Susceptible Peanut Cultivar Research Articles

Comparative proteomic analysis of the moderately resistant and susceptible peanut cultivars during infestation by Cylindrocladium parasiticum

Cylindrocladium parasiticum Crous, Wingfield & Alfenas, the causal agent of Cylindrocladium black rot of peanut, has led to economic losses in China. However, the response of peanut plant to C. parasiticum is poorly understood. Here, two contrasting peanut cultivars, T09 (C. parasiticum-moderately resistant) and P562 (C. parasiticum-susceptible) were used for comparative analysis of protein profiles in root segment of plant tissues in responses to C. parasiticum infection. Proteomic profiling identified 1647 and 391 differentially accumulated proteins (DAPs) in P562 and T09 relative to uninoculated control, respectively. A total of 350 and 1095 DAPs were identified between P562 and T09 after 9 day(s) post infection (dpi), respectively. Functional categorization showed that C. parasiticum-responsive proteins were mainly involved in catalytic activity and binding functions. KEGG pathway analysis indicated both moderately resistant and susceptible peanut cultivars can regulate gene expression in the phenylpropanoid pathway, terpenoid backbone biosynthesis, SA, and JA pathways to induce defensive genes and protein expression which enhances plant defence or susceptibility capacity. However, MAPK signal pathway was more pronounced in moderately resistant peanut cultivar T09. We also observed an increase of CYP73A100 involved in phenylpropanoid and flavonoid biosynthesis pathways in susceptible peanut ecotype P562, while decrease in moderately resistant interaction. Additionally, there was a marked activation of brassinosteroid biosynthesis in T09, which indicated a possible involvement of activation of plant immune response in moderately resistant responses of peanut to C. parasiticum. This study provides some insights into molecular networks involved in cellular and physiological responses to C. parasiticum in peanut.

A Proof-of-Principle Study of Non-invasive Identification of Peanut Genotypes and Nematode Resistance Using Raman Spectroscopy.

Identification of peanut cultivars for distinct phenotypic or genotypic traits whether using visual characterization or laboratory analysis requires substantial expertise, time, and resources. A less subjective and more precise method is needed for identification of peanut germplasm throughout the value chain. In this proof-of-principle study, the accuracy of Raman spectroscopy (RS), a non-invasive, non-destructive technique, in peanut phenotyping and identification is explored. We show that RS can be used for highly accurate peanut phenotyping via surface scans of peanut leaves and the resulting chemometric analysis: On average 94% accuracy in identification of peanut cultivars and breeding lines was achieved. Our results also suggest that RS can be used for highly accurate determination of nematode resistance and susceptibility of those breeding lines and cultivars. Specifically, nematode-resistant peanut cultivars can be identified with 92% accuracy, whereas susceptible breeding lines were identified with 81% accuracy. Finally, RS revealed substantial differences in biochemical composition between resistant and susceptible peanut cultivars. We found that resistant cultivars exhibit substantially higher carotenoid content compared to the susceptible breeding lines. The results of this study show that RS can be used for quick, accurate, and non-invasive identification of genotype, nematode resistance, and nutrient content. Armed with this knowledge, the peanut industry can utilize Raman spectroscopy for expedited breeding to increase yields, nutrition, and maintaining purity levels of cultivars following release.

Metabolite profiling identified pipecolic acid as an important component of peanut seed resistance against Aspergillus flavus infection

In a previous study, we identified a halotolerant rhizobacterium belonging to the genus Klebsiella (MBE02) that protected peanut seeds from Aspergillus flavus infection. Here, we investigated the mechanisms underlying the effect of MBE02 against A. flavus via untargeted metabolite profiling of peanut seeds treated with MBE02, A. flavus, or MBE02+A. flavus. Thirty-five metabolites were differentially accumulated across the three treatments (compared to the control), and the levels of pipecolic acid (Pip) were reduced upon A. flavus treatment only. We validated the function of Pip against A. flavus using multiple resistant and susceptible peanut cultivars. Pip accumulation was strongly associated with the resistant genotypes that also accumulated several mRNAs of the ALD1-like gene in the Pip biosynthesis pathway. Furthermore, exogenous treatment of a susceptible peanut cultivar with Pip reduced A. flavus infection in the seeds. Our findings indicate that Pip is a key component of peanut resistance to A. flavus.

