Next Article in Journal
Postharvest Storage Techniques and Quality Evaluation of Fruits and Vegetables for Reducing Food Loss
Next Article in Special Issue
Tomato Graft Union Failure Is Associated with Alterations in Tissue Development and the Onset of Cell Wall Defense Responses
Previous Article in Journal
Comparison of Organic and Inorganic Mulching for Weed Suppression in Wheat under Rain-Fed Conditions of Haripur, Pakistan
Previous Article in Special Issue
Evaluation of Cell Wall Modification in Two Strawberry Cultivars with Contrasted Softness
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:

Relationships between Stalk Resistance and Corn Borers, Agronomic Traits, and Cell Wall Hydroxycinnamates in a Set of Recombinant Inbred Lines from a Maize MAGIC Population

Ana López-Malvar
Zoila Reséndiz
Rogelio Santiago
José Cruz Jiménez-Galindo
4 and
Rosa Ana Malvar
Facultad de Biología, Departamento de Biología Vegetal y Ciencias del Suelo, Universidad de Vigo, As Lagoas Marcosende, 36310 Vigo, Spain
Agrobiología Ambiental, Calidad de Suelos y Plantas (UVIGO), Unidad Asociada a la MBG (CSIC), 36310 Vigo, Spain
Misión Biológica de Galicia (CSIC), Pazo de Salcedo, Carballeira 8, 36143 Pontevedra, Spain
National Institute of Forestry Agriculture and Livestock Research (INIFAP), Ave. Hidalgo 1213, Cd. Cuauhtémoc, Chihuahua 31500, Mexico
Author to whom correspondence should be addressed.
Agronomy 2021, 11(6), 1132;
Submission received: 23 April 2021 / Revised: 27 May 2021 / Accepted: 28 May 2021 / Published: 2 June 2021
(This article belongs to the Special Issue Advances in Cell Wall Research of Crop Plants)


Corn borers are the most important pest affecting maize. Resistance to corn borer attack may compromise plant fitness being detrimental for some important agronomic traits such as yield. Against the attack of this pest, cell wall-bound hydroxycinnamates have been previously described as a possible defense mechanism. In this study, agronomic characterization and cell wall-bound hydroxycinnamates quantification was performed in a subset of Recombinant Inbred Lines (RILs) from a Multiparent Advanced Generation Intercross (MAGIC) population that showed contrasting behavior against corn borer attack. Resistant lines showed greater concentration of p-coumaric acid, the only hydroxycinnamate that could have a role in the resistance in these particular materials. In addition, results indicated that resistant lines showed precocity, low grain moisture at harvest, and reduced plant height, thus, selecting for resistance may be detrimental for yield. In this way, a breeding strategy directly targeting grain yield in order to tolerate corn borer attack would be the recommended one.

