Resistant Maize Inbred Lines Research Articles

Assessment of Resistance Mechanisms to Fall Armyworm, Spodoptera frugiperda in Tropical Maize Inbred Lines

The fall armyworm (FAW), Spodoptera frugiperda, a pest of maize native to the Americas first reported in West and Central Africa in 2016, severely threatens maize production and food security in Sub-Saharan Africa. Native genetic resistance is one of the best methods of control of insect pests as it is contained in the seed making it more amenable for use by farmers compared to other interventions and it is also compatible with other integrated pest management (IPM) options. An intensive screening against FAW was carried out by artificial infestation in greenhouse conditions in Kenya between 2017 and 2018 on about 3000 inbred lines available in the germplasm collection of the International Maize and Wheat Improvement Center (CIMMYT). Among these lines, only four showed to be resistant to FAW, but the mechanisms of resistance are not yet known. The objective of this study was to determine the resistance mechanisms specifically non-preference and antibiosis to S. frugiperda in these four selected resistant inbred lines. The studies were conducted under laboratory and net house conditions in Kenya from April 2020 to November 2021. Non-preference was assessed estimating the feeding preference by counting the number of FAW neonates found on each leaf portion, silk portion and grain using binary and multiple choice methods under laboratory conditions, while antibiosis was assessed through the relative growth rate (RGR) and developmental time of FAW larvae on leaves, silks and grains under both laboratory and net house conditions. Among the four resistant maize inbred lines tested, two, namely CML71 and CKSBL10008, exhibited the highest level of antibiosis resistance on leaves. Under laboratory conditions, the larval RGR reduced from 13 mg/d on the most susceptible line to 8 mg/d on CML71. CML71 also showed a good non-preference on leaves compared to other tested lines. Only 6% of neonates choose to feed on CML71 whereas more than 10% choose to feed on the other lines (and 15% on the most susceptible) in multiple choice tests. The non-preference for feeding and lower RGR of larvae on CML71 suggest a biochemical involvement resistance to FAW. Through this study, CML71 is revealed as a highly promising line for use in breeding for native genetic resistance to FAW in tropical maize.

Comparative Proteomic Analyses of Susceptible and Resistant Maize Inbred Lines at the Stage of Enations Forming following Infection by Rice Black-Streaked Dwarf Virus.

Rice black-streaked dwarf virus (RBSDV) is the main pathogen causing maize rough dwarf disease (MRDD) in China. Typical enation symptoms along the abaxial leaf veins prevail in RBSDV-infected maize inbred line B73 (susceptible to RBSDV), but not in X178 (resistant to RBSDV). Observation of the microstructures of epidermal cells and cross section of enations from RBSDV-infected maize leaves found that the increase of epidermal cell and phloem cell numbers is associated with enation formation. To identify proteins associated with enation formation and candidate proteins against RBSDV infection, comparative proteomics between B73 and X178 plants were conducted using isobaric tags for relative and absolute quantitation (iTRAQ) with leaf samples at the enation forming stage. The proteomics data showed that 260 and 316 differentially expressed proteins (DEPs) were identified in B73 and X178, respectively. We found that the majority of DEPs are located in the chloroplast and cytoplasm. Moreover, RBSDV infection resulted in dramatic changes of DEPs enriched by the metabolic process, response to stress and the biosynthetic process. Strikingly, a cell number regulator 10 was significantly down-regulated in RBSDV-infected B73 plants. Altogether, these data will provide value information for future studies to analyze molecular events during both enation formation and resistance mechanism to RBSDV infection.

