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Dendrocalamus brandisii (Munro) Kurz is a sympodial bamboo species with a wide distribution in tropical and subtropical regions. Due to its remarkable regenerative ability and exceptional flavor, this species plays a pivotal role in bolstering the economies of numerous nations across these regions. We recently published a high-quality genome of this species. To date, no study results have identified the optimal reference genes for quantitative real-time polymerase chain reaction (qRT-PCR) normalization in Dendrocalamus brandisii. qRT-PCR offers a highly accurate and effective approach to analyzing gene expression. However, the precision of the resulting quantitative data hinges on the correct choice of reference genes. Twenty-one potential reference genes were identified from the D. brandisii transcriptomes. Their expression in 23 samples, including 8 different tissue organs and 15 samples of D. brandisii under various treatment conditions, were evaluated through qRT-PCR. Subsequently, four software programs-Delta CT, geNorm, NormFinder, and RefFinder-were employed to compare their expression stability. The results revealed that the selection of optimal reference genes varied based on the particular organ and condition being examined. EF-1-α-2 consistently exhibits the most stable expression across diverse tissues, while ACTIN-1, TUBULIN-1, and EF-1-α-2 were the most consistent reference genes in roots, culms, and leaves under various treatments, respectively. In this study, we identified and characterized appropriate internal genes utilized for calibrating qRT-PCR analyses of D. brandisii across different tissue organs and under various treatments. This research will provide key insights for advancing the study of gene functionality and molecular biology in D. brandisii and related species.

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The common clownfish, Amphiprion ocellaris, is an iconic coral reef fish, ubiquitous in the marine aquarium hobby and useful for studying a variety of biological processes (e.g., mutual symbiosis, ultraviolet vision, and protandrous sex change). Recently, CRISPR/Cas9 methods were developed for knocking out specific genes for mechanistic studies. Here, we expand the genetic toolkit for A. ocellaris by creating the first transgenic line using the Tol2 transposon system. Fertilized eggs were co-injected with Tol2 transposase mRNA and a plasmid encoding an elongation factor-1α (Ef1α): green fluorescent protein (GFP) cassette at various concentrations, needle tip dimensions, and timepoints post-fertilization. We compared various injection parameters and sterilization methods to maximize the survival of injected eggs. F0s (n = 10) that were genotyped GFP + were then raised to 6 months of age and crossed with wild-type (WT) females to confirm germline transmission. F1 offspring were also raised and crossed in the same manner. The highly efficient Tol2 transposon system resulted in a 37% rate of transgenesis for surviving eggs amounting to a 2.7% yield of all injected eggs surviving and being GFP + (n = 160). Of these, 10 were raised to adulthood, 8 spawned, and 5/8 (62.5%) produced GFP + offspring. Further, two F1s crossed with WT females produced 54.2% and 44.6% GFP + offspring respectively, confirming the creation of a stable line. This is, to our knowledge, the first generation of a transgenic line in any coral reef fish. The ability to express transgenes of interest in the iconic anemonefish opens the door to a new era of exploration into their fascinating biology.

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Glycoside hydrolases (GHs) are pivotal in the hydrolysis of the glycosidic bonds of sugars, which are the main carbon and energy sources. The genome of Marinomonas sp. ef1, an Antarctic bacterium, contains three GHs belonging to family 3. These enzymes have distinct architectures and low sequence identity, suggesting that they originated from separate horizontal gene transfer events. M-GH3_A and M-GH3_B, were found to differ in cold adaptation and substrate specificity. M-GH3_A is a bona fide cold-active enzyme since it retains 20 % activity at 10 °C and exhibits poor long-term thermal stability. On the other hand, M-GH3_B shows mesophilic traits with very low activity at 10 °C (< 5 %) and higher long-term thermal stability. Substrate specificity assays highlight that M-GH3_A is a promiscuous ß-glucosidase mainly active on cellobiose and cellotetraose, whereas M-GH3_B is a ß-xylosidase active on xylan and arabinoxylan. Structural analysis suggests that such functional differences are due to their differently shaped active sites. The active site of M-GH3_A is wider but has a narrower entrance compared to that of M-GH3_B. Genome-based prediction of metabolic pathways suggests that Marinomonas sp. ef1 can use monosaccharides derived from the GH3-catalyzed hydrolysis of oligosaccharides either as a carbon source or for producing osmolytes.

