[show abstract][hide abstract] ABSTRACT: Drought is a major abiotic stress limiting crop productivity worldwide especially in rain-fed areas. Plant physiologists and crop breeders have aimed to better understand the mechanisms underlying drought resistance to increase the success in breeding for drought resistant crops. The objectives of this study were to (i) determine the effects of different soil water availability on stomatal behavior and water use efficiency (WUE) of diverse peanut genotypes and ii) to investigate associations between surrogate physiological traits for drought tolerance. Eleven peanut genotypes and three soil moisture levels (field capacity, 2/3 available water and 1/3 available water) were assigned in a split plot design with four replications for two experiments. WUE, stomatal traits and root distribution (indicated by root length density, RLD, in the soil layers at 0-40 cm and 40-100 cm depths) were measured at 37, 67 and 97 days after sowing (DAS). Drought reduced stomatal aperture and stomatal conductance but increased WUE. Differential responses among peanut genotypes were observed for WUE, and stomatal conductance. ICGV 98300 and Tifton 8 had high stomatal conductance and WUE under moderate water stress. Stomatal conductance and %RLD (40-100cm) play an important roles in increasing WUE under mild drought stress.
[show abstract][hide abstract] ABSTRACT: Peanut seeds contain high amounts of oil and protein as well as some useful bioactive phytochemicals which can contribute to human health. The U.S. peanut mini-core collection is an important genetic resource for improving seed quality and developing new cultivars. Variability of seed chemical composition within the mini-core was evaluated from freshly harvested seeds for two years. Oil, fatty acid composition, and flavonoid/ resveratrol content were quantified by NMR, GC, and HPLC, respectively. Significant variability was detected in seed chemical composition among accessions and botanical varieties. Accessions were further genotyped with a functional SNP marker from the FAD2A gene using real-time PCR and classified into three genotypes with significantly different O/L ratios: wild type (G/G with a low O/L ratio <1.7), heterozygote (G/A with O/L ratio >1.4 but <1.7), and mutant (A/A with a high O/L ratio >1.7). The results from real-time PCR genotyping and GC fatty acid analysis were consistent. Accessions with high amounts of oil, quercetin, high seed weight, and O/L ratio were identified. The results from this study may be useful not only for peanut breeders, food processors, and product consumers to select suitable accessions or cultivars, but also for curators to potentially expand the mini-core collection.
Journal of Agricultural and Food Chemistry 02/2013; · 2.91 Impact Factor
[show abstract][hide abstract] ABSTRACT: Peanut kernels are susceptible to colonization by some species of Aspergillus which, under conditions of drought and high temperatures, can produce aflatoxins prior to harvest. The objective of this research was to determine the mechanism by which the peanut root-knot nematode (Meloidogyne arenaria) increases aflatoxin contamination in peanut. Research determined 1) the role of nematode infection of roots vs. pods in increased aflatoxin contamination and 2) whether increased aflatoxin production in nematode-infected peanut is due to a greater percentage of small or immature kernels. An additional objective was to determine whether a peanut cultivar with resistance to M. arenaria would reduce the risk of preharvest aflatoxin contamination. In the greenhouse, researchers physically separated root growth from pod set and inoculated each location with M. arenaria or a water control in a 2 × 2 factorial design with 12–15 replications. Of the six trials conducted, data indicated that pod and root infection by M. arenaria was associated with elevated aflatoxin concentrations in one and three trials, respectively. This suggests that root infection by the nematode can increase aflatoxin concentrations in the peanut kernel. Another 2 × 2 factorial experiment was conducted with two peanut genotypes (Tifguard and TifGP-2) and two nematode treatments (with and without M. arenaria) with six replications. The cultivar Tifguard is resistant to M. arenaria and TifGP-2 is susceptible. The experiment was carried out in 24 field microplots equipped with a rainout shelter. The experiment was conducted five times from 2006 to 2010. Infection of TifGP-2 by M. arenaria did not lead to greater percentages of small kernels. In only one year (2007), nematodes appeared to increase the percentage of damaged kernels, though aflatoxin concentrations were not affected by nematodes in that year. In the rainout shelter experiment, 2006 was the only year where nematode infection of peanut increased aflatoxin concentrations. In that year, there were lower aflatoxin concentrations in the nematode-resistant cultivar Tifguard than the susceptible germplasm TifGP-2 (12 vs. 136 ng/g).
