Brian T. Scully

Monitoring the Expression of Maize Genes in Developing Kernels under Drought Stress using Oligo‐microarray

Preharvest aflatoxin contamination of grain grown on the US southeastern Coast Plain is provoked and aggravated by abiotic stress. The primary abiotic stress is drought along with high temperatures. The objectives of the present study were to monitor gene expression in developing kernels in response to drought stress and to identify drought-responsive genes for possible use in germplasm assessment. The maize breeding line Tex6 was used, and gene expression profiles were analyzed in developing kernels under drought stress verses well-watered conditions at the stages of 25, 30, 35, 40, 45 d after pollination (DAP) using the 70 mer maize oligo-arrays. A total of 9 573 positive array spots were detected with unique gene IDs, and 7 988 were common in both stressed and well-watered samples. Expression patterns of some genes in several stress response-associated pathways, including abscisic acid, jasmonic acid and phenylalanine ammonia-lyase, were examined, and these specific genes were responsive to drought stress positively. Real-time quantitative polymerase chain reaction validated microarray expression data. The comparison between Tex6 and B73 revealed that there were significant differences in specific gene expression, patterns and levels. Several defense-related genes had been downregulated, even though some defense-related or drought responsive genes were upregulated at the later stages.

Environmental influences on maize-Aspergillus flavus interactions and aflatoxin production

Since the early 1960s, the fungal pathogen Aspergillus flavus (Link ex Fr.) has been the focus of intensive research due to the production of carcinogenic and highly toxic secondary metabolites collectively known as aflatoxins following pre-harvest colonization of crops. Given this recurrent problem and the occurrence of a severe aflatoxin outbreak in maize (Zea mays L.), particularly in the Southeast U.S. in the 1977 growing season, a significant research effort has been put forth to determine the nature of the interaction occurring between aflatoxin production, A. flavus, environment and its various hosts before harvest. Many studies have investigated this interaction at the genetic, transcript, and protein levels, and in terms of fungal biology at either pre- or post-harvest time points. Later experiments have indicated that the interaction and overall resistance phenotype of the host is a quantitative trait with a relatively low heritability. In addition, a high degree of environmental interaction has been noted, particularly with sources of abiotic stress for either the host or the fungus such as drought or heat stresses. Here, we review the history of research into this complex interaction and propose future directions for elucidating the relationship between resistance and susceptibility to A. flavus colonization, abiotic stress, and its relationship to oxidative stress in which aflatoxin production may function as a form of antioxidant protection to the producing fungus.

Resistance to Spodoptera frugiperda (Lepidoptera: Noctuidae) and Euxesta stigmatias (Diptera: Ulidiidae) in Sweet Corn Derived from Exogenous and Endogenous Genetic Systems

Abstract Field trials using Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae) and Euxesta stigmatias Loew (Diptera: Ulidiidae) were conducted to evaluate resistance and potential damage interactions between these two primary corn, Zea mays L., pests against Lepidoptera-resistant corn varieties derived from both endogenous and exogenous sources. The endogenous source of resistance was maysin, a C-glycosyl flavone produced in high concentrations in varieties ‘Zapalote Chico 2451′ and ‘Zapalote Chico sh2’. The exogenous resistance source was the Bacillus thuringiensis (Bt)11 gene that expresses CryIA(b) insecticidal protein found in ‘Attribute GSS-0966′. Damage by the two pests was compared among these resistant varieties and the susceptible ‘Primetime’. Single-species tests determined that the Zapalote Chico varieties and GSS-0966 effectively reduced S. frugiperda larval damage compared with Primetime. E. stigmatias larval damage was less in the Zapalote Chico varieties than the other varieties in single-species tests. E. stigmatias damage was greater on S. frugiperda-infested versus S. frugiperda-excluded ears. Ears with S. frugiperda damage to husk, silk and kernels had greater E. stigmatias damage than ears with less S. frugiperda damage. Reversed phase high-performance liquid chromatography analysis of nonpollinated corn silk collected from field plots determined that isoorientin, maysin, and apimaysin plus 3′-methoxymaysin concentrations followed the order Zapalote Chico sh2 > Zapalote Chico 2451 > Attribute GSS-0966 = Primetime. Chlorogenic acid concentrations were greatest in Zapalote Chico 2451. The two high maysin Zapalote Chico varieties did as well against fall armyworm as the Bt-enhanced GSS-0966, and they outperformed GSS-0966 against E. stigmatias.

