Resistance Alleles Research Articles

Identification of Genetic Loci for Grain Traits and Disease Resistance Reveals a Potential Trade-Off in Rice

Understanding how plants utilize limited resources for reproductivity and disease resistance can improve the efficiency of plant breeding. In the present study, we mapped genes for grain traits, including grain length and width and thousand grain weight (TGW). A previously constructed linkage map of a recombinant inbred line population (MHM) was used, which was derived from a cross between an indica restorer rice variety (Minghui 63) and a temperate japonica rice variety (M-202). The MHM population was grown in replicated field plots, and grain trait data were collected. Upon a 3-year field evaluation, a total of 16 loci were mapped on 9 of 12 chromosomes, including 7 for grain length, 4 for grain weight, and 5 for TGW. A TGW locus ( qTGW2) and a grain width locus ( qWIG2.2) were mapped at the known blast resistance locus qBLAST2 on chromosome 2 that harbors a major blast resistance allele Pi-b. Another TGW locus ( qTGW3.3) and a grain length locus ( qLNG3.3) were mapped at qBLAST3 on chromosome 3. These results reveal potential trade-offs between disease resistance and productivity, which are important for breeders to develop high-yielding and disease-resistant rice varieties. [Formula: see text] The author(s) have dedicated the work to the public domain under the Creative Commons CC0 “No Rights Reserved” license by waiving all of his or her rights to the work worldwide under copyright law, including all related and neighboring rights, to the extent allowed by law, 2024.

A single-nucleotide insertion in Rxp confers durable resistance to bacterial pustule in soybean.

The soybean Rxp gene, encoding a bHLH transcription factor and an ACT-like domain, has an rxp allele producing a truncated protein that confers resistance to pustule-causing Xanthomonas axonopodis pv. glycines. In soybean, bacterial pustules caused by Xanthomonas axonopodis pv. glycines lead to premature defoliation and decreased yield in warm, wet climates. In the USA, approximately 70years ago, bacterial pustules were eliminated by introducing a recessive resistance allele, rxp, of the Rxp gene, representing the first example of successful soybean breeding for durable disease resistance in North America. In this study, we isolated this historical Rxp gene from resistant soybean varieties using positional cloning. The 1.06Mb region where Rxp was reported to reside was narrowed down to an 11.1kb region containing a single gene, Glyma.17g090500. The resistance allele, rxp, contains a T insertion. A complementation test of the Rxp allele in resistant plants confirmed the identification of the Rxp gene. The product of the susceptible wild-type allele, Rxp, is presumed to be a basic helix-loop-helix (bHLH) transcription factor with an aspartate kinase, chorismate mutase, and TyrA (ACT)-like domain. This gene was mainly expressed in extended leaves, and its homologs were identified to be distributed in angiosperms. A total of six alleles were obtained: four from spontaneous variation, including the wild-type and three mutant alleles that encoded truncated proteins, and two from ethyl methanesulfonate mutants, including an allele that encoded a truncated protein and a missense allele. By evaluating the resistance of these six alleles, we found that the loss of function of RXP decreased the bacterial pustule lesions. This study provides important insights into the soybean rxp allele, which confers durable resistance to bacterial pustules.

DISTRIBUTION OF PLASMODIUM FALCIPARUM CHLOROQUINE RESISTANCE TRANSPORTER (PFCRT) AND PLASMODIUM FALCIPARUM MULTIDRUG RESISTANCE 1(PFMDR-1) GENES IN ILORIN NORTH-CENTRAL NIGERIA

Globally, malaria is a peculiar health challenge particularly in continent of Africa. The ease of developing resistance to anti-malaria drugs by Plasmodium falciparum is a serious impediment towards programs aim at control and elimination of malaria. Consequently, drug efficacy surveillance is desirable to institute adequate and effective treatment policies. In this study, distribution of these genes in Plasmodium falciparum isolates in Ilorin metropolis were investigated. One hundred and three samples collected randomly from 5 hospitals in Ilorin metropolis. The samples were amplified at codon 76 and 86 for Pfcrt and Pfmdr-1 respectively using PCR / Restricted Fragment Length Polymorphism (RFLP). The most prevalent alleles were drug susceptible alleles in the study area (K76 and N86) with 31% and 40% respectively, while drug resistance alleles (86Y) have 13% and the least prevalence 2% was the mixed alleles K76T. UITH has the highest number of susceptible alleles followed by CSC 38% and 46% for K76 and N86 respectively. There are significant differences in the distribution of these alleles and the study sites (P< 0.05). The prevalence of resistance alleles in the area is a welcome development that can be employed for possibility of re-introduction of CQ for the treatment of malaria in the study area which will serve as an advantage over expensive ACT due to fact that it is safe cheap and readily affordable.