Optimizing use of U.S. Ex-PVP inbred lines for enhancing agronomic performance of tropical Striga resistant maize inbred lines

BackgroundTemperate maize inbred lines with expired Plant Variety Protection Act certificates (Ex-PVP) are potential sources of desirable alleles for tropical germplasm improvement. Up to now, the usefulness of the Ex-PVP inbred lines as a potential source of novel beneficial alleles for Striga hermonthica resistance breeding to enhance genetic gain in tropical maize has not been reported.ResultsThis study was thus conducted to characterize the combining ability of 24 Ex-PVP inbred lines in crosses with two tropical Striga resistant inbred testers under Striga-infested and non-infested conditions and across three locations for 2 years. Many testcrosses between Ex-PVP inbred lines and the first tester (T1) produced competitive or significantly higher grain yields compared to the hybrid between the two resistant testers under Striga infested and non-infested conditions and across multiple test locations. Also, most of the testcrosses with positive heterosis for grain yield and negative heterosis for Striga damage and emerged Striga count involved T1 as a tester. Our study identified six Ex-PVP inbred lines with positive GCA effects for grain yield under Striga infested and non-infested conditions and across multiple test locations. Amongst these, inbred lines HB8229-1 and WIL900-1 also displayed negative GCA effects for emerged Striga count and Striga damage rating. The inbred line HB8229-1 showed positive SCA effects for grain yield with T2, whereas WIL900-1 had positive SCA effects for grain yield with T1. Over 70% of the Ex-PVP inbred lines were consistently assigned to specific heterotic groups using yield-based classifying methods (mean grain yield and SCA effects).ConclusionsThese results could facilitate systematic introgression of the Ex-PVP inbred lines into the existing Striga resistant heterotic groups in IITA. The Ex-PVP inbred lines with positive GCA effects and producing high grain yields in hybrid combinations could be useful parents for enhancing Striga resistance and agronomic performance of tropical maize hybrids.

Development of an inoculation technique for rapidly evaluating maize inbred lines for resistance to stalk rot caused by Fusarium spp. in the field.

Maize (Zea mays L.) stalk rot, caused primarily by the soil-borne fungal pathogen Fusarium spp., reduces maize quality and yield worldwide. This study was undertaken to develop and utilize a rapid continuous injection inoculation technique to evaluate maize inbred lines for resistance to Fusarium spp. under field conditions, which could facilitate the identification and development of new sources of host resistance to manage the disease. Continuous injection inoculation is a rapid, stable, and simple method that can evaluate the resistance of maize inbred lines to Fusarium stalk rot (FSR) within 20 days. To verify the feasibility and reliability of inoculation method, Fusarium graminearum, F. proliferatum, and F. subglutinans were isolated, identified, and inoculated into maize at the six-leaf stage (V6) by a veterinary adjustable bottle continuous vaccination syringe. Our results showed that out of a total of 97 inbred maize lines, six (6.2%) showed high resistance to maize stalk rot, 20 showed resistance (20.6%), 32 were susceptible (33.0%), and 39 were very susceptible (40.2%). Based on simple sequence repeat (SSR) markers, an analysis of molecular variance indicated a significant correlation between population of the inbred maize line and resistance to FSR (P = 0.001). Overall, this study provided a systematic, rapid, stable, and simple identification method for maize inbred lines resistant to FSR in the field. At the same time, this method was also suitable for genetic diversity analysis of maize inbred lines resistant to FSR.

Identifying suitable tester for evaluating Striga resistant lines using DArTseq markers and agronomic traits.

A desirable tester that elicits greater genetic difference in Striga resistance among test crosses in a breeding program has not been reported. Therefore, this study was conducted to characterize 30 Striga resistant yellow endosperm maize inbred lines and three testers with varying resistance levels to Striga using DArTseq SNP markers and agronomic traits to identify a suitable tester for resistance hybrid breeding. Marker-based and agronomic trait-based genetic distances were estimated for yellow endosperm maize inbred lines and testers with varying resistance levels to Striga. The Marker-based cluster analysis separated the Striga resistant lines and testers into two distinct groups. Although the susceptible tester (T3) was the most distantly related to the 30 Striga resistant inbred lines, it exhibited a narrower range in genetic distance estimates and poor agronomic performance under Striga infestation in crosses with the resistant lines. In contrast, the resistant tester (T2) showed a broader range in genetic distance estimates in pairs with the 30 resistant lines. Also, it formed many high yielding hybrids with desirable traits under parasite pressure. Furthermore, the most significant positive association between agronomic trait-based and marker-based distance estimates (r = 0.389, P = 0.01) was observed when T2 has paired with the Striga resistant maize inbred lines. It thus appears that T2 may be used as a suitable tester to determine the breeding value of lines in hybrid maize resistance breeding programs. T2 was the most suitable tester, with a tolerant tester (T1) as an alternative tester to characterize the combining ability of Striga resistant maize inbred lines. This result can also encourage other breeders to investigate testers relative discriminating ability with varying levels of resistance in hybrid breeding for resistance to diseases, pests, and other parasitic plants.

