Combining Ability and Heterotic Grouping of Turcicum-Resistant Early-Maturing Maize Inbreds

Baffour Badu-Apraku1*, Faith A. Bankole1,2, Morakinyo A. B. Fakorede2, Gregory Ogbe1, Ranajit

Bandyopadhyay1, Alejandro Ortega-Beltran1

1
International Institute of Tropical Agriculture (IITA), Ibadan, Nigeria

2
Obafemi Awolowo University, Ile-Ife, Nigeria

*Corresponding author: [email protected]

Faith A. Bankole: [email protected]

Morayinko Abiodun Bamidele Fakorede: [email protected]

Gregory Ogbe: [email protected]

Ranajit Bandyopadhyay: [email protected]

Ortega-Beltran, Alejandro: [email protected]

Abstract

Maize, an important source of calories and nutrients in sub-Saharan Africa is threatened by northern

corn leaf blight (NCLB) caused by Exserohilum turcicum. This study examined combining ability and

heterotic patterns of early-maturing (EM) maize inbreds, gene action conditioning NCLB resistance,

performance of derived hybrids across environments, and identified testers. Fifteen each of white and

yellow inbreds, were intercrossed using North Carolina Design II to obtain 75 hybrids per endosperm

color. Hybrids plus six checks were inoculated with a virulent isolate of E. turcicum four weeks after

planting in six inoculated and three non-inoculated environments in Nigeria, 2018 and 2019. Inbreds

were assigned to heterotic groups using general combining ability (GCA) of multiple traits method.

Specific combining ability (SCA), GCA, and genotype × environment interactions (G × E) were

significant for grain yield (GYLD), disease severity, and other traits. NCLB caused 46% GYLD

reduction. GCA effects were preponderant over SCA for GYLD and NCLB severity across

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environments indicating that hybridization and recurrent selection would enhance genetic gains and

hybrid performance. White and yellow inbreds were placed in two and three heterotic groups,

respectively. High-yielding NCLB resistant testers identified could be used to classify other inbreds yet

to be field-tested into heterotic groups. TZEI 32 × TZEI 5 and TZEI 124 × TZEI 134 were identified as

single-cross testers. TZEI 32 × TZEI 5, TZEI 5 × TZEI 75 (white) and TZEI 124 × TZEI 134, TZEI

124 × TZEI 11 (yellow) were identified for on-farm testing and possible commercialization.

Keywords: host-plant resistance, heterosis, testers.

Abbreviations: AMMI: additive main effects and multiplicative interaction; ASI: anthesis-silking

interval; BI: base index; DA: days to 50% anthesis; DS: days to 50% silking; EASP: ear aspect; EHT:

ear height; EM: early maturing; EPP: number of ears per plant; GCA: general combining ability; G × E:

genotype by environment; GYLD: grain yield; HGCAMT: heterotic grouping based on GCA of

multiple traits; HUSK: husk cover; PASP: plant aspect; PHT: plant height; RL: root lodging; SCA:

specific combining ability; SL: stalk lodging; SSA: sub-Saharan Africa; TURC2WAI: disease score two

weeks after inoculation; TURC6WAI: disease score six weeks after inoculation; TURC: average disease

severity score; WCA: West and Central Africa.

Core Ideas

 Gene action for grain yield and turcicum resistance was predominantly additive in early-

maturing (EM) inbreds.

 Three heterotic groups identified for planned hybrid crosses involving EM inbreds.

 Inbred and single-cross testers identified for grouping and testing for turcicum resistance.

 Identified testers should be used to classify EM inbreds which are yet to be field-tested.

 Ten high-yielding and stable hybrids with turcicum resistance identified for commercialization.

Introduction

Maize (Zea mays L.), a cereal crop widely cultivated for high source of calories, minerals, and other

nutrients for populations in sub-Saharan Africa (SSA) (Badu-Apraku and Fakorede, 2017), also plays

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a strategic role in the emerging brewery and livestock feed industries as a raw material (Badu-Apraku

et al., 2013; Badu-Apraku and Fakorede, 2017). However, maize productivity in SSA is constrained

by several biotic and abiotic stresses. Among the prominent biotic stresses are the parasitic weed

Striga hermonthica, the fall armyworm (Spodoptera frugiperda), stemborers, and several foliar

diseases, including the northern corn leaf blight (NCLB). The NCLB caused by the fungus,

Exserohilum turcicum (Pass.) Leonard & Suggs., is a destructive foliar disease accounting for over

30% yield loss and complete loss if infection occurs at the seedling stage (Hooda et al., 2016; Human

et al., 2016). Recently, there has been increased incidence and severity of NCLB in the lowland

tropics of SSA calling for urgent disease control strategies to manage the disease (Akinwale and

Oyelakin, 2018). Several methods could be adopted to manage NCLB; however, breeding for

resistance is the most appropriate, environmentally friendly, and most feasible and sustainable control

method (Ayiga-Aluba et al., 2015; Chen et al., 2016; Wiesner-Hanks and Nelson, 2016). Maize

varieties, inbred lines, and hybrids vary in susceptibility to NCLB and there are no immune

accessions. Susceptible germplasm has poor physiological development and growth, reduced quality

of produce and low yield. Contrarily, tolerant germplasm support disease development but exhibits

little or no reduction in physiological development, growth, quality of produce and yield (Badu-

Apraku and Fakorede, 2017). Both qualitatively and quantitatively inherited resistance have been

reported for the control of NCLB (Welz and Geiger, 2000). However, repeated emergence of

physiologically diverse races renders qualitative resistance unstable. Therefore, the development of

quantitatively inherited resistance has been the goal of breeders (Sibiya et al., 2013).

Increased maize production by farmers in West and Central Africa (WCA) has been possible through

cultivation of open-pollinated varieties (OPV); only a small area (~5%) in WCA is planted with

hybrids (Abdoulaye et al., 2009). However, use of hybrids holds a great promise for maize production

in WCA (Oyekunle and Badu-Apraku, 2014), but the hybrids need to be resistant to major diseases to

have increased usage. Early maturing (EM), high-yielding, and stable maize hybrids could contribute

to increased hybrid preference in the region. Therefore, during the past two decades development of

such hybrids has received considerable attention in the International Institute of Tropical Agriculture

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maize improvement program (IITA-MIP). The major emphases of the program, apart from high grain

yield, has been on Striga and drought tolerance. Breeding for resistance to NCLB was recently

included as a target trait due to severe incidence of the disease in the region. Development and

commercialization of high-yielding EM hybrids with stable resistance to NCLB across multiple

environments would contribute to increased adoption of hybrids in WCA. Additionally, information

on the heterotic patterns of parental lines is needed to determine the potential usefulness of the inbred

lines in the hybrid program. This would facilitate better understanding of the genetic relationships

among the inbreds and classification into appropriate heterotic groups for effective utilization in the

IITA maize improvement program for the development of multiple-stress tolerant hybrids, synthetic

varieties, as well as heterotic populations.

The combining ability of maize inbreds and their classification into heterotic groups provide

information on their potentials for the development of productive hybrids. Panhwer et al. (2008)

defined combining ability as the potential of inbreds to hybridize so that favourable genes could be

transmitted to their progenies. Studies on combining ability are of utmost importance for designing

breeding programs for hybrid development and comparison of the performance of hybrids developed

from inbreds. Information on combining ability of inbreds in hybrid combinations is, therefore,

important for successful maize hybrid improvement (Panhwer et al., 2008; Amegbor et al., 2017). An

assessment of the general combining ability (GCA) effects of inbreds for grain yield (GYLD) and

other agronomic traits relative to the specific combining ability (SCA) effects of the derived hybrids is

crucial for identifying outstanding hybrids for commercial production (Badu-Apraku and Oyekunle,

2012). Genetic studies have demonstrated the predominance of GCA over SCA effects for NCLB and

grain yield, indicating that genetic gains in breeding for resistance to NCLB could be maximized

through population improvement methods (Vivek et al., 2010; Sibiya et al., 2013). Contrarily,

Schechert et al. (1997) reported the predominance of SCA over GCA effects of inbreds studied

indicating that hybridization of inbreds would be the best approach.

