T. G. Isleib

High Levels of Field Resistance to Tomato spotted wilt virus in Peanut Breeding Lines Derived from hypogaea and hirsuta Botanical Varieties

Tomato spotted wilt, caused by Tomato spotted wilt tospovirus (TSWV) is a major problem in peanut (Arachis hypogaea L.) producing areas of the southeastern U.S. The integrated program used to manage spotted wilt relies heavily on cultivars with field resistance to TSWV, and finding new sources and greater levels of resistance to TSWV is highly desirable. Field tests were conducted in 2003 and 2004 in Marianna, FL and Tifton, GA to compare three peanut breeding lines, F NC94022-1-2-1-1-b3-B, C 11-2-39, and C 11-186 to that of standard moderately resistant cultivar Georgia Green for field response to TSWV. F NC94022-1-2-1-1-b3-B was of particular interest because it was developed from a cross between lines of A. hypogaea subsp. hypogaea var. hirsuta Kohler and A. hypogaea subsp. hypogaea var. hypogaea. In all tests, final spotted wilt ratings for breeding lines F NC94022-1-2-1-1-b3-B, C 11-2-39, and C 11-186 were lower and pod yields were higher than for Georgia Green. In three tests, final spotted wilt intensity ratings did not differ among F NC94022-1-2-1-1-b3-B, C 11-2-39, and C 11-186. At Marianna in 2004, spotted wilt intensity ratings were lower and pod yields were higher in F NC94022-1-2-1-1-b3-B than in any other entry. The high level of field resistance to TSWV in F NC94022-1-2-1-1-b3-B is presumably derived from its hirsuta type parent, PI 576638.

Evaluation of peanut (Arachis hypogaea L.) leaflets from mature zygotic embryos as recipient tissue for biolostic gene transfer

Leaflets from mature peanut embryos are a useful recipient tissue for biolistic DNA transfer. Fertile plants were regenerated from leaflets from genotypes representing all botanical types of peanut. Regeneration frequency was strongly influenced by genotype. NPT II and GUS chimaeric gene fusions, driven by the CaMV 35S promoter, were expressed transiently following biolistic delivery to unexpanded leaflets. Bombardment conditions affecting transient expression frequency were determined using a prototype of the Bio Rad PDS 1000/He helium-powered particle acceleration apparatus. Stably transformed calli were derived routinely from leaflet tissue bombarded with the NPT II gene and subsequently cultured on kanamycin. Several plants have been regenerated from treated explants under kanamycin selection. Thus far, none of these has been stably transformed. The occurrence of escapes suggests that kanamycin is an inefficient selective agent for the recovery of transgenic peanuts from this explant. Experiments designed to regenerate plants using published regeneration protocols from stably transformed calli, devoid of primary explant tissue, have been unsuccessful.

Genome-wide SNP Genotyping Resolves Signatures of Selection and Tetrasomic Recombination in Peanut

Peanut (Arachis hypogaea; 2n = 4x = 40) is a nutritious food and a good source of vitamins, minerals, and healthy fats. Expansion of genetic and genomic resources for genetic enhancement of cultivated peanut has gained momentum from the sequenced genomes of the diploid ancestors of cultivated peanut. To facilitate high-throughput genotyping of Arachis species, 20 genotypes were re-sequenced and genome-wide single nucleotide polymorphisms (SNPs) were selected to develop a large-scale SNP genotyping array. For flexibility in genotyping applications, SNPs polymorphic between tetraploid and diploid species were included for use in cultivated and interspecific populations. A set of 384 accessions was used to test the array resulting in 54 564 markers that produced high-quality polymorphic clusters between diploid species, 47 116 polymorphic markers between cultivated and interspecific hybrids, and 15 897 polymorphic markers within A. hypogaea germplasm. An additional 1193 markers were identified that illuminated genomic regions exhibiting tetrasomic recombination. Furthermore, a set of elite cultivars that make up the pedigree of US runner germplasm were genotyped and used to identify genomic regions that have undergone positive selection. These observations provide key insights on the inclusion of new genetic diversity in cultivated peanut and will inform the development of high-resolution mapping populations. Due to its efficiency, scope, and flexibility, the newly developed SNP array will be very useful for further genetic and breeding applications in Arachis.

