transgenic Potato Expressing a Rice GLP Superoxide Dismutase Gene

Germin like proteins (GLPs) are a large group of related and ubiquitous plant proteins which are considered to be involved in different processes important for plant development and defense. Multiple functional copies of this gene family have been reported in a number of species (wheat, barley, rice, soybean mosses and liverwort), and their role is being evaluated by gene regulation studies and transgenic approaches. To analyze the role of a rice (Oryza sativa) root expressed germin like protein1 OsRGLP1, for its antifungal activity, transgenic potato plants were developed. These transgenic potato plants were molecularly characterized and biologically assessed after inoculation with Fusarium oxysporum f. sp. tuberosi. Functional analysis showed high accumulation of H₂O₂, increased Superoxide Dismutase (SOD) activity and no oxalate oxidase activity (OxO) in transgenics in comparison to nontransformed control. This increased SOD activity, resistance to heat and sensitivity to H₂O₂ suggest it is a Fe-like SOD. OsRGLP1 expression in potato plants exhibited enhanced resistance in comparison to nontransformed wild type plants suggesting its role in providing protection against Fusarium oxysporum f. sp. tuberosi through elevated SOD level. Overall, results suggest that OsRGLP1 is a candidate for the engineering of potato plants with increased fungal tolerance however, the greater height and tuber number was observed. This phenotype associated with the resistance needs to be evaluated to determine if this is a positive or negative feature.     

Divergent evolution of rice blast resistance <Emphasis Type="Italic">Pi54</Emphasis> locus in the genus <Emphasis Type="Italic">Oryza</Emphasis>

Background

The rice blast resistance gene Pi54 was cloned from Oryza sativa ssp. indica cv. Tetep, which conferred broad-spectrum resistance against Magnaporthe oryzae. Pi54 allelic variants have been identified in not only domesticates but also wild rice species, but the majority of japonica and some indica cultivars lost the function.

Results

We here found that Pi54 (Os11g0639100) and its homolog Os11g0640600 (named as #11) were closely located on a 25 kbp region in japonica cv. Sasanishiki compared to a 99 kbp region in japonica cv. Nipponbare. Sasanishiki lost at least six genes containing one other R-gene cluster (Os11g0639600, Os11g0640000, and Os11g0640300). Eight AA-genome species including five wild rice species were classified into either Nipponbare or Sasanishiki type. The BB-genome wild rice species O. punctata was Sasanishiki type. The FF-genome wild rice species O. brachyantha (the basal lineage of Oryza) was neither, because Pi54 was absent and the orientation of the R-gene cluster was reversed in comparison with Nipponbare-type species. The phylogenetic analysis showed that #11gene of O. brachyantha was on the root of both Pi54 and #11 alleles. All Nipponbare-type Pi54 alleles were specifically disrupted by 143 and 37/44?bp insertions compared to Tetep and Sasanishiki type. In addition, Pi54 of japonica cv. Sasanishiki lost nucleotide-binding site and leucine-rich repeat (NBS–LRR) domains owing to additional mutations.

Conclusions

These results suggest that Pi54 might be derived from a tandem duplication of the ancestor #11 gene in progenitor FF-genome species. Two divergent structures of Pi54 locus caused by a mobile unit containing the nearby R-gene cluster could be developed before domestication. This study provides a potential genetic resource of rice breeding for blast resistance in modern cultivars sustainability.

     

Amplification of the <Emphasis Type="Italic">Phytophthora infestans</Emphasis> RG57 loci Facilitates <Emphasis Type="Italic">in planta</Emphasis> T-RFLP Identification of Late Blight Genotypes