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Since 2016, there have been serious outbreaks of NCLB in the lowlands of WCA leading to large

yield losses. Fortunately, numerous drought, low soil nitrogen tolerant, and Striga resistant EM and

extra-early-maturing (EEM) inbreds have been developed in the IITA-MIP. In an effort to mitigate the

damage caused by NCLB, the EM and EEM multiple stress tolerant inbreds were screened for

resistance to NCLB (Badu-Apraku et al., 2021a). Several resistant inbreds were identified thus paving

the way for the development of high-yielding hybrids resistant to NCLB and other stresses. In order to

develop multiple stress tolerant hybrids with resistance/tolerance to NCLB, it is necessary to study the

combining ability of traits and patterns of inheritance of the inbreds to identify hybrids with potential

for commercialization (Badu-Apraku et al., 2013; Badu-Apraku and Oyekunle, 2012). Additionally,

there is a critical need for accurate evaluation of the crosses under high disease pressure, which could

be achieved through artificial E. turcicum inoculation. Additionally, there is the need to examine the

heterotic patterns of the inbreds and identify testers in selected IITA-MIP EM inbreds with resistance

to E. turcicum.

The present study was conducted with the objectives of i) estimating the combining abilities of sets of

EM white and yellow inbreds, ii) determining the gene action conditioning resistance to NCLB, iii)

classifying the inbreds into distinct heterotic groups, iv) identifying EM testers for maize breeding

programs in SSA, and v) determining stability of EM hybrids in contrasting test environments.

Materials and Methods

Generation of crosses

Thirty EM maize inbred lines developed by IITA-MIP were used for the present study. The inbreds

were selected based on their performance under high NCLB disease pressure provided by artificial

inoculation at Ile-Ife, Ikenne, and Zaria, Nigeria (Badu-Apraku et al., 2021a). There were 15 yellow

and 15 white endosperm types which were inter-crossed using the NCD II mating scheme (Comstock

and Robinson, 1948). The inbreds were classified into six sets (three sets per grain color) with five

inbreds in each set. Inbreds within each grain color were crossed with one set used as female and

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another set as male; a total of 150 single-cross hybrids (75 each of white and yellow endosperm

colors) were developed in 2017 in IITA, Ibadan.

Inoculum preparation for field studies

Inoculum for each location was prepared using a total of 100 Erlenmeyer flasks containing sorghum

grains, inoculated with E. turcicum. Briefly, 100 g of white sorghum (Sorghum bicolor) grains were

weighed into each Erlenmeyer flask and soaked overnight. The grains were washed in distilled water

and sterilized at 121°C for 1 hr in an autoclave. After cooling in aseptic conditions, the grains were

inoculated with a 4 ml spore suspension (105 spores ml-1) of virulent E. turcicum isolate NGIB16-13

and incubated for 4 d at room temperature (~26°C) (Badu-Apraku et al., 2021a). The colonized grains

were stored in a refrigerator (4°C) for less than 48 h before field inoculations.

Field evaluations

One hundred and fifty EM single-cross hybrids plus six checks (81 hybrids) each of white and yellow

endosperm color were evaluated under artificial E. turcicum inoculation at Ikenne (6°53’ N, 3°42’ E),

Ile-Ife 7 8 E and Zaria (11°7’ N, 7°45’ E) in Nigeria during the 2018 growing season. The

experiment was repeated in 2019 growing season at the three sites but, in addition non-inoculated

trials were evaluated at the three test locations. The characteristics of the test sites are described in

Table 1. The test sites are in target environments of the IITA-MIP, where severe NCLB outbreaks

have occurred recently. The hybrids were evaluated using 9 × 9 lattice design with two replicates per

grain color. A plot consisted of a single-cross hybrid planted in 4 m single-row plot, each spaced 0.75

m × 0.40 m. Three seeds of each hybrid were planted per hill and thinned to two seedlings per hill 2

weeks after planting (WAP), resulting in 22 plants per plot. Basal application of NPK 15-15-15

fertilizer was done 3 WAP and top-dressed with urea at 5 WAP. Artificial inoculation of the trials was

carried out within each plot at 4 WAP by placing 10 to 15 E. turcicum colonized sorghum grains (at a

rate of 40 kg ha-1) into the whorl using a sterile scoop. Non-inoculated trials were treated with 0.4 ml

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l-1 of a combination of pyraclostrobin and dimethomorph emulsified concentrate at an interval of 2

weeks until 10 WAP to prevent the spread of the pathogen from the inoculated fields to non-

inoculated trials.

Disease scoring method

Whole plots of each experiment were visually scored twice during crop growth and development for

disease severity: i) two weeks after inoculation (TURC2WAI, 6 WAP) to determine the initial

response of the hybrids at the early plant growth stage, and ii) six weeks after inoculation

(TURC6WAI, 10 WAP) to determine the reaction of the hybrids as they matured. The plots were

scored using a 1 to 9 scale (Badu-Apraku et al., 2021a).

Measurement of other agronomic traits

Observations on days to pollen shed was recorded as the days when half of the plants per plot had

started producing pollen (days to anthesis, DA), while days to silking (DS) was determined by

counting the number of days from planting to incipient silk extrusion of 50% of plants in a plot.

Anthesis-silking interval (ASI) was obtained as days to silking minus days to pollen shed. Plant

(PHT) and ear (EHT) heights were obtained as the distance measured from the base of the plant to the

height of the first tassel branch and to the node bearing the upper ear, respectively. Root lodging (RL)

was computed as the number of plants leaning >30° from the upright position. Stalk lodging (SL) was

recorded as the number of stalks broken below the ear few days before harvesting. Ears per plant

(EPP) was calculated by counting the total number of ears per plot and dividing by the number of

plants harvested. Plant aspect (PASP) was scored based on the overall plant appeal, considering

factors such as relative uniformity of plant height and ear placement, reaction to diseases and insect

damage, and lodging resistance, using a scale of 1 to 9 where 1 = excellent phenotypic appeal and 9 =

poor phenotypic appeal. Ear aspect (EASP) was measured as the extent of disease and insect damage

of ears, ear size, and uniformity. The EASP was measured using a scale of 1 to 9; where 1 = clean,

uniform, large, and well-filled ears and 9 = ears with undesirable features (Badu-Apraku et al., 2013).

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Husk cover was based on a scale of 1 to 9, with 1 = ears with tightly arranged husks which extended

beyond the ear tip, while 9 = ears with the tips greatly exposed. Field weight represented weight of

dehusked ears per plot. GYLD (kg ha-1) was estimated based on 80% shelling. The moisture content

of the grains was adjusted to 150 g kg-1 and computed as previously described (Badu-Apraku et al.,

2012).

