F3 Families Research Articles

Ppe.RPT/SSC-1: from QTL mapping to a predictive KASP test for ripening time and soluble solids concentration in peach

Genomic regions associated with ripening time (RPT) and soluble solids concentration (SSC) were mapped using a pedigreed population including multiple F1 and F2 families from the Clemson University peach breeding program (CUPBP). RPT and SSC QTLs were consistently identified in two seasons (2011 and 2012) and the average datasets (average of two seasons). A target region spanning 10,981,971–11,298,736 bp on chromosome 4 of peach reference genome used for haplotype analysis revealed four haplotypes with significant differences in trait values among different diplotype combinations. Favorable alleles at the target region for both RPT and SSC were determined and a DNA test for predicting RPT and SSC was developed. Two Kompetitive Allele Specific PCR (KASP) assays were validated on 84 peach cultivars and 163 seedlings from the CUPBP, with only one assay (Ppe.RPT/SSC-1) needed to predict between early and late-season ripening cultivars and low and high SSC. These results advance our understanding of the genetic basis of RPT and SSC and facilitate selection of new peach cultivars with the desired RPT and SSC.

Adaptive Responses of Biofortified Common Bean Lines to Acidic Soil and High Temperatures in the Colombian Amazon Region

One of the strategies to combat micronutrient malnutrition is by developing biofortified common bean lines (Phaseolus vulgaris L.) capable of tolerating different stress conditions. In this study, the adaptive responses of different biofortified bean lines grown under combined stress of acidic soil and high-temperatures were evaluated in the Colombian Amazon. A total of 247 common bean lines from the Mesoamerican gene pool were used to determine the adaptive response in terms of phenological, physiological, and agronomic behavior under combined stress conditions. The lines tested were obtained from different single crosses, double crosses, and backcrosses between different bean materials, of which 146 were obtained from F4 families with high iron (Fe) content in seed and 99 common bean lines from F5 families. Different bean lines had grain yields (GY) higher than 1400 kg ha−1 from the F5 (lines: 859, 805, 865, and 657) and F4 (lines: 2853 and 2796) families. The superior performance of these lines was related to a higher photosynthate partitioning that has allowed an increase in pod formation (pod partitioning index, PPI) from the canopy biomass (CB) and grain filling (pod harvest index, PHI; harvest index, HI), resulting in higher values of GY. Values of GY were correlated with CB (r = 0.36), PPI (r = 0.6), PHI (r = 0.68), and HI (r = 0.8, p < 0.001). This increase in agronomic performance is due to a greater allocation of energy to the photosynthetic machinery (ΦII) and its dissipation in the form of heat (ΦNPQ), with increases in the leaf temperature difference (LTD). Based on the results obtained, six biofortified lines of common bean (lines F5: 859, 805, 865, and 657; lines F4: 2853 and 2796) showed traits of tolerance to combined stress and can serve as progenitors to increase Fe and Zn concentration in the seeds of lines that tolerate the combined stress from acidic soil and high temperature in the Colombian Amazon region.

Methods for evaluating effects of transgenes for quantitative traits

AbstractTransgenes that improve quantitative traits have traditionally been evaluated in one or a few genetic backgrounds across multiple environments. However, testing across multiple genetic backgrounds can be equally important to accurately quantify the value of a transgene for breeding objectives. Creating near‐isogenic lines across a wide germplasm space is costly and time consuming, which renders it impractical during early stages of testing. In this experiment, we evaluate three approaches to sample the genetic space while concurrently testing across environments. We created both transgenic and non‐transgenic doubled haploid maize (Zea mays L.) lines, F2:3 lines, and bulk F3 families to determine if all methods resulted in similar estimation of transgene value and to identify the number of yield trial plots from each method necessary to obtain a stable estimate of the transgene value. With one exception, the three methods consistently estimated a similar effect of the transgene. We suggest that bulked F3 lines topcrossed to a tester inbred is the most effective method to estimate the value of a transgene across both genetic space and environments.