A major falling number locus on chromosome 4B governs resistance to pre-harvest sprouting in bread wheat line Soru#1

Pre-harvest sprouting (PHS) is a serious threat to wheat quality and occurs when maturing seeds on plants encounter substantial rain before harvest. In the present study, the genetics of PHS resistance were studied using a population of 131 recombinant inbred lines (RILs) from a cross between Soru#1 and Naxos. The population was genotyped with the Illumina 90 K and Axiom 35 K wheat SNP arrays and tested in field trials across two locations in Norway and one in China. Seed dormancy as a measure of sprouting damage was evaluated using germination index (GI) and falling number (FN). Days to heading, days to maturity, and plant height were also recorded to evaluate their potential impact on PHS. Thirteen PHS-related quantitative trait loci (QTL) were detected. The most consistent QTL for FN, contributed by Soru#1, on the long arm of chromosome 4B explained 10.0–19.2% of the phenotypic variation across trials. Other important PHS resistance QTL from Soru#1 detected on chromosome arms 2AL, 2BL, and 4BS, affected both GI and FN. Naxos carried favorable QTL alleles for PHS resistance on chromosome arm 4AS and the Phs1 allele on 4AL. The reduced height Rht-D1 and vernalization Vrn-A1 alleles were the major determinants of plant height and earliness; however, these traits had minimal impact on the genetic control of PHS resistance in this population. Thus, both parents contributed valuable alleles for improving PHS resistance in breeding programs.

Allelic interactions defining Raphanus raphanistrum AHAS resistance level: strong vs weak target-site AHAS resistance alleles.

Chlorsulfuron resistance and genetic dominance was evaluated in Raphanus raphanistrum genotypes homozygous (122-RR, 376-RR), heterozygous (122-RS, 376-RS) and compound heterozygous (122-R/376-R) for the target-site resistance mutations Ala-122-Tyr and Asp-376-Glu in the AHAS (acetohydroxyacid synthase) gene. At the AHAS level, 122-RR and 122-RS plants exhibited significantly higher I50 values than 376-RR and 376-RS plants, respectively. However, plants of the compound heterozygous genotype (122-R/376-R), showed no difference in AHAS activity compared to the 122-RS genotype but lower activity than the 122-RR genotype, and showed a nearly 400-fold greater I50 value than both genotypes (376-RR and 376-RS) carrying the 376-Glu allele. At the whole-plant level, 100% survival was observed for 122-RR plants at the highest chlorsulfuron dose of 640 g ha-1, yet 376-RR plants showed no survival at 380 g ha-1. Thus, this survival difference resulted in different median lethal dose (LD50) estimates [>640 (122-RR) versus 330 g ha-1(376-RR)]. The effect of chlorsulfuron in reducing aboveground growth of surviving plants also was markedly lower for the homozygous 122-RR (GR50 = 566 g ha-1) than for 376-RR plants (GR50 = 66). Heterozygous plants carrying the 122-Tyr allele (122-RS) exhibited two- and five-fold higher LD50 values than both homozygous and heterozygous plants carrying the 376-Glu allele (376-RR, 376-RS), respectively. Along the difference in plant survival, 122-RS plants also showed four-fold higher GR50 than both 376-RR and 376-RS plants. Survival of plants with the compound heterozygous genotype (122-R/376-R) under increasing chlorsulfuron doses was similar to 122-RR or 122-RS genotypes. However, this compound heterozygous genotype showed two- and six-fold higher LD50 values than 376-RR or 376-RS genotypes, respectively. However, both resistance 122-Tyr and 376-Glu alleles were dominant or nearly dominant over the wild-type susceptible alleles (ALA-122 and ASP-376), and the resistance 122-Tyr allele was dominant over the 376-Glu allele. These results broaden our understanding of AHAS target-site resistance in R. raphanistrum and strengthens the hypothesis that the AHAS 122-Tyr allele corresponds to a stronger target-site resistance allele than the 376-Glu allele. © 2024 Society of Chemical Industry.