Graphical Abstract

1. Introduction

Maize is consumed by a large variety of herbivorous insects that have diverse feeding habits and consume many plant parts. Stem corn borers include diverse lepidopteran insects that feed on the pith of maize producing tunnels that can cause 30% yield losses, equivalent to world-wide losses of 311.3 million tons every year [1]. In the northwest of Spain, the average yield loss is around 15% and is mainly caused by the Mediterranean corn borer, Sesamia nonagrioides Lef. (MCB); the damage produced by the European corn borer, Ostrinia nubilalis (ECB), being less important. Both borers show similar life cycles and herbivorous behavior, but the MCB is more voracious than the ECB [2].
MCB infestation on maize begins at an early phenological stage in this area. After completing their first generation, that usually coincides with the plant juvenile stage, stem borers of the second generation attack the plants during the reproductive stage. The damage produced by second generation larvae causes the most important losses, decreasing forage, stover and grain yields as a consequence. Tunnels produced in the stalk pith interfere with nutrient assimilation moving toward the developing ear and increases the lodging rate [1,3]. Corn borers attack the ears by causing direct damage to kernels and increasing infections by mycotoxin-producing fungus [4,5].
Breeding for increased resistance to borers has been effective for reducing the length of tunnels made by borers but has provoked undesirable changes in agronomic traits [6,7,8]. Previous research indicated that recurrent selection for resistance to the European corn borer (ECB) resulted in decreased yielding ability [6,9] and changes in important agronomic traits [9]. Klenke et al. [6] found that four selection cycles reduced the damage from attacks by both the first and second generation ECB but decreased grain yield, suggesting that yield should be included in the selection criteria in a selection program. Likewise, recurrent selection to improve resistance to MCB also led to a decrease in yield, even when yield was included in the selection criteria [8].
On the other hand, several cell wall components have been associated with maize resistance to corn borers. Many studies have focused on studying the cell wall as a biochemical and structural barrier against insects [10,11,12]. In particular, the role of cell wall-bound hydroxycinnamates in cell wall strengthening, stiffening, and fortification has been deeply evaluated. The most important hydroxycinnamates are p-coumaric acid (PCA) and ferulic acid (FA). The accumulation of PCA is considered a relevant indicator of lignin deposition, one of the main components of cell wall fibers, and lignin has been directly related with corn borer resistance by increasing the rigidity of the cell wall [13,14]. On the other hand, FA can undergo dehydrodimerization, and the resulting dimers (diferulates, DFAs) crosslink heteroxylans that enhance cell wall stiffening and strengthening. Additionally, during lignification, FA and diferulic esters form crosslinks through the etherification of the phenolic hydroxyl group to lignin polymers, producing a polysaccharide-lignin matrix [15,16,17]. Taking into account those functions, several studies have pointed out the role of hydroxycinnamates in plant resistance to pests and diseases [10,18,19,20]. Furthermore, negative correlations were observed between larval weight and DFA concentration in the leaf-sheath of maize [21], and between stem tunnelling by ECB and MCB, and total PCA and DFAs [19,22].
In addition, recurrent selection to improve resistance to corn borer attack has been demonstrated to influence cell wall-bound hydroxycinnamates concentration. Higher concentrations of total DFAs were associated with shorter tunnel lengths and fewer larvae per stem over cycles of selection to MCB resistance in the maize synthetic EPS12 [23]. The same relationship between resistance and DFA concentration was found when DFAs were the primary selection criteria [24].
Studies on the relationship between resistance and agronomic or biochemical traits have usually been carried out using a diverse set of genotypes (hybrids, inbred lines, populations) with different levels of resistance [18,25]. One of the problems of this approach is that each genotype comes from different genetic backgrounds and the relationships between them are not known, which can lead to false associations [26]. On the other hand, to study the relationships between resistance to corn borer and other traits, Recombinant Inbred Lines (RILs) populations derived from biparental crosses have also been used [27,28,29]. The derived RILs have a common genetic background and are developed randomly, by the single seed descending method, without selection or drift. The main disadvantage of this approach is the limited variability of the biparental populations, with only two parents, and great linkage disequilibrium [30]. To solve these problems, we have evaluated RILs from a MAGIC (Multiparent Advanced Generation Intercross) population of eight parents, with enough variability and reduced linkage disequilibrium, since six recombination cycles have been performed before starting with the RILs development.
Hence, in the current research, agronomic characterization, and cell wall-bound hydroxycinnamates quantification was performed in a subset of Recombinant Inbred Lines derived from a MAGIC population chosen for their contrasting behavior against corn borer attack. The main goals are to deepen the knowledge of: (1) the relationship between resistance to corn borer and the main agronomic traits, and (2) the role of cell wall-bound hydroxycinnamates as a defense mechanism against corn borers using RILs from the same genetic background that have been randomly developed from the MAGIC population.

2. Materials and Methods

2.1. Experimental Design, Plant and Insect Material

Misión Biológica de Galicia’s Maize Genetics and Breeding Group has developed a MAGIC population with 700 RILs (currently, there are approximately 600 lines that can be purchased on request). Details of further developing the MAGIC population was described in Butrón et al. [31] and Jiménez-Galindo et al. [32]. Jiménez-Galindo et al. [32] evaluated 680 RILs for resistance to corn borers and agronomic traits. Based on that evaluation and the seed availability for future evaluations, 56 RILs, presenting extremes values for tunnel length, were selected and classified into two groups according to their resistance to corn borer attack: (1) short tunnel length (resistant); (2) large tunnel length (susceptible).
Selected RILs, along with eight inbred lines chosen as corn borer attack checks, were evaluated for resistance to corn borer and agronomic traits (Supplementary Table S1). The inbred checks included five of the eight MAGIC population founders that are partially resistant to corn borer attack (A509, EP125, EP17, EP86, F473) and inbreds EP42, EP47, and EP80, which are susceptible to corn borer attack.
Selected RILs and checks were evaluated across two years (2016 and 2017) in Pontevedra (42°26′01″ N 8°38′51″ O). Each year, two adjacent trials were conducted with an 8 × 8 simple lattice design. The experimental plot consisted of 15 plants on an area of 2.2 m2 and a density of 70,000 plants/ha.
One trial was protected with insecticide, treating the plants every 21 days from approximately 45 days after sowing until harvest; the other was infested with S. nonagrioides eggs prior to flowering. Standard procedures were followed for the insect rearing (carried out in Misión Biológica de Galicia) and production of S. nonagrioides eggs [33]. Before flowering, 10 plants from each plot were infested with ~40–80 S. nonagrioides eggs placed between the sheath and the stem in the internode below the main ear.