Inheritance of resistance to S-metolachlor in a waterhemp (Amaranthus tuberculatus) population from central Illinois

AbstractWaterhemp [Amaranthus tuberculatus (Moq.) Sauer] is a dioecious weed that has evolved resistance to very-long-chain fatty-acid elongase (VLCFAE)–inhibiting herbicides via rapid metabolism. Although detoxification enzyme activities are associated with S-metolachlor resistance in two multiple herbicide–resistant (MHR) A. tuberculatus populations from Illinois, the genetic basis of resistance is unknown. Therefore, our goal was to investigate inheritance of S-metolachlor resistance in the Stanford, Illinois–resistant (SIR) population. Specifically, our research objectives were to: (1) generate a uniformly resistant, full-sib near-inbred line (DK3-2) via three generations of recurrent selection for resistance using preemergence S-metolachlor; (2) develop A. tuberculatus populations segregating for S-metolachlor resistance via reciprocal single-plant (one male × one female) full-sib mating of DK3-2 and a VLCFAE-inhibiting herbicide-sensitive population, SEN; (3) quantify S-metolachlor resistance levels in parental lines and their F1 progenies via greenhouse dose–response analysis; and (4) evaluate inheritance of S-metolachlor resistance in F2 progenies. Dose–response analysis using six to eight S-metolachlor concentrations (0.015 to 15.0 μM, varying per population) generated lethal dose (LD) estimates of 50% (LD50) and 90% (LD90) for SIR, SEN, DK3-2, and F1 progenies. LD estimates indicated DK3-2 has a higher magnitude of S-metolachlor resistance than the SIR population, demonstrating single crosses significantly increased S-metolachlor resistance in DK3-2. Levels of S-metolachlor resistance in F1 populations were intermediate compared with DK3-2 and SEN. Segregation of S-metolachlor resistance in F2 families from the paternal-derived lines fit a single-gene model (R:S = 3:1), indicating a single, dominant gene confers S-metolachlor resistance in SIR. However, F2 segregation results from the maternal-derived lines fit a duplicate recessive epistasis model (R:S = 9:7), indicating a second recessive gene may also modify S-metolachlor resistance in SIR. Results and germplasm derived from this research can assist in identifying the gene(s) conferring resistance to S-metolachlor in A. tuberculatus.

Genetic bases of resistance to the rice hoja blanca disease deciphered by a quantitative trait locus approach.

Rice hoja blanca (RHB) is one of the most serious diseases in rice-growing areas in tropical Americas. Its causal agent is RHB virus (RHBV), transmitted by the planthopper Tagosodes orizicolus Müir. Genetic resistance is the most effective and environment-friendly way of controlling the disease. So far, only 1 major quantitative trait locus (QTL) of Oryza sativa ssp. japonica origin, qHBV4.1, that alters the incidence of the virus symptoms in 2 Colombian cultivars has been reported. This resistance has already started to be broken, stressing the urgent need for diversifying the resistance sources. In the present study, we performed a search for new QTLs of O. sativa indica origin associated with RHB resistance. We used 4 F2:3-segregating populations derived from indica-resistant varieties crossed with a highly susceptible japonica pivot parent. Besides the standard method for measuring disease incidence, we developed a new method based on computer-assisted image processing to determine the affected leaf area (ALA) as a measure of symptom severity. Based on the disease severity and incidence scores in the F3 families under greenhouse conditions and SNP genotyping of the F2 individuals, we identified 4 new indica QTLs for RHB resistance on rice chromosomes 4, 6, and 11, namely, qHBV4.2WAS208, qHBV6.1PTB25, qHBV11.1, and qHBV11.2, respectively. We also confirmed the wide-range action of qHBV4.1. Among the 5 QTLs, qHBV4.1 and qHBV11.1 had the largest effects on incidence and severity, respectively. These results provide a more complete understanding of the genetic bases of RHBV resistance in the cultivated rice gene pool and can be used to develop marker-aided breeding strategies to improve RHB resistance. The power of joint- and meta-analyses allowed precise mapping and candidate gene identification, providing the basis for positional cloning of the 2 major QTLs qHBV4.1 and qHBV11.1.