Evaluating Isolate Aggressiveness and Host Resistance from Peanut Leaflet Inoculations with Sclerotinia minor

Sclerotinia minor is a major pathogen of peanut in North Carolina, Virginia, Oklahoma, and Texas. Partial resistance to S. minor has been reported based on field screening, but field performance is not always correlated with laboratory or greenhouse evaluations of resistance. More efficient screening methods and better understanding of the mechanisms contributing to Sclerotinia blight resistance are needed, and a detached leaf assay was developed and evaluated. Detached leaflets of 12 greenhouse-grown peanut lines were inoculated on the adaxial surface with a 4-mm-diameter mycelial plug of a single isolate of S. minor. Leaflets were incubated in the dark at 20°C in Nalgene utility boxes containing moistened sand. Lesion length 3 days after inoculation ranged from 11 to 24 mm, with a mean of 19 mm. Lengths differed significantly among the entries, with GP-NC WS 12, an advanced breeding line derived from a cross of NC 6 × (NC 3033 × GP-NC WS 1), being the most resistant. Forty-eight isolates of S. minor obtained from peanut were inoculated on leaflets of the susceptible cultivar NC 7 and aggressiveness was assessed by measuring lesion-length expansion. Three days after inoculation, lesion length differed among the isolates and ranged from 2 to 24 mm, with a mean of 15 mm. Finally, the potential for specific interactions between peanut lines and S. minor isolates was evaluated. A subset of S. minor isolates was selected to represent the observed range of aggressiveness and a subset of peanut entries was selected to represent the range of resistance or susceptibility. Nine-week-old greenhouse- or field-grown plants were compared for five peanut entries. Main effects of isolates and entries were highly significant, but isolate-entry interactions were not significant. The most resistant peanut entry (GP-NC WS 12) performed consistently with all isolates regardless of plant source.

Genotypic effect of ahFAD2 on fatty acid profiles in six segregating peanut (Arachis hypogaea L) populations.

BackgroundFatty acid composition of oil extracted from peanut (Arachis hypogaea L.) seed is an important quality trait because it may affect the flavor and shelf life of resulting food products. In particular, a high ratio of oleic (C18:1) relative to linoleic (C18:2) fatty acid (O/L ≥ 10) results in a longer shelf life. Previous reports suggest that the high oleic (~80%) trait was controlled by recessive alleles of ahFAD2A and ahFAD2B, the former of which is thought to have a high frequency in US runner- and virginia-type cultivars. Functional mutations, G448A in ahFAD2A and 442insA in ahFAD2B eliminate or knock down desaturase activity and have been demonstrated to produce peanut oil with high O/L ratios. In order to employ marker assisted selection (MAS) to select a high oleic disease resistant peanut and to evaluate genotypic and phenotypic variation, crosses were made between high oleic (~80%) and normal oleic (~50%) peanuts to produce segregating populations.ResultsA total of 539 F2 progenies were randomly selected to empirically determine each ahFAD2 genotype and the resulting fatty acid composition. Five of the six crosses segregated for the high oleic trait in a digenic fashion. The remaining cross was consistent with monogenic segregation because both parental genotypes were fixed for the ahFAD2A mutation. Segregation distortion was significant in ahFAD2A in one cross; however, the remaining crosses showed no distortion. Quantitative analyses revealed that dominance was incomplete for the wild type allele of ahFAD2, and both loci showed significant additive effects. Oleic and linoleic acid displayed five unique phenotypes, based on the number of ahFAD2 mutant alleles. Further, the ahFAD2 loci did exhibit pleiotropic interactions with palmitic (C16:0), oleic (C18:1), linoleic (C18:2) acids and the O/L ratio. Fatty acid levels in these progeny were affected by the parental genotype suggesting that other genes also influence fatty acid composition in peanut. As far as the authors are aware, this is the first study in which all of the nine possible ahFAD2 genotypes were quantitatively measured.ConclusionsThe inheritance of the high oleic trait initially was suggested to be controlled by dominant gene action from two homoeologous genes (ahFAD2A and ahFAD2B) exhibiting complete recessivity. Analyzing the ahFAD2 genotypes and fatty acid compositions of these segregating peanut populations clearly demonstrated that the fatty acid contents are quantitative in nature although much of the variability in the predominant fatty acids (oleic, linoleic, and palmitic) is controlled by only two loci.