Proteomics unravels new candidate genes for Meloidogyne resistance in wild Arachis

Arachis stenosperma is a wild peanut relative exclusive to South America that harbors high levels of resistance against several pathogens, including the peanut root-knot nematode (RKN) Meloidogyne arenaria. In this study, a proteomic survey of A. stenosperma-M. arenaria interaction using 2-DE and LC-MS/MS identified approximately 1400 proteins, out of which 222 were differentially abundant (DAPs) when RKN inoculated root samples were compared to the control. Most of these DAPs were assigned to functional categories related to plant responses to pathogens including stress, glycolysis, redox and tricarboxylic acid cycle. The comparison between the transcriptome (RNA-Seq) and proteome expression changes, showed that almost 55% of these DAPs encode genes with a similar expression trend to their protein counterparts. Most of these genes were induced during RKN infection and some were related to plant defense, such as MLP-like protein 34 (MLP34), cinnamoyl-CoA reductase 1 (CCR1), enolase (ENO), alcohol dehydrogenase (ADH) and eukaryotic translation initiation factor 5A (eIF5A). The overexpression of AsMLP34 in Agrobacterium rhizogenes transgenic roots in a susceptible peanut cultivar showed a reduction in the number of M. arenaria galls and egg masses, indicating that AsMLP34 is a promising candidate gene to be exploited in breeding programs for RKN control in peanut. SignificanceThe use of an integrated approach to compare plant-nematode transcriptional and translational data enabled the identification of a new gene, AsMLP34, for Meloidogyne resistance.

Disease and Yield Response of a Stem-rot-resistant and -Susceptible Peanut Cultivar under Varying Fungicide Inputs.

Peanut (Arachis hypogaea L.) producers rely on costly fungicide programs to manage stem rot, caused by Sclerotium rolfsii. Planting disease-resistant cultivars could increase profits by allowing for the deployment of less-expensive, lower-input fungicide programs. Field experiments were conducted to characterize stem rot and early and late leaf spot (caused by Passalora arachidicola and Nothopassalora personata, respectively), yield, and overall profitability of cultivars Georgia-06G (stem-rot-susceptible) and Georgia-12Y (stem-rot-resistant) as influenced by seven commercial fungicide programs. Stem rot incidence was consistently lower on Georgia-12Y for all fungicides when compared with Georgia-06G and was lowest for both cultivars in plots treated with prothioconazole plus a tank mixture of penthiopyrad and tebuconazole. Leaf spot severity was similar for both the resistant and susceptible cultivars, and the greatest reduction occurred in plots treated with prothioconazole plus a tank mixture of penthiopyrad and tebuconazole. Fungicide programs gave similar yield and net return on Georgia-12Y; however, plots of Georgia-06G treated with prothioconazole plus a tank mixture of penthiopyrad and tebuconazole had the greatest yield and net return. Yields and economic return from the highest level of fungicide inputs on Georgia-06G were numerically less than those of Georgia-12Y treated with only chlorothalonil. These results show the value of fungicides in peanut disease management with susceptible cultivars, as well as the benefits of planting stem-rot-resistant cultivars in high-risk situations.

Overexpression of Wild Arachis Lipocalin Enhances Root-Knot Nematode Resistance in Peanut Hairy Roots

Plant parasitic root-knot nematodes (RKN) have evolved sophisticated strategies for exploiting plants, and they cause devastating yield losses in many susceptible crops. The RKN Meloidogyne arenaria is the most predominant pathogenic nematode affecting cultivated peanut (Arachis hypogaea L.). Resistance against the M. arenaria has been identified in the wild peanut relative, Arachis stenosperma. Transcriptome studies on A. stenosperma–M. arenaria interaction revealed candidate genes potentially involved in the first stages of this resistance response, including a temperature-induced lipocalin (TIL) protein. Plant TILs have a protective role against harmful molecules produced in response to a number of stresses. In this study, the characterization of the RKN-responsive TIL from A. stenosperma (AsTIL) provides new insights into the role of plant lipocalins in nematode resistance. The AsTIL gene has a 2-exon/1-intron structure and encodes a 21.4 kDa polypeptide. It contains three structurally conserved regions, which are a signature for plant lipocalins. Overexpression of AsTIL in transgenic hairy roots from a susceptible peanut cultivar led to the reduction of M. arenaria galls and egg masses 60 days after inoculation. This is the first report of a possible involvement of plant lipocalins in RKN resistance. It also reveals a promising candidate gene for peanut breeding to produce cultivars resistant to M. arenaria.

Overexpression of a novel peanut NBS-LRR gene AhRRS5 enhances disease resistance to Ralstonia solanacearum in tobacco.