Genetic variation among cocoa (Theobroma cacao L.) families for resistance to black pod disease under field and laboratory conditions

The laboratory detached black pod test method has been suggested as the most economical and effective screening procedure to complement field black disease resistance selection. General combining ability and association between black pod disease incidence under field and laboratory conditions among 24 cocoa F1 families were investigated over a 7-year period. The 24 F1 families were derived by crossing three seed garden parental clones with eight GU clones as males in a 3 x 8 North Carolina II (NC II) mating design. The 24 families together with four standard varieties planted in a randomized complete-block design with four replicates of 25 trees per plot were monitored for percentage black pod disease incidence (PBDI). Selfed pods from the 24 NC families and four standard varieties were arranged in a completely randomized design with four replicates of five pods each in the laboratory and subsequently inoculated with Phytophthora megakarya isolates. Percentage detached pod disease incidence (PDDI) was determined after seven days of incubation. Family effects was significant (p < .01) for PBDI and PDDI, and the association between them was positively and moderately correlated (r = 0.71; p < .01). Broad-sense heritability was high but narrow-sense heritability was low for both traits. General combining ability effects of eight of the 11 parental clones and specific combining ability effects of 17 of the 24 crosses were consistent (positive or negative) for PDDI and PBDI. Results from the study indicate that there is a considerable association between both traits, and clones GU 125/C and GU 255/V and their corresponding crosses PA 150 × GU 255/V, T60/887 × GU 125/C, and PA7/808 × GU 255/V had favorable effects for resistance to black pod disease expression. It is concluded that the detached black pod test represents a suitable method to support the field selection of black pod-resistant cocoa varieties.

The first linkage map for Australo-Papuan Treefrogs (family: Pelodryadidae) reveals the sex-determination system of the Green-eyed Treefrog (Litoria serrata)

Amphibians represent a useful taxon to study the evolution of sex determination because of their highly variable sex-determination systems. However, the sex-determination system for many amphibian families remains unknown, in part because of a lack of genomic resources. Here, using an F1 family of Green-eyed Treefrogs (Litoria serrata), we produce the first genetic linkage map for any Australo-Papuan Treefrogs (family: Pelodryadidae). The resulting linkage map contains 8662 SNPs across 13 linkage groups. Using an independent set of sexed adults, we identify a small region in linkage group 6 matching an XY sex-determination system. These results suggest Litoria serrata possesses a male heterogametic system, with a candidate sex-determination locus on linkage group 6. Furthermore, this linkage map represents the first genomic resource for Australo-Papuan Treefrogs, an ecologically diverse family of over 220 species.

Mapping of quantitative trait loci for growth traits around the first overwintering period in Songpu mirror carp (Cyprinus carpio L.) cultured in Northeast China