Genetics of resistance to Fusarium stalk rot caused by Fusarium verticilloides in maize (Zea mays L.)

Fusarium stalk rot disease (FSR), incited by Fusarium verticilloides, is becoming an important biotic production constraint in many major maize growing areas causing substantial yield losses. The present investigation was conducted to understand the genetics of resistance to FSR through six generation means and variances, as a first step in addressing the problem. Five crosses were developed by crossing four FSR susceptible inbreds (VL1043, VL108867, VL121096 and VL1218) with two resistant inbreds (CM202 and CM212). Six generations of the five crosses (VL1043 × CM212, VL108867 × CM202, VL121096 × CM212, VL1218 × CM202 and VL1218 × CM212) were evaluated through artificial disease inoculation during post rainy season of 2018 and summer, 2019. The scaling tests and joint scaling tests indicated the inadequacy of additive-dominance model and showed the presence of epistatic gene effects in all the five crosses for FSR resistance. The study further revealed the importance of additive, dominance and additive × additive gene effects in the expression of FSR. The magnitude and direction of the additive genetic effects [a], dominance genetic effects [d], magnitudes of additive genetic variance (2A) and dominance genetic variance (2D) varied with the genetic background of the crosses over seasons. Duplicate gene interaction was evident in the inheritance of FSR resistance. Both, additive and non-additive components were found important thus reciprocal recurrent selection would be more effective in obtaining FSR resistant maize inbred lines.

Phenotypic characterization of mycotoxin resistant maize (Zea mays L.) inbred lines

Understanding the genetic variation among ear rot/mycotoxin resistant maize inbred lines would greatly aid in breeding high yielding and stable ear rot and mycotoxin resistant hybrids. This study was executed with the objective to characterize ear rot resistant maize inbred lines using phenotypic traits. Experiments were conducted at Ukulinga and Makhathini Research Stations in South Africa. Genetic variability study revealed a significant difference among the mycotoxin resistant inbred lines for Aspergillus ear rot, Fusarium ear rot and other selected secondary traits except for husk cover. Plant height, ear height and primary tassel branches recorded higher heritability estimates (81.6, 81.2 and 82.8%, respectively) compared to all the traits. The principal component analysis clustered the ear rot resistant inbred lines into five groups over the quadrants based on their phenotypic characteristics within the family. The information generated from this study will assist in developing local hybrids with high resistance to ear rots and mycotoxin contaminations, which attribute better agronomic performance.

Response of Selected Maize Inbred Germplasm to Maize Lethal Necrosis Disease and Its Causative Viruses (Sugarcane Mosaic Virus and Maize Chlorotic Mottle Virus) in Kenya