SummaryBacterial wilt caused by Ralstonia solanacearum is a ruinous soilborne disease affecting more than 450 plant species. Efficient control methods for this disease remain unavailable to date. This study characterized a novel nucleotide‐binding site‐leucine‐rich repeat resistance gene AhRRS5 from peanut, which was up‐regulated in both resistant and susceptible peanut cultivars in response to R. solanacearum. The product of AhRRS5 was localized in the nucleus. Furthermore, treatment with phytohormones such as salicylic acid (SA), abscisic acid (ABA), methyl jasmonate (MeJA) and ethephon (ET) increased the transcript level of AhRRS5 with diverse responses between resistant and susceptible peanuts. Abiotic stresses such as drought and cold conditions also changed AhRRS5 expression. Moreover, transient overexpression induced hypersensitive response in Nicotiana benthamiana. Overexpression of AhRRS5 significantly enhanced the resistance of heterogeneous tobacco to R. solanacearum, with diverse resistance levels in different transgenic lines. Several defence‐responsive marker genes in hypersensitive response, including SA, JA and ET signals, were considerably up‐regulated in the transgenic lines as compared with the wild type inoculated with R. solanacearum. Nonexpressor of pathogenesis‐related gene 1 (NPR1) and non‐race‐specific disease resistance 1 were also up‐regulated in response to the pathogen. These results indicate that AhRRS5 participates in the defence response to R. solanacearum through the crosstalk of multiple signalling pathways and the involvement of NPR1 and R gene signals for its resistance. This study may guide the resistance enhancement of peanut and other economic crops to bacterial wilt disease.

Gene Expression Profiling and Identification of Resistance Genes to Aspergillus flavus Infection in Peanut through EST and Microarray Strategies

Aspergillus flavus and A. parasiticus infect peanut seeds and produce aflatoxins, which are associated with various diseases in domestic animals and humans throughout the world. The most cost-effective strategy to minimize aflatoxin contamination involves the development of peanut cultivars that are resistant to fungal infection and/or aflatoxin production. To identify peanut Aspergillus-interactive and peanut Aspergillus-resistance genes, we carried out a large scale peanut Expressed Sequence Tag (EST) project which we used to construct a peanut glass slide oligonucleotide microarray. The fabricated microarray represents over 40% of the protein coding genes in the peanut genome. For expression profiling, resistant and susceptible peanut cultivars were infected with a mixture of Aspergillus flavus and parasiticus spores. The subsequent microarray analysis identified 62 genes in resistant cultivars that were up-expressed in response to Aspergillus infection. In addition, we identified 22 putative Aspergillus-resistance genes that were constitutively up-expressed in the resistant cultivar in comparison to the susceptible cultivar. Some of these genes were homologous to peanut, corn, and soybean genes that were previously shown to confer resistance to fungal infection. This study is a first step towards a comprehensive genome-scale platform for developing Aspergillus-resistant peanut cultivars through targeted marker-assisted breeding and genetic engineering.

Differential gene expression in roots of nematode-resistant and -susceptible peanut ( Arachis hypogaea) cultivars in response to early stages of peanut root-knot nematode ( Meloidogyne arenaria) parasitization

The peanut root-knot nematode (RKN, Meloidogyne arenaria) can cause significant yield losses in cultivated peanut ( Arachis hypogaea). However, molecular events underlying successful RKN infection and host responses in peanut are sparsely understood. Using suppression subtractive hybridization (SSH), cDNA libraries, enriched with differentially expressed ESTs, were constructed from RKN-challenged root tissues in the pre-penetration and early infection stages from near-isogenic nematode-resistant and -susceptible peanut cultivars NemaTAM and Florunner. Following an initial screen of 960 expressed sequence tags (ESTs) for at least three-fold differential expression between the two libraries, 70 ESTs (36 from the NemaTAM-specific library and 34 from the Florunner-specific library) were identified and annotated into seven functional categories (stress responses, metabolism, transcriptional regulation, protein synthesis and/or modification, transport functions, cellular architecture and proteins with unknown functions). Discreet gene tag clusters primarily including pathogenesis related (PR), patatin-like proteins and universal stress related proteins (USPs), as well as those implicated in alleviation of oxidative stress were primarily represented in RKN-infected NemaTAM roots, reflective of a basal level of resistance operative against invading nematodes. However, significant transcriptional reprogramming and upregulation of genes implicated in modification of cellular architecture, adhesion, and proliferation marked an early onset of compatible host–pathogen interactions discernible in Florunner roots.

First Report of Sclerotinia minor Infecting Ipomoea hederacea and I. coccinea in Texas.

