A Histological Study of Aspergillus flavus Colonization of Wound Inoculated Maize Kernels of Resistant and Susceptible Maize Hybrids in the Field.

Aspergillus flavus colonization in developing kernels of maize single-cross hybrids resistant (Mp313E × Mp717) and susceptible (GA209 × T173) to aflatoxin accumulation was determined in the field over three growing seasons (2012-2014). Plants were hand pollinated, and individual kernels were inoculated with a needle dipped in a suspension of A. flavus conidia 21 days after pollination. Kernels were harvested at 1- to 2-day intervals from 1 to 21 days after inoculation (DAI). Kernels were placed in FAA fixative, dehydrated, embedded in paraffin, sectioned, and stained with toluidine blue. Kernels were also collected additional kernels for aflatoxin analyses in 2013 and 2014. At 2 DAI, A. flavus hyphae were observed among endosperm cells in the susceptible hybrid, but colonization of the endosperm in the resistant hybrid was limited to the wound site of the resistant hybrid. Sections of the scutellum of the susceptible hybrid were colonized by A. flavus by 5 DAI. Fungal growth was slower in the resistant hybrid compared to the susceptible hybrid. By 10 DAI, A. flavus had colonized a large section of the embryo in the susceptible hybrid; whereas in the resistant hybrid, approximately half of the endosperm had been colonized and very few cells in the embryo were colonized. Fungal colonization in some of the kernels of the resistant hybrid was slowed in the aleurone layer or at the endosperm-scutellum interface. In wounded kernels with intact aleurone layers, the fungus spread around the kernel between the pericarp and aleurone layer with minimal colonization of the endosperm. Aflatoxin B1 was first detected in susceptible kernel tissues 8 DAI in 2013 (14 µg/kg) and 2014 (18 µg/kg). The resistant hybrid had significantly lower levels of aflatoxin accumulation compared to the susceptible hybrid at harvests 10, 21, and 28 DAI in 2013, and 20 and 24 DAI in 2014. Our study found differential A. flavus colonization of susceptible and resistant kernel tissues, and that the aleurone and the outer layer of the scutellum slowed the rate of colonization by A. flavus.

Expression Profiling Coupled with In-silico Mapping Identifies Candidate Genes for Reducing Aflatoxin Accumulation in Maize.

Aflatoxin, produced by Aspergillus flavus, is hazardous to health of humans and livestock. The lack of information about large effect QTL for resistance to aflatoxin accumulation is a major obstacle to employ marker-assisted selection for maize improvement. The understanding of resistance mechanisms of the host plant and the associated genes is necessary for improving resistance to A. flavus infection. A suppression subtraction hybridization (SSH) cDNA library was made using the developing kernels of Mp715 (resistant inbred) and B73 (susceptible inbred) and 480 randomly selected cDNA clones were sequenced to identify differentially expressed genes (DEGs) in response to A. flavus infection and map these clones onto the corn genome by in-silico mapping. A total of 267 unigenes were identified and majority of genes were related to metabolism, stress response, and disease resistance. Based on the reverse northern hybridization experiment, 26 DEGs were selected for semi-quantitative RT-PCR analysis in seven inbreds with variable resistance to aflatoxin accumulation at two time points after A. flavus inoculation. Most of these genes were highly expressed in resistant inbreds. Quantitative RT-PCR analysis validated upregulation of PR-4, DEAD-box RNA helicase, and leucine rich repeat family protein in resistant inbreds. Fifty-six unigenes, which were placed on linkage map through in-silico mapping, overlapped the QTL regions for resistance to aflatoxin accumulation identified in a mapping population derived from the cross between B73 and Mp715. Since majority of these mapped genes were related to disease resistance, stress response, and metabolism, these should be ideal candidates to investigate host pathogen interaction and to reduce aflatoxin accumulation in maize.

Foliar herbivory triggers local and long distance defense responses in maize.

Many studies have documented the induction of belowground defenses in plants in response to aboveground herbivory and vice versa, but the genes and signaling molecules mediating systemic induction are not well understood. We performed comparative microarray analysis on maize whorl and root tissues from the insect resistant inbred Mp708 in response to foliar feeding by fall armyworm (Spodoptera frugiperda) caterpillars. Although Mp708 has elevated jasmonic acid (JA) levels prior to herbivory, genes involved in JA biosynthesis were up-regulated in whorls in response to fall armyworm feeding. Alternatively, genes possibly involved in regulating ethylene (ET) perception and signaling were up-regulated in roots following foliar herbivory. Transcript levels of genes encoding proteins involved in direct defenses against herbivores were enhanced both in roots and leaves, but transcriptional factors and genes involved in various biosynthetic pathways were selectively down-regulated in the whorl. The results indicate that foliar herbivory by fall armyworm changes root gene expression pathways suggesting profound long distance signaling. Tissue specific induction and suppression of JA and ET signaling pathway genes provides a clue to their possible roles in signaling between the two distant tissue types that eventually triggers defense responses in the roots in response to foliar herbivory.

Systemic infection of stalks and ears of corn hybrids by Aspergillus parasiticus.