[show abstract][hide abstract] ABSTRACT: Drought affects yield of peanut, but its effect on oleic and linoleic acids that influence its oil quality of peanut genotypes with different levels of drought resistance has not been clearly investigated. Therefore, the aims of this research were to determine whether soil water levels could affect oil quality by changes in fatty acid compositions of peanut, and to investigate the changes in oil characters in peanut genotypes with their potential drought resistance under different water regimes. Field experiments were conducted in split-plot designs with four replications during dry season for two years (2003/04 and 2004/05). Three water regimes [field capacity (FC), 2/3 available soil water (2/3 AW) and 1/3 available soil water (1/3 AW)] were assigned as main-plots, and six peanut genotypes were assigned as sub-plots. The data were recorded at maturity for fatty acid compositions and % oil. Seed samples were analyzed for % oil by Soxtec System HT, and fatty acid compositions were analyzed by gas liquid chromatography. Differences among water regimes and peanut genotypes were significant for oleic and linoleic acids content and their ratio (O/L ratio), unsaturated to saturated fatty acid ratio (U/S ratio) and iodine value (IV). Genotype × water regime interactions were also significant for all characters. Drought improved the oil quality by significant increase in oleic acid and O/L ratio, and reduced the linoleic acid, IV and U/S ratio. Peanut genotypes with different levels of drought resistance displayed similar tendency in fatty acid characters under drought conditions.
[show abstract][hide abstract] ABSTRACT: Many plant ESTs have been sequenced as an alternative to whole genome sequences, including peanut because of the genome size and complexity. The US peanut research community had the historic 2004 Atlanta Genomics Workshop and named the EST project as a main priority. As of August 2011, the peanut research community had deposited 252,832 ESTs in the public NCBI EST database, and this resource has been providing the community valuable tools and core foundations for various genome-scale experiments before the whole genome sequencing project. These EST resources have been used for marker development, gene cloning, microarray gene expression and genetic map construction. Certainly, the peanut EST sequence resources have been shown to have a wide range of applications and accomplished its essential role at the time of need. Then the EST project contributes to the second historic event, the Peanut Genome Project 2010 Inaugural Meeting also held in Atlanta where it was decided to sequence the entire peanut genome. After the completion of peanut whole genome sequencing, ESTs or transcriptome will continue to play an important role to fill in knowledge gaps, to identify particular genes and to explore gene function.
Comparative and Functional Genomics 01/2012; 2012:373768. · 0.92 Impact Factor
[show abstract][hide abstract] ABSTRACT: Construction and improvement of a genetic map for peanut (Arachis hypogaea L.) continues to be an important task in order to facilitate quantitative trait locus (QTL) analysis and the development of tools for marker-assisted breeding. The objective of this study was to develop a comparative integrated map from two cultivated × cultivated recombinant inbred line (RIL) mapping populations and to apply in mapping Tomato spotted wilt virus (TSWV) resistance trait in peanut. A total of 4,576 simple sequence repeat (SSR) markers from three sources: published SSR markers, newly developed SSR markers from expressed sequence tags (EST) and from bacterial artificial chromosome end-sequences were used for screening polymorphisms. Two cleaved amplified polymorphic sequence markers were also included to differentiate ahFAD2A alleles and ahFAD2B alleles. A total of 324 markers were anchored on this integrated map covering 1,352.1 cM with 21 linkage groups (LGs). Combining information from duplicated loci between LGs and comparing with published diploid maps, seven homoeologous groups were defined and 17 LGs (A1-A10, B1-B4, B7, B8, and B9) were aligned to corresponding A-subgenome or B-subgenome of diploid progenitors. One reciprocal translocation was confirmed in the tetraploid-cultivated peanut genome. Several chromosomal rearrangements were observed by comparing with published cultivated peanut maps. High consistency with cultivated peanut maps derived from different populations may support this integrated map as a reliable reference map for peanut whole genome sequencing assembling. Further two major QTLs for TSWV resistance were identified for each RILs, which illustrated the application of this map.