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The translation elongation factor 1α (EF1α) protein is a highly conserved G protein that is crucial for protein translation in all eukaryotic organisms. EF1α quickly became insoluble at temperatures 42 °C treatment for 2h in vitro, but generally remained soluble in vivo even after being exposed to temperatures as high as 45 °C for an extended period, which suggests that protective mechanisms exist for keeping EF1α soluble in plant cells under heat stress. EF1α had fast in vivo insolubilization when exposed to 45 °C, resulting in about 40% of the protein aggregating after 9 h. Given its established role in protein translation, heat-induced aggregation is most likely to impact the function of the elongation factor. Overexpression of constitutive mutants in both GTP-bound and GDP-bound forms of EF1α resulted in significantly decreased heat tolerance. These findings provide evidence to support the critical role of EF1α, a thermosensitive protein, in the heat tolerance of plants.

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The nivicolous species of the genus Diderma are challenging to identify, and there are several competing views on their delimitation. We analyzed 102 accessions of nivicolous Diderma spp. that were sequenced for two or three unlinked genes to determine which of the current taxonomic treatments is better supported by molecular species delimitation methods. The results of a haplotype web analysis, Bayesian species delimitation under a multispecies coalescent model, and phylogenetic analyses on concatenated alignments support a splitting approach that distinguishes six taxa: Diderma alpinum, D. europaeum, D. kamchaticum, D. meyerae, D. microcarpum and D. niveum. The first two approaches also support the separation of Diderma alpinum into two species with allopatric distribution. An extended dataset of 800 specimens (mainly from Europe) that were barcoded with 18S rDNA revealed only barcode variants similar to those in the species characterized by the first data set, and showed an uneven distribution of these species in the Northern Hemisphere: Diderma microcarpum and D. alpinum were the only species found in all seven intensively sampled mountain regions. Partial 18S rDNA sequences serving as DNA barcodes provided clear signatures that allowed for unambiguous identification of the nivicolous Diderma spp., including two putative species in D. alpinum.

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First-phase ejection fraction （EF1） is the volume change rate of left ventricle （LV） from end-diastole to the time of peak aortic velocity. This article reviewed the research progress of EF1 in detecting early left ventricular systolic dysfunction （LVSD） of patients with aortic stenosis （AS）， stable coronary artery disease （SCAD）， Coronavirus Disease 2019 （COVID-19） and so on， analyzed the advantages and limitations of EF1 in clinical application， and envisioned the future development of EF1 as a novel predictor of early LVSD in clinical use.

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Maple syrup urine disease (MSUD) is rare autosomal recessive metabolic disorder caused by the dysfunction of the mitochondrial branched-chain 2-ketoacid dehydrogenase (BCKD) enzyme complex leading to massive accumulation of branched-chain amino acids and 2-keto acids. MSUD management, based on a life-long strict protein restriction with nontoxic amino acids oral supplementation represents an unmet need as it is associated with a poor quality of life, and does not fully protect from acute life-threatening decompensations or long-term neuropsychiatric complications. Orthotopic liver transplantation is a beneficial therapeutic option, which shows that restoration of only a fraction of whole-body BCKD enzyme activity is therapeutic. MSUD is thus an ideal target for gene therapy. We and others have tested AAV gene therapy in mice for two of the three genes involved in MSUD, BCKDHA and DBT. In this study, we developed a similar approach for the third MSUD gene, BCKDHB. We performed the first characterization of a Bckdhb-/- mouse model, which recapitulates the severe human phenotype of MSUD with early-neonatal symptoms leading to death during the first week of life with massive accumulation of MSUD biomarkers. Based on our previous experience in Bckdha-/- mice, we designed a transgene carrying the human BCKDHB gene under the control of a ubiquitous EF1α promoter, encapsidated in an AAV8 capsid. Injection in neonatal Bckdhb-/- mice at 1014 vg/kg achieved long-term rescue of the severe MSUD phenotype of Bckdhb-/- mice. These data further validate the efficacy of gene therapy for MSUD opening perspectives towards clinical translation.