2.2. Resistance Trait (Tunnel Length)

At harvest, ten infested plants per plot were collected in infested trials. The stalks were split lengthwise, and the lengths of the tunnels (cm) produced by the larvae were measured.

2.3. Agronomic Traits

Agronomic traits were recorded in both the infested and protected trials.

2.4. Days to Silking/Anthesis

Considered as the time passed from the day of sowing until approximately 50% of the plants showed either pollen (male anthesis) or silks (female silking). They were recorded periodically by the time each plot started to show pollen and silks.

2.5. Plant Height

Considered as the mean plant height (in cm) of five plants per plot. Plant height was measured from the base to the tip of flag leaf after flowering and the average of five plants per plot was presented in this study.

2.6. Lodging

This was calculated as the sum of broken and leaning plants divided by the total number of plants in the plot, expressed in percentage. A plant was considered broken when it was split underneath the main ear and leaned when the stem formed an angle with the ground at less than 45°. It was recorded at harvest.

2.7. Grain Moisture

Expressed as a percentage, was recorded at harvest using a moisture meter Kett (model PM-400) in a sample of 240 cm3.

2.8. Grain Yield

Grain yield was calculated as the weight of grain in Mg ha−1 and adjusted at 14% grain moisture. It was determined by the following Equation (1).
G r a i n   Y i e l d   ( M g h a ) = P l o t   w e i g h t   ( k g ) × ( 100 H u m i d i t y ) × G r a i n   w e i g h t   o f   5   e a r s   ( g ) × 10 S u r f a c e   ( m 2 ) × 86 × T o t a l   w e i g h t   o f   5   e a r s   ( g )
where grain moisture was estimated as described above; surface was calculated as the number of plants per plot multiplied by the space between rows (0.80 m) and the space between plants (0.18 m). The value 86 on the equation corresponds to a constant to adjust the yield at 14% humidity. Yields were calculated per plot and then transformed to Mg ha−1; thus, this refers to a maximum potential yield.

2.9. Biochemical Analysis

The second internode below the main ear was collected from five plants in each plot in protected trials. Samples were collected at 55 days after silking. For each harvested internode, the pith was manually detached and frozen at −20 °C. Then, samples were lyophilized and ground in a Wiley (Arthur H. Thomas, Philadelphia, PA, USA) mill with a 0.75 mm screen before being analyzed. A recently optimized protocol was used for hydroxycinnamate quantification [34]. Phenolic standards ferulic acid and p-coumaric acid were purchased from Sigma-Aldrich Química SL, Madrid, Spain Sigma. The identities of FA dimers were confirmed by a comparison with the authentic 5−5 standard or published retention times and UV spectra. The total diferulate content (DFAT) was calculated as the sum of the following three identified and quantified DFA isomers: DFA 8–O–4, DFA 5–5, and DFA 8–5. The DFA 8–5 concentrations were calculated as the sum of 8−5-cyclic (or benzofuran)-DFA and 8–5-noncyclic (or open).

2.10. Statistical Analysis

Combined analyses across years were performed for each trait for protected and infested trials using the mixed model procedure of the SAS program (version 9.4) [35]. Means for each trait were calculated based on combined data across years. Lines were considered as fixed effects and years, replicates, and blocks as random effects. The comparison of means was carried out using least significant difference (LSD).

2.11. Contrast Analysis

After a variance analysis, the RILs were, again, qualitatively classified according to their BLUEs (Best Linear Unbiased Estimators) for tunnel length (under infestation) in resistant, susceptible, or intermediate. Resistant and susceptible groups were formed by 20 RILs in each. With the qualitative dataset, including the checks, mean comparisons were performed in order to determine the existence of significant differences for agronomic traits and cell wall-bound hydroxycinnamates between groups of RILs with contrasting values for resistance to corn borer. Discussion will focus on differences among resistant (short tunnel length) and susceptible (large tunnel length) groups.

2.12. Correlation Analysis

Genotypic and phenotypic correlations were performed among tunnel length and agronomic traits using Restricted Maximum Likelihood (REML) according to a published SAS mixed model procedure [36].

2.13. Multiple Linear Regression Analysis

In order to understand the role of cell wall-bound hydroxycinnamates as one of the defense mechanisms against corn borer, BLUES estimates were used to build multiple linear regression models. The stepwise method following the PROC REG procedure in SAS was used [35]. We have considered as dependent variables the trait involved in stalk resistance to corn borer (tunnel length). On the other side, cell wall-bound hydroxycinnamates were considered as independent variables.

3. Results

As a starting point, in means comparison analyses, RILs differ significantly for every trait under study in both infestation conditions with the exception of grain moisture under infestation (Supplementary Tables S1 and S2). Among others, the RILs differed significantly in tunnel length under infestation conditions and that allowed the classification in resistant, medium, or susceptible for contrast analysis (Supplementary Tables S1 and S2).