Genotype-by-Environment Interactions for Seed Composition Traits of Breeding Lines in the Uniform Peanut Performance Test

Peanut composition is influenced by several groups of factors: environmental, genetic, and their interaction. This study evaluated the relative contributions of these factors using data from the USDA-ARS quality testing program using samples from the multi-state Uniform Peanut Performance Tests (UPPT). Data were subjected to restricted maximum likelihood estimation of variance components reflecting the main effects of year, production region, location within region, genotype (cultivar or breeding line), and kernel grade (‘‘seed size’’) within genotype, and the interactions among these main effects. Genetic variation in oil content was low (9% of total variation); however, fatty acid composition of the oil was highly influenced by genotype (34–77%) with the exception of lignoceric acid (1%). Genetic influence on tocopherols was generally less than that of fatty acids. Environmental variation of tocopherols was greater than the variation attributable to genotype-by-environment interaction. The lowest genetic variation was observed in sugar content; however, environmental variation was high (68%). The magnitude of genetic influence on oil content and fatty acid concentrations suggests that these traits are amenable to improvement through breeding.

Screening of Virginia-Type Peanut Breeding Lines for Resistance to Cylindrocladium Black Rot and Sclerotinia Blight in the Greenhouse

Abstract Cylindrocladium black rot (CBR) caused by Cylindrocladium parasiticum and Sclerotinia blight caused by Sclerotinia minor are two economically important diseases of peanut (Arachis hypogaea...

Allergenicity attributes of different peanut market types.

Four different market classes of peanut (Runner, Virginia Spanish, and Valencia) are commonly consumed in Western countries, but for some consumers peanuts are a main cause of food-induced anaphylaxis. Limited information is available on the comparative allergenicity of these distinct market classes. The aim of this study was to compare allergenicity attributes of different peanut cultivars. The protein content and protein profiles were highly comparable for all tested cultivars. All cultivar samples contained the major allergens Ara h 1, Ara h 2, Ara h 3 and Ara h 6, as assessed by SDS-PAGE and RP-HPLC, although some minor differences in major allergen content were found between samples. All samples were reactive in commercial ELISAs for detection and quantification of peanut protein. IgE-binding potency differed between samples with a maximum factor of 2, indicating a highly comparable allergenicity. Based on our observations, we conclude that peanuts from the main market types consumed in Western countries are highly comparable in their allergenicity attributes, indicating that safety considerations with regard to peanut allergy are not dependent on the peanut cultivar in question.

Development and Phenotyping of Recombinant Inbred Line (RIL) Populations for Peanut (Arachis hypogaea)

ABSTRACT The identification of molecular markers for economically significant traits should greatly improve the speed and efficiency of all peanut (Arachis hypogaea L.) breeding programs. Developme...

A Site-Specific, Weather-Based Disease Regression Model for Sclerotinia Blight of Peanut

In North Carolina, losses due to Sclerotinia blight of peanut, caused by the fungus Sclerotinia minor, are an estimated 1 to 4 million dollars annually. In general, peanut (Arachis hypogaea) is very susceptible to Sclerotinia blight, but some partially resistant virginia-type cultivars are available. Up to three fungicide applications per season are necessary to maintain a healthy crop in years highly favorable for disease development. Improved prediction of epidemic initiation and identification of periods when fungicides are not required would increase fungicide efficiency and reduce production costs on resistant and susceptible cultivars. A Sclerotinia blight disease model was developed using regression strategies in an effort to describe the relationships between modeled environmental variables and disease increase. Changes in incremental disease incidence (% of newly infected plants of the total plant population per plot) for the 2002-2005 growing seasons were statistically transformed and described using 5-day moving averages of modeled site-specific weather variables (localized, mathematical estimations of weather data derived at a remote location) obtained from SkyBit (ZedX, Inc.). Variables in the regression to describe the Sclerotinia blight disease index included: mean relative humidity (linear and quadratic), mean soil temperature (quadratic), maximum air temperature (linear and quadratic), maximum relative humidity (linear and quadratic), minimum air temperature (linear and quadratic), minimum relative humidity (linear and quadratic), and minimum soil temperature (linear and quadratic). The model explained approximately 50% of the variability in Sclerotinia blight index over 4 years of field research in eight environments. The relationships between weather variables and Sclerotinia blight index were independent of host partial resistance. Linear regression models were used to describe progress of Sclerotinia blight on cultivars and breeding lines with varying levels of partial resistance. Resistance affected the rate of disease progress, but not disease onset. The results of this study will be used to develop site- and cultivar-specific spray advisories for Sclerotinia blight.