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Many groups of parasites lack basic information on biodiversity and host associations, which poses challenges for conservation and understanding the ecological relationships between hosts and their parasites. This gap in knowledge is particularly relevant for parasitic species with obscure lifestyles. Ectoparasitc bird lice (Insecta: Psocodea: Phthiraptera) are a group of parasites that has received a relatively substantial research focus, yet patterns of bird-louse relationships and louse diversity remain understudied in many geographic regions, including in parts of the southeastern United States. In this study, we assessed the diversity, prevalence, abundance, and intensity of lice from live and salvaged birds in northeastern Arkansas. We also focused on the frequency of co-occurrence of lice and symbiotic feather mites. Finally, we used nuclear and mitochondrial genes to assess the phylogenic relationships among the most common genera of lice in our sample. We found a total louse prevalence of 10.57% with the highest prevalence on the Passeriformes families Turdidae, Passerellidae, and Parulidae. We also found the louse genera Myrsidea and Brueelia to be the most prevalent and abundant in our sample. Additionally, we reported several novel associations among well-studied bird species. We also found that louse phylogenic patterns tend to reflect host taxonomy and/or ecology. Overall, our results provide important insight into the biodiversity, community structure, and host interactions of parasitic lice from North American birds.

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Eimeria intestinalis is the most pathogenic species of rabbit coccidiosis, causing weight loss, diarrhea, and even acute death. The currently used anticoccidial drugs against E. intestinalis in rabbits are associated with drug resistance and residues. Immunological control might be a potential alternative. We cloned and expressed the E. intestinalis recombinant EF1α and EFG (rEi-EF1α and rEi-EFG, respectively). Rabbits were immunized subcutaneously every 14 days with 100 µg of rEi-EF1α and rEi-EFG and followed by 5 × 104 E. intestinalis sporulated oocysts orally challenge. Serum samples were collected every 7 days to measure the levels of specific antibodies and cytokines. On post-challenge day 14, rabbits were sacrificed and the anticoccidial index was evaluated. The rabbits of PBS challenged groups exhibited anorexia, diarrhea, marked intestinal wall thickening, and white nodules that formed patches, while rabbits from the rEi-EF1α or rEi-EFG challenged group exhibited milder symptoms. The rEi-EF1α group showed a 75.18% oocyst reduction and 89.01%wt gain; the rEi-EFG group had a 60.58% oocyst reduction and 56.04%wt gain. After vaccination, specific IgG levels increased and stayed high (P < 0.05). The IL-4 and IL-2 levels of rEi-EF1α immunized groups showed a significant increase after immunization (P < 0.05). Both rEi-EF1α and rEi-EFG could induce humoral and cellular immune responses. In contrast, rabbits immunized with rEi-EF1α were better protected from challenge by E. intestinalis than rEi-EFG.

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BACKGROUND: Prime editing (PE) is the most recent gene editing technology able to introduce targeted alterations to the genome, including single base pair changes, small insertions, and deletions. Several improvements to the PE machinery have been made in the past few years, and these have been tested in a range of model systems including immortalized cell lines, stem cells, and animal models. While double nicking RNA (dncRNA) PE systems PE3 and PE5 currently show the highest editing rates, they come with reduced accuracy as undesired indels or SNVs arise at edited loci. Here, we aimed to improve single ncRNA (sncRNA) systems PE2 and PE4max by generating novel all-in-one (pAIO) plasmids driven by an EF-1α promoter, which is especially suitable for human-induced pluripotent stem cell (hiPSC) models. RESULTS: pAIO-EF1α-PE2 and pAIO-EF1α-PE4max were used to edit the voltage gated potassium channel gene KCNQ2 and voltage gated sodium channel gene SCN1A. Two clinically relevant mutations were corrected using pAIO-EF1α-PE2 including the homozygous truncating SCN1A R612* variant in HEK293T cells and the heterozygous gain-of-function KCNQ2 R201C variant in patient-derived hiPSC. We show that sncRNA PE yielded detectable editing rates in hiPSC ranging between 6.4% and 9.8%, which was further increased to 41% after a GFP-based fluorescence-activated cell sorting (FACS) cell sorting step. Furthermore, we show that selecting the high GFP expressing population improved editing efficiencies up to 3.2-fold compared to the low GFP expressing population, demonstrating that not only delivery but also the number of copies of the PE enzyme and/or pegRNA per cell are important for efficient editing. Edit rates were not improved when an additional silent protospacer-adjacent motif (PAM)-removing alteration was introduced in hiPSC at the target locus. Finally, there were no genome-wide off-target effects using pAIO-EF1α-PE2 and no off-target editing activity near the edit locus highlighting the accuracy of snc prime editors. CONCLUSION: Taken together, our study shows an improved efficacy of EF-1α driven sncRNA pAIO-PE plasmids in hiPSC reaching high editing rates, especially after FACS sorting. Optimizing these sncRNA PE systems is of high value when considering future therapeutic in vivo use, where accuracy will be extremely important.