3.1. Contrast Analysis

RILs were classified in three groups according to their tunnel length under infestation conditions: (1) short tunnel length (resistant, 5–26 cm); (2) medium tunnel length (26–44 cm); (3) large tunnel length (susceptible, 45–63 cm). The RIL of the resistant group with the shortest tunnels (26 cm) differed significantly (p < 0.05) from the RIL of the susceptible group with the largest tunnels (45 cm). Even though values for the checks and the medium RILS are included, for the discussion, we will only focus on differences among resistant (short tunnel length) and susceptible (large tunnel length) RILs. According to this classification, significant differences for agronomic traits between groups are also shown in Table 1.
RILS classified as resistant to corn borer attack, under control conditions, were the earliest (63–64 days to anthesis, silking), the smallest (157 cm) and the driest (16.4%), in addition to producing a lower yield (4 t ha−1). Furthermore, at a biochemical level, resistant lines presented higher concentrations of PCA (10 mg/g).
Under infestation with MCB eggs, resistant lines were also the earliest, showing similar flowering dates as in controlled conditions, and the smallest (10 cm smaller than the plants in control conditions). Nevertheless, resistant and susceptible RILs did not differ in grain yield and moisture.

3.2. Correlation Analysis

The relationships among agronomic traits, under both infestation conditions, were evaluated through genotypic and phenotypic correlations. Correlation coefficients are shown in Table 2. In this section and further in the discussion, we will focus on correlations above 0.50 (absolute value) highlighted in bold in Table 2. Apart from days to anthesis and silking (rp 0.87 c, 0.93 ui), we did not find any other important phenotypic correlation under both infestation conditions: control (c), under infestation (ui). On the other hand, we found a strong positive genotypic correlation between tunnel length and grain yield (rg 0.56 c, 0.68 ui) and moisture (rg 0.54 c, 1 ui) and between tunnel length and plant height (rg 0.57 c, 0.60 ui) both in infested and control trials. Under infestation, we found strong genotypic correlations between flowering time and grain moisture (rg 0.71, 0.55) and between plant height and grain yield (rg 0.56).

3.3. Multiple Linear Regression Analysis

In order to understand the role of cell wall-bound hydroxycinnamates as a mechanism of defense against corn borers, we performed multiple linear regression models. We considered tunnel length (under infestation) as a dependent variable and cell wall-bound hydroxycinnamates as independent variables. The best model for tunnel length explained 53% of the variance, mainly by PCA (15%), FA (28%) and DFA 5-5 (5%) and DFAT (4%) (Table 3).

4. Discussion

In order to achieve an accurate phenotyping, we repeated the evaluation of MAGIC RILs performed by Jiménez-Galindo et al. [32] selecting those presenting extreme values for tunnel length and according to their seed availability for future evaluations, resulting in a total of 56 RILS [32]. From those 56, another selection according to tunnel length under infestation was performed for contrast analysis, resulting in three groups: resistant, intermediate, and susceptible. Further on we will discuss differences between resistant and susceptible RILs.