QTL mapping studies of growth traits based on high-density linkage maps in Northeast China has great significance for local enterprises in achieving MAS and improving the selection accuracy of parent fish. Here, we constructed a high-density genetic linkage map with 11,445 single nucleotide polymorphism (SNP) markers in a full-sib F1 family consisting of 120 progenies in Songpu mirror carp (Cyprinus carpio) reared in Northeast China. The consensus map covered 5471.93 centimorgans (cM) across the 50 linkage groups with an average resolution of 0.54 cM. Around the first overwintering period, a total of 15 QTLs for growth traits were identified on five LGs (LG7, LG8, LG14, LG30, and LG41). All of them were responsible for body weight (BW) at both 150 (before overwintering) and 350 (after overwintering) days, explaining 9.8–17.9 % of the phenotypic variation (PV). Two major-effect QTLs were detected on LG30, which explained 16.5 % and 16.4 % (qTL9–30), and 15.8 % and 17.9 % (qTL10–30) of the PV at 150 and 350 days, respectively. In addition, 15 loci were identified for 4 growth traits other than BW at both time points, including 6 loci for body length (BL), 7 loci for body height (BH), 3 loci for body thickness (BT) and 5 loci for head length (HL). These loci explained PV ranging from 15.9 % to 21.2 % for BL, 9.3–19.6 % for BH, 9.6–15.5 % for BT, and 10.6–18.1 % for HL. Nine candidate genes (IGF1b, LEPb, PACAP1a, GHSR1a, PPSS2, IRS1, APOAIb, IRS2b and ADSS1) were identified, five of which were directly related to growth hormone (GH), including IGF1b, PACAP1a, GHSR1a, PPSS2 and IRS1. Notably, three genes (GHSR1a, PPSS2 and IRS1) were derived from two major-effect QTLs (qTL9–30 and qTL10–30). In conclusion, these novel findings may shed light on early selection of growth traits of common carp cultured in Northeast China and other relatively high-latitude regions.

Combining ability analysis of yield and biomass allocation related traits in newly developed wheat populations

Increasing biomass allocation to the root system may increase soil-organic carbon stocks and confer drought adaptation in water-limited environments. Understanding the genetic bases and inheritance of biomass allocation is fundamental for drought tolerance breeding and soil health. The objective of this study was to determine the general and specific combining ability, maternal effects and the mode of gene action controlling the major yield and biomass allocation related traits in wheat to identify good combiners for breeding and enhanced carbon sequestration. Ten selected wheat genotypes were crossed in a full diallel mating design, and 90 F2 families were generated and evaluated in the field and greenhouse under drought-stressed and non-stressed conditions. Significant differences were recorded among the tested families revealing substantial variation for plant height (PH), kernels per spike (KPS), root biomass (RB), shoot biomass (SB), total plant biomass (PB) and grain yield (GY). Additive gene effects conditioned PH, SB, PB and GY under drought, suggesting the polygenic inheritance for drought tolerance. Strong maternal and reciprocal genetic effects were recorded for RB across the testing sites under drought-stressed conditions. Line BW162 had high yield and biomass production and can be used to transfer favourable genes to its progeny. The parental line LM75 maintained the general combining ability (GCA) effects in a positive and desirable direction for SB, PB and GY. Early generation selection using PH, SB, PB and GY will improve drought tolerance by exploiting additive gene action under drought conditions. Higher RB production may be maintained by a positive selection of male and female parents to capture the significant maternal and reciprocal effects found in this study.

Deciphering Biochemical Traits in Selected RILs of Mungbean [Vigna radiata (L.) Wilczek] against Bruchids (Callosobruchus maculatus F.)

Background: Mungbean [Vigna radiata (L.) Wilczek] is a vital pulse crop having a colossal nutritionary and gastronomic significance. The bruchid, Callosobruchus maculatus is recognised as the most heinous storage pest of leguminous crops that causes drastic economic loss. Crop breeders may use the nutritional and antinutritional traits of seeds to successfully combat storage pests. Methods: The present investigation has been framed to compare the degree of seed damage and weight loss with the biochemical parameters, viz., starch, sugar, protein, phenol, tannin and alpha-amylase inhibitor, of mungbean seeds from parental and selected lines of F5 populations for bruchid resistance. Result: The resistant parent, V2802BG and seven F5 families (BSR-GG-1-49-2, BSR-GG-1-49-5, BSR-GG-1-56-5, BSR-GG-1-170-5, BSR-GG-1-198-2, BSR-GG-1-198-3, BSR-GG-1-160-1) were completely resistant to bruchid infestation with no seed damage. One of the probable reasons for the stunted development of Callosobruchus maculatus might be the high level of phenol. Seed weight loss and damage by bruchids were shown to be positively correlated with high levels of protein, starch and sugar; however, alpha-amylase inhibitors and tannins did not reach statistical significance at the 5% level.