Background: Maize lethal necrosis (MLN) disease continues to reduce the productivity of maize drastically threatening food security in the affected regions. It continues to cause yield loss of 30–100 percent in farmers’ fields, depending on the time of infestation which is valued at $198 million in Kenya. This has not only threatened regional trade, but also seed industry. It has been reported in the major maize belts of Uasin Gishu, Trans-Nzoia, Bomet, Narok and Nandi Counties. MLN is caused by the synergistic interaction between Sugarcane Mosaic Virus (SCMV) and Maize Chlorotic Mottle Virus (MCMV). The disease has then spread to other Eastern and Central African countries with devastating food security and economic consequences. Objectives: This study highlights result after screening selected maize inbred lines for resistance to MLN, SCMV and MCMV in identifying promising lines for integration into the breeding program for MLN resistance. Methods: Sixty-five (65) maize genotypes were artificially inoculated using virus strains collected from Bomet County in Kenya at 3-4 leaf stage. Data on disease severity and incidence, AUDPC and flowering were recorded. Results: From the result, the inbred lines had significant differences for SCMV, MCMV and MLN reactions. Based on Area Under Disease Progress Curve (AUDPC) score and ELISA analysis, genotypes MLN001 and MLN006 have the lowest score of 270, whereas OH28 had a maximum at 1259 under MCMV. Genotypes MLN042 and MLN041 were identified as the most promising sources of resistant against SCMV. However, no genotype was identified to have acceptable levels of tolerance to MLN, but MLN001 and MLN013 were identified as the best performers under MLN. This study also validated the presence of MLN tolerance in MLN013 (CKDHL120312) and MLN001 (CKDHL120918) as earlier reported by CIMMYT. These tolerant genotypes are now serving as donors in the introgression of the tolerance into the Kenyan adapted maize backgrounds and development of improved MLN tolerant varieties. This will go a long way in restoring and ensuring sustainable maize productivity in improving the livelihoods of the smallholder farmers who form 75% of the major maize producers in Kenya. Conclusion: The identified inbred lines would be recommended for use in varietal development, MLN management and to enhance maize productivity, in the MLN endemic regions and further research in understanding the mode of gene action for MLN tolerance.

Heterotic affinity and combining ability of exotic maize inbred lines for resistance to aflatoxin accumulation

Aflatoxin accumulation in maize (Zea mays L.) kernels is a serious economic and health problem that reduces grain quality and nutritional values and causes death to livestock and humans. Understanding the genetic parameters and heterotic responses of exotic maize inbred lines can facilitate their use for developing aflatoxin resistant parents of hybrids in Africa. This study was designed to (1) determine the heterotic affinities of aflatoxin resistant exotic lines, (2) identify exotic inbreds with good combining ability, and (3) determine the mode of inheritance of resistance to aflatoxin contamination in these lines. A line × tester mating design was used to determine combining ability of 12 yellow and 13 white inbreds and classify them into heterotic groups. The inbreds were crossed to two adapted testers representing two African heterotic groups and the resulting testcrosses along with hybrid checks were evaluated in separate trials at two locations for 2 years in Nigeria. General combining ability (GCA) effects were more important than specific combining ability effects for aflatoxin and grain yield. Among 15 exotic inbred lines having negative GCA effects for aflatoxin and 13 with positive GCA effects for grain yield, six combined the two desired traits. Five white and six yellow endosperm testcrosses were found to be good specific combiners for the two desired traits. The exotic lines with negative GCA effects for aflatoxin accumulation will be used as donor parents to develop backcross populations for generating new inbred lines with much higher levels of resistance to aflatoxin accumulation.

Dissection of mechanisms of resistance toAspergillus flavus and aflatoxin using tropical maize germplasm

Aspergillus flavus induced ear rots and subsequent contamination of maize (Zea mays L.) by aflatoxin is a serious food safety issue, especially in developing countries where the crop is mostly cultivated by smallholder famers for own consumption and income generation. A better understanding of the mechanisms of resistance could help breeders to develop resistant maize varieties. In this study, a set of six tropical maize inbred lines previously identified as resistant or susceptible under natural field conditions were evaluated for response toA. flavus colonisation and aflatoxin contamination. Fungal biomass was significantly higher (P<0.05) in susceptible than resistant maize inbred lines, and this was highly correlated (P=0.001) to aflatoxin levels. Maize inbred lines MRI, MR2 and MR3 had low fungal biomass and low aflatoxin levels, suggesting that resistance in these lines was mediated through restricted fungal colonisation and establishment. Among the three putatively resistant inbred lines mentioned above, MR2 had a relatively high colonisation compared to the other two lines, revealing thatA. flavus could establish and colonise kernels that were injured during inoculation, but did not contain high levels of aflatoxin. This could signify the presence of host genes that interfere with the aflatoxin biosynthetic pathway.