Identification of genetic loci for powdery mildew resistance in common wheat.

Powdery mildew (PM) poses an extreme threat to wheat yields and quality. In this study, 262 recombinant inbred lines (RILs) of Doumai and Shi 4185 cross were used to map PM resistance genes across four environments. High-density genetic linkage map of the Doumai/Shi 4185 RIL population was constructed using the wheat Illumina iSelect 90K single-nucleotide polymorphism (SNP) array. In total, four stable quantitative trait loci (QTLs) for PM resistance, QPm.caas-2AS, QPm.caas-4AS, QPm.caas-4BL, and QPm.caas-6BS, were detected and explained 5.6%-15.6% of the phenotypic variances. Doumai contributed all the resistance alleles of QPm.caas-2AS, QPm.caas-4AS, QPm.caas-4BL, and QPm.caas-6BS. Among these, QPm.caas-4AS and QPm.caas-6BS overlapped with the previously reported loci, whereas QPm.caas-2AS and QPm.caas-4BL are potentially novel. In addition, six high-confidence genes encoding the NBS-LRR-like resistance protein, disease resistance protein family, and calcium/calmodulin-dependent serine/threonine-kinase were selected as the candidate genes for PM resistance. Three kompetitive allele-specific PCR (KASP) markers, Kasp_PMR_2AS for QPm.caas-2AS, Kasp_PMR_4BL for QPm.caas-4BL, and Kasp_PMR_6BS for QPm.caas-6BS, were developed, and their genetic effects were validated in a natural population including 100 cultivars. These findings will offer valuable QTLs and available KASP markers to enhance wheat marker-assisted breeding for PM resistance.

The impact of lethal and sub-lethal exposure of emamectin benzoate on populations of Spodoptera litura (Lepidoptera: Noctuidae) under laboratory conditions

Emamectin benzoate is an avermectin bio-insecticide commonly used for managing several insect pests including Spodoptera litura (Fabricius) (Lepidoptera: Noctuidae), a major polyphagous pest of many cultivated crops. The current study was conducted to evaluate the effects of emamectin benzoate on the fitness of S. litura populations exhibiting differential susceptibility to insecticide. The selection process and all the bioassays were carried out using 6-day-old 2nd instar larvae of S. litura. A field-collected population of S. litura was divided into two groups: one selected with emamectin benzoate for eight generations (EB-Sel) and the other kept unexposed (Unsel-Lab) to insecticide in the laboratory. An increase in resistance ratio from 1.71-fold in the F1 generation to 22.54-fold in the F8 generation of the EB-Sel population was observed compared to the Unsel-Lab (F8) population. The EB-Sel and Unsel-Lab populations were treated with their respective lethal and sub-lethal concentrations which resulted in an extended development period, decreased larval survival, and adult emergence along with increased morphological abnormalities in adults. Significant reductions were observed in both male and female longevity, fecundity, egg hatching, net reproductive rate (R0), intrinsic rate of increase (rm), and finite rate of increase (λ) in EB-Sel and Unsel-Lab populations. Higher concentrations of the insecticide also reduced the relative fitness (Rf) of S. litura larvae, with maximum effect at LC50 of the EB-Sel population where the Rf value was 0.32 compared to the Unsel-Lab population. Both populations have been affected by emamectin benzoate exposure, however, the impact was more pronounced in the EB-Sel population indicating fitness costs. Our results suggested the fitness cost linked to emamectin benzoate resistance in S. litura which might favor managing insecticide resistance by reducing the frequency of resistant alleles by removing selection pressure. Consequently, our research provides significant insights to devise better pest management strategies for S. litura.