Late blight, caused by the oomycete Phytophthora infestans (Mont.) de Bary, is a devastating disease in potato and tomato and causes yield and quality losses worldwide. The disease first emerged in central America and has since spread in North America including the United States and Canada. Several new genotypes of P. infestans have recently emerged, including US-22, US-23 and US-24. Due to significant economic and environmental impacts, there has been an increasing interest in the rapid identification of P. infestans genotypes. In addition to providing details regarding the various phenotypic characteristics such as fungicide resistance, host preference, and pathogenicity associated with various P. infestans genotypes, information related to pathogen movement and potential recombination may also be determined from the genetic analyses. Restriction fragment length polymorphism (RFLP) analysis with the RG57 loci is one of the most reliable procedures used to genotype P. infestans. However, the RFLP procedure requires propagation and isolation of the pathogen and relatively large amounts of DNA. Isolation of the late blight pathogen is sometimes impossible due to the poor condition of the infected tissues or the presence of fungicide residues. In this study, we describe a procedure to identify P. infestans at the molecular level in planta using terminal restriction fragment length polymorphism (T-RFLP) of the RG57 loci. This T-RFLP assay is sufficiently sensitive to detect and differentiate P. infestans genotypes directly in planta without propagation and isolation of the pathogen, to facilitate the timely implementation of best management practices.     

Association of Potato Psyllid (<Emphasis Type="Italic">Bactericera cockerelli</Emphasis>; Hemiptera: Triozidae) with <Emphasis Type="Italic">Lycium</Emphasis> spp. (Solanaceae) in Potato Growing Regions of Washington,Idaho, and Oregon

Potato psyllid, Bactericera cockerelli (?ulc), causes economic damage to potato crops throughout the major potato growing regions of western North America. When cultivated crops are not available, potato psyllid often occurs on non-crop hosts. In the southern U.S. and northern Mexico, native species of Lycium (Solanaceae) are important non-crop hosts for the psyllid. We determined whether Old World species of Lycium now widespread in the Pacific Northwest are reservoirs of potato psyllid in this growing region. We examined Lycium spp. across a wide geographic region in Washington, Oregon, and Idaho at irregular intervals during three growing seasons. Potato psyllids were present at all locations. To determine whether Lycium is also a host during intervals of the year in which the potato crop is not available, we monitored a subset of these sites over the entire year. Six sites were monitored at 1- to 3-week intervals from June 2014 to June 2016. Psyllids were present on Lycium throughout the year at all sites, including during winter, indicating that Lycium is also a host when the potato crop is seasonally not available. Psyllid populations included a mixture of Northwestern and Western haplotypes. We observed well-defined spring and fall peaks in adult numbers, with peaks separated by long intervals in which psyllid numbers were very low. Seasonal patterns in psyllid numbers on these non-native Lycium hosts were very similar to what has been observed on native Lycium in the desert southwest region of the U.S. Our findings demonstrate that potato psyllid associates with Lycium across a broad geographic region within the Pacific Northwest. These results will assist in predicting sources of potato psyllid colonizing potatoes in this important growing region.     

Genetic Diversity of Pathogenic <Emphasis Type="Italic">Fusarium semitectum</Emphasis> Isolates from Potato Tubers,Using PCR-IGS-RFLP Markers

Fusarium semitectum is one of the important causal agents of dry rot of potato tubers in the world. In order to determine genetic variability among 41 isolates of F. semitectum, morphological and molecular studies were carried out. All F. semitectum isolates were recovered from infected potato tubers with dry rot symptoms collected from four provinces in Iran. According to macroscopic and microscopic characteristics, 41 isolates of F. semitectum were classified in two morphotypes (morphotypes I and II). All 41 isolates were evaluated for their pathogenicity on healthy potato tubers. Tuber rot symptoms were observed on the 21st day after inoculation of Fusarium isolates on the tubers tested. The measurement was done by comparing the depth and width of lesion expansion among the isolates. Molecular characterization through PCR-IGS-RFLP analysis by six restriction enzymes (AluI, BsuRI, Eco88I, MspI, TaqI and PstI) divided the 41 isolates of F. semitectum into two separated clusters that were in accordance with the morphological characterization.     

Seasonal deviation effects foliar endophyte assemblage and diversity in <Emphasis Type="Italic">Asparagus racemosus</Emphasis> and <Emphasis Type="Italic">Hemidesmus indicus</Emphasis>

Background

Fungal endophytes are the living symbionts which cause no apparent damage to the host tissue. The distribution pattern of these endophytes within a host plant is mediated by environmental factors. This study was carried out to explore the fungal endophyte community and their distribution pattern in Asparagus racemosus and Hemidesmus indicus growing in the study area.