Sclerotinia blight, caused by the soilborne fungus Sclerotinia minor Jagger, is a major disease of peanut (Arachis hypogaea L.) in parts of west Texas. Previous reports have indicated that annual weed species may serve as collateral hosts for S. minor (2). Several Ipomoea spp. are commonly found in peanut fields throughout the region. In September of 2007, Ipomoea hederacea and I. coccinea plants with bleached, shredded stems, and signs of black sclerotia were collected from a field known to be infested with S. minor. Symptomatic stem sections were rinsed in tap water, surface disinfested in 0.5% sodium hypochlorite for 1 min, air dried, and plated on potato dextrose agar (PDA). Pure cultures of S. minor consisting of white, fluffy mycelia and small (<2 mm), black, irregular sclerotia were consistently recovered. Pathogenicity tests were conducted by wound-inoculating healthy I. hederacea and I. coccinea transplants (n = 3) with agar plugs obtained from the edges of actively growing S. minor cultures. Plants were incubated in a dew chamber at 20°C and 95% relative humidity for 5 days. Plants inoculated with sterile PDA plugs served as controls (n = 3). A similar test was conducted using the susceptible peanut cultivar Flavorunner 458. Characteristic symptoms of Sclerotinia blight (3) were observed on all inoculated weed and peanut plants; whereas, the controls remained healthy. Pathogenicity tests were repeated with similar results. Cultures of S. minor were obtained from all symptomatic tissues, fulfilling Koch's postulates. These results indicate that I. hederacea and I. coccinea are additional hosts of S. minor and that sclerotia produced on infected plants can significantly augment soil inoculum. S. minor has been observed to infect I. batatas seedlings in New Jersey (1); however, this to our knowledge is the first report of S. minor infecting Ipomoea spp. in Texas. Therefore, weed management should inevitability be a part of disease management strategies for the control of Sclerotinia blight in peanut.

Sources of Resistance to Tobacco streak virus in Wild Arachis (Fabaceae: Papilionoidae) Germplasm.

Stem necrosis disease caused by Tobacco streak virus (TSV), first recognized in 2000, has emerged as a potential threat to peanut (Arachis hypogaea) in southern states of India. The virus induces severe necrosis of shoots leading to death of the plant, and plants that survive are malformed, with severe reduction in pod yield. All the currently grown peanut cultivars in India are highly susceptible to the virus. Therefore, wild relatives of peanut were evaluated to identify potential sources of resistance to TSV infection. In all, 56 germplasm accessions from 20 wild Arachis spp. in four sections (Arachis, Erectoides, Procumbente, and Rhizomatosae), along with susceptible peanut cultivars (JL 24 and K 1375), were evaluated for resistance to TSV under greenhouse conditions using mechanical sap inoculations. Systemic virus infection, determined by enzyme-linked immunosorbent assay (ELISA), in the test accessions ranged between 0 and 100%. Twenty-four accessions in section Arachis that had 0 to 35% systemically infected plants were retested, and systemic infection was not detected in eight of these accessions in repeated trials in the greenhouse. These are International Crops Research Institute for the Semi-Arid Tropics groundnut (ICG) accession nos. 8139, 8195, 8200, 8203, 8205, and 11550 belonging to A. duranensis; ICG 8144 belonging to A. villosa; and ICG 13210 belonging to A. stenosperma. Even though the resistant accessions had 0 to 100% TSV infection in inoculated leaves, TSV was not detected in the subsequently emerged leaves. This is the first report of TSV resistance in Arachis spp. The eight TSV resistant accessions are cross compatible with A. hypogaea for utilization in breeding for stem necrosis disease resistance.

Effect of Sclerotinia minor Infection Loci on Peanut Production Parameters

Fungal diseases of peanut, such as Sclerotinia blight caused by Sclerotinia minor, are responsible for increased production costs and yield losses of up to 50% for peanut producers in the Southwestern U.S., North Carolina, and Virginia. The literature is replete with information on the pathology, epidemiology, and control of S. minor infection of peanut, however little is known about the physical location of infection on the plant and its effect on plant production. In a 2-yr field trial, the susceptible peanut cultivar Okrun was subjected to high disease intensity without fungicide application for control of S. minor. Location of initial S. minor infection was noted to occur either on the ‘‘crown’’ or ‘‘limb’’ of infected plants, and the date of initial onset was recorded. In general, plants with initial crown infections had reduced yield and seed quality compared to those with initial limb infections. Initial infections occurring in early- and mid-season were more detrimental to plant productivity than those infections occurring late in the season. Mid-season onset of a crown infection had the greatest impact on plant productivity, causing a severe decrease in seed quality and pod yield. This study demonstrates that both infection location and date of disease onset affect plant productivity, and the results may be used to improve Sclerotinia blight management strategies.