Theoretical and Applied Genetics 11/2011; 124(4):653-64. · 3.66 Impact Factor
[show abstract][hide abstract] ABSTRACT: Peanut (Arachis hypogaea L.) is one of the most important oilseed and nutritional crops in the world. To efficiently utilize the germplasm collection, a peanut mini-core containing 112 accessions was established in the United States. To determine the population structure and its impact on marker-trait association, this mini-core collection was assessed by genotyping 94 accessions with 81 SSR markers and two functional SNP markers from fatty acid desaturase 2 (FAD2). Seed quality traits (including oil content, fatty acid composition, flavonoids, and resveratrol) were obtained through nuclear magnetic resonance (NMR), gas chromatography (GC), and high-performance liquid chromatography (HPLC) analysis. Genetic diversity and population structure analysis identified four major subpopulations that are related to four botanical varieties. Model comparison with different levels of population structure and kinship control was conducted for each trait and association analyses with the selected models verified that the functional SNP from the FAD2A gene is significantly associated with oleic acid (C18:1), linoleic acid (C18:2), and oleic-to-linoleic (O/L) ratio across this diverse collection. Even though the allele distribution of FAD2A was structured among the four subpopulations, the effect of FAD2A gene remained significant after controlling population structure and had a likelihood-ratio-based R ( 2 ) (R ( LR ) ( 2 ) ) value of 0.05 (oleic acid), 0.09 (linoleic acid), and 0.07 (O/L ratio) because the FAD2A alleles were not completely fixed within subpopulations. Our genetic analysis demonstrated that this peanut mini-core panel is suitable for association mapping. Phenotypic characterization for seed quality traits and association testing of the functional SNP from FAD2A gene provided information for further breeding and genetic research.
Theoretical and Applied Genetics 08/2011; 123(8):1307-17. · 3.66 Impact Factor
[show abstract][hide abstract] ABSTRACT: Allergic reactions to peanuts (Arachis hypogaea L.) can cause severe symptoms and in some cases can be fatal, but avoidance is difficult due to the prevalence of peanut-derived products in processed foods. One strategy of reducing the allergenicity of peanuts is to alter or eliminate the allergenic proteins through mutagenesis. Other seed quality traits could be improved by altering biosynthetic enzyme activities. Targeting Induced Local Lesions in Genomes (TILLING), a reverse-genetics approach, was used to identify mutations affecting seed traits in peanut.
Two similar copies of a major allergen gene, Ara h 1, have been identified in tetraploid peanut, one in each subgenome. The same situation has been shown for major allergen Ara h 2. Due to the challenge of discriminating between homeologous genes in allotetraploid peanut, nested PCR was employed, in which both gene copies were amplified using unlabeled primers. This was followed by a second PCR using gene-specific labeled primers, heteroduplex formation, CEL1 nuclease digestion, and electrophoretic detection of labeled fragments. Using ethyl methanesulfonate (EMS) as a mutagen, a mutation frequency of 1 SNP/967 kb (3,420 M2 individuals screened) was observed. The most significant mutations identified were a disrupted start codon in Ara h 2.02 and a premature stop codon in Ara h 1.02. Homozygous individuals were recovered in succeeding generations for each of these mutations, and elimination of Ara h 2.02 protein was confirmed. Several Ara h 1 protein isoforms were eliminated or reduced according to 2D gel analyses. TILLING also was used to identify mutations in fatty acid desaturase AhFAD2 (also present in two copies), a gene which controls the ratio of oleic to linoleic acid in the seed. A frameshift mutation was identified, resulting in truncation and inactivation of AhFAD2B protein. A mutation in AhFAD2A was predicted to restore function to the normally inactive enzyme.
This work represents the first steps toward the goal of creating a peanut cultivar with reduced allergenicity. TILLING in peanut can be extended to virtually any gene, and could be used to modify other traits such as nutritional properties of the seed, as shown in this study.