Molecular Marker and Test Cross Information Aid Selective Advancement of F4 Generation of CB174R/Azucena: An inter Sub-specific Cross in Rice for Restorer Development

The present investigation was aimed to develop restorer lines for three-line hybrid rice using indica/tropical japonica derivatives to exploit the inter sub-specific heterosis. From 75 F4 families of CB174R/Azucena, two plants were randomly selected and screened using simple sequence repeat markers DRRM-RF 3 -10 for Rf3 gene and RM6100 for Rf4 gene. One hundred and five plants possessing either or both of the genes were test crossed with CMS line COMS 23A. In 67 hybrids evaluated, the mean pollen fertility ranged from 97.3% (CB174R/Azucena 177-4-9) to 13.7% (CB174R/Azucena 13-2-4). The frequency of restorers was high (49.25%) followed by partial restorers (29.85%) and partial maintainers (20.90%). The selection efficiency for DRRM- RF3-10 and RM6100 markers were 75.75% and 54.54% respectively. Segregation for fertility restorer genes and pollen fertility among individual plants within a family was witnessed from molecular and phenotypic data. Based on phenotypic and marker information, it was concluded to advance 53.3% of plants to F5 generation to isolate stable restorer lines that can be exploited in future to produce highly heterotic three-line hybrids in rice.

Water Stress and Seed Color Interacting to Impact Seed and Oil Yield, Protein, Mucilage, and Secoisolariciresinol Diglucoside Content in Cultivated Flax (Linum usitatissimum L.).

Flaxseed (Linum usitatissimum L.) is a plant with a wide range of medicinal, health, nutritional, and industrial uses. This study assessed the genetic potential of yellow and brown seeds in thirty F4 families under different water conditions concerning seed yield, oil, protein, fiber, mucilage, and lignans content. Water stress negatively affected seed and oil yield, while it positively affected mucilage, protein, lignans, and fiber content. The total mean comparison showed that under normal moisture conditions, seed yield (209.87 g/m2) and most quality traits, including oil (30.97%), secoisolariciresinol diglucoside (13.89 mg/g), amino acids such as arginine (1.17%) and histidine (1.95%), and mucilage (9.57 g/100 g) were higher in yellow-seeded genotypes than the brown ones ((188.78 g/m2), (30.10%), (11.66 mg/g), (0.62%), (1.87%), and (9.35 g/100 g), respectively). Under water stress conditions, brown-seeded genotypes had a higher amount of fiber (16.74%), seed yield (140.04 g/m2), protein (239.02 mg. g-1), methionine (5.04%), and secondary metabolites such as secoisolariciresinol diglucoside (17.09 mg/g), while their amounts in families with yellow seeds were 14.79%, 117.33 g/m2, 217.12 mg. g-1, 4.34%, and 13.98 mg/g, respectively. Based on the intended food goals, different seed color genotypes may be appropriate for cultivation under different moisture environments.

Sex-Determined Growth of Yearling Pacific Oysters (Crassostrea gigas)

The sexes of sequentially hermaphroditic cupped oysters are both environmentally and genetically determined. Sex is also associated with growth or body size, females being larger than males of the same age on average. Whether sex determines size or size determines sex has not been resolved. To examine the relationship between growth and sex, 1,360 tagged individuals from six F2 families of Pacific oyster Crassostrea gigas were reared through their second summer in Thorndyke Bay, Washington; individual live weights were recorded for 5 consecutive months, starting in June. At the last weighing, microscopic examination of gonadal tissue revealed four “sexes” in the cohort, females (43% of cohort), males (29%), hermaphrodites (2%), and sex-undetermined individuals with no gametes (26%). Live weights were fit to various sigmoidal growth curves (logistic, Gompertz, and Richards) with fixed and random parameters to account for individual variation. The best fitting curves suggested that growth of the cohort was exponential until late August, when temperatures declined and more than 70% of final weight had been gained. Individual live weights, growth-curve parameters, and growth measures were fit to general linear models with family and sex classification variables. Family-by-sex interaction was significant in many such analyses. Females were larger than males on average and appeared to attain their size advantage from superior growth characteristics rather than from initial size. Males were strikingly either like females or like the smaller sex-undetermined oysters with respect to growth comparisons within family. Sex-undetermined oysters may have been “true” males that were too small to mature as yearlings, though a size-threshold for maturation was not apparent. Genetically determined sex and family genetics appeared to control growth.