A regression approach for comparing field resistance of peanut cultivars to tomato spotted wilt tospovirus

Relationships between one TSWV susceptible peanut cultivar (Georgia Runner), one moderately resistant peanut cultivar (C-99R), and the current standard cultivar (Georgia Green) in the southeastern United States were examined. Both moderately resistant cultivars had greater field resistance and higher yields than the susceptible cultivar, Georgia Runner, regardless of the level of disease pressure. Linear regression revealed that under heavy disease pressure, C-99R appears to exhibit greater disease resistance than has been previously reported for Georgia Green and other currently available cultivars. Only 13% and 33% of the variation in yield difference between cultivars could be explained by differences in TSWV incidence for C-99R and Georgia Runner, respectively, when compared by linear regression with Georgia Green. Although the correlations were weak for these relationships, the relationships were still significant ( P<0.05). Linear regression equations allow us to designate TSWV risk assesment index scores for cultivars relative to scores for the standard Georgia Green. Such a comparison allows for an examination of the relative levels of resistance observed among cultivars under varying levels of disease pressure. This type of analysis would be most useful under heavy disease pressure situations, when the predictive power of this analysis is greater and differences in field resistance between cultivars would be most significant.

Variation in Aggressiveness and Virulence Among Isolates of Cercospora arachidicola

Abstract Forty-three isolates of Cercospora arachidicola, incitant of early leaf spot of peanut, were collected from peanut fields in Florida, Georgia, North Carolina, and Texas. Variation in aggressiveness among these isolates using the susceptible peanut cultivar Spanco was examined in greenhouse tests. Variation (P ≤ 0.05) among the isolates was observed for the parameters of incubation period (IP), the reciprocal of the latent period (an estimate of sporulation rate), and number of lesions per leaflet (LN). Additionally, as a group, the isolates from Texas were more aggressive in terms of having a shorter latent period and greater number of lesions per leaflet than isolates from other states. When a subset of eight isolates, which differed in aggressiveness based on IP and LN, were tested on Spanco and three early leaf spotresistant breeding lines (TX957910–5, TX957910–27, TX957910–68), the breeding lines were resistant to all isolates and, except for lesion diameter, no host genotype-by-isolate interaction was detected (P &amp;gt; 0.05). These observations suggest that success of efforts to identify resistance to early leaf spot can be affected by the aggressiveness of the pathogen isolate. Additionally, the resistance of the breeding lines tested is likely to be effective against a wide array of isolates of C. arachidicola.

Spatial and Temporal Patterns of Spotted Wilt Epidemics in Peanut

The spatio-temporal patterns of spotted wilt disease in peanut, caused by thrips-vectored tomato spotted wilt tospovirus (TSWV), were analyzed by several methods. The spatial distributions of symptomatic plants were mapped at 2-wk intervals during two consecutive years in fields of the susceptible peanut cultivar Florunner and in the resistant cultivar Southern Runner. The disease frequency data were subjected to quadrat analyses, including indices of dispersion (variance/mean ratio and the negative binomial k parameter), comparison of frequency distributions of symptomatic plants to expected distributions derived from several models of spatial dispersion (binomial, Poisson, and negative binomial), and two-dimensional distance class analysis at each sampling interval. Although significant spatial aggregation of diseased plants was detected in most samples, random or nearly random clusters of infected plants that apparently arose from a continuous immigration of viruliferous vectors dominated spatial aspects of epidemic progress. Aside from smaller incidence of disease in the resistant compared to the susceptible cultivar, no effect of cultivar was noted in relation to spatial or temporal patterns of disease development in this pathosystem. Spotted wilt temporal progress was well described by monomolecular models of disease progress in each cultivar and year. The data and analyses were consistent with the hypothesis that most infections arise as a result of primary transmission and that there is limited secondary spread of TSWV after it becomes established in the field.

Peanut Web Blotch: II Symptoms and Host Range of Pathogen1

Abstract Web blotch, caused by Phoma arachidicola, has varied in severity from year to year in Texas, depending on survival of inoculum on peanut crop residue, presence of susceptible peanut cultivars and suitable environmental conditions, especially favorable temperature and high relative humidity. The web and blotch symptoms may develop in sequence on the same leaflet or independently on different leaflets. At relative humidities below 80% within the plant canopy, the fungal hyphae grew under the leaf cuticle causing a web-type symptom. At relative humidities above 90%, hyphal strands grew more repidly and branched extensively in the leaf resulting in the blotch-type symptom. P. arachidicola isolates from South Africa, Argentina, and the USA were all pathogenic and caused similar symptoms. Of 32 legumes, peanut, sweetclover, hairy vetch and alfalfa were the most susceptible. Symptoms on hosts other than peanuts consisted of small necrotic spots. A hypersensitive reaction developed on eight legumes. Typical web blotch symptoms were observed only on peanuts in the field.