This study was conducted to explore systemic infection by the Aspergillus flavus group into corn ears via the stalk. An A. parasiticus mutant which produces norsolorinic (NOR) acid (a visible orange intermediate of the aflatoxin biosynthetic pathway) was used in field studies to monitor systemic infection of corn stalk and ear tissues. Corn hybrids resistant and susceptible to aflatoxin contamination were grown in the field and inoculated prior to tasseling by inserting A. parasiticus infested toothpicks into stalks between the 5th and 6th node below the lowest ear shoot. Beginning 2 weeks after inoculation, systemic infection by the NOR mutant was assessed weekly by collecting ear shank tissue and stalk tissue from the nodes between the infection sites and the developing ears. Ears were collected at the end of the growing season to determine the level of kernel infection by the NOR mutant. In two separate studies, the A. parasiticus NOR mutant was isolated from stalk tissues at all of node positions and ear shank tissue from several susceptible corn hybrid plants at the first harvest date 2 weeks after inoculation. The NOR mutant was also isolated from stalk and ear tissue of a resistant hybrid. The NOR mutant was only isolated from kernels of susceptible hybrids in 2003 and 2004. Infection rates of kernels in infected ears were very low (<1%). In 2005, the fungus was found in only one kernel from an ear of the resistant hybrid. The NOR mutant was not isolated from stalks, ears, or kernels from control (uninoculated) plants grown in the plots with inoculated plants. Although infection levels of corn kernels were low, systemic movement of the A. parasiticus up the stalk appears to be another possible route to infection of developing corn ears.

Heritability and correlation among some selected morphological traits and their relationship with fall armyworm damage in sweet corn.

Fall armyworm [Spodoptera frugiperda J. E. Smith] (FAW) is a serious pest in field corn and sweet corn [Zea mays L.] in many parts of the world. Sweet corn germplasm with effective levels of resistance to damage by the fall armyworm is needed to transfer resistance to commercial hybrids. The objectives of this study were to estimate heritability and to estimate the correlation among some selected morphological traits and their relationship with FAW damage. Seven shrunken-2 (sh2) inbred lines and four commercial sh2 hybrids of sweet corn were crossed to Mp708, a FAW-resistant field corn line. The F2 populations were subdivided with one half being selected for the sh2 trait and the other half was left unselected. Parent, F2, and F3 populations were artificially infested with FAW and evaluated for leaf damage caused by FAW. Heritability estimates for FAW resistance ranged from 0.22 to 0.61 depending on method of estimation used. The highest correlations occurred between silk color and anther color (0.70) and silk color and glume color (0.49). There were no consistent correlations of most morphological traits with FAW damage. A linkage between white silk and shrunken-2 was observed.

Heritability and correlation among some selected morphological traits and their relationship with fall armyworm damage in sweet corn.

Fall armyworm [Spodoptera frugiperda J. E. Smith] (FAW) is a serious pest in field corn and sweet corn [Zea mays L.] in many parts of the world. Sweet corn germplasm with effective levels of resistance to damage by the fall armyworm is needed to transfer resistance to commercial hybrids. The objectives of this study were to estimate heritability and to estimate the correlation among some selected morphological traits and their relationship with FAW damage. Seven shrunken-2 (sh2) inbred lines and four commercial sh2 hybrids of sweet corn were crossed to Mp708, a FAW-resistant field corn line. The F2 populations were subdivided with one half being selected for the sh2 trait and the other half was left unselected. Parent, F2, and F3 populations were artificially infested with FAW and evaluated for leaf damage caused by FAW. Heritability estimates for FAW resistance ranged from 0.22 to 0.61 depending on method of estimation used. The highest correlations occurred between silk color and anther color (0.70) and silk color and glume color (0.49). There were no consistent correlations of most morphological traits with FAW damage. A linkage between white silk and shrunken-2 was observed.

Heritability and correlation among some selected morphological traits and their relationship with fall armyworm damage in sweet corn

ABSTRACT: Fall armyworm [Spodoptera frugiperda J. E. Smith] (FAW) is a serious pest in field corn and sweet corn [Zea mays L.] in many parts of the world. Sweet corn germplasm with effective levels of resistance to damage by the fall armyworm is needed to transfer resistance to commercial hybrids. The objectives of this study were to estimate heritability and to estimate the correlation among some selected morphological traits and their relationship with FAW damage. Seven shrunken-2 (sh2) inbred lines and four commercial sh2 hybrids of sweet corn were crossed to Mp708, a FAW-resistant field corn line. The F2 populations were subdivided with one half being selected for the sh2 trait and the other half was left unselected. Parent, F2, and F3 populations were artificially infested with FAW and evaluated for leaf damage caused by FAW. Heritability estimates for FAW resistance ranged from 0.22 to 0.61 depending on method of estimation used. The highest correlations occurred between silk color and anther color (0.70) and silk color and glume color (0.49). There were no consistent correlations of most morphological traits with FAWdamage. A linkage between white silk and shrunken-2 was observed.

Heritability and correlation among some selected morphological traits and their relationship with fall armyworm damage in sweet corn

Fall armyworm [Spodoptera frugiperda J. E. Smith] (FAW) is a serious pest in field corn and sweet corn [Zea mays L.] in many parts of the world. Sweet corn germplasm with effective levels of resistance to damage by the fall armyworm is needed to transfer resistance to commercial hybrids. The objectives of this study were to estimate heritability and to estimate the correlation among some selected morphological traits and their relationship with FAW damage. Seven shrunken-2 (sh2) inbred lines and four commercial sh2 hybrids of sweet corn were crossed to Mp708, a FAW-resistant field corn line. The F2 populations were subdivided with one half being selected for the sh2 trait and the other half was left unselected. Parent, F2, and F3 populations were artificially infested with FAW and evaluated for leaf damage caused by FAW. Heritability estimates for FAW resistance ranged from 0.22 to 0.61 depending on method of estimation used. The highest correlations occurred between silk color and anther color (0.70) and silk color and glume color (0.49). There were no consistent correlations of most morphological traits with FAW damage. A linkage between white silk and shrunken-2 was observed