Data analysis

Each location × year combination was regarded as a test environment and analysis of variance

(ANOVA) was conducted per test environment to estimate the means for each entry adjusted for

block effects (Cochran and Cox, 1960). The MIXED MODEL procedure of Statistical Analysis

System (SAS) package version 9.13 (SAS Institute, 2011) was used, with blocks nested in replication

× environments. Genotype effect was fixed while replicates within environments were considered as

random factors. The PROC GLM with RANDOM statement and a TEST option was employed for

ANOVA. The model used for the ANOVA is:

Yijkg= μ + Ei + Rj(i) + Bk(ij) + Gg + EGig + εijkg

Yijkg represents the measurement for the gth genotype in the ith environment, in the kth block within

jth replicate; μ is the grand mean; Ei is main effect of environment; Rj(i) is the effect of replication

within environment effect; Bk(ij) is the block effect nested in replicate j by environment i; Gg are the

effects due to genotypes; EGig is the interaction between genotypes and environments and εijkg is the

error term.

ANOVA for NCD II pooled across sets and test environments was done with the component of

variation due to the hybrids partitioned into female (sets), male (sets), and interaction between female

× male (sets). General combining ability (GCA) was represented by female (sets) and male (sets)

main effects while specific combining ability (SCA) was the interaction between female × male (sets)

(Hallauer et al., 2010). The F tests were calculated for male (sets), female (sets), and female × male

(sets) mean squares employing the respective mean squares for interaction with environment (E).

Selected inbreds were classified into heterotic groups using the GCA of multiple traits (HGCAMT)

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method proposed by Badu-Apraku et al. (2013). A brief description of the method using a small data

set is presented in Supplementary Table 1. The statistical model of the HGCAMT method used to

group the inbreds is as follows:

∑ ( ) ɛij

where Y represents the HGCAMT, which denotes genetic value that measures the relationship among

genotypes used to determine the GCA of multiple traits i to n: Yi is the individual GCA effects of

genotypes for trait i. μ is the average GCA effects across Y genotypes for trait i. Si represents the

standard deviation of the GCA effects of trait i. ɛij denotes the residual of the model associated with
the combination of inbred line i and trait j. Characters with significant mean squares under artificial

inoculations were identified and GCA effects of the traits were standardized to reduce the effects of

various rating scales employed to measure the characters. The GCA standardized values were

employed for cluster analysis using Ward’s minimum variance method SAS Institute 20 to

classify the inbreds. Furthermore, a base index (BI) for selecting the resistant and susceptible hybrids

was computed using the following formula: BI= GYLD˗0.7 PASP -0.9(EASP)-0.3(TURC)+0.4(EPP)

(Badu-Apraku et al., 2021a)

To reduce the effects of contrasting scales for the BI, the parameters of the BI were standardized with

means equal to zero and standard deviation equal to one. Genotypes with positive BI values were

identified as resistant to E. turcicum while genotypes with negative BI values were considered as

susceptible to the pathogen.

Based on the BI, 35 (including the best 20, middle 5, and the worst 10 genotypes) white and yellow

single-cross hybrids were selected for stability analysis. The additive main effects and multiplicative

interaction (AMMI) biplot was used to study the genotype (G), environment (E), and G × E

interaction effects of GYLD of hybrids as well as the stability of performance of the hybrids across

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the test environments (Crossa, 1990; Badu-Apraku et al., 2015). Following the principal component

analysis, the AMMI model divided the G × E interactions into different principal component axes

(IPCAs) (Yan, 2001). Subsequently, the AMMI model equation described by Sadeghi et al. (2011)

was applied. We adopted the AMMI biplot to examine the performance of the hybrids across test

environments under artificial E. turcicum inoculation.

Results

Analysis of variance, GCA and SCA of GYLD and NCLB disease severity scores of early

maturing parents under NCLB disease.

For the white EM single-cross hybrids artificially inoculated with E. turcicum, environment mean

squares were highly significant for all measured traits (Table 2). Similar trends occurred for male in

set, female in set, female × male in set, environment × male in set, environment × female in set, and

environment × female × male in set for all measured traits with the exception of a few cases such as

EPP (Table 2). For the artificially inoculated yellow EM single-cross hybrids, similar trends occurred

for all measured traits, apart from EPP which showed a few more highly significant traits, and PHT

with more non-significant cases compared with the white EM hybrids (Table 3). Therefore, by

implication, all these effects were highly significant for all measured traits except in a few cases,

especially EPP, PASP, and EASP. For the two scenarios (Tables 2 and 3), the R2 values and

heritabilities were mostly high apart from ASI and TURC2WAI with relatively low R2 values. The

CVs also were relatively low except that for ASI that was very high in both cases.

Inbred lines with significant positive GCA (male and female) effects, having significant negative

GCA effects for TURC6WAI are the most desirable in this study. Among the white lines, TZEI 5,

TZEI 89, TZEI 32, TZEI 60, and TZEI 75 satisfied that condition (Table 4) and were, therefore,

considered as good general combiners usable either as female or male parents under high E. turcicum

pressure. TZEI 40 had significant positive GCA-male for GYLD and significant negative GCA

(female and male) for TURC6WAI. Contrarily, TZEI 39, TZEI 51, TZEI 56, and TZEI 269 had

significant negative GCA (male and female) effects for GYLD and positive GCA for TURC6WAI.

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They are good combiners for E. turcicum resistance but poor general combiners for GYLD. Among

the yellow EM inbred lines, TZEI 134 and TZEI 124 were the only lines that satisfied the condition

and were, therefore, identified as good combiners usable either as male or female for GYLD and

TURC6WAI under high E. turcicum pressure (Table 4). Furthermore, TZEI 128 possessed significant

positive GCA-male effects for GYLD and significant negative GCA (male and female) effects for

TURC6WAI. However, TZEI 9 and TZEI 23 had negative GCA for GYLD but had significant

positive GCA-female and GCA-male effects for TURC6WAI.

Of the total variation for the measured traits under artificial inoculation, the sum of squares (SS) of

GCA (female and male) for the white EM hybrids ranged from 43 to 83% while 14 to 57% was

attributed to SCA (Table 5). The contributions of the GCA SS to the variation observed among the

white EM hybrids were greater than the SCA SS for the measured traits except for EPP. In addition,

the GCA-male SS was greater than GCA-female SS for GYLD, DA, DS, EASP, TURC6WAI, and

TURC while GCA-female SS was higher for PASP, TURC2WAI, and EPP. Similar results (with few

exceptions) were generally recorded for the measured traits under non-inoculated conditions. Under

the non-inoculated condition, the GCA SS made larger contributions to GYLD, DA, DS, PLHT, EHT,

PASP, and EASP while SCA SS contributions were higher than GCA SS for ASI, TURC2WAI,

TURC6WAI, TURC, and EPP for the white EM hybrids (Table 5). In contrast, 49 to 76% of the total

variation for measured traits under artificial inoculation for the EM yellow hybrids was explained by

GCA (female and male) while the SCA explained 24 to 55 % (Table 6). The contribution of the GCA

SS among the hybrids was greater than the SCA SS for the measured traits except for EHT and EPP.

In addition, the GCA-female SS was greater than GCA-male SS for GYLD, DA, DS, PASP, EASP,

TURC2WAI, TURC6WAI, TURC, and EPP. Under non-inoculated conditions, the GCA SS was

larger for GYLD, DA, DS, PLHT, EHT, PASP, and EASP. However, the contributions of SCA SS

were higher than the GCA SS for ASI, TURC2WAI, TURC6WAI, TURC, and EPP (Table 6). For the

white hybrids, SCA made larger SS contribution for only one trait (EPP) under inoculation whereas

SCA was more important for five traits under non-inoculation treatment. For the yellow hybrids, SCA

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made larger SS contribution to the two traits (EHT and EPP) under inoculation and five traits under

non-inoculation treatment.