Impact of Brown Stink Bug (Heteroptera: Pentatomidae) Feeding on Corn Grain Yield Components and Quality

ABSTRACT Brown stink bug, Euschistus servus (Say) (Heteroptera: Pentatomidae), damage on developing corn, Zea mays L., ears was examined in 2005 and 2006 by using eight parameters related to its yield and kernel quality. Stink bug infestations were initiated when the corn plants were at tasseling (VT), mid-silking (R1), and blister (R2) stages by using zero, three, and six in 2005 or zero, one, two, and four bugs per ear in 2006, and maintained for 9 d. The percentage of discolored kernels was affected by stink bug number in both years, but not always affected by plant growth stage. The growth stage effect on the percentage of discolored kernels was significant in 2006, but not in 2005. The percentage of aborted kernels was affected by both stink bug number and plant growth stage in 2005 but not in 2006. Kernel weight was significantly reduced when three E. servus adults were confined on a corn ear at stage VT or R1 for 9 d in 2005, whereas one or two adults per ear resulted in no kernel weight loss, but four E. servus adults did cause significant kernel weight loss at stage VT in 2006. Stink bug feeding injury at stage R2 did not affect kernel damage, ear weight or grain weight in either year. The infestation duration (9 or 18 d) was positively correlated to the percentage of discolored kernels but did not affect kernel or ear weight. Based on the regression equations between the kernel weight and stink bug number, the gain threshold or economic injury level should be 0.5 bugs per ear for 9 d at stage VT and less for stage R1. This information will be useful in developing management guidelines for stink bugs in field corn during ear formation and early grain filling stages.

Genetic Mapping and Quantitative Trait Loci Analysis for Disease Resistance Using F 2 and F 5 Generation-based Genetic Maps Derived from 'Tifrunner' × 'GT-C20' in Peanut

One mapping population derived from Tifrunner × GT‐C20 has shown great potential in developing a high density genetic map and identifying quantitative trait loci (QTL) for important disease resistance, tomato spotted wilt virus (TSWV) and leaf spot (LS). Both F2 and F5 generation‐based genetic maps were previously constructed with 318 and 239 marker loci, respectively. Higher map density could be achieved with the F2 map (5.3 cM per locus) as compared to the F5 (5.7 cM per locus). Quantitative trait loci analysis using multi‐environment phenotyping data from F8 and higher generations for disease resistance identified 54 QTL in the F2 map including two QTL for thrips (12.14–19.43% phenotypic variation explained [PVE]), 15 for TSWV (4.40–34.92% PVE), and 37 for LS (6.61–27.35% PVE). Twenty‐three QTL could be identified in the F5 map including one QTL for thrips (5.86% PVE), nine for TSWV (5.20–14.14% PVE), and 13 for LS (5.95–21.45% PVE). Consistent QTL identified in each map have shown higher phenotypic variance than nonconsistent QTL. As expected, the number of QTL and their estimates of phenotypic variance were lower in the F5 map. This is the first QTL study reporting novel QTL for thrips, TSWV, and LS in peanut (Arachis hypogaea L.), and therefore, future studies will be conducted to refine these QTL.

Strategies in Prevention of Preharvest Aflatoxin Contamination in Peanuts: Aflatoxin Biosynthesis, Genetics and Genomics

Peanut (Arachis hypogaea L.), or groundnut, is an important crop economically and nutritionally in many tropical and subtropical areas of the world. It is also one of the most susceptible host crops to Aspergillus flavus resulting in aflatoxin contamination. The prevention or elimination of aflatoxin contamination in preharvest and postharvest crops is a serious challenge facing scientists. The recent International Conference on Groundnut Aflatoxin Management and Genomics held in Guangzhou, China, provided an international forum for discussions on the latest accomplishments, the development of strategies, and the initiation of cooperative research for the prevention of aflatoxin contamination. This review summarizes the progress in genetic and genomic research of peanuts and the toxinproducing fungus A. flavus. In particular, the pathway for production and the genetic regulation of afaltoxin, and the peanut-Aspergillus interaction are discussed. The use of a peanut-Aspergillus microarray will help scientists to study the croppathogen interaction; aids in the identification of genes involved in both fungal invasion and crop resistance, and ultimately enhance research to find solutions that prevent aflatoxin contamination in agricultural commodities.