Genetic analysis and mapping of dwarf gene without yield penalty in a γ-ray-induced wheat mutant.

Plant height is one of the most important agronomic traits that affects yield in wheat, owing to that the utilization of dwarf or semi-dwarf genes is closely associated with lodging resistance. In this study, we identified a semi-dwarf mutant, jg0030, induced by γ-ray mutagenesis of the wheat variety 'Jing411' (wild type). Compared with the 'Jing411', plant height of the jg0030 mutant was reduced by 7%-18% in two years' field experiments, and the plants showed no changes in yield-related traits. Treatment with gibberellic acid (GA) suggested that jg0030 is a GA-sensitive mutant. Analysis of the frequency distribution of plant height in 297 F3 families derived from crossing jg0030 with the 'Jing411' indicated that the semi-dwarf phenotype is controlled by a major gene. Using the wheat 660K SNP array-based Bulked Segregant Analysis (BSA) and the exome capture sequencing-BSA assay, the dwarf gene was mapped on the long arm of chromosome 2B. We developed a set of KASP markers and mapped the dwarf gene to a region between marker PH1 and PH7. This region encompassed a genetic distance of 55.21 cM, corresponding to a physical distance of 98.3 Mb. The results of our study provide a new genetic resource and linked markers for wheat improvement in molecular breeding programs.

Homozygosity mapping identified loci and candidate genes responsible for freezing tolerance in Camelina sativa.

Homozygosity mapping is an effective tool for detecting genomic regions responsible for a given trait when the phenotype is controlled by a limited number of dominant or co-dominant loci. Freezing tolerance is a major attribute in agricultural crops such as camelina. Previous studies indicated that freezing tolerance differences between a tolerant (Joelle) and susceptible (CO46) variety of camelina were controlled by a small number of dominant or co-dominant genes. We performed whole genome homozygosity mapping to identify markers and candidate genes responsible for freezing tolerance difference between these two genotypes. A total of 28 F3 RILs were sequenced to ∼30× coverage, and parental lines were sequenced to >30-40× coverage with Pacific Biosciences high fidelity technology and 60× coverage using Illumina whole genome sequencing. Overall, about 126k homozygous single nucleotide polymorphism markers were identified that differentiate both parents. Moreover, 617 markers were also homozygous in F3 families fixed for freezing tolerance/susceptibility. All these markers mapped to two contigs forming a contiguous stretch of chromosome 11. The homozygosity mapping detected 9 homozygous blocks among the selected markers and 22 candidate genes with strong similarity to regions in or near the homozygous blocks. Two such genes were differentially expressed during cold acclimation in camelina. The largest block contained a cold-regulated plant thionin and a putative rotamase cyclophilin 2 gene previously associated with freezing resistance in arabidopsis (Arabidopsis thaliana). The second largest block contains several cysteine-rich RLK genes and a cold-regulated receptor serine/threonine kinase gene. We hypothesize that one or more of these genes may be primarily responsible for freezing tolerance differences in camelina varieties.