Heterotic patterns and identification of testers

The white EM inbreds were assigned into two distinct groups at 38% level of dissimilarity (R-squared

value) (Figure 1). Eight inbreds (TZEI 1, TZEI 89, TZEI 35, TZEI 40, TZEI 5, TZEI 32, TZEI 75, and

TZEI 60) were placed in group 1 while seven (TZEI 39, TZEI 51, TZEI 86, TZEI 56, TZEI 83, TZEI 269,

and TZEI 4) were assigned to the second group. According to the criteria proposed by Pswarayi and

Vivek (2008) for the identification of a tester, inbreds TZEI 60, TZEI 75, and TZEI 32 classified into

group 1, possessed significant positive GCA (female and male) effects for GYLD, as well as high per se

GYLD. Furthermore, the three inbreds possessed significant negative GCA (male and female) effects for

NCLB severity scores. However, the per se GYLD of TZEI 60 was significantly higher than that of TZEI

75 and TZEI 32 (Table 7) and was therefore selected as a tester for group 1. The single-cross hybrid TZEI

32 × TZEI 5 had the highest GYLD under artificial inoculation and was identified as a single-cross hybrid

tester in group 1. None of the inbreds in group 2 met the criteria for selection as a tester. At 40% level of

dissimilarity (R-squared value of 40%), three distinct heterotic groups were identified for the yellow EM

inbreds (Figure 2). The inbreds TZEI 9, TZEI 24, TZEI 23 and TZEI 120 were classified into group 1;

TZEI 17, TZEI 13, TZEI 14, and TZEI 11 were assigned to group 2 while TZEI 128, TZEI 135, TZEI

182, TZEI 219, TZEI 161, TZEI 134 and TZEI 124 were assigned to group 3 (Figure 2). None of the

inbreds in groups 1 and 2 satisfied the criteria for a good tester. However, inbreds TZEI 134 and TZEI

124 from group 3, combined significant positive GCA effects for GYLD and significant negative GCA

(female and male) effects for NCLB disease severity scores. The per se GYLD of TZEI 124 was

significantly higher than that of TZEI 134 (Table 7). Therefore, TZEI 124 was selected as a tester for

group 3. Additionally, TZEI 124 and TZEI 134 belonged to the same heterotic group and had the highest

GYLD in hybrid combinations. Therefore, TZEI 124 × TZEI 134 was chosen as single-cross tester (Table

9).

Performance of the hybrids under artificial inoculation and non-infected conditions

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The average GLYD under artificial inoculation and natural conditions, GYLD reduction, and the BI

values of the white and yellow EM single-cross hybrids are presented in Tables 8 and 9. The white

EM single-cross hybrids produced GYLD ranging from 3,221 kg ha-1 for TZEI 39 × TZEI 4 to 7,201

kg ha-1 for TZEI 32 × TZEI 5 under artificial inoculation. Contrarily, under natural conditions, TZEI

75 × TZEI 60 had the highest GYLD of 8,409 kg ha-1 and a GYLD reduction of 22% under artificial

inoculation (Table 8). Overall, artificial inoculation reduced GYLD from 1% for TZEI 32 × TZEI 5 to

40% for TZEI 39 × TZEI 269 (Table 8). The yellow EM single-cross hybrids produced GYLD

ranging from 1,961 kg ha-1 for TZEI 14 × TZEI 11 to 7,020 kg ha-1 for TZEI 124 × TZEI 134 under

artificial inoculation (Table 9). In contrast, TZEI 124 × TZEI 11 had a GYLD of 8,852 kg ha -1 under

natural conditions with a yield reduction of 27% when artificially inoculated. Artificial inoculation of

the yellow EM hybrids resulted in GYLD reduction of 1% for TZEI 161 × TZEI 219 to 46% for TZEI

23 × TZEI 182 (Table 9). Using the BI, 47 of the white EM hybrids were classified as resistant to the

disease while 28 were susceptible. Similarly, 50 and 25 of the yellow EM hybrids were classified as

resistant and susceptible to the disease, respectively.

In the AMMI biplots displayed in Figures 3 and 4, the vertical dotted lines represent the grand mean

of GYLD while the horizontal line depicts the zero IPC1 values. Hybrids close to the zero IPC1 line

had stable GYLD across the test environments and had little interactions with the test environments

(Yan, 2001). Hybrids farther from the zero IPC1 line were less stable. In addition, hybrids that fell on

the negative side of the main effect axis of the biplots were characterized by GYLD lower than the

mean while those on the positive side had GYLD greater than the mean. Furthermore, hybrids that fell

on the upper right quadrant of the biplots (i.e., above the zero IPC1 line) were positively correlated

with the IPC1 scores and were adapted to favourable environments, while hybrids on the lower right

quadrant (i.e., below the IPC1 line) were negatively associated with the IPC1 scores and were adapted

to low-yield environments (Yan, 2001). A wide range of GYLD and large variability across the test

environments were observed. The principal component analyses explained 87.6% and 88.1% of the

GYLD SS for the white and yellow EM hybrids, respectively. This indicates that the biplots were

adequate in describing the G × E interactions across the test environments under NCLB (Figure 3).

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EM white hybrids 3, 13, and 47 possessed GYLD higher than the average and were the most stable

across the test environments. These hybrids had little interaction with test environments as denoted by

their proximity to the zero IPC1 score. However, hybrid 17 was the highest-yielding across the test

environments and was positively correlated with the IPC1 axis suggesting that it was adapted to high-

yielding environments. Also, hybrids 10, 16, 46, 55, and 63 had average GYLD greater than the mean

and were positively correlated with the IPC1 axis indicating adaptation to high-yielding environments.

Contrarily, hybrids 12, 31, 35, 50, 51, and 60 had GYLD higher than the mean but were negatively

correlated with the IPC1 scores implying adaptation to low-yield test environments. Furthermore, EM

yellow hybrids 84, 116, and 134 were the most stable as they had little association with the test

environments and had GYLD greater than the average (Figure 4). However, hybrid 118 had the

highest yield across test environments and was positively correlated with the IPC1 axis suggesting

adaptation to high-yielding environments. Similarly, hybrids 119, 120, 127, 128, 130, 140, and 145

had GYLD greater than the average and were adapted to favourable test-environments. However, the

GYLD of hybrids 93, 117, 125, and 138, were higher than the grand average but were negatively

correlated with the IPC1 scores implying adaptation to low-yield research environments (Figure 4).

Discussion

Recently, our research group screened yellow and white endosperm EM maize inbreds under artificial

E. turcicum inoculation across contrasting agro-ecologies in Nigeria and identified several resistant

inbreds (Badu-Apraku et al., 2021a). Outstanding inbreds with combined resistance to E. turcicum

and other agronomically desirable traits identified in that study were used in the present study to

develop high-yielding EM white and yellow single-cross hybrids with resistance to NCLB. The

different responses of the hybrids to contrasting environments should facilitate the selection of

outstanding hybrids adapted to different environments. The significant genotype and environment

effects detected for most measured traits, especially GYLD and NCLB disease severity scores for

both white and yellow EM hybrids, indicates that the performance of the hybrids varied under

contrasting environmental conditions. This result also implied that the test environments were

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different, and that the observed genetic variability could facilitate selection for improved GYLD and

resistance to NCLB. Vivek et al. (2010) reported similar results for intermediate to late maturing

hybrids evaluated under NCLB pressure.

Additionally, significant G × E interactions observed in the present study for GYLD, NCLB disease

severity scores, and other measured traits indicated varying responses of the genotypes to the

contrasting test environments and that it would be necessary to identify genotypes characterized by

high-yield and stable performance across research environments (Moghaddam and Pourdad, 2009).

Therefore, there is a need for extensive testing of the hybrids in contrasting environments before

making hybrid recommendations.