Genetic analysis of tropical maize inbred lines for resistance to maize lethal necrosis disease

Maize lethal necrosis (MLN) disease is a recent outbreak in eastern Africa and has emerged as a significant threat to maize production in the region. The disease is caused by the co-infection of Maize chlorotic mottle virus and any member of potyviridae family. A total of 28 maize inbred lines with varying levels of tolerance to MLN were crossed in a half-diallel mating design, and the resulting 340 F1 crosses and four commercial checks were evaluated under MLN artificial inoculation at Naivasha, Kenya in 2015 and 2016 using an alpha lattice design with two replications. The objectives of the study were to (i) investigate the magnitude of general combining ability variance (σGCA2) and specific combining ability variance (σSCA2) and their interaction with years; (ii) evaluate the efficiencies of GCA based prediction and hybrid performance by means of a cross-validation procedure; (iii) estimate trait correlations in the hybrids; and (iv) identify the MLN tolerant single cross hybrids to be used as female parents for three-way cross hybrids. Results of the combined analysis of variance revealed that both GCA and SCA effects were significant (P < 0.05) for all traits except for ear rot. For MLN scores at early and late stages, GCA effects were 2.5–3.5 times higher than SCA effects indicating that additive gene action is more important than non-additive gene action. The GCA based prediction efficiency for MLN resistance and grain yield accounted for 67–90% of the variations in the hybrid performance suggesting that GCA-based prediction can be proposed to predict MLN resistance and grain yield prior to field evaluation. Three parents, CKDHL120918, CML550, and CKLTI0227 with significant GCA effects for GY (0.61–1.21; P < 0.05) were the most resistant to MLN. Hybrids “CKLTI0227 × CML550”, “CKDHL120918 × CKLTI0138”, and “CKDHL120918 × CKLTI0136” ranked among the best performing hybrids with grain yield of 6.0–6.6 t/ha compared with mean yield of commercial check hybrids (0.6 t/ha). The MLN tolerant inbred lines and single cross hybrids identified in this study could be used to improve MLN tolerance in both public and private sector maize breeding programs in eastern Africa.

Construction of high-density linkage map and identification of QTLs for resistance to sorghum downy mildew in maize (Zea mays L.)

Sorghum downy mildew (SDM), caused by obligate biotrophic fungi Peronosclerospora sorghi, is an economically important disease of maize. The genetics of resistance was reported to be polygenic thereby necessitating identification of QTLs for resistance to SDM to initiate effective marker-assisted selection programs. During post-rainy and winter season of 2012, 645 F2:3 progeny families from the cross CML153 (susceptible) × CML226 (resistant) were screened for their reaction to SDM. Characterization of QTLs affecting resistance to SDM was undertaken using the genetic linkage map with 319 polymorphic SSR and SNP marker loci and the phenotypic data of F2:3 families. Three QTLs conferring resistance to SDM were consistently identified on chromosomes 2, 3 and 6 in both seasons. The resistant parent CML226 contributed all the QTL alleles conferring resistance to SDM. The major QTL located on chromosome 2 explained 38.68% of total phenotypic variation in the combined analysis with a LOD score of 9.12. All the three QTL showed partially dominant gene effects in combined analysis. The detection of more than one QTL supports the hypothesis that quantitative genes control resistance to P. sorghi. The generation was advanced to F6 using markers linked to major QTLs on chromosomes 2 and 3 to derive 33 SDM resistant maize inbred lines.