[show abstract][hide abstract] ABSTRACT: Germin-like proteins (GLPs) play diversified roles in plant development and defense response. Here, we identified 36 expressed
sequence tags (ESTs) encoding GLPs from peanut (Arachis hypogaea L.). After assembly, these ESTs were integrated into eight unigenes named AhGLP1 to AhGLP8, of which, three (AhGLP1-3) were comprised 14, ten, and seven EST clones, respectively, whereas the remaining ones were associated with one single
clone. The length of the deduced amino acid (AA) residues ranged from 208 to 223 AAs except for AhGLP6 and AhGLP8, which were
incomplete at the carboxyl terminus. All of the AhGLPs contained a possible N-terminal signal peptide that was 17 to 24 residues
in length excluding AhGLP7, where there is likely a non-cleavable amino terminus. Phylogenetic analysis showed that these
AhGLPs were classified into three subfamilies. Southern blot analysis indicated that AhGLP1 and AhGLP2 likely have multiple copies in the peanut genome. The recombinant mature AhGLP1 and AhGLP2 proteins were successfully expressed
in Escherichia coli. The purified AhGLP2 has superoxide dismutase (SOD) activity in enzymatic assay, but not oxalate oxidase activity. The SOD
activity of AhGLP2 was stable up to 70°C and resistant to hydrogen peroxide, suggesting that AhGLP2 might be a manganese-containing
SOD. Furthermore, AhGLP2 could confer E. coli resistance to oxidative damage caused by paraquat, suggesting that the AhGLP2 likely protects peanut plants from reactive
oxygen metabolites. Thus, information provided in this study indicates the diverse nature of the peanut GLP family and suggests
that some of AhGLPs might be involved in plant defense response.
KeywordsPeanut–GLP family–Superoxide dismutase–Oxalate oxidase–Disease resistance
[show abstract][hide abstract] ABSTRACT: With 1 figure and 4 tablesAbstractTerminal drought induces preharvest aflatoxin contamination (PAC) in peanut. Drought resistance traits are promising as indirect selection tools for improving resistance to PAC. The objectives of this study were to determine the effects of terminal drought on PAC and to investigate the associations between surrogate traits for drought tolerance and PAC. Field tests under rainout shelters were conducted in the dry seasons 2004/2005 and 2005/2006. Eleven peanut genotypes were evaluated under irrigated and terminal drought conditions. Data were recorded for physiological traits, total biomass, pod yield, Aspergillus flavus colonization and PAC. ICGV 98305, ICGV 98330, ICGV 98348, ICGV 98353 and Tifton-8 had low aflatoxin contamination in both years. Traits related to drought resistance were associated well with those related to PAC under drought conditions. Specific leaf area, relative water content, chlorophyll density and drought stress ratings are the best traits for use as indirect selection tools for lower PAC. Breeding for drought tolerance using these traits as selection criteria may help to accelerate progress in developing resistance to PAC.
[show abstract][hide abstract] ABSTRACT: More rapid progress in breeding peanut for reduced aflatoxin contamination should be achievable with a better understanding of the inheritance of, aflatoxin trait and physiological traits that are associated with reduced contamination. The objectives of this study were to estimate the heritability of aflatoxin traits and genotypic (rG) and phenotypic (rP) correlations between drought resistance traits and aflatoxin traits in peanut. One hundred-forty peanut lines in the F4:6 and F4:7 generations were generated from four crosses, and tested under well-watered and terminal drought conditions. Field experiments were conducted under the dry seasons 2006/2007 and 2007/2008. Data were recorded for biomass (BIO), pod yield (PY), drought tolerance traits [harvest index (HI), drought tolerance index (DTI) of BIO and PY, specific leaf area (SLA), and SPAD chlorophyll meter reading (SCMR)], and aflatoxin traits [seed infection and aflatoxin contamination]. Heritabilities of A. flavus infection and aflatoxin contamination in this study were low to moderate. The heritabilities for seed infection and aflatoxin contamination ranged from 0.48 to 0.58 and 0.24 to 0.68, respectively. Significant correlations between aflatoxin traits and DTI (PY), DTI (BIO), HI, biomass and pod yield under terminal drought conditions were found (rP = −0.25** to 0.32**, rG = −0.57** to 0.53**). Strong correlations between SLA and SCMR with A. flavus infection and aflatoxin contamination were also found. Positive correlations between SLA at 80, 90, and 100 DAP and aflatoxin traits were significant (rP = 0.13** to 0.46**, rG = 0.26** to 0.81**). SCMR was negatively correlated with aflatoxin traits (rP = −0.10** to −0.40**, rG = −0.11** to −0.66**). These results indicated that physiological-based selection approaches using SLA and SCMR might be effective for improving aflatoxin resistance in peanut.