In combining ability studies, if GCA effect is the largest component, then breeding schemes that

capitalize on additive gene effects of the parents are appropriate. Therefore, it is advisable to employ

breeding schemes that would lead to accumulation of additive genes. This calls for the use of

recurrent selection methods such as the S1, half-sib, or full-sib family selection. Contrarily, if the SCA

effects are preponderant, then a back-cross breeding program or hybridization are the preferred

choices for introgression of beneficial genes into agronomically desirable parents. However, if both

GCA and SCA effects are important, then there is a need to employ a recurrent selection method to

increase the frequency of the favorable alleles (additive genes), followed by hybridization to take

advantage of the SCA effects. The significant SCA and GCA (male/female) effects observed for most

measured traits especially GYLD and TURC6WAI suggested that the inheritance of GYLD and

NCLB resistance were controlled by both non-additive and additive genetic effects. This finding is

similar to those of Vivek et al. (2010) who reported significant GCA and SCA effects for GYLD and

NCLB resistance. Our results indicated that hybrids, synthetics and populations with high GYLD and

resistance to NCLB could be developed through recurrent selection, hybridization, and back-crossing

methods (Badu-Apraku et al., 2015). Additionally, our results suggested that inbreds with outstanding

combining abilities as well as efficient testers could be identified and assigned to distinct groups

under NCLB infection. The preponderance of GCA (female/male) effects compared to the SCA

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effects observed for measured traits such as GYLD, NCLB disease severity scores, PASP, EASP, and

EPP implied that additive genetic effects were largely responsible for the variations observed among

the parents of the hybrids tested in the present study. This result is consistent with those of Badu-

Apraku et al. (2015) who reported that the additive genetic effects were more important for GYLD

and other agronomic characters in the stress environments. Therefore, early generation testing would

be effective and selection of hybrids with high GYLD and NCLB resistance would be successful

through predictions based on additive genetic effects in both the yellow and white inbreds studied.

The identified inbreds with higher GCA-female could serve as female parents while those with higher

GCA-male could be used as male parents in crosses for improving resistance to NCLB. Additionally,

significant SCA × environment interactions for GYLD and the disease severity scores implied that the

GYLD and NCLB resistance of the white and yellow EM hybrids were not the same in the contrasting

test environments. This result suggested the possibility of selecting different inbreds for the

development of hybrids under each test environment (Badu-Apraku et al., 2015). Single-cross hybrids

are sensitive to changes in environmental conditions (Hallauer et al., 2010). This justified the need for

evaluating hybrids across contrasting test environments to identify stable and high yielding hybrids

(Akinwale et al., 2014). GCA effects of an inbred play an important role in improving a particular

trait of a genotype. The outstanding inbreds identified based on GCA (male and female) for GYLD

and NCLB disease severity scores could be used for the development of heterotic populations from

which NCLB resistant and high-yielding hybrids and synthetics could be extracted. In the present

study, EM white inbreds, TZEI 60, TZEI 75, TZEI 32, TZEI 5, TZEI 89 and the yellow inbreds TZEI

124, and TZEI 134 had significant positive GCA (female / male) effects for GYLD. These inbreds

possessed favorable alleles for GYLD that had high probability of being transferred to the progenies.

Furthermore, inbreds such as TZEI 60, TZEI 75, TZEI 5, TZEI 89, TZEI 124, TZEI 134, TZEI 135,

and TZEI 182 which had significant negative GCA (male and female) effects for disease severity

score are invaluable resources for breeding for resistance to NCLB.

Information on the heterotic groups of inbreds is important for successful commercial hybrid

programs. Therefore, classifying inbreds into appropriate heterotic groups is invaluable for

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determining the usefulness of inbreds as well as improved understanding of the relationships among

white and yellow EM inbreds. This would help speed-up the development of synthetics, hybrids, and

heterotic populations and NCLB resistant inbreds could be extracted from such populations for

development of high-yielding and NCLB resistant hybrids. Based on the reports of Fan et al. (2009),

an efficient grouping method should facilitate identification of groups which permit inter-heterotic

group crosses to display higher heterosis than intra-heterotic group crosses. In the present study, the

HGCAMT method classified the white and yellow inbreds with positive GCA effects for GYLD and

negative GCA effects for NCLB disease severity scores into the same group while white inbreds with

negative GCA effects for GYLD and positive GCA effects for NCLB scores were assigned to a

different group. The choice of potential testers is crucial to the successful development of different

hybrid types. A tester should discriminate between inbreds within its group and other groups (Badu-

Apraku et al., 2013). The testers identified in this study could be used to classify the large number of

EM white and yellow inbreds in the IITA-MIP which are yet to be tested in the field. In addition, the

single-cross testers identified in the present study could be used for developing three-way and double-

cross hybrids.

An important objective of the present study was to identify high-yielding and NCLB resistant white

and yellow EM hybrids for commercialization in SSA. Several white and yellow hybrids with

outstanding GYLD under artificial inoculations with a virulent E. turcicum isolate were identified. For

example, TZEI 32 × TZEI 5, TZEI 60 × TZEI 89, TZEI 5 × TZEI 75, TZEI 60 × TZEI 5, TZEI 124 ×

TZEI 134, and TZEI 134 × TZEI 17, had GYLD of more than 7,000 kg ha-1. Despite the outstanding

performance of these hybrids, GYLD reduction due to NCLB recorded for the white and yellow EM

hybrids such as TZEI 39 × TZEI 269, TZEI 86 × TZEI 83 and TZEI 23 × TZEI 182, was very high

ranging from 40% to 46% and may be attributed to loss of photosynthetic apparatus of the infected

plants (Hooda et al., 2016; Hurni et al., 2015; Weems and Bradley, 2018). Therefore, it should be

possible to improve these hybrids for NCLB resistance by introgressing resistant genes from selected

NCLB resistant inbreds identified in this study.

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The AMMI biplots revealed that TZEI 5 × TZEI 75, TZEI 60 × TZEI 5, TZEI 1 × TZEI 75 (white),

and TZEI 124 × TZEI 11, TZEI 219 × TZEI 17, TZEI 11 × TZEI 128 (yellow) were high-yielding

and stable hybrids across the contrasting test environments. These hybrids should be urgently tested

extensively in multi-location trials and on farmers’ fields to confirm their outstanding performance for

possible commercialization.

Conclusions

The present study revealed the preponderance of the GCA effects over the SCA effects for inheritance

of GYLD and NCLB resistance in the yellow and white EM maize hybrids. The white inbreds were

classified into two groups while the yellow EM inbreds were placed in three groups. Promising testers

were identified for some of the groups. In addition, single-cross testers were identified for both white

and yellow EM hybrids. There were both white and yellow hybrids with high yield under both

artificial inoculation and natural conditions with stable performance across contrasting test

environments. The inbred testers identified in the present study could be used to classify other inbreds

into appropriate groups and introgressed into tropical breeding populations to enhance GYLD and

NCLB resistance. Additionally, the single-cross testers could be used for the development of NCLB

resistant three-way hybrids for commercialization in SSA. The commercialization of improved EM

hybrids with NCLB resistance would contribute to increased maize production and productivity and

lead to increased incomes and poverty alleviation in SSA.

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Supplementary Table 1 Caption: General combining ability estimate of grain yield of white early

maturing inbred lines artificially inoculated with Exserohilum turcicum in six environments in Nigeria,

for two years using the HGCAMT method.

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Table 1. Characteristics of locations where early maturing maize hybrids were artificially inoculated
with Exserohilum turcicum during 2018 and 2019. The same locations were used for the evaluation of
the hybrids under non-inoculated conditions in 2019.