Evaluation of maize inbred lines for resistance to pre-harvest aflatoxin and fumonisin contamination in the field

Two important mycotoxins, aflatoxin and fumonisin, are among the most potent naturally occurring carcinogens, contaminating maize (Zea mays) and affecting crop yield and quality. Resistance of maize to pre-harvest mycotoxin contamination, specifically aflatoxin produced by Aspergillus flavus and fumonisin produced by Fusarium verticillioides, is a goal in breeding programs that screen for these important traits with the aim of developing resistant commercial hybrids. We conducted two years of field evaluations on 87 inbred lines originating primarily in China and Mexico and not previously screened for resistance. The objectives of our study were to identify resistant germplasm for breeding purposes and to examine possible relationships between resistances to the two mycotoxins. Aflatoxin and fumonisin were present in samples harvested from all lines in both years. Concentrations of total aflatoxin ranged from 52.00±20.00 to 1524.00±396.00μgkg−1, while those of fumonisin ranged from 0.60±0.06 to 124.00±19.50mgkg−1. The inbred lines TUN15, TUN61, TUN37, CY2, and TUN49 showed the lowest aflatoxin accumulation and CN1, GT601, TUN09, TUN61, and MP717 the lowest fumonisin accumulation. TUN61 showed the lowest accumulation of both mycotoxins. This study confirmed previous observations that high levels of aflatoxin can coexist with fumonisin, with 55 maize lines showing a positive correlation coefficient between the concentrations of aflatoxin and fumonisin and 32 lines showing a negative correlation coefficient. These selected lines, particularly TUN61, may provide sources of resistance to mycotoxin contamination in breeding programs. However, the mechanism of resistance in this germplasm remains to be identified. Future research should also address factors that influence the fungus–plant interaction, such as herbivory and environmental stress.

Multi-environment Evaluation of Maize Inbred Lines for Resistance to Fusarium Ear Rot and Fumonisins.

Fusarium verticillioides causes Fusarium ear rot (FER) of maize and produces fumonisins, which affects grain quality. Host-plant resistance can reduce both FER and fumonisins in maize. In this study, 18 maize inbred lines were evaluated for resistance to F. verticillioides and fumonisin accumulation at five localities in South Africa. Additive main effects and multiplicative interaction analyses revealed significant environment × genotype interactions, with inbred lines CML 390, US 2540W, RO 424W, and VO 617y-2 consistently exhibiting low FER severity (≤5.4%), fungal target DNA (≤0.1 ng μl-1), and fumonisin levels (≤5.6 ppm). Genotype main effect and genotype × environment biplots showed that inbred lines CML 390, US 2540W, and RO 424W were most resistant to FER, fungal colonization, and fumonisin accumulation, respectively, while inbred line RO 424W was most stable in its resistance response over environments. These inbred lines also demonstrated broad adaptability by consistently exhibiting resistance to FER, fungal colonization, and fumonisins across localities. The identified lines could serve as valuable sources of resistance against F. verticillioides and its fumonisins in local breeding programs.

English

Stem borers, Busseola fusca (Fuller) Noctuidae, Lepidoptera and Chilo partellus (Swinhoe) Pyralidae, Lepidoptera, are serious insect pests of maize. However, genotypes showing exclusive resistance to each of these borers and with resistance to both species have not been identified in Kenya. The objective of this study was to evaluate tropical maize inbred lines for resistance to the two species. 112 maize inbred lines were artificially infested with the stem borers at three sites in Kenya. Each row of each line received three different treatments in different parts, namely infestation with B. fusca larvae, infestation with C. partellus larvae, and protection with beta 25 g/L cyfluthrin pesticide. Data was collected on leaf feeding damage rating, cumulative stem tunnel length, number of exit holes, number of dead-hearts, stalk strength and selected agronomic traits. There were significant differences among the test genotypes, (p&lt;0.01) for resistance to B. fusca and C.&nbsp;partellus, for all the traits measured. The results also showed that most of the test genotypes were susceptible to B. fusca and less so to C. partellus. Twenty one (21) lines showed resistance to both B. fusca and C. partellus in at least two sites, and only four lines showed resistance to both species across the locations. Among all the test genotypes, 26 lines showed resistance to C. partellus only, while five entries had resistance to B. fusca only. Furthermore, 84 and 28 entries showed susceptibility to B.&nbsp;fusca and C.&nbsp;partellus, respectively. The others were categorized as either moderately resistant or moderately susceptible to either species. The identified inbred lines variously resistant to B.&nbsp;fusca and C.&nbsp;partellus may be used as parents in hybrid breeding programmes that emphasize stem borer resistance or as sources of resistance in breeding programs. &nbsp; Key words: Busseola fusca, Chilo partellus, combined resistance, tropical maize inbred lines.