Molecular diversiry studies among blackgram genotypes for yellow mosaic virus resistance

Yellow mosaic disease (YMD), incited by a geminivirus. Mungbean Yellow Mosaic Virus (MYMV), is transmitted by whiteflies, poses a significant threat to various leguminous crops in India. This study focuses on identifying resistant sources and validating molecular markers for their efficacy in detecting YMV resistance in blackgram. In the present study, 150 blackgram genotypes were evaluated to identify resistant sources for YMD under natural field conditions and through molecular markers. Among the 150 genotypes, 22 (both resistant and susceptible) were selected based on field screening. These genotypes were validated using 15 molecular markers, including 8 simple sequence repeats (SSRs) and 7 gene-specific markers linked to MYMV resistance. The markers amplified a total of 48 alleles, with polymorphic information content (PIC) values ranging from 0.82 (CEDG67) to 0.21 (HSP), with an average value of 0.55. The highest PIC values were exhibited by markers CEDG67 (0.82), CEDG115 (0.81), and CEDG20 (0.74). Cluster analysis using DARWIN revealed that most phenotypically resistant genotypes grouped together, while susceptible genotypes formed separate clusters. The genotypes LBG 904, LBG 932, LBG 884, PU 1815, PU 1808, PU 1803, MASH 1008, MASH 114, VBN 10, and PU 31 exhibited resistance to YMV phenotypically. Among these, LBG 904, LBG 932, PU 1815, PU 31, and MASH 1008 produced a 162 bp resistant allele using the marker CEDG180. Similarly, the gene-specific marker DEF produced a 190 bp resistant allele in all the resistant genotypes, whereas the susceptible genotypes produced both 190 and 250 bp alleles. These identified genotypes can serve as valuable sources for MYMV resistance in blackgram breeding programs. Keywords: SSR markers, Blackgram, Diversity, MYMV

Distribution of Plasmodium falciparum Chloroquine Resistance Transporter (Pfcrt) and Plasmodium falciparum Multidrug Resistance 1(Pfmdr-1) Genes in Ilorin North-central Nigeria

Malaria remains a major health problem globally, especially in Sub-Saharan Africa. The ease of developing resistance to anti-malaria drugs by Plasmodium falciparum is a serious challenge to malaria control and elimination programs. Consequently, continuous monitoring of drug efficacy is necessary to ascertain effective treatment policies. The study investigated the distribution pattern of these genes in Plasmodium falciparum isolates in Ilorin metropolis. One hundred and three samples were collected randomly from 5 hospitals in Ilorin metropolis. The samples were amplified at codon 76 and 86 for Pfcrt and Pfmdr-1 respectively using PCR/Restricted Fragment Length Polymorphism (RFLP). Drug susceptible alleles were most prevalent in the study area (K76 and N86) with 31% and 40% respectively, followed by drug resistance alleles (86Y) with 13%, and the least prevalent (2%) was the mixed alleles K76T. UITH has the highest number of susceptible alleles followed by CSC 38% and 46% for K76 and N86 respectively. There are significant differences in the distribution of these alleles and the study sites (P55< 0.05). The prevalence of resistance alleles in the area is a welcome development that can be employed for the possible re-introduction of CQ to treat malaria in the study area. It will be an advantage over expensive ACT because it is safe, cheap, and readily affordable.

Marker Assisted Selection (MAS) and It's Application in Vegetable Crop Improvement

Vegetable crop breeding has changed dramatically since the early 1980s due to technological breakthroughs, especially high-throughput sequencing and molecular marker technologies, which have made it possible to sequence many plant genomes, create dense genetic maps, and identify important genes and QTLs linked to traits like yield and disease resistance. ..These instruments have expedited genetic research and breeding programs in crops including tomato, pepper, eggplant, and cucumber, establishing marker-assisted selection (MAS) as a crucial component of contemporary breeding. The use of these technologies has improved crop characteristics significantly and is still influencing the development of vegetable crops in the future. .. For instance, in the majority of private and public tomato breeding projects, major-gene disease resistance traits like verticillium wilt and anthracnose are selected for using PCR-based markers such as SCAR and CAPS.Bs5 and Bs6 have broad-spectrum resistance against Xanthomonas spp. against chilli leaf spot thanks to marker-assisted gene pyramiding. Major QTLs associated to SSR marker CAMS451 on chromosome 1 (from Capsicum accession LS2341) and marker ID10-194305124 on chromosome 10 (from C. annuum BVRC1) have been mapped for resistance to Ralstonia bacterial wilt.Constantly searching crop germplasm for new resistance genes or alleles is essential to diversify the gene pool and stop resistance from breaking down. By using marker-assisted selection (MAS), it is possible to combine genes for resistance to different pathogens or several sources of resistance to the same pathogen to create novel cultivars.