Location Coordinates Agro-ecological zone Elevation Average Annual rainfal

(m) humidity (%) (mm)
Ikenne 65 ’ 2’ E Rain Forest 60 81 1,800
Ile-Ife 7 8 E Rain Forest 244 81 1,600
Zaria 7 7 5E Northern Guinea Savanna 640 68 1,500
Table 2: Mean squares derived from the analysis of variance for grain yield, Exserohilum turcicum infection
ratings and other agronomic traits of the white early maturing single-cross hybrids artificially inoculated with E.
turcicum in six environments in Nigeria.

Ant
hesi Anthesis TURC
Grain s silking Ears
Degree yield Day Silkinginterv Plant Ear Plant 2 TU per
Ear
Source of of (kg ha- s Days al height height aspec aspectW TURCRC pla
1
Variation freedom ) (days) (cm) (cm) td e AIf h
6WAIg nt
Env 5 19066 653. 715 35.1* 2551 1736 35.7* 8. 47. 16. 18. 0.1
2832.1 7** .9* 8.0* 3.6* * 4* 3* 2* 0** **
** * * * * * *
Set 2 1.8* 5.3 2. 0.0
25561 31.2 28. 482. 109. 4* 2.9 0.2 0.9 1
47.6* ** 3** 8* 1 * ** **
Env × Set 10 0.3 1.5 0. 0.0
12508 4.1* 3.4 350. 91.8 6 0.5 0.6 0.4 2
59.5 * * 8** ** **
Rep (Env × 12 0.2 0.7 0. 0.0
Set) 11502 1.0 1.5 104. 42.4 5 0.3 0.2 0.1 1
83.3 2
Block (Env 96 0.3 2.7 0. 0.0
× Rep) 13629 3.5* 4.0 230. 104. 5* 0.3 0.6 0.3 2
28.8 * ** 3** 5** ** * **
Male (Set) 12 1.5** 4. 0.0
18312 36.8 36. 3468 1457 6.5** 2* 1.1 8.7 3.6 2
710.4* ** 4** .7** .6** * ** ** **
*
Female (Set) 12 1.1** 3. 0.0
15495 34.4 3.0 2916 1089 8.4** 6* 1.5 6.4 3.0 4*
796.2* ** .2** .1** * ** ** ** *
*
Female × 48 0.4 3.5 0. 0.0
Male (Set) 20097 3.6* 3.4 350. 192. 6* 0.4 1.4 0.6 1
30.3** * ** 6** 4* * ** ** **
Env × Male 60 0.6** 0. 0.0
(Set) 14866 2.6* 2.8 220. 101. 4.2** 8* 0.5 1.3 0.6 2
50.2** * ** 0** 8** * ** ** **
Env × 60 0.5* 2.7 0. 0.0
Female (Set) 18592 2.7* 2.9 249. 77.5 6* 0.5 1.1 0.5 1

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 83.0** * ** 8** * ** ** **
Env × 240 0.3 2.7 0. 0.0
Female × 99948 2.0 2.0 171. 79.9 4 0.3 0.6 0.3 1
Male (Set) 9.1* * 8** ** * ** **
Error 336 0.3 2.6 0. 0.0
79330 1.7 1.6 120. 60.2 4 0.2 0.4 0.2
6.0 9
92. 80.0 69.7 80. 89. 86. 89. 67.
2a
R 90.4 92.3 8 90.7 91.1 1 1 2 3 3
2.4 242.9 34.2 12. 16. 16. 11. 14.
b
CV (%) 17.0 2.4 6.7 10.0 7 3 1 9 5
53. 0.2 4.8 4.8
5253.2 53.1 4 163. 77.8 2.7 4.0 3.4 0.9
c
GM 9
Heritability 70 67 59 76 89 53 89 84 73
(%) 78 83 75
a 2
R (%): Coefficient of determination.
b
CV (%): coefficient of variation.
c
GM: grand mean.
d
Plant aspect recorded on a scale of 1-9 based on plant type, where 1 = excellent and 9 = poor.
e
Ear aspect rated on a scale of 1 – 9, where 1 = clean, uniform, large, and well-filled ears and 9 = ears with undesirable
features.
f
TURC2WAI: disease score two weeks after inoculation rated on a scale of 1-9, where 1 = complete resistance, no visible
NCLB symptoms and 9 = abundant and coalescing lesions on all leaves covering >80% of the leaf surface with some plants
prematurely dead.
g
TURC6WAI: disease score six weeks after inoculation rated as above.
h
TURC: average disease severity score.
* Significance at P <0.05.
** Significance at P < 0.01.

Table 3: Means square derived from the analysis of variance for grain yield, Exserohilum turcicum infection
ratings and other agronomic traits for yellow early maturing single-cross hybrids artificially inoculated with
Exserohilum turcicum in six environments in Nigeria.
TURCTURC
Anthesis
Degree Grain Silkin silking Plant Ear Ear 2 6 Ears
of yield Anthe g interv height height Plant As W W TU per
Source of freedo (kg sis (Days al (cm (cm Aspe pe AI AI RC pl
-1
Variation m ha ) (Days) ) (days) ) ) ctd cte f g h
ot
Env 5 11177 515.5* 565.5 56.6* 181 131 11.8* 17. 33. 14. 6.6 0.
6271. * ** * 99.7 60.7 * 9* 8* 1* ** 1*
1** ** ** * * * *
Set 2 0.4 0.
28237 47.5** 61.6* 1.7** 405 344. 5.4** 6.6 1.2 0.2 03
12.2* * 9.5* 5 ** **
* *
Env × Set 10 2.2 1.1 0.8* 0.5 0.3 0.
19134 77.7 369. 1.1 0.4 0.7 02
84.9* 3 * ** 9
*
Rep (Env × 12 1.1 1.5 0.6 0.3 0.1 0.
Set) 55108 120. 132 0.5 0.1 0.4 02
1.8 3 4.9*
*

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 Block (Env 96 0.4 0.2 0.
×Rep) 19021 2.8** 2.9** 299. 656. 0.6** 1.2 0.2 0.6 03
97.1* 6** 9 ** * **
*
Male (Set) 12 3.1 0.
16404 44.7** 53.1* 2.3** 477 956. 4.4** 6.2 0.7 7.9 ** 05
654.0 * 7.0* 2 ** ** ** **
** *
Female 12 3.8 0.
(Set) 23936 61.2** 73.3* 1.9** 482 119 4.8** 6.9 1.0 8.8 ** 06
194.8 * 3.8* 6.8* ** ** ** **
** *
Female × 48 0.5 0.6 0.
Male (Set) 62732 9.3** 9.9** 740. 566. 1.4** 2.1 0.3 1.6 ** 03
42.0* 2** 9 ** ** ** **
*
Env × Male 60 0.4 0.
(Set) 19540 2.6** 3.1** 0.6** 181. 572. 0.7** 0.8 0.4 1.0 ** 02
71.6* 1 8 ** ** **
*
Env × 60 0.4 0.6 0.
Female 14332 2.7** 3.2** 0.5** 235. 609. 0.6 0.3 1.5 ** 02
(Set) 97.3* 2** 8 ** **
*
Env × 240 1.9 2.0 0.4 0.3 0.
Female × 96811 182. 632. 0.5** 0.6 0.2 0.7 ** 02
Male (Set) 7.0* 1* 6 * * **
Error 336 1.7 1.7 0.4 0.4 0.2 0.
78913 144. 612. 0.5 0.2 0.4 01
5.0 1 8
92.1 92.8 85.5 84.3 82. 87. 85. 87. 71
2a
R 90.5 90.3 69.9 7 6 8 2 .1
2.5 2.5 146.6 12.3 15. 17. 19. 14. 14
b
CV (%) 18.3 7.9 34.9 2 8 5 0 .9
52.7 53.1 0.4 4.8
4867. 152. 70.9 4.7 2.3 3.4 2.8 0.
c
GM 0 2 9
Heritability 80 81 60 87 86 51 90 90 77
(%) 83 89 67
a 2
R (%): Coefficient of determination.
b
CV (%): coefficient of variation.
c
GM: grand mean.
d
Plant aspect recorded on a scale of 1-9 based on plant type, where 1 = excellent and 9 = poor.
e
Ear aspect rated on a scale of 1 – 9, where 1 = clean, uniform, large, and well-filled ears and 9 = ears with undesirable
features.
f
TURC2WAI: disease score two weeks after inoculation rated on a scale of 1-9, where 1 = complete resistance, no visible
NCLB symptoms and 9 = abundant and coalescing lesions on all leaves covering >80% of the leaf surface with some plants
prematurely dead.
g
TURC6WAI: disease score six weeks after inoculation rated as above.
h
TURC: average disease severity score.
* Significance at P <0.05.
** Significance at P < 0.01.