4.1. Relationship between Resistance to Corn Borer and Main Agronomic Traits

Resistant and susceptible RILs differed in agronomic performance under both infestation conditions. Statistical analyses indicated that RILs presenting greater tunnel length, hence classified as susceptible to corn borer attack, were taller, presented with delayed maturity and had greater grain moisture at harvest. Besides, susceptible RILs yielded significantly more than resistant RILs under controlled conditions. These results are in agreement with previous findings observed in diverse genetic backgrounds [5,37,38,39,40].
The relationship between flowering time and resistance to corn borer attack is complex, and results from experimentation may be contradictory since this is a trait dependent on a lot of factors, such as the genetic background or the infestation time. Thereby, our results agree with those obtained by Jimenez-Galindo et al. [41], Santiago et al. [18], and Krakowsky et al. [42], but are opposite to those presented by Ordás et al. [27], Samayoa et al. [39], and Bohn et al. [43].
Late flowering genotypes are related to greater plant height, vigor, and yield; traits that have been negatively correlated to resistance to corn borer attack [27,44]. In northwestern Europe, the damage produced by second generation larvae causes the most important losses, damaging the plants during the reproductive stage [1,3]. An agronomic strategy proposed in order to alleviate the larvae damage is to advance the sowing time producing similar effects of those of early flowering. Early genotypes, with fast growth rate, would be in an advanced developmental stage by the time the MCB infestation peak occurs. Therefore, precocity would favor resistance to corn borer. Along with this, it has been demonstrated that tissue toughness, and cell wall lignification and fortification increase with plant maturity, interfering with the larvae progress.
The relationship between tunnel length and flowering time is clear in the contrast analysis, but it was not observed in correlation analysis in the infection condition. We identified genetic correlations above 0.5 between flowering time and grain moisture. Those results indicate that tunnels produced by the larvae under infestation conditions cause a delay in grain maturation [45] and could suggest that the period between flowering until grain maturation is more decisive in resistance than the period before flowering.
Moving on in our results, we found a high genetic correlation between plant height and tunnel length. Velasco et al. [46] hypothesized that taller plants would provide larger damage extent to the larvae. In an F2 derived from the cross of B73 and Dc811, Krakowsky et al. [42] found positive genetic correlations between stalk tunneling and ear height, which is highly correlated with plant height, indicating that the length of the stalk may be a limiting issue in the larvae-lant interaction. The above is supported to a greater extent if the positive relationship between plant height and days to flowering is taken into account: tall plants presented delayed maturity and greater susceptibility. The same correlations between plant height and tunnel length were found by Samayoa et al. [39] in bi-parental populations derived from the cross of resistant and susceptible inbred lines, and from crosses of a tolerant and sensible line [27,41]. In relation to these observations, several authors have also found QTL co-localization between plant height and stalk tunneling, reporting that favorable alleles increased the values for both traits, indicating an important genomic region in relation to resistance and its influence in agronomic traits [27,39,47]. This association between traits need to be considered in order to breed for borer resistance.
Finally, susceptible RILs yielded significantly more than resistant RILs under protected conditions, and we observed a strong, positive genetic correlation between grain yield and tunnel length under both infestation conditions. Breeding for increased resistance to corn borer attack has been related to decreases in grain yield [3,4,5,6], which agrees with our results. In contrast analysis, under infestation, we did not observe differences in grain yield between resistant and susceptible groups, which was contrary to controlled conditions. This could be explained because resistant RILs invest in constitutive defenses against the pest, which may imply a reduction of its yield. Under protected conditions, susceptible RILs do not suffer the attack of the plague producing significantly greater grain yield than resistant RILs, since they do not produce constitutive defenses. However, these differences are no longer significant when the RILs are exposed to the attack of the larvae: resistant RILs maintain their yield because of their defense mechanism and, in contrast, susceptible RILs see their grain yield compromised.
To sum up, greater tunnel length has been related to delayed maturity, which corresponds to a lower development of the plant tissues at the time of the larvae attack and greater grain moisture at the time of harvest. Delayed maturity also correlates with taller plants that present a greater extent to be consumed by the larvae. Lastly, susceptible RILs presented greater grain yield than resistant RILs when not exposed to the pest.

4.2. Role of Cell Wall Hydroxycinnamates as Defence Mechanism against Corn Borers

Resistant and susceptible RILs differed in cell wall-bound hydroxycinnamates content under both infestation conditions. From a biochemical point of view, the cell wall fortification mediated by hydroxycinnamates has been proposed as a defense mechanism against diverse pests [48]. Specifically, Santiago et al. [7] observed significant variation for cell wall phenylpropanoids in maize inbred lines presenting a wide range of susceptibility against S. nonagrioides attack. They observed significant differences in FA and DFATs pith concentrations, being greater in resistant inbreds. Subsequently, a successful selection for higher DFATs in maize pith resulted in increased resistance to corn borer attack, as a result of increases in the cell wall stiffening and strengthening by crosslinking hemicellulose chains [24]. However, in the current study, we did not observe an influence of DFATs on tunnel length. Even so, we did find in the regression analysis a positive influence of FA in greater tunnel length, the opposite of what was found by Santiago et al. [7]. There is a dependency relationship between FA and DFAT, since DFAT are a consequence of FA monomer dimerization, which is supported by strong genotypic and phenotypic correlations between those traits [49,50]. In this sense, greater concentrations of FA could indicate lower dimerization, hence lower crosslink, and stiffening. This would explain the major positive effect of FA in the tunnel length regression model.
On the other hand, a higher concentration of PCA monomer in the pith of maize was related to higher resistance to corn borer damage in both the regression model and contrast analysis. Increases in PCA concentration have previously been observed in both pith [18,19] and rind tissues [18] of maize resistant inbreds. In subsequent studies, Santiago et al. [51] concluded that alleles for increased ester hydroxycinnamates content, affecting one or more hydroxycinnamate compounds, could be associated with increased stem resistance to MCB; PCA being the hydroxycinnamate with the highest contribution to it. Furthermore, in a recent study, Gesteiro et al. [52] selected 10 F2:3 families with contrasting values for PCA, DFAT, and tunnel length in order to elucidate how cell wall-bound phenolic affects borer resistance. They observed a negative correlation between tunnel length and PCA concentration, in agreement with our results.
In maize, lignins (primarily syringyl units) are acylated at the γ-position by p-coumarates [53]. Most PCA accretion occurs in tandem with lignification and its accumulation could be considered a relevant indicator of lignin deposition. The fact that, in Gesteiro et al. [8], families showing the highest levels of PCA also showed the highest proportion of subunit S, suggests that the role of PCA in resistance could be associated not only to the lignin content but also to the lignin composition and structure [2]. S lignin, indirectly associated to more PCA acetylation, has been noted in resistance against different biotic stresses [52,54,55].
To sum up, the present study using MAGIC RILS adds to the list of evidence for the potential role of PCA in pest resistance. However, in this vegetal material, stiffening and crosslinking of DFAs does not take a part in the resistance to corn borer attack.