Genetic diversity of naked barley accessions from the VIR collection for resistance to powdery mildew in the North-West Region of the Russian Federation

BACKGROUND: Barley is an important grain crop characterized by adaptability to various growing conditions. In recent years close attention has been paid to groups of naked barley which compared to covered barley is more susceptible to the influence of pathogens, that decreases the yield and quality of grain. Powdery mildew is the most harmful diseases of barley (caused by the biotrophic fungus Blumeria graminis (DC.) Golovin ex Speer f. sp. hordei Marchal). The search for new promising sources of resistance to powdery mildew is necessary for barley breeding. AIM: The aim is studying the powdery mildew resistance of naked spring barley accessions from the world collection of VIR and identifying promising sources for economically valuable traits. MATERIALS AND METHODS: In field and laboratory conditions 271 of naked barley accessions of different ecological and geographical origin were studied for powdery mildew resistance and other economically valuable traits (lodging resistance, duration of the vegetative period, yield). The pathogen inoculum for laboratory work was represented by the north-western population of the pathogen. The resistance of barley samples was assessed in the field using the VIR point scale, in the laboratory using the E.B. Mains, S.M. Dietz point scale. The highly effective to powdery mildew resistance allele mlo11 was identified using PCR. RESULTS: The diversity of barley of different origins in resistance to powdery mildew is shown. 16 samples mainly from Europe, local forms from Ethiopia, Japan and India, were found to be resistant in field condition. According to the results of the assessment at the juvenile stage, 1 highly resistant sample k-31058 and 5 moderately resistant (k-20923, k-25801, k-27165, k-29911, k-30231) were identified. The naked forms k-20923 from Germany and k-29911 from Poland were resistant at all stages of plant development. The mlo11 allele was detected in the Arabische variety (k-20923) and heterogeneous pathogen resistance accessions k-20921, k-25793, k-27080, k-29440, k-29820. CONCLUSIONS: As a result of the work the sources of resistance to powdery mildew with a complex of economically valuable traits were identified, which can be involve in the breeding.

Study on the cross-resistance of Aedes albopictus (Skuse) (Diptera: Culicidae) to deltamethrin and pyriproxyfen

BackgroundInsecticide resistance poses a significant challenge in the implementation of vector-borne disease control strategies. We have assessed the resistance levels of Aedes albopictus to deltamethrin and pyriproxyfen (PPF) in Fujian Province (China) and investigated the correlation between these resistance levels and mutations in the voltage-gated sodium channel (VGSC).MethodsThe WHO bioassay protocol was used to evaluate the resistance coefficient of Ae. albopictus to deltamethrin and PPF, comparing a susceptible population from the Foshan (FS) area with wild populations from the Sanming (SM), Quanzhou (QZ), Zhangzhou (ZZ), Putian (PT) and Fuzhou (FZ) areas in Fujian Province. Genomic DNA was analyzed by PCR and sequencing to detect knockdown resistance (kdr) in the VGSC, specifically at the pyrethroid resistance alleles V1016V, I1532I and F1534F. Molecular docking was also performed to analyze the binding interactions of PPF and its metabolite 4'-OH-PPF to cytochrome P450 (CYP) 2C19, 2C9 and 3A4 and Ae. albopictus methoprene-tolerant receptors (AeMet), respectively.ResultsThe analysis of resistance to deltamethrin and PPF among Ae. albopictus populations from the various regions revealed that except for the sensitive population in FS and the SM population, the remaining four regional populations demonstrated resistance levels ranging from 4.31- to 18.87-fold for deltamethrin and from 2.85– to 3.62-fold for PPF. Specifically, the FZ and PT populations exhibited high resistance to deltamethrin, whereas the ZZ and QZ populations approached moderate resistance levels. Also, the resistance of the FZ, PT and ZZ populations to PPF increased slowly but consistently with the increasing trend of deltamethrin resistance. Genomic analysis identified multiple non-synonymous mutations within the VGSC gene; the F1534S and F1534L mutations showed significant resistance to deltamethrin in Ae. albopictus. Molecular docking results revealed that PPF and its metabolite 4'-OH-PPF bind to the Ae. albopictus AeMet receptor and CYP2C19.ConclusionsThe wild Ae. albopictus populations of Fujian Province showed varying degrees of resistance to deltamethrin and PPF and a trend of cross-resistance to deltamethrin and PPF. Increased vigilance is needed for potential higher levels of cross-resistance, especially in the PT and FZ regions.Graphical

Improvement of Quality and Disease Resistance for a Heavy-Panicle Hybrid Restorer Line, R600, in Rice (Oryza sativa L.) by Gene Pyramiding Breeding.