Gene Editing of the Follicle-Stimulating Hormone Gene to Sterilize Channel Catfish, Ictalurus punctatus, Using a Modified Transcription Activator-like Effector Nuclease Technology with Electroporation

Follicle-stimulating hormone (fsh) plays an important role in sexual maturation in catfish. Knocking out the fsh gene in the fish zygote should suppress the reproduction of channel catfish (Ictalurus punctatus). In this study, transcription activator-like effector nuclease (TALEN) plasmids targeting the fsh gene were electroporated into fertilized eggs with the standard double electroporation technique. Targeted fsh cleavage efficiency was 63.2% in P1fsh-knockout catfish. Ten of fifteen (66.7%) control pairs spawned, and their eggs had 32.3-74.3% average hatch rates in 2016 and 2017. Without hormone therapy, the spawning rates of P1 mutants ranged from 33.3 to 40.0%, with an average egg hatching rate of 0.75%. After confirmation of the low fertility of P1 mutants in 2016, human chorionic gonadotropin (HCG) hormone therapy improved the spawning rates by 80% for female mutants and 88.9% for male mutants, and the mean hatch rate was 35.0% for F1 embryos, similar to that of the controls (p > 0.05). Polymerase chain reaction (PCR) identification showed no potential TALEN plasmid integration into the P1 channel catfish genome. Neither the P1 nor the F1 mutant fish showed any noticeable changes in in body weight, survival rate, and hatching rate when the reproductive gene was knocked out. F1 families had a mean inheritance rate of 50.3%. The results brought us one step closer to allowing implementation of certain genetic techniques to aquaculture and fisheries management, while essentially eliminating the potential environment risk posed by transgenic, hybrid, and exotic fish as well as domestic fish.

Resistance Allele Frequency of Helicoverpa zea to Vip3Aa Bacillus thuringiensis Protein in the Southeastern U.S.

Helicoverpa zea is a major target pest of Bt crops expressing Cry and/or Vip3Aa proteins in the U.S.A. Widespread practical resistance of H. zea to the Cry1 and Cry2 proteins makes Vip3Aa the only effective Bt protein against this pest. Understanding the frequency of resistance alleles against Vip3Aa in field populations of H. zea is crucial for resistance management and the sustainability of Vip3Aa technology. Using a modified F2 screen method by crossing susceptible laboratory female moth with feral male moth of H. zea, we successfully screened a total of 24,576 neonates from 192 F2 families of H. zea collected from Arkansas, Louisiana, Mississippi, and Tennessee during 2019-2020. We found five F2 families containing ≥3rd instar survivors on the diagnostic concentration of 3.0 µg/cm2 Vip3Aa39. Dose-response bioassays confirmed the high levels of Vip3Aa resistance in these F2 families, with an estimated resistance ratio of >909.1-fold relative to the susceptible strain. The estimated resistance allele frequency against Vip3Aa in H. zea for these four southern states is 0.0155 with a 95% CI of 0.0057-0.0297. These data should provide critical information for understanding the risks of Vip3Aa resistance in H. zea and help design appropriate resistance management strategies for the sustainability of the Vip3Aa technology.

Selection Procedures for Improving Some Economic Characters and Correlated Responses in Cotton (Gossypium barbadense L.) Cross

A study was done at Sakha Agricultural Research Station Farm, Kafr El-Sheikh, Egypt during 2019-2021 growing seasons. The study aimed at assessing the efficacy and usability of various selection procedures, determining the effectiveness of selection for superior families and estimating the response to selection. The materials utilized for selection of promising families in early segregating generations included F2, F3 and F4 generations of Giza 94 × S106 cotton cross.. Findings revealed better F4 means than the F3 and F2 means for all studied characters due to the selection procedures used. High heritability values were recorded for most studied characters. Significant desirable correlations were noticed between seed cotton yield with lint yield, bolls/plant, lint/seed, seeds/boll and boll weight. Canonical discriminant analysis among F3 families showed that the first five canonical functions accounted for 100% of total variances. While first canonical discriminant function represented 46% of the total variance among genotypes with the greatest Eigen value and prevailed by great loading from most yield characters and micronaire reading. Ten of the selection indices surpassed direct selection in improving lint yield. The highest actual advance in lint yield for F3 obtained from selection index comprising lint yield/plant in addition to bolls/plant, seeds per boll and lint per seed. In F4 generation, the maximum actual gains were obtained for most yield traits with indices involved lint yield/plant and seeds/boll with lint/seed. Most of the selected families in F4 scored high values for almost all the studied characters and surpassed the corresponding means of selected families in F3 and F2 generations; these selected families might be used to improve cotton yield and fiber quality in breeding programs.