Results

Foliar endophytes were isolated for 2 years from A. racemosus and H. indicus at four different seasons (June–August, September–November, December–February, March–May). A total of 5400 (675/season/year) leaf segments harbored 38 fungal species belonging to 17 genera, 12 miscellaneous mycelia sterile from 968 isolates and 13 had yeast like growth. In A. racemosus, Acremonium strictum and Phomopsis sp.1, were dominant with overall relative colonization densities (RCD) of 7.11% and 5.44% respectively, followed by Colletotrichum sp.3 and Colletotrichum sp.1 of 4.89% and 4.83% respectively. In H. indicus the dominant species was A. strictum having higher overall RCD of 5.06%, followed by Fusarium moniliforme and Colletotrichum sp.2 with RCD of 3.83% and 3%, respectively. Further the overall colonization and isolation rates were higher during the wet periods (September–November) in both A. racemosus (92.22% and 95.11%) and H. indicus (82% and 77.11%).

Conclusion

Study samples treated with 0.2% HgCl₂ and 75% EtOH for 30 s and 1 min, respectively, confirmed most favorable method of isolation of the endophytes. Owing to high mean isolation and colonization rates, September–November season proved to be the optimal season for endophyte isolation in both the study plants. Assessing the bioactive potential of these endophytes, may lead to the isolation of novel natural products and metabolites.

     

Phenotypic and Genomic Modifications Associated with <Emphasis Type="Italic">Globodera pallida</Emphasis> Adaptation to Potato Resistances

Studying phenotypic and genomic modifications associated with pathogen adaptation to resistance is a crucial step to better understand and anticipate resistance breakdown. This short review summarizes recent results obtained using experimentally evolved populations of the potato cyst nematode Globodera pallida. In a first step, the variability of resistance durability was explored in four different potato genotypes carrying the resistance quantitative trait loci (QTL) GpaV_vrn originating from Solanum vernei but differing by their genetic background. The consequences of the adaptation to resistance in terms of local adaptation, cross-virulence and virulence cost were then investigated. Finally, a genome scan approach was performed in order to identify the genomic regions involved in this adaptation. Results showed that nematode populations were able to adapt to the QTL GpaV_vrn, and that the plant genetic background has a strong impact on resistance durability. A trade-off between the adaptations to different resistant potato genotypes was detected, and we also showed that adaptation to the resistance QTL GpaV_vrn from S. vernei did not allow adaptation to the colinear locus from S. sparsipilum (GpaV_spl). Unexpectedly, the adaptation to resistance led to an increase of virulent individual’s fitness on a susceptible host. Moreover, the genome scan approach allowed the highlighting of candidate genomic regions involved in adaptation to host plant resistance. This review shows that experimental evolution is an interesting tool to anticipate the adaptation of pathogen populations and could be very useful for identifying durable strategies for resistance deployment.     

Transmission Efficiency of <Emphasis Type="Italic">Potato Leafroll Virus</Emphasis> by Four Potato Colonizing Aphid Species in Tunisian Potato Fields

This study investigated the transmission efficiency of Potato leafroll virus (PLRV) by four potato colonizing aphid species, Myzus persicae, Macrosiphum euphorbiae, Aphis gossypii and Aphis fabae, reported from leaves and yellow water trap. Physalis floridana was used as a test plant for virus transmission. DAS-ELISA was used for virus screening of samples as well as virus detection on the test plant after transmission experiment. A 2-h period was sufficient for the tested aphids to acquire PLRV virions. However, a difference in the transmission potential occurred according to the aphid species. The highest potential was recorded for M. persicae and M. euphorbiae, at 90 and 80%, respectively. For the first time, the study revealed the PLRV transmission efficiency of A. fabae, estimated at 50%. The lowest potential rate of 30% was recorded for A. gossypii. This study highlights the PLRV transmission capacities of four potato colonizing aphids suspected to play a key role in the spread of PLRV in potato seed production sites.     