Root Morphology and Gene Expression Analysis in Response to Drought Stress in Maize (Zea mays)

Water-deficit stress tolerance is a complex trait, and water deficit results in various physiological and chemical changes in maize (Zea mays L.) and exacerbates pre-harvest aflatoxin contamination. The objective of this study was to characterize the variations in morphology, physiology, and gene expression in two contrasting inbred lines, Lo964 and Lo1016, in order to understand the differences in response to water-deficit stress. The results revealed that Lo964 was less sensitive to water-deficit stress, and had a strong lateral root system and a higher root/shoot ratio in comparison to Lo1016. In response to water-deficit stress by comparing stressed versus well-watered conditions, abscisic acid syntheses were increased in leaves, roots, and kernels of both Lo964 and Lo1016, but by different magnitudes. Indole-3-acetic acid (IAA) was undetectable in the leaves and roots of either genotype regardless of treatments, but increases of 58% and 8% in IAA concentration were observed in 20 DAP kernels, in response to water-deficit stress, respectively. The expression of the MIPS was up-regulated 7-fold in leaf tissues of Lo964 compared to Lo1016 at watered conditions, but decreased significantly to similar levels in both genotypes at water-deficit conditions. ZmPR10 and ZmFer1 expressions tended to up-regulate although ZmPR10 was expressed higher in root tissue while ZmFer1 was expressed higher in leaf tissue. Further study is needed to confirm if Lo964 has reduced aflatoxin contamination associated with the drought tolerance in the field in order to utilize the resistant trait in breeding.

Spatial patterns of aflatoxin levels in relation to ear-feeding insect damage in pre-harvest corn.

Key impediments to increased corn yield and quality in the southeastern US coastal plain region are damage by ear-feeding insects and aflatoxin contamination caused by infection of Aspergillus flavus. Key ear-feeding insects are corn earworm, Helicoverpa zea, fall armyworm, Spodoptera frugiperda, maize weevil, Sitophilus zeamais, and brown stink bug, Euschistus servus. In 2006 and 2007, aflatoxin contamination and insect damage were sampled before harvest in three 0.4-hectare corn fields using a grid sampling method. The feeding damage by each of ear/kernel-feeding insects (i.e., corn earworm/fall armyworm damage on the silk/cob, and discoloration of corn kernels by stink bugs), and maize weevil population were assessed at each grid point with five ears. The spatial distribution pattern of aflatoxin contamination was also assessed using the corn samples collected at each sampling point. Aflatoxin level was correlated to the number of maize weevils and stink bug-discolored kernels, but not closely correlated to either husk coverage or corn earworm damage. Contour maps of the maize weevil populations, stink bug-damaged kernels, and aflatoxin levels exhibited an aggregated distribution pattern with a strong edge effect on all three parameters. The separation of silk- and cob-feeding insects from kernel-feeding insects, as well as chewing (i.e., the corn earworm and maize weevil) and piercing-sucking insects (i.e., the stink bugs) and their damage in relation to aflatoxin accumulation is economically important. Both theoretic and applied ramifications of this study were discussed by proposing a hypothesis on the underlying mechanisms of the aggregated distribution patterns and strong edge effect of insect damage and aflatoxin contamination, and by discussing possible management tactics for aflatoxin reduction by proper management of kernel-feeding insects. Future directions on basic and applied research related to aflatoxin contamination are also discussed.

Preharvest aflatoxin contamination of corn and other grain crops grown on the U.S. Southeastern Coastal Plain

Preharvest aflatoxin contamination of grain grown on the U.S. Southeastern Coastal Plain is provoked and aggravated by both biotic and abiotic stress factors that influence infection by the Asperigillus group. Asperigillus flavus, Link ex Fr., is one of the principal toxigenic fungi of summer grains grown in the region, and the hot, humid weather patterns along with suboptimal summer rainfall favor the development of this organism. An array of arthropod species also contributes to the dispersal of this fungus as they attack and feed on the developing grain. Research on summer grains grown on the Coastal Plain has the expressed goal of reducing, and perhaps eliminating aflatoxin contamination in adapted germplasm using classical crop improvement methods to deploy host plant resistance. This research is complimented and enhanced by molecular techniques that have proven invaluable in the identification and development of superior germplasm. It also emphasizes the need to fully understand the biological interactions between fungus, arthropods, crops, and the environmental conditions that govern the aflatoxin contamination. Alternative cropping systems that avoid contamination are also integrated into this summary of this research progress.

Identifying and developing maize germplasm with resistance to accumulation of aflatoxins

Efforts to identify maize germplasm with resistance to Aspergillus flavus infection and subsequent accumulation of aflatoxins were initiated by the US Department of Agriculture, Agricultural Research Service at several locations in the late 1970s and early 1980s. Research units at four locations in the south-eastern USA are currently engaged in identification and development of maize germplasm with resistance to A. flavus infection and accumulation of aflatoxins. The Corn Host Plant Resistance Research Unit, Mississippi State, MS, developed procedures for screening germplasm for resistance to A. flavus infection and accumulation of aflatoxins. Mp313E, released in 1990, was the first line released as a source of resistance to A. flavus infection. Subsequently, germplasm lines Mp420, Mp715, Mp717, Mp718, and Mp719 were released as additional sources of resistance. Quantitative trait loci associated with resistance have also been identified in four bi-parental populations. The Crop Protection and Management ...