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The accurate identification of plant pathogens is a critical step to prevent their spread and attenuate their impact. Among the wide range of methods available, DNA-barcoding, i.e., the identification of an organism through the PCR amplification and sequencing of a single locus, remains one of the most straightforward and accurate plant-pathogen identification techniques that can be used in a generic molecular biology lab. This chapter provides a detailed protocol for the isolation of genomic DNA of fungal and oomycete pathogens from fresh field samples and the amplification and sequencing of the internal transcribed spacer (ITS) locus for DNA-barcoding purpose. Amendments to the protocol are provided to help in resolving issues related to the analysis of complicated samples and to the lack of species resolution that can be encountered with ITS barcodes.

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The new genus Bryorutstroemia is established for the red-brown, stipitate, bryoparasitic discomycete Helotium fulvum Boud. Combined phylogenetic analysis of ITS and LSU rDNA and EF1α revealed that Bryorutstroemia fulva belongs to the sclerotiniaceous clade, which comprises the paraphyletic families Rutstroemiaceae and Sclerotiniaceae. Bryorutstroemia formed with Clarireedia a supported clade (Rutstroemiaceae s.l.), though with high distance. Bryorutstroemia closely resembles other Rutstroemiaceae in having uninucleate ascospores with high lipid content and an ectal excipulum of textura porrecta, but is unique because of its bryophilous lifestyle and is extraordinary with its thick-walled inamyloid ascus apex. Although B. fulva was described in 1897, very few records came to our notice. The present study summarizes the known distribution of the species, including 25 personal collections from the years 2001-2022. Bryorutstroemia fulva was most often encountered on Dicranella heteromalla, and rarely on other members of Dicranales or Grimmiales, while inducing necrobiosis of the leaves. A detailed description based on mainly fresh apothecia is provided together with a rich photographic documentation. Six new combinations are proposed based on our phylogenetic results and unpublished personal morphological studies: Clarireedia asphodeli, C. calopus, C. gladioli, C. henningsiana, C. maritima, and C. narcissi.

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This study was conducted to investigate the synergistic effects of orally delivered B. subtilis-cNK-2 on vaccination with rEF-1α against E. maxima infection in broiler chickens. Chickens were assigned into the following five groups: control (CON, no Eimeria infection), non-immunized control (NC, PBS), component 1 (COM1, rEF-1α only), component 2 (COM2, rEF-1α plus B. subtilis empty vector), and component 3 (COM3, rEF-1α plus B. subtilis-NK-2). The first immunization was administered intramuscularly on day 4, and the second immunization was given one week later with the same concentration of components as the primary immunization. The immunization of B. subtilis spores (COM2 and COM3) was performed by oral administration given for 5 consecutive days a week later than the second immunization. On day 19, all the chickens except the CON group were orally challenged with E. maxima oocysts (1.0 × 104/chicken). The results of the in vivo vaccination showed that all the chickens immunized with rEF-1α (COM1, COM2, and COM3) produced higher (p < 0.05) serum antibodies against EF-1α on 12 days post-E. maxima infection (dpi). The COM3 group showed a significantly (p < 0.05) higher average body weight gain (BWG) on 0-6, 6-9, and 0-12 dpi compared to those of the non-immunized chickens (NC). Immunization with rEF-1α alone (COM1) reduced the gut lesion score on 6 dpi and the fecal oocyst shedding on 9 dpi, whereas co-administration with B. subtilis spores (COM2 or COM3) led to further reduction in the lesion score. E. maxima infection increased the expression levels of IFN-γ and IL-17ß in the jejunum, but these expressions were downregulated in the rEF-1α immunized (COM1) group and in the groups immunized with rEF-1α and orally treated with B. subtilis spores (COM2 or COM3) at 4 dpi. A reduced gene expression of occludin in the jejunum of the E. maxima-infected chickens on 4 dpi was upregulated following the immunization with COM2. Collectively, rEF-1α vaccination induced significant protection against E. maxima infection in the broiler chickens, and the efficacy of rEF-1α vaccination was further enhanced by co-administration with orally delivered B. subtilis spores expressing cNK-2.