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Table 4: Estimates of general combining ability effects for grain yield and disease severity of white and
yellow early maturing maize inbred lines artificially inoculated with Exserohilum turcicum, in six
environments in Nigeria.
White inbred lines Yellow inbred lines
Grain yield (kg ha-1) TURC6WAIa Grain yield (kg ha-1) TURC 6WAIa
GENOTYP GCA- GCA- GCA- GCA GENOTYP GCA- GCA- GCA- GCA
E female male femal - E female male femal -
e male e male
TZEI 9 - - 0.2*
TZEI 1 258.0 66.9 0.2 -0.1 968.4** 898.1** 0.4**
- TZEI 17 301.4 464.9* -0.1 -0.2*
- 0.6*
TZEI 5 411.6* 360.5* 0.5** *
- TZEI 120 86.5 -131.5
952.0* 0.5* 0.5** 0.7*
TZEI 39 -967.8** * 0.5** * *
TZEI 86 -114.0 73.5 0.01 0.01 TZEI 128 413.0* 144.5 -0.2* -0.2*
TZEI 135 167.5 420.1* - -
451.0* 0.7** 0.6*
TZEI 89 412.19* * -0.2* -0.2* *
TZEI 11 166.3 -628.3* -0.1
608.23* 598.6* 0.6*
TZEI 32 * * -0.3* -0.2* *
TZEI 35 -246.10 -236.4 -0.04 0.2 TZEI 24 -472.9 -448.5 0.04 0.2*
TZEI 134 1006.4* 1056.8* - -
- * * 0.7** 0.4*
TZEI 51 -696.5** 410.1* 0.5 0.5 *
- TZEI 182 -265.7 -214.5 - -
568.1* 0.6** 0.4*
TZEI 56 -467.9** * 0.3 0.2 *
- TZEI 219 -434.2* 234.4 -0.1 0.1
615.6* - 0.7*
TZEI 60 802.3** * 0.5** *
- TZEI 13 -302.6 -169.2 0.3*
704.7* 0.6**
TZEI 4 -276.8* * 0.1 0.1
- TZEI 14 -343.2 497.2 -0.1 -0.1
541.9* 0.4*
TZEI 40 269.2 * -0.2* *
TZEI 23 - - 0.2*
975.4* 905.0** 631.2** 0.4*
TZEI 75 693.4** * -0.2 -0.3* *
- TZEI 124 1533.9* - -
559.7* 0.4* * 710.9** 0.6** 0.4*
TZEI 83 -448.7* * 0.4** * *
- TZEI 161 16.9 -407.7* -0.04 -0.1
TZEI 269 -237.1* 252.9* -0.1 0.3*
SE 140.8 157.4 0.1 0.1 SE 161.4 138.2 0.1 0.1
a
TURC6WAI: disease score six weeks after inoculation rated on a scale of 1-9, where 1 = complete
resistance, no visible NCLB symptoms and 9 = abundant and coalescing lesions on all leaves covering
>80% of the leaf surface with some plants prematurely dead.
* Significance at P <0.05.
** Significance at P < 0.01.

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Table 5: Percentage contribution of the combining ability effects of white early maturing inbred lines
for grain yield, Exserohilum turcicum infection ratings and other agronomic traits under artificial
inoculation and non-inoculated conditions, in six environments in Nigeria.
Artificially inoculated Non-inoculated
Agronomic GCAb GCA GCA GCA
a c
traits Male Female SCA Male Female SCA
(%) (%) (%) (%) (%) (%)
GYLD 44 37 19 36 35 30
DA 43 40 17 23 30 47
DS 45 38 17 21 31 48
ASI 37 27 36 13 24 63
PLHT 45 37 18 32 41 27
EHT 44 33 23 35 30 35
PASP 22 29 49 32 30 38
EASP 41 35 24 42 23 35
TURC2WAI 37 50 14 12 13 75
TURC6WAI 34 30 36 12 31 58
TURC 40 32 28 11 32 57
EPP 13 30 57 10 13 77
a
GYLD: grain yield; DA: days to anthesis; DS: days to silking; ASI: anthesis-silking interval; PLHT:
plant height; EHT: ear height; PASP: plant aspect; EASP: ear aspect; TURC2WAI: disease severity
scores two weeks after inoculation; TURC6WAI: disease severity scores six weeks after inoculation;
TURC: average disease severity score; EPP: ears per plant.
b
GCA: General combining ability.
c
SCA: Specific combining ability.

Table 6: Percentage contribution of the combining ability effects of yellow early maturing inbred lines
for grain yield, Exserohilum turcicum infection ratings and other agronomic traits under artificial
inoculation and non-inoculated conditions, in six environments in Nigeria.

Artificially inoculated Non-inoculated

Agronomic GCAb GCA GCA GCA
traitsa Male Female SCA c
Male Female SCA
(%) (%) (%) (%) (%) (%)
GYLD 25 37 38 22 40 38
DA 31 43 26 30 29 41
DS 32 44 24 31 29 39
ASI 37 32 31 12 25 64
PLHT 38 38 24 34 39 27
EHT 22 27 51 22 32 46
PASP 30 33 37 18 37 45
EASP 29 32 39 17 41 42
TURC2WAI 26 36 38 14 16 71
TURC6WAI 34 38 28 18 20 62
TURC 33 41 26 18 24 58
EPP 20 25 55 12 33 55
a
GYLD: grain yield; DA: days to anthesis; DS: days to silking; ASI: anthesis-silking interval; PLHT:
plant height; EHT: ear height; PASP: plant aspect; EASP: ear aspect; TURC2WAI: disease severity
scores two weeks after inoculation; TURC6WAI: disease severity scores six weeks after inoculation;
TURC: average disease severity score; EPP: ears per plant.

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b
GCA: General combining ability.
c
SCA: Specific combining ability.
Table 7: Per se grain yield and disease severity score six weeks after inoculation of early maturing
inbreds evaluated under artificial Exserohilum turcicum inoculation, in six environments in Nigeria.