Aflatoxin Resistance in Maize: What Have We Learned Lately?

Aflatoxin contamination of maize grain is a huge economic and health problem, causing death and increased disease burden in much of the developing world and income loss in the developed world. Despite the gravity of the problem, deployable solutions are still being sought. In the past 15 years, much progress has been made in creating resistant maize inbred lines; mapping of genetic factors associated with resistance; and identifying possible resistance mechanisms. This review highlights this progress, most of which has occurred since the last time a review was published on this topic. Many of the needs highlighted in the last reviews have been addressed, and several solutions, taken together, can now greatly reduce the aflatoxin problem in maize grain. Continued research will soon lead to further solutions, which promise to further reduce and even eliminate the problem completely.

Identification of maize genes associated with host plant resistance or susceptibility to Aspergillus flavus infection and aflatoxin accumulation.

Background Aspergillus flavus infection and aflatoxin contamination of maize pose negative impacts in agriculture and health. Commercial maize hybrids are generally susceptible to this fungus. Significant levels of host plant resistance have been observed in certain maize inbred lines. This study was conducted to identify maize genes associated with host plant resistance or susceptibility to A. flavus infection and aflatoxin accumulation.ResultsGenome wide gene expression levels with or without A. flavus inoculation were compared in two resistant maize inbred lines (Mp313E and Mp04∶86) in contrast to two susceptible maize inbred lines (Va35 and B73) by microarray analysis. Principal component analysis (PCA) was used to find genes contributing to the larger variances associated with the resistant or susceptible maize inbred lines. The significance levels of gene expression were determined by using SAS and LIMMA programs. Fifty candidate genes were selected and further investigated by quantitative RT-PCR (qRT-PCR) in a time-course study on Mp313E and Va35. Sixteen of the candidate genes were found to be highly expressed in Mp313E and fifteen in Va35. Out of the 31 highly expressed genes, eight were mapped to seven previously identified quantitative trait locus (QTL) regions. A gene encoding glycine-rich RNA binding protein 2 was found to be associated with the host hypersensitivity and susceptibility in Va35. A nuclear pore complex protein YUP85-like gene was found to be involved in the host resistance in Mp313E.ConclusionMaize genes associated with host plant resistance or susceptibility were identified by a combination of microarray analysis, qRT-PCR analysis, and QTL mapping methods. Our findings suggest that multiple mechanisms are involved in maize host plant defense systems in response to Aspergillus flavus infection and aflatoxin accumulation. These findings will be important in identification of DNA markers for breeding maize lines resistant to aflatoxin accumulation.