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Flax (Linum usitatissimum L.) is a self-pollinating, annual, diploid crop grown for multi-utility purposes for its quality oil, shining bast fiber, and industrial solvent. Being a cool (Rabi) season crop, it is affected by unprecedented climatic changes such as high temperature, drought, and associated oxidative stress that, globally, impede its growth, production, and productivity. To precisely assess the imperative changes that are inflicted by drought and associated oxidative stress, gene expression profiling of predominant drought-responsive genes (AREB, DREB/CBF, and ARR) was carried out by qRT-PCR. Nevertheless, for normalization/quantification of data obtained from qRT-PCR results, a stable reference gene is mandatory. Here, we evaluated a panel of four reference genes (Actin, EF1a, ETIF5A, and UBQ) and assessed their suitability as stable reference genes for the normalization of gene expression data obtained during drought-induced oxidative stress in flax. Taking together, from the canonical expression of the proposed reference genes in three different genotypes, we report that EF1a as a stand-alone and EF1a and ETIF5A in tandem are suitable reference genes to be used for the real-time visualization of cellular impact of drought and oxidative stress on flax.

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The synergistic effects of orally-delivered chicken NK-lysin peptide 2 (cNK-2) or recombinant chicken IL-7 (rchIL-7) on vaccination with recombinant Eimeria elongation factor-1α (rEF-1α) against Eimeria maxima (E. maxima) infection was investigated in broiler chickens. Chickens were divided into six groups: control (CON, no Eimeria infection), non-immunized control (NC, PBS), Vaccination 1 (VAC 1, rEF-1α plus cNK-2), Vaccination 2 (VAC 2, rchIL-7 plus cNK-2), Vaccination 3 (VAC 3, rEF-1α/rchIL-7 plus cNK-2), and Vaccination 4 (VAC 4, rEF-1α/rchIL-7 plus cNK-2). All groups, except the CON and NC, were orally treated with cNK-2 for 5 days. The first immunization, except for the VAC 4 group, was performed intramuscularly on day 4, and the second immunization was given with the same concentration of components as the primary immunization one week later. The immunization of the VAC 4 group was carried out by an oral inoculation on the same days. On day 19, all chickens except the CON group, were orally challenged with E. maxima (1.0 × 104 oocysts/chicken). The in vivo vaccination results showed that the VAC 1 and VAC 3 groups produced high (p < 0.05) levels of serum antibody titers to rEF-1α, and the VAC 3 showed enhanced (p < 0.05) levels of serum IL-7. Furthermore, the VAC 3 group showed significantly (p < 0.01) greater body weight gains at 6- and 9-days post-E. maxima infection (dpi) with reduced oocyst shedding at 6 dpi. The average jejunal lesion score of the NC group was 2.5 whereas the VAC 1 group showed a significantly (p < 0.05) lower lesion scores at 6 dpi. E. maxima infection significantly (P < 0.05) up-regulated the expression levels of cytokines (IL-6, IL-10 and IFN-γ) in the jejunum at 4 dpi, but those expressions were down-regulated in VAC 1 or VAC 3 groups. Moreover, the gene expression levels of Jam 2 and Occludin, were significantly (P < 0.05) decreased following E. maxima infection in jejunum at 4 dpi (NC), but their expressions were increased in the VAC 3 group. Collectively, these results showed that the efficacy of rEF-1α vaccination was significantly enhanced when rEF-1α vaccine co-immunized with chIL-7 or cNK-2.

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Ophiocordyceps sinensis Berk. is a fungal parasite that parasitizes the larvae of Hepialidae and is endemic to the Qinghai-Tibet Plateau (QTP). The phylogeny and divergence time of O. sinensis and its host insects were analyzed for 137 individuals from 48 O. sinensis populations based on the elongation factor 1 alpha (EF-1α) gene. Lower nucleotide variation, with only 7 and 16 EF-1α haplotypes, was detected in O. sinensis and its host insects, respectively. The isolated and broad distribution patterns coexisted in both O. sinensis and its host insects on the QTP. The divergence time estimates show that O. sinensis and its host insects originated later than 14.33 million years (Myr) and earlier than 23.60 Myr in the Miocene period, and the major differentiation occurred later than 4 Myr. Their origin and differentiation match well with the second and third uplifts of the QTP, respectively. The host insects from the O. sinensis populations distributed around Qinghai Lake are inferred as an ancient and relict species that has survived various geological events of the QTP. It is suitable to estimate the divergence times of both O. sinensis and its host insects from the same individuals using one gene: EF-1α. Our findings of the origin, phylogeny, and evolution of the endemic species also support the epoch of geological events on the QTP.