5. Conclusions

Corn borer resistant RILs showed precocity, low grain moisture at harvest, and reduced plant height. We observed that the negative correlation between yield and tunnel length, previously observed in other vegetal materials, is maintained after all of the recombination events in this MAGIC population. Therefore, breeding using the tunnel length criteria should be carried out with caution because this may compromise plant fitness and grain yield. In this case, we recommend a breeding strategy directly targeting grain yield.
Furthermore, taking account of a higher level of PCA acetylation in the selected materials as a secondary criteria could be advisable in future breeding programs, although negative correlations between plant fitness and p-coumarilation need to be addressed.

Supplementary Materials

The following are available online at, Supplementary Table S1: Means for agronomic traits and cell wall-bound hydroxycinnamates in protected from infestation and under infestation conditions of the RILs evaluated. Supplementary Table S2: Means for agronomic traits under infestation conditions of the RILs evaluated.

Author Contributions

R.A.M., R.S. conceived the study and participated in its design; R.S., R.A.M., A.L.-M., J.C.J.-G. and Z.R. carried out the field trial and sample collection; A.L.-M., R.A.M., Z.R. performed data analysis; A.L.-M. wrote the manuscript. Z.R. performed and assisted in the laboratory analysis. All authors have read and agreed to the published version of the manuscript.


This research has been developed in the frame of the Agri-Food Research and Transfer Centre of the Water Campus (CITACA) at the University of Vigo (Spain), which is economically supported by the Galician Government and in the Misión Biológica de Galicia-CSIC. It was funded by the “Plan Estatal de Ciencia y Tecnología de España” (projects RTI2018–096776-B-C21, RTI2018–096776-B-C22, and PID2019-108127RB-I00 co-financed with European Union funds under the FEDERprogram). A. López-Malvar’s contract was charged to the project RTI2018–096776-B-C22. J.C. Jiménez Galindo’s contract was financed by a PhD scholarship #314685 from the National Council for Science and Technology (CONACYT), Mexico. Zoila Resendiz’s contracts were financed by a Scholarship from the National Council of Science and Technology (CONACYT) to carry out Postdoctoral studies.

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

The data presented in this study are available on request from the corresponding author.


We thank Ana Carballeda for her technical assistance in the laboratory analysis.

Conflicts of Interest

The authors declare no conflict of interest.