White endosperm inbred lines Yellow endosperm inbred lines

a
Genotype Grain yield TURC 6WAI Genotype Grain yield TURC 6WAIa
-1 -1
(kg ha ) (kg ha )
TZEI 60 2,260 2.4 TZEI 135 2,490 2.8
TZEI 4 1,983 3.9 TZEI 14 2,131 3.4
TZEI 1 1,890 5.5 TZEI 124 2,104 3.4
TZEI 86 1,766 4.0 TZEI 17 1,966 4.1
TZEI 32 1,764 3.0 TZEI 219 1,872 5.0
TZEI 75 1,760 3.2 TZEI 13 1,644 5.1
TZEI 5 1,756 3.8 TZEI 23 1,570 4.3
TZEI 89 1,619 4.6 TZEI 11 1,500 4.4
TZEI 56 1,564 4.1 TZEI 24 1,481 4.5
TZEI 40 1,525 3.8 TZEI 161 1,303 3.9
TZEI 269 1,418 4.9 TZEI 128 1,260 4.1
TZEI 83 1,406 3.4 TZEI 182 1,144 4.0
TZEI 39 1,146 4.6 TZEI 120 958 4.8
TZEI 35 1,095 5.0 TZEI 134 937 4.1
TZEI 51 818 5.0 TZEI 9 897 4.2
LSD (0.05) 479.9 0.8 LSD 479.9 0.8
a
TURC6WAI: disease score six weeks after inoculation. Rated on a scale of 1-9, where 1 = complete
resistance, no visible NCLB symptoms and 9 = abundant and coalescing lesions on all leaves covering
>80% of the leaf surface with some plants prematurely dead.

Table 8: Grain yield (kg ha-1) and yield reduction of selected (the best 20, middle 5, and the worst 10
based on grain yield) white grain early maturing single-cross maize hybrids artificially inoculated
with Exserohilum turcicum and under non-inoculated conditions at Ikenne, Ile-Ife, and Zaria in
2018/2019.

Entry Pedigree GYLDa GYLD Reduction BIc

artificially inoculated natural conditions (%) b
13 TZEI 5 × TZEI 75 6,890 8,333 17 4
47 TZEI 60 × TZEI 5 6,750 7,062 4 4
50 TZEI 60 × TZEI 89 7,189 7,337 2 4
3 TZEI 1 × TZEI 75 6,115 7,865 22 3
17 TZEI 32 × TZEI 5 7,206 7,294 1 3
35 TZEI 40 × TZEI 60 6,027 7,500 20 3
52 TZEI 75 × TZEI 35 5,976 7,449 20 3
51 TZEI 75 × TZEI 32 6,440 6,620 3 3
49 TZEI 60 × TZEI 86 5,884 7,228 19 3
46 TZEI 60 × TZEI 1 6,065 7,685 21 3
10 TZEI 4 × TZEI 60 6,058 6,602 8 2
55 TZEI 75 × TZEI 60 6,571 8,409 22 2

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62 TZEI 86 × TZEI 40 5,981 6,106 2 2
68 TZEI 89 × TZEI 75 6,479 6,745 4 2
31 TZEI 40 × TZEI 32 6,329 7,070 11 2
16 TZEI 32 × TZEI 1 5,904 7,032 16 2
75 TZEI 269 × TZEI 60 5,219 5,757 9 1
12 TZEI 5 × TZEI 40 5,900 7,200 18 1
60 TZEI 83 × TZEI 60 5,982 6,830 12 1
27 TZEI 39 × TZEI 40 5,271 5,475 4 1
71 TZEI 269 × TZEI 32 5,886 6,825 14 1
5 TZEI 1 × TZEI 269 5,246 5,524 5 0
65 TZEI 86 × TZEI 269 5,260 5,533 5 0
29 TZEI 39 × TZEI 83 3,891 4,461 13 -1
14 TZEI 5 × TZEI 83 5,185 5,757 10 -1
63 TZEI 86 × TZEI 75 5,978 6,063 1 -1
59 TZEI 83 × TZEI 56 4,211 4,503 7 -2
74 TZEI 269 × TZEI 56 4,205 5,827 28 -2
23 TZEI 35 × TZEI 39 3,904 5,736 32 -2
8 TZEI 4 × TZEI 51 4,053 6,047 33 -3
11 TZEI 5 × TZEI 4 4,185 6,012 30 -3
38 TZEI 51 × TZEI 39 3,986 5,125 22 -4
64 TZEI 86 × TZEI 83 3,679 6,117 40 -4
26 TZEI 39 × TZEI 4 3,221 4,119 22 -4
30 TZEI 39 × TZEI 269 3,363 5,572 40 -5
LSD (0.05) 735 1,623
a
GYLD: grain yield.
b
Percentage GYLD reduction .
c
BI: base index.
Table 9: Grain yield (kg ha-1) and yield reduction of selected (the best 20, middle 5, and the worst 10
based on grain yield) yellow grain early maturing single-cross maize hybrids artificially inoculated with
Exserohilum turcicum and under non-inoculated condition at Ikenne, Ile-Ife, and Zaria in 2018/2019.
Entry Pedigree GYLDa GYLDa Reductionb BIc
artificially inoculated natural conditions (%)
118 TZEI 124 × TZEI 134 7,020 8,124 14 4
120 TZEI 124 × TZEI 219 6,246 8,010 22 4
128 TZEI 134 × TZEI 120 6,268 7,480 16 3
127 TZEI 134 × TZEI 17 6,768 7,182 6 3
119 TZEI 124 × TZEI 182 6,223 6,968 11 3
116 TZEI 124 × TZEI 11 6,474 8,852 27 3
130 TZEI 134 × TZEI 135 6,279 8,056 22 3
129 TZEI 134 × TZEI 128 5,853 6,756 13 3
88 TZEI 13 × TZEI 134 5,985 7,442 20 2
136 TZEI 161 × TZEI 11 5,781 6,466 11 2
84 TZEI 11 × TZEI 128 5,674 6,859 17 2
138 TZEI 161 × TZEI 134 6,054 6,218 3 2
93 TZEI 14 × TZEI 134 6,063 7,298 17 2
147 TZEI 219 × TZEI 17 4,824 6,189 22 1
134 TZEI 135 × TZEI 124 6,142 6,914 11 1

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107 TZEI 24 × TZEI 17 5,557 6,028 8 1
145 TZEI 182 × TZEI 135 5,599 7,486 25 1
125 TZEI 128 × TZEI 161 5,011 5,779 13 1
117 TZEI 124 × TZEI 24 5,697 6,630 14 1
140 TZEI 161 × TZEI 219 5,650 5,706 1 1
99 TZEI 17 × TZEI 124 5,576 6,150 9 1
100 TZEI 17 × TZEI 161 5,581 6,738 17 0
82 TZEI 11 × TZEI 17 4,813 5,787 17 0
115 TZEI 120 × TZEI 161 4,938 6,777 27 0
150 TZEI 219 × TZEI 135 4,805 5,573 14 -1
146 TZEI 219 × TZEI 9 3,825 5,842 35 -2
104 TZEI 23 × TZEI 182 3,710 6,889 46 -3
105 TZEI 23 × TZEI 219 3,772 4,964 24 -3
106 TZEI 24 × TZEI 9 2,775 3,433 19 -4
102 TZEI 23 × TZEI 24 3,230 3,510 8 -4
78 TZEI 9 × TZEI 23 2,340 3,551 34 -4
86 TZEI 13 × TZEI 11 2,032 3,219 37 -5
80 TZEI 161 × TZEI 24 2,289 2,948 22 -6
91 TZEI 14 × TZEI 11 1,961 3,296 41 -6
80 TZEI 9 × TZEI 161 2,122 2,842 25 -7
LSD (0.05) 689 1,461
a
GYLD: grain yield.
b
Percentage GYLD reduction .
c
BI: base index.

This article is protected by copyright. All rights reserved.

This article is protected by copyright. All rights reserved.
This article is protected by copyright. All rights reserved.
This article is protected by copyright. All rights reserved.
This article is protected by copyright. All rights reserved.