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Sucking lice live in intimate association with their hosts and often display a high degree of host specificity. The present study investigated sucking lice of the genus Lemurpediculus from six mouse lemur (Microcebus) and two dwarf lemur (Cheirogaleus) species endemic to the island of Madagascar, considered a biodiversity hotspot. Louse phylogenetic trees were created based on cytochrome C oxidase subunit I (COI), elongation factor 1α (EF1α) and internal transcribed spacer 1 (ITS1) sequences. While clustering according to host species was generally observed for COI and ITS1, suggesting high host specificity of the examined lice, EF1α sequences alone did not distinguish between lice of different Microcebus species, possibly due to rather recent divergence. As bootstrap support for basal tree structure was rather low, further data are necessary to resolve the evolutionary history of louse-mouse lemur associations. Three new species of sucking lice are described: Lemurpediculus zimmermanni sp. Nov. From Microcebus ravelobensis, Lemurpediculus gerpi sp.nov. from Microcebus gerpi, and Lemurpediculus tsimanampesotsae sp. nov. from Microcebus griseorufus. These new species are compared with all known congeneric species and identifying features are illustrated for all known species of Lemurpediculus.

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Phlebotomus (Ph.) sergenti is the main vector of Leishmania (L.) tropica (Trypanosomatida: Trypanosomatidae), the causative agent of anthroponotic cutaneous leishmaniasis in Morocco. This species has an extended geographical distribution, wider than that of the parasite. The main objective of our study was to analyze the genetic diversity of Ph. sergenti collected in four foci in Morocco: Taza, Foum Jemâa, El Hanchane, and Ouarzazate. We studied a set of diversity and population structure indices by sequencing two markers; nuclear EF-1α and mitochondrial Cyt b from 175 individual sand flies. Our results showed a considerable degree of intraspecific polymorphism with a high number of haplotypes identified in both genes. Many polymorphic sites detected in the Cyt b sequences (SCyt b = 45) indicate that it is the most polymorphic marker showing a distinct distribution of haplotypes according to their geographical origin, whereas the EF-1α marker showed no geographical isolation. Analysis by Tajima's D and Fu's Fs tests revealed a possible recent expansion of the populations, especially with the EF-1α marker, showing significant values in Taza and Ouarzazate sequences. The present study revealed significant genetic diversity within Ph. sergenti populations in Morocco. The results warrant further research using a combination of more than two markers including mitochondrial and non-mitochondrial markers, which may provide more information to clarify the genetic status of Ph. sergenti.

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Population genetic studies can reveal clues about the evolution of adaptive strategies of aphid species in agroecosystems and demonstrate the influence of environmental factors on the genetic diversity and gene flow among aphid populations. To investigate the genetic diversity of two Rhopalosiphum aphid species from different geographical regions, 32 populations (n = 535) of the bird cherry-oat aphid (Rhopalosiphum padi Linnaeus) and 38 populations (n = 808) of the corn leaf aphid (Rhopalosiphum maidis Fitch) from China and Europe were analyzed using one nuclear (elongation factor-1 alpha) and two mitochondrial (cytochrome oxidase I and II) genes. Based on the COI-COII sequencing, two obvious clades between Chinese and European populations and a low level of gene flow (Nm = 0.15) were detected in R. padi, while no geographical-associated genetic variation was found for EF-1α in this species. All genes in R. maidis had low genetic variation, indicating a high level of gene flow (Nm = 5.31 of COI-COII and Nm = 2.89 of EF-1α). Based on the mitochondrial result of R. padi, we concluded that the long distance between China and Europe may be interrupting the gene flow. The discordant results of nuclear gene analyses in R. padi may be due to the slower evolution of nuclear genes compared to mitochondrial genes. The gene exchange may occur gradually with the potential for continuous migration of the aphid. This study facilitates the design of control strategies for these pests.

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