The utilization of heavy-panicle hybrid rice exemplifies the successful integration of architectural enhancement and heterosis, which has been widely adopted in the southwest rice-producing area of China. Iterative improvement in disease resistance and grain quality of heavy-panicle hybrid rice varieties is crucial to promote their sustainable utilization. Here, we performed a molecular design breeding strategy to introgress beneficial alleles of broad-spectrum disease resistance and grain quality into a heavy-panicle hybrid backbone restorer line Shuhui 600 (R600). We successfully developed introgression lines through marker-assisted selection to pyramid major genes (Wxb + ALKA-GC + Pigm + Xa23) derived from three parents (Huanghuazhan, I135, I488), which significantly enhance grain quality and confer resistance to rice blast and bacterial blight (BB). The improved parental R600 line (iR600) exhibited superior grain quality and elevated disease resistance while maintaining the heavy-panicle architecture and high-yield capacity of R600. Moreover, the iR600 was crossed with male sterility line 608A to obtain a new heavy-panicle hybrid rice variety with excellent eating and cooking quality (ECQ) and high yield potential. This study presents an effective breeding strategy for rice breeders to expedite the improvement of grain quality and disease resistance in heavy-panicle hybrid rice.

Identification of novel bovine leukocyte antigen alleles and association of BoLA-DRB3.2*020:02:01 with resistance to Theileria orientalis infection in crossbred Kedah-Kelantan cattle: a pilot study.

The bovine leukocyte antigen (BoLA) gene is a significant genetic part of the immune system and has been used as a disease marker in cattle. In this study, we detected Theileria orientalis, T. sinensis, Anaplasma marginale, Anaplasma platys, Candidatus Mycoplasma haemobos and Trypanosoma evansi by PCR amplification and sequencing of the amplicons. The allelic association of the BoLA-DRB3.2 gene with blood pathogen disease resistance and susceptibility in 87 Kedah-Kelantan x Brahman (KKB) and 38 Bali cattle was determined by Fisher's exact test and Cochran Mantel Haenszel (CMH) correction test. Sequence-based typing of the BoLA-DRB3.2 gene identified 43 alleles (27 previously reported alleles and 16 novel alleles) across the two cattle breeds. Alignment analysis of the 16 novel alleles revealed 90.7-95.8% and 85-92% nucleotide and amino acid identities, with the reference allele, BoLA-DRB3*016:01 cDNA clone NR-1. BoLA-DRB3*009:02 (25.6%) and BoLA-DRB3*036:01 (36%) were the most frequent alleles in KKB and Bali cattle, respectively. In KKB cattle, BoLA-DRB3*020:02:01 was significantly associated with resistance to T. orientalis whereas *007:01 and *009:02 were significantly associated with resistance to C. Mycoplasma haemobos. Also, DRB3*017:01 was associated with susceptibility to T. orientalis in KKB cattle. In the Bali cattle, BoLA-DRB3*015:01 was found to be a genetic marker of susceptibility to C. Mycoplasma haemobos infection. Therefore, this study identified BoLA-DRB3.2 alleles associated with resistance and susceptibility to T. orientalis infection in KKB cattle and susceptibility to C. Mycoplasma haemobos infection in Bali cattle for the first time. Therefore, this study suggests that these BoLA-DRB3 resistance alleles could be used as candidate markers for selection, whereas susceptibility alleles could be used as candidate markers for culling in the beef industry.