  1. Meissle, M.; Mouron, P.; Musa, T.; Bigler, F.; Pons, X.; Vasileiadis, V.P.; Otto, S.; Antichi, D.; Kiss, J.; Pálinkás, Z.; et al. Pests, pesticide use and alternative options in European maize production: Current status and future prospects. J. Appl. Entomol. 2010, 134, 357–375. [Google Scholar] [CrossRef]
  2. Butrón, A.; Malvar, R.A.; Cartea, M.E.; Ordás, A.; Velasco, P. Resistance of maize inbreds to pink stem borer. Crop Sci. 1999, 39, 102–107. [Google Scholar] [CrossRef]
  3. López, C.; Sans, A.; Asin, L.; EizaGuirre, M. Phenological Model for Sesamia nonagrioides (Lepi-doptera: Noctuidae). Environ. Entomol. 2001, 30, 23–30. [Google Scholar] [CrossRef]
  4. Avantaggiato, G.; Quaranta, F.; Desiderio, E.; Visconti, A. Fumonisin contamination of maize hybrids visibly damaged by Sesamia. J. Sci. Food Agric. 2003, 83, 13–18. [Google Scholar] [CrossRef]
  5. Butrón, A.; Santiago, R.; Mansilla, P.; Pintos-Varela, C.; Ordás, A.; Malvar, R.A. Maize (Zea mays L.) genetic factors for preventing fumonisin contamination. J. Agric. Food Chem. 2006, 54, 6113–6117. [Google Scholar] [CrossRef] [PubMed]
  6. Klenke, J.R.; Russell, W.A.; Guthrie, W.D. Recurrent selection for resistance to European corn borer in corn synthetic and correlated effects on agronomic traits. Crop Sci. 1986, 26, 864–868. [Google Scholar] [CrossRef]
  7. Sandoya, G.; Malvar, R.A.; Santiago, R.; Alvarez, A.; Revilla, P.; Butrón, A. Effects of selection for resistance to Sesamia nonagrioides on maize yield, performance and stability under infestation with Sesamia nonagrioides and Ostrinia nubilalis in Spain. Ann. Appl. Biol. 2010, 156, 377–386. [Google Scholar] [CrossRef] [Green Version]
  8. Butrón, A.; Romay, M.C.; Peña-Asín, J.; Alvarez, A.; Malvar, R.A. Genetic relationship between maize resistance to corn borer attack and yield. Crop Sci. 2012, 52, 1176–1180. [Google Scholar] [CrossRef] [Green Version]
  9. Russell, W.A.; Wa, R. Effects of recurrent selection for European corn borer resistance on other agronomic characters in synthetic cultivars of maize. Maydica 1979, 24, 33–47. [Google Scholar]
  10. Bergvinson, D.J.; Arnason, J.T. Phytochemical changes during recurrent selection for resistance to the European corn borer. Crop Sci. 1997, 37, 1567–1572. [Google Scholar] [CrossRef]
  11. Buendgen, M.R.; Coors, J.G.; Grombacher, A.W.; Russell, W.A. European corn borer resistance and cell wall composition of tree maize populations. Crop Sci. 1990, 30, 505–510. [Google Scholar] [CrossRef]
  12. Ostrander, B.M.; Coors, J.G. Relationship between plant composition and European corn borer resistance in three maize populations. Crop Sci. 1997, 37, 1741–1745. [Google Scholar] [CrossRef]
  13. Fontaine, A.S.; Briand, M.; Barrière, Y. Genetic variation and QTL mapping of para-coumaric and ferulic acid contents in maize stover at silage harvest. Maydica 2003, 48, 75–84. [Google Scholar]
  14. Méchin, V.; Argillier, O.; Menanteau, V.; Barrière, Y.; Mila, I.; Rollet, B.; Lapierre, C. Relationship of cell wall composition to in vitro cell wall digestibility of maize inbred line stems. J. Sci. Food Agric. 2000, 80, 574–580. [Google Scholar] [CrossRef]
  15. Scalbert, A.; Monties, B.; Lallemand, J.Y.; Guittet, E.; Rolando, C. Ether linkage between phenolic acids and lignin fractions from wheat straw. Phytochemistry 1985, 26, 1359–1362. [Google Scholar] [CrossRef]
  16. Iiyama, K.; Lam, T.; Stone, B.A. Covalent crosslinks in the cell wall. Plant Physiol. 1994, 104, 315–320. [Google Scholar] [CrossRef] [Green Version]
  17. Grabber, J.H.; Mertens, D.R.; Kim, H.; Funk, C.; Lu, F.; Ralph, J. Cell wall fermentation kinetics are impacted more by lignin content and ferulate cross-linking than by lignin composition. J. Sci. Food Agric. 2009, 89, 122–129. [Google Scholar] [CrossRef]
  18. Santiago, R.; Butron, A.; Arnason, J.T.; Reid, L.M.; Souto, X.C.; Malvar, R.A. Putative role of pith cell wall phenylpropanoids in Sesamia nonagrioides (Lepidoptera: Noctuidae) resistance. J. Agric. Food Chem. 2006, 54, 2274–2279. [Google Scholar] [CrossRef] [PubMed]
  19. Barros-Rios, J.; Malvar, R.A.; Jung, H.J.G.; Santiago, R. Cell wall composition as a maize defense mechanism against corn borers. Phytochemistry 2011, 72, 365–371. [Google Scholar] [CrossRef] [Green Version]
  20. Assabgui, R.A. Correlation of kernel (E)ferulic acid content of maize with resistance to fusarium graminearum. Phytopathology 1993, 83, 949. [Google Scholar] [CrossRef]