Field Crops Research 08/2010; 118(2):169–176. · 2.47 Impact Factor
[show abstract][hide abstract] ABSTRACT: The construction of genetic linkage maps for cultivated peanut (Arachis hypogaea L.) has and continues to be an important research goal to facilitate quantitative trait locus (QTL) analysis and gene tagging for use in a marker-assisted selection in breeding. Even though a few maps have been developed, they were constructed using diploid or interspecific tetraploid populations. The most recently published intra-specific map was constructed from the cross of cultivated peanuts, in which only 135 simple sequence repeat (SSR) markers were sparsely populated in 22 linkage groups. The more detailed linkage map with sufficient markers is necessary to be feasible for QTL identification and marker-assisted selection. The objective of this study was to construct a genetic linkage map of cultivated peanut using simple sequence repeat (SSR) markers derived primarily from peanut genomic sequences, expressed sequence tags (ESTs), and by "data mining" sequences released in GenBank.
Three recombinant inbred lines (RILs) populations were constructed from three crosses with one common female parental line Yueyou 13, a high yielding Spanish market type. The four parents were screened with 1044 primer pairs designed to amplify SSRs and 901 primer pairs produced clear PCR products. Of the 901 primer pairs, 146, 124 and 64 primer pairs (markers) were polymorphic in these populations, respectively, and used in genotyping these RIL populations. Individual linkage maps were constructed from each of the three populations and a composite map based on 93 common loci were created using JoinMap. The composite linkage maps consist of 22 composite linkage groups (LG) with 175 SSR markers (including 47 SSRs on the published AA genome maps), representing the 20 chromosomes of A. hypogaea. The total composite map length is 885.4 cM, with an average marker density of 5.8 cM. Segregation distortion in the 3 populations was 23.0%, 13.5% and 7.8% of the markers, respectively. These distorted loci tended to cluster on LG1, LG3, LG4 and LG5. There were only 15 EST-SSR markers mapped due to low polymorphism. By comparison, there were potential synteny, collinear order of some markers and conservation of collinear linkage groups among the maps and with the AA genome but not fully conservative.
A composite linkage map was constructed from three individual mapping populations with 175 SSR markers in 22 composite linkage groups. This composite genetic linkage map is among the first "true" tetraploid peanut maps produced. This map also consists of 47 SSRs that have been used in the published AA genome maps, and could be used in comparative mapping studies. The primers described in this study are PCR-based markers, which are easy to share for genetic mapping in peanuts. All 1044 primer pairs are provided as additional files and the three RIL populations will be made available to public upon request for quantitative trait loci (QTL) analysis and linkage map improvement.
[show abstract][hide abstract] ABSTRACT: A large root system can be an important character for drought tolerance. Measuring root characteristics in soil medium is tedious, time consuming, and labor intensive. The objective of this study was to determine the association between root characteristics of peanut (Arachis hypogaea L.) grown in hydroponics and in pot experiments. Twelve peanut genotypes were planted in a randomized complete block design with four replications in a hydroponics study, a small pot, and a large pot experiment. The studies were conducted in two years during 2004–2005. Shoot dry weight, root dry weight, root-to-shoot ratio, root length, root surface, average diameter of roots, and root volume were measured. Tifton-8 and ‘KK 60-3’ had consistently higher values for all root characteristics compared to the other genotypes. Root characteristics of peanut genotypes grown in hydroponics were positively correlated with those of peanut genotypes grown in pot conditions. The results suggested that assessment of root characteristics of peanut in hydroponics for possible drought tolerance could replace the assessment in soil medium conditions.