Detection of mutations in the voltage-gated sodium channel gene of Varroa destructor (Acari: Varroidae) populations

The honeybee (Apis mellifera) ectoparasite, Varroa destructor, is one of the most important honeybee pests worldwide. Acaricides, including the pyrethroids (tau-fluvalinate, flumethrin), and organophosphate (coumaphos) have been applied to control this mite within apiaries, still the long-term, constant, and excessive use of these products has led to the development of resistance in many populations. Three different mutations (L925V, L925I, L925M) at position 925 and one mutation (M918L) at 918 position of the V. destructor voltage-gated sodium channel (VGSC) have been associated with the resistance to these compounds. In the present study, we examined the presence of resistance mutations in the VGSC gene, encoding the target of pyrethroids, in the V. destructor population collected from the Kayseri and Sivas provinces of Central Anatolia Region of Türkiye. A total of 200 V. destructor samples were collected from 20 apiaries in two provinces throughout 2023. To investigate the mutations in Varroa samples, the domain II region of the VGSC gene was amplified using PCR and sequenced. The nucleotide sequencing of the IIS4-IIS5 linker region of the VGSC gene revealed one amino acid change at position 925: a leucine to isoleucine substitution (L925I). No mutations at other positions were identified. Homozygous resistant alleles were detected in 20 (40 %) of the sequenced 50 samples in the study areas. However, we detected the homozygous sensitive allele (wild-type allele, L925) in the remaining samples (50/30, 60 %). The result shows that this status may indicate a problem for Varroa control in the future. Thus, alternative acaricide with a mode of action of different pyrethroids should be considered in the control of V. destructor populations in these provinces.

Population suppression by release of insects carrying a dominant sterile homing gene drive targeting doublesex in Drosophila

CRISPR homing gene drives can suppress pest populations by targeting female fertility genes, converting wild-type alleles into drive alleles in the germline of drive heterozygotes. fsRIDL (female-specific Release of Insects carrying a Dominant Lethal) is a self-limiting population suppression strategy involving continual release of transgenic males carrying female lethal alleles. Here, we propose an improved pest suppression system called “Release of Insects carrying a Dominant-sterile Drive” (RIDD), combining performance characteristics of homing drive and fsRIDL. We construct a split RIDD system in Drosophila melanogaster by creating a 3-gRNA drive disrupting the doublesex female exon. Drive alleles bias their inheritance in males, while drive alleles and resistance alleles formed by end-joining cause dominant female sterility. Weekly releases of RIDD males progressively suppressed and eventually eliminated cage populations. Modeling shows that RIDD is substantially stronger than SIT and fsRIDL. RIDD is also self-limiting, potentially allowing targeted population suppression.

A novel mutation in SoIAA20 confers cross-resistance to 2,4-Dichlorophenoxyacetic acid and other auxinic herbicides in Sonchus oleraceus.

2,4-Dichlorophenoxyacetic acid (2,4-D) and other auxinic herbicides are important for weed control in cropping systems globally. Weeds with resistance to 2,4-D and other auxinic herbicides have evolved, including several populations of Sonchus oleraceus from multiple sites in Australia. We report the underlying mechanism in these populations that gives rise to auxinic herbicide resistance. We studied a total of three susceptible and eight resistant Sonchus oleraceus populations. All resistant populations had a deletion of three amino acids flanking the degron sequence of an Aux/IAA gene, SoIAA20, which was not found in the three susceptible populations. The eight populations with the resistant allele were also resistant to dicamba, fluroxypyr and clopyralid. The resistant plants also had reduced movement of 2,4-D out of the treated tissues compared to susceptible plants. The paired deletion flanking the degron region of SoIAA20 likely provides resistance to 2,4-D by restricting the movement of 2,4-D from the treated tissue to the rest of the plant. We hypothesise that this deletion keeps the 2,4-D bound to the target site. © 2024 The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Population suppression with dominant female-lethal alleles is boosted by homing gene drive

BackgroundMethods to suppress pest insect populations using genetic constructs and repeated releases of male homozygotes have recently been shown to be an attractive alternative to older sterile insect techniques based on radiation. Female-specific lethal alleles have substantially increased power, but still require large, sustained transgenic insect releases. Gene drive alleles bias their own inheritance to spread throughout populations, potentially allowing population suppression with a single, small-size release. However, suppression drives often suffer from efficiency issues, and the most well-studied type, homing drives, tend to spread without limit.ResultsIn this study, we show that coupling female-specific lethal alleles with homing gene drive allowed substantial improvement in efficiency while still retaining the self-limiting nature (and thus confinement) of a lethal allele strategy. Using a mosquito model, we show the required release sizes for population elimination in a variety of scenarios, including different density growth curves, with comparisons to other systems. Resistance alleles reduced the power of this method, but these could be overcome by targeting an essential gene with the drive while also providing rescue. A proof-of-principle demonstration of this system in Drosophila melanogaster was effective in both biasing its inheritance and achieving high lethality among females that inherit the construct in the absence of antibiotic.ConclusionsOverall, our study shows that substantial improvements can be achieved in female-specific lethal systems for population suppression by combining them with various types of gene drive.