  21. Santiago, R.; Butrón, A.; Reid, L.M.; Arnason, J.T.; Sandoya, G.; Souto, X.C.; Malvar, R.A. Diferulate content of maize sheaths is associated with resistance to the Mediterranean corn borer Sesamia nonagrioides (Lepidoptera: Noctuidae). J. Agric. Food Chem. 2006, 54, 9140–9144. [Google Scholar] [CrossRef] [PubMed]
  22. Barros-Rios, J.; Malvar, R.A.; Jung, H.J.G.; Bunzel, M.; Santiago, R. Divergent selection for ester-linked diferulates in maize pith stalk tissues. Effects on cell wall composition and degradability. Phytochemistry 2012, 83, 43–50. [Google Scholar] [CrossRef] [PubMed]
  23. Santiago, R.; Sandoya, G.; Butrón, A.; Barros, J.; Malvar, R.A. Changes in phenolic concentrations during recurrent selection for resistance to the Mediterranean corn borer (Sesamia nonagrioides Lef.). J. Agric. Food Chem. 2008, 56, 8017–8022. [Google Scholar] [CrossRef]
  24. Barros-Rios, J.; Santiago, R.; Jung, H.J.G.; Malvar, R.A. Covalent cross-linking of cell-wall polysaccharides through esterified diferulates as a maize resistance mechanism against corn borers. J. Agric. Food Chem. 2015, 63, 2206–2214. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  25. Cartea, M.E.; Malvar, R.A.; Revilla, P.; Ordás, A.; Alvarez, A. Seasonal occurrence and response of maize inbred lines to pink stem borer in the northwest of Spain. Maydica 1994, 39, 191–196. [Google Scholar]
  26. Santiago, R.; Souto, X.C.; Sotelo, J.; Butrón, A.; Malvar, R.A. Relationship between maize stem structural characteristics and resistance to pink stem borer (Lepidoptera: Noctuidae) attack. J. Econ. Entomol. 2003, 96, 1563–1570. [Google Scholar] [CrossRef]
  27. Ordas, B.; Malvar, R.A.; Santiago, R.; Butron, A. QTL mapping for Mediterranean corn borer resistance in European flint germplasm using recombinant inbred lines. BMC Genom. 2010, 11, 174. [Google Scholar] [CrossRef] [Green Version]
  28. Cardinal, A.J.; Lee, M.; Sharopova, N.; Woodmanclikeman, W.L.; Long, M.J.; European, T.; Hu, O. Tunneling by the European Corn Borer in Maize. Crop Sci. 2001, 41, 835–845. [Google Scholar] [CrossRef]
  29. Papst, C.; Bohn, M.; Utz, H.F.; Melchinger, A.E.; Klein, D.; Eder, J. QTL mapping for European corn borer resistance (Ostrinia nubilalis Hb.), agronomic and forage quality traits of testcross progenies in early-maturing European maize (Zea mays L.) germplasm. Theor. Appl. Genet. 2004, 108, 1545–1554. [Google Scholar] [CrossRef]
  30. Glowinski, A.; Flint-Garcia, S. Germplasm resources for mapping quantitative traits in maize. In The Maize Genome; Bennetzen, J., Flint-Garcia, S., Hirsch, C., Tuberosa, R., Eds.; Springer International Publishing: Cham, Switzerland, 2018; pp. 143–159. ISBN 978-3-319-97427-9. [Google Scholar]
  31. Butrón, A.; Santiago, R.; Cao, A.; Samayoa, L.; Malvar, R. QTLs for Resistance to fusarium ear rot in a multiparent advanced generation intercross (MAGIC) maize population. Plant Dis. 2019, 103, 897–904. [Google Scholar] [CrossRef]
  32. Jiménez-Galindo, J.C.; Malvar, R.A.; Butrón, A.; Santiago, R.; Samayoa, L.F.; Caicedo, M.; Ordás, B. Mapping of resistance to corn borers in a MAGIC population of maize. BMC Plant Biol. 2019, 19, 1–17. [Google Scholar] [CrossRef] [Green Version]
  33. Eizaguirre, M.; Albajes, R. Diapause induction in the stem corn borer, Sesamia nonagrioides (Lepidoptera: Noctuidae). Entomol. Gen. 1992, 17, 277–283. [Google Scholar] [CrossRef]
  34. Santiago, R.; López-Malvar, A.; Souto, C.; Barros-Ríos, J. Methods for determining cell wall-bound phenolics in maize stem tissues. J. Agric. Food Chem. 2018, 66, 1279–1284. [Google Scholar] [CrossRef]
  35. SAS Institute. Base SAS 9.4 Procedures Guide; SAS Institute: Cary, NC, USA, 2015. [Google Scholar]
  36. Holland, J.B. Estimating genotypic correlation and their standard errors using multivariate restricted maximum likelihood estimation with SAS Prco MIXED. Crop Sci. 2006, 46, 642–656. [Google Scholar] [CrossRef] [Green Version]
  37. Butrón, A.; Revilla, P.; Sandoya, G.; Ordás, A.; Malvar, R.A. Resistance to reduce corn borer damage in maize for bread, in Spain. Crop Prot. 2009, 28, 134–138. [Google Scholar] [CrossRef] [Green Version]
  38. Sandoya, G.; Santiago, R.; Malvar, R.A.; Butrón, A. Evaluation of structural and antibiosis resistance mechanisms during selection against Mediterranean corn borer (Sesamia nonagrioides Lef) in the maize synthetic EPS12. Crop Prot. 2010, 29, 7–10. [Google Scholar] [CrossRef]