A Modified F2 Screen for Estimating Cry1Ac and Cry2Ab Resistance Allele Frequencies in Helicoverpa zea (Lepidoptera: Noctuidae).

Evaluating the frequency of resistance alleles is important for resistance management and sustainable use of transgenic crops that produce insecticidal proteins from Bacillus thuringiensis. Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae) is a major crop pest in the United States that has evolved practical resistance to the crystalline (Cry) proteins in Bt corn and cotton. The standard F2 screen for estimating resistance allele frequency does not work well for H. zea because successful single-pair matings are rare. In this study, we developed and implemented a modified F2 screen for H. zea that generates F1 progeny by crossing three laboratory susceptible female moths with one feral male moth instead of single-pair crosses. During 2019-2020, we used this modified method to establish 192 F2 families from 623 matings between susceptible females and feral males from Arkansas, Louisiana, Mississippi, and Tennessee. From each F2 family, we screened 128 neonates against discriminating concentrations of Cry1Ac and Cry2Ab in diet overlay bioassays. Based on these discriminating concentration bioassays, families were considered positive for resistance if at least five larvae survived to second instar, including at least one to third instar. The percentage of positive families was 92.7% for Cry1Ac and 38.5% for Cry2Ab, which yields an estimated resistance allele frequency (with 95% confidence interval) of 0.722 (0.688-0.764) for Cry1Ac and 0.217 (0.179-0.261) for Cry2Ab. The modified F2 screen developed and implemented here may be useful for future resistance monitoring studies of H. zea and other pests.

Genetic Diversity and Differences among Three F1 Families and Two Wild Populations of Genus Scylla Using Microsatellite Markers

Genetic diversity is the determinant of the allocation of germplasm resources in the genetic improvement of aquaculture species. In this study, three F1 families, including a hybrid Scylla family (S. paramamosain ♂ × S. serrata ♀), a paternal family of S. paramamosain, a maternal family of S. serrata, and two wild populations, including a paternal population of S. paramamosain and a maternal population of S. serrata, were used to investigate the genetic diversity and genetic difference. The results indicated that 98 alleles of nine microsatellites loci were observed in five Scylla populations. The highest average value of Ho (observed heterozygosity), He (expected heterozygosity), and PIC (polymorphic information content) of the wild S. paramamosain population were 0.790, 0.799, and 0.771, respectively, suggesting the wild paternal population has high genetic diversity. The comparative analysis of PIC, Fst (fixation index), and HWE (Hardy–Weinberg equilibrium) indicated that the paternal S. paramamosain may be more suitable for artificial breeding than the maternal S. serrata from the perspective of allele frequency. Analysis of molecular variance analysis (AMOVA) showed that the total genetic variation mainly occurred within populations (73.28%), demonstrating that artificial breeding may induce genetic differentiation of the family groups of Scylla. The results of the analysis of Fst value, UPGMA (unweighted pair-group mean analysis) dendrogram, and genetic diversity indicated that the F1 hybrid offspring had a close genetic distance and high genetic identity with the paternal S. paramamosain populations. It indicated that the F1 hybrid offspring showed potential paternal genetic affinities and a similar potential for artificial breeding with S. paramamosain. The study will provide valuable information to evaluate the difference in the genetic diversity and population structure between hybrid offspring and distinct parental populations of Scylla.