Novel resistance loci for quantitative resistance to Septoria tritici blotch in Asian wheat (Triticum aestivum) via genome-wide association study

BackgroundSeptoria tritici blotch (STB) disease causes yield losses of up to 50 per cent in susceptible wheat cultivars and can reduce wheat production. In this study, genomic architecture for adult-plant STB resistance in a Septoria Association Mapping Panel (SAMP) having 181 accessions and genomic regions governing STB resistance in a South Asian wheat panel were looked for.ResultsField experiments during the period from 2019 to 2021 revealed those certain accessions, namely BGD52 (CHIR7/ANB//CHIR1), BGD54 (CHIR7/ANB//CHIR1), IND92 (WH 1218), IND8 (DBW 168), and IND75 (PBW 800), exhibited a high level of resistance. Genetic analysis revealed the presence of 21 stable quantitative trait nucleotides (QTNs) associated with resistance to STB (Septoria tritici blotch) on all wheat chromosomes, except for 2D, 3A, 3D, 4A, 4D, 5D, 6B, 6D, and 7A. These QTNs were predominantly located in chromosome regions previously identified as associated with STB resistance. Three Quantitative Trait Loci (QTNs) were found to have significant phenotypic effects in field evaluations. These QTNs are Q.STB.5A.1, Q.STB.5B.1, and Q.STB.5B.3. Furthermore, it is possible that the QTNs located on chromosomes 1A (Q.STB.1A.1), 2A (Q.STB_DH.2A.1, Q.STB.2A.3), 2B (Q.STB.2B.4), 5A (Q.STB.5A.1, Q.STB.5A.2), and 7B (Q.STB.7B.2) could potentially be new genetic regions associated with resistance.ConclusionOur findings demonstrate the importance of Asian bread wheat as a source of STB resistance alleles and novel stable QTNs for wheat breeding programs aiming to develop long-lasting and wide-ranging resistance to Zymoseptoria tritici in wheat cultivars.

Effectiveness of benzimidazole treatments against Haemonchus contortus in sheep and goats – Do they produce similar responses?

The primary aim of this study was to compare the in vivo responses to orally administered doses of albendazole (5 mg/kg body weight) between experimentally infected sheep and goats. Fifty-four Improved Valachian lambs and 54 Saanen goat kids were split into six groups of nine animals. The sheep and goats were infected with larvae of the gastrointestinal nematode parasite Haemonchus contortus containing 10, 20, 30, 40, 60, and 80 % of the isotype-1 β-tubulin gene codon 200 alleles previously shown to be associated with benzimidazole (BZ)-resistance. All groups of goats generally had higher mean eggs per gram (EPG) before treatment, which was significant (p<0.05) only for the group with 80 % resistance alleles. An in vivo faecal egg reduction test (FECRT) was used to determine the efficacy of albendazole (ABZ) eight days after treatment. Anthelmintic treatment significantly reduced the EPGs in the groups with 10, 20, and 80 % resistance alleles in sheep and with 10, 20, 30, and 40 % resistance alleles in goats. Differences in efficacy between the sheep and goats after the application of doses of ABZ recommended for sheep mostly ranged from 2 % to 10 %. The largest variation was in the group infected with worms containing 60 % resistance alleles, where the efficacy was 13 % higher in goats. Our secondary aims were to evaluate the data obtained from an in vitro egg hatch test (EHT) in sheep and goats and to compare these data with the results from the isotype-1 β-tubulin gene codon 200 pyrosequencing and the FECRT. The percentages of the BZ-resistance alleles were comparable with the mean hatching obtained in the EHT and were also supported by the FECRT data for all groups. The results of the in vivo tests should be verified in the future using in vivo surveys conducted in mixed breeds and infections in multiple species.