Effective Management of the <Emphasis Type="Italic">Phthorimaea operculella</Emphasis> (Zeller) Using PVA Nanofibers Loaded with <Emphasis Type="Italic">Cinnamomum zeylanicum</Emphasis> Essential Oil

Phthorimaea operculella (Zeller) is one of the most common insect pests of cultivated potato in tropical and subtropical regions. In this research, a potential strategy to improve the insecticidal activity of plant essential oils for the effective management of P. operculella was studied. The insecticidal and residual effects of nanofiber oil (NFO) and pure essential oil (PEO) of Cinnamomum zeylanicum were assessed on PTM under laboratory conditions. The nanofibers were made by the electrospinning method using polyvinyl alcohol (PVA) polymer. The morphological characteristics of the nanofibers were evaluated by scanning electron microscopy and Fourier transform infrared spectroscopy. The chemical constituents of cinnamon essential oil (EO) were detected by GC/MS. Fumigant toxicity of NFO and PEO were evaluated on different growth stages (egg, male and female adults) of P. operculella. SEM and FTIR analyses confirmed the presence of EO on the nanofiber structure. The yield of the EO from C. zelanicum on the nanofibers was 1.86%. GC/MS analysis showed that cinnamaldehyde was the primary constituent (69.88%) of cinnamon EO. LC₅₀ values of C. zelanicum EO and NFO were 4.92 and 1.76 μl/l air for eggs, 0.444 and 0.212 μl/l air for female adults, and 0.424 and 0.192 μl/l air for male adults, respectively. Fumigant bioassays revealed that NFO was more toxic than C. zeylanicum oil against at all stages of P. operculella. The residual effect of PEO and NFO was evaluated against the egg stage of the P. operculella. NFO lost insecticidal effectiveness 47 days after application, while the efficacy of PEO decreased 15 days after application. Our results suggest that NFO of C. zeylanicum can be used as an effective new tool for the management of P. operculella.     

Effects of Different Soil Treatments on the Development of <Emphasis Type="Italic">Spongospora subterranea</Emphasis> f. sp. <Emphasis Type="Italic">subterranea</Emphasis> in Potato Roots and Tubers in the Greenhouse

Powdery scab caused by Spongospora subterranea f. sp. subterranea (Sss) causes extensive losses in potato production systems globally. Two pot experiments were established in the greenhouse in summer 2013 and winter 2014 to evaluate the effectiveness of different soil chemicals, fumigant, amendments and biological control agents (BCAs) against Sss in the rhizospheric soil, potato roots and tubers. The study used visual assessment methods to assess the effect of treatments on root galling and zoosporangia production, and qPCR to measure Sss concentration in the soil and in the potato roots and tubers. All six soil treatments, namely metam sodium, fluazinam, ZincMax, calcium cyanamide, Biocult and a combination of Bacillus subtilis and Trichoderma asperellum recorded significantly (P < 0.05) lower numbers of zoosporangia in the roots compared to the untreated control. The same effect was observed on the concentration of Sss DNA in the roots at tuber initiation. A more diverse picture was obtained when root gall scores at tuber initiation and Sss DNA in the rhizospheric soil at tuber initiation and harvesting were compared. Significant differences (P < 0.05) were also noted in disease severity, disease incidence, and tuber yield between metam sodium, fluazinam, ZincMax, calcium cyanamide and the untreated control. Calcium cyanamide gave the highest tuber yield. The study demonstrated the potential of soil treatments such as metam sodium, fluazinam, ZincMax and calcium cyanamide in managing Sss in potatoes by reducing the pathogen both in the rhizospheric soil and the roots of the potato plant.     

<Emphasis Type="Italic">Lupinus albus</Emphasis> Conglutin Gamma Modifies the Gene Expressions of Enzymes Involved in Glucose Hepatic Production <Emphasis Type="Italic">In Vivo</Emphasis>