Field evaluation of maize inbred lines for resistance to Exserohilum turcicum

Nine maize inbred lines with excellent grey leaf spot resistance and good combining ability were evaluated for their reaction to infection by Exserohilum turcicum. The lines were compared to a differential set consisting of Oh43 and B73 (susceptible controls) and Oh43Ht1B, Oh43Ht2, Oh43Ht3, B37HtN (containing resistant genes) in both growth chamber and field trials. All nine lines possessed seedling resistance to northern corn leaf blight (NCLB). Two field trials were planted over two growing seasons (2007/08 and 2008/09) as randomized complete block designs with three replicates. Trials were inoculated twice (at the 4–5 and 8–12 leaf stages) with NCLB and disease assessments made at growth stages R1, R2, R3, R4 and R5. Disease progress curves were created and total severity (yt ), diseased plant severity (ydp ) and AUDPC were determined for each replicate. Linearised forms of the exponential, logistic and Gompertz models were fitted to the disease-progress data and the best model selected for each trial. Rate of disease increase (r) and level of disease at the onset of the epidemic (y0 ; time=0) were obtained from which y0 * (back-transformed from y0 ) were determined. Canonical vari-ate analysis (CVA) indicated that yt, ydp , sAUDPC and y0 * were responsible for 73.68% of the variation observed between the lines, with r explaining 15.44% of the variation. Ranking of lines were based on their yt, ydp , sAU-DPC and y0 * performance. GLS resistant lines 185–2, 182–2, 72–2A and 182–1 demonstrated higher levels of resistance to NCLB than that of lines Oh43Ht1B, Oh43Ht2, Oh43Ht3 and B37HtN while 122–2, 578 and 72–3 showed higher levels of resistance than that of Oh43Ht2 and B37HtN.

Combining ability among twenty insect resistant maize inbred lines resistant to Chilo partellus and Busseola fusca stem borers.

A partial diallel design was used among 20 maize inbred lines to form 110 F1 hybrids to generate information on the values of these lines for developing insect resistant maize varieties during the short rains season of 2006. The hybrids were evaluated for resistance to the C. partellus and B. fusca, and for agronomic performance over two seasons during long and short rains of 2007 at a mid-altitude dry early maturity site at KARI Kiboko, and the moist mid-altitude medium maturity site at KARI Embu. leaf damage score (using a scale of 1-9 where: 1= No damage and 9= extremely damaged), number of exit holes, cumulative tunnel length, and grain yield were measured as resistance traits. The genotype sum of square was partitioned into general combining ability (GCA) and specific combining ability (SCA) effects. Maize inbred lines with good general combining ability for insect resistance including maize inbred lines with significant and negative GCAs for leaf damage were identified as lines 12,16,18,19 and 20 at Kiboko and lines 8, 17, 18, and 20 in Embu. Results showed that the problem of stem borers intensified by over 40% within four years in the experimental region of Eastern Kenya; currently mean yield loss due to stem borers was assessed to be about 56%. Several hybrids had significant negative SCA for leaf damage and significant positive SCA for grain yield. The maize inbred lines studied revealed their potential for use in breeding programs for insect resistance that could result in a correlated response for increased grain yield. Recurrent selection would be the best option to develop high yielding insect resistant germplasm for this region of Kenya considering that additive gene action were predominant. Evidently, it would be more difficult to develop host plant resistance to B. fusca than to C. partellus.

Evaluation of Maize Inbred Lines for Resistance to Aspergillus and Fusarium Ear Rot and Mycotoxin Accumulation

Mycotoxin contamination in corn (Zea mays L.) grain is a worldwide threat to safety of both human food and animal feed. A select group of inbred corn lines was evaluated in field trials for ear rot caused by Aspergillus flavus and Fusarium verticillioides and mycotoxin accumulation in grain. Our goal was to identify lines resistant to both fungi. In separate tests, 20 inbred lines were inoculated with either A. flavus or F. verticillioides After harvest, ears were rated for rot and evaluated for levels of aflatoxin or fumonisin contamination. Inbred line Mp717 exhibited low grain aflatoxin contamination and it also had the lowest levels of fumonisin. Inbred line Mp317 has been shown previously to have low levels of F. verticillioides kernel infection and fumonisin contamination. Mp317 also had low levels of aflatoxin contamination in this study. Area of the ear rotted by F. verticillioides and A. flavus was significantly correlated to toxin production for both fumonisin (P = 0.0002; r = 0.74) and aflatoxin (P = 0.004; r = 0.61), indicating that inbreds exhibiting aflatoxin resistance may also be good sources of fumonisin resistance. Our method of quantifying ear rot may also be used to rapidly screen lines for A. flavus resistance and subsequent aflatoxin accumulation in preliminary evaluations.