[show abstract][hide abstract] ABSTRACT: Twelve peanut genotypes were tested under three water regimes in two greenhouses to investigate the effects of drought on biomass production and N<SUB>2</SUB> fixation. Drought reduced biomass production from 36.5 to 56.0% and reduced nitrogen fixation from 26.8 to 68.8%. Most genotypes with high biomass production under Field Capacity (FC) had high reduction in biomass production under drought conditions, but fewer genotypes with high N<SUB>2</SUB> fixed under FC showed high reduction in N<SUB>2</SUB> fixed. Biomass production under FC in general gave more contribution to biomass production under drought conditions than did the reduction. N<SUB>2</SUB> fixed under FC and the reduction in N<SUB>2</SUB> fixed contributed similarly to N<SUB>2</SUB> fixed under drought conditions. Positive and significant correlations between N<SUB>2 </SUB>fixed and biomass production were found at FC and 2/3 available soil water (AW), but the correlation was not significant at 1/3 AW. Tifton-8 was the best genotype for high N<SUB>2</SUB> fixed under FC and KK 60-3 was the best genotype for low reduction. Correlations between N<SUB>2</SUB> fixed and nodule dry weight and shoot dry weight were high and consistent across water regimes. This information is important for breeders to develop peanut cultivars with reasonably high nitrogen fixation under drought conditions.
[show abstract][hide abstract] ABSTRACT: Peanut is vulnerable to a range of foliar diseases such as spotted wilt caused by Tomato spotted wilt virus (TSWV), early (Cercospora arachidicola) and late (Cercosporidium personatum) leaf spots, southern stem rot (Sclerotium rolfsii), and sclerotinia blight (Sclerotinia minor). In this study, we report the generation of 17,376 peanut expressed sequence tags (ESTs) from leaf tissues of a peanut cultivar (Tifrunner, resistant to TSWV and leaf spots) and a breeding line (GT-C20, susceptible to TSWV and leaf spots). After trimming vector and discarding low quality sequences, a total of 14,432 high-quality ESTs were selected for further analysis and deposition to GenBank. Sequence clustering resulted in 6,888 unique ESTs composed of 1,703 tentative consensus (TCs) sequences and 5185 singletons. A large number of ESTs (5717) representing genes of unknown functions were also identified. Among the unique sequences, there were 856 EST-SSRs identified. A total of 290 new EST-based SSR markers were developed and examined for amplification and polymorphism in cultivated peanut and wild species. Resequencing information of selected amplified alleles revealed that allelic diversity could be attributed mainly to differences in repeat type and length in the SSR regions. In addition, a few additional INDEL mutations and substitutions were observed in the regions flanking the microsatellite regions. In addition, some defense-related transcripts were also identified, such as putative oxalate oxidase (EU024476) and NBS-LRR domains. EST data in this study have provided a new source of information for gene discovery and development of SSR markers in cultivated peanut. A total of 16931 ESTs have been deposited to the NCBI GenBank database with accession numbers ES751523 to ES768453.
International Journal of Plant Genomics 02/2009; 2009:715605.
[show abstract][hide abstract] ABSTRACT: Cylindrocladium black rot (CBR), caused by Cylindrocladium parasiticum, and root-knot nematode, Meloidogyne arenaria, both infect and cause damage to the roots of peanut. Greenhouse and microplot experiments were conducted with the runner type peanut genotypes C724-19-15, C724-19-25 and Georgia-02C with different levels of resistance to nematode and CBR to better understand the interactions between the two pathogens. In the greenhouse, inoculation of 500–3000 eggs per plant of M. arenaria did not affect the level of root rot induced by 1·0 to 5·0 microsclerotia of C. parasiticum per g soil. In microplots, the root rot ratings from Georgia-02C and C724-19-25 were higher in plots infested with M. arenaria (0·4–2·0 eggs per cm3 soil) and C. parasiticum than in plots with C. parasiticum alone; however, M. arenaria did not increase the root rot ratings on the nematode resistant C724-19-15. This was inconsistent with results in the greenhouse. Gall indices were not affected by C. parasiticum inoculations in the greenhouse or microplots. In both 2006 and 2007, a significant interaction between C. parasiticum inoculum densities and nematode level was observed on plant mortality. CBR inoculum greatly increased mortality on C724-19-25 and Georgia-02C, but not on C724-19-15, in the presence of M. arenaria. The mortality increase was more apparent at lower inoculum levels of both pathogens, but on the nematode-susceptible cultivars plant mortality was more with co-inoculations of the two pathogens than from either alone. Simultaneous inoculation with the two pathogens decreased yield of C724-19-25 and Georgia-02C as C. parasiticum inoculum levels increased, but even the largest inoculum of M. arenaria (2·0 eggs per cm3 soil) did not decrease yield of C724-19-15.