Lupinus albus seeds contain conglutin gamma (Cγ) protein, which exerts a hypoglycemic effect and positively modifies proteins involved in glucose homeostasis. Cγ could potentially be used to manage patients with impaired glucose metabolism, but there remains a need to evaluate its effects on hepatic glucose production. The present study aimed to analyze G6pc, Fbp1, and Pck1 gene expressions in two experimental animal models of impaired glucose metabolism. We also evaluated hepatic and renal tissue integrity following Cγ treatment. To generate an insulin resistance model, male Wistar rats were provided 30% sucrose solution ad libitum for 20 weeks. To generate a type 2 diabetes model (STZ), five-day-old rats were intraperitoneally injected with streptozotocin (150 mg/kg). Each animal model was randomized into three subgroups that received the following oral treatments daily for one week: 0.9% w/v NaCl (vehicle; IR-Ctrl and STZ-Ctrl); metformin 300 mg/kg (IR-Met and STZ-Met); and Cγ 150 mg/kg (IR-Cγ and STZ-Cγ). Biochemical parameters were assessed pre- and post-treatment using colorimetric or enzymatic methods. We also performed histological analysis of hepatic and renal tissue. G6pc, Fbp1, and Pck1 gene expressions were quantified using real-time PCR. No histological changes were observed in any group. Post-treatment G6pc gene expression was decreased in the IR-Cγ and STZ-Cγ groups. Post-treatment Fbp1 and Pck1 gene expressions were reduced in the IR-Cγ group but increased in STZ-Cγ animals. Overall, these findings suggest that Cγ is involved in reducing hepatic glucose production, mainly through G6pc inhibition in impaired glucose metabolism disorders.     

<Emphasis Type="Italic">OsLAP6/OsPKS1</Emphasis>, an orthologue of <Emphasis Type="Italic">Arabidopsis PKSA/LAP6</Emphasis>, is critical for proper pollen exine formation

Background

Male fertility is crucial for rice yield, and the improvement of rice yield requires hybrid production that depends on male sterile lines. Although recent studies have revealed several important genes in male reproductive development, our understanding of the mechanisms of rice pollen development remains unclear.

Results

We identified a rice mutant oslap6 with complete male sterile phenotype caused by defects in pollen exine formation. By using the MutMap method, we found that a single nucleotide polymorphism (SNP) variation located in the second exon of OsLAP6/OsPKS1 was responsible for the mutant phenotype. OsLAP6/OsPKS1 is an orthologous gene of Arabidopsis PKSA/LAP6, which functions in sporopollenin metabolism. Several other loss-of-function mutants of OsLAP6/OsPKS1 generated by the CRISPR/Cas9 genomic editing tool also exhibited the same phenotype of male sterility. Our cellular analysis suggested that OsLAP6/OsPKS1 might regulate pollen exine formation by affecting bacula elongation. Expression examination indicated that OsLAP6/OsPKS1 is specifically expressed in tapetum, and its product is localized to the endoplasmic reticulum (ER). Protein sequence analysis indicated that OsLAP6/OsPKS1 is conserved in land plants.

Conclusions

OsLAP6/OsPKS1 is a critical molecular switch for rice male fertility by participating in a conserved sporopollenin precursor biosynthetic pathway in land plants. Manipulation of OsLAP6/OsPKS1 has potential for application in hybrid rice breeding.

     

The Effects of <Emphasis Type="Italic">Pimpinella anisum</Emphasis> Essential Oils on Young Larvae <Emphasis Type="Italic">Leptinotarsa decemlineata</Emphasis> Say (Coleoptera: Chrysomelidae)

The effect of essential oil (EO) from anise (Pimpinellia anisum) on the mortality of young larvae of Colorado potato beetles has been studied. In our bioassays, P. anisum EO significantly increased the mortality of the second instar larvae of L. decemlineata. Significantly different values of LD₅₀ and LD₉₀ were established for acute (LD₅₀ = 1.76, and LD₉₀ = 8.29) as well as chronic toxicity (LD₅₀ = 0.45, and LD₉₀ = 1.01). Decrease of both values over experimental period was evident, which showed that the larval mortality was slow and cumulative. The composition of EO used for biological experiments was also assessed. The main component detected in EO from P. anisum was anethole (79.87%), followed by anisaldehyde (7.74%), estragole (5.88%) and β-linalool (1.07%). Within five days, residual concentration of EO decreased from 3.87 mg/g of dry weight immediately after foliar applications to 0.9 mg per g of dry weight. The effect of this slow evaporation could be explained by dominant presence of anethole or by the type of formulation and the addition of oil and tween. Results of our study demonstrate that EO from P. anisum has insecticidal properties that may lead to the development of new organic products for the control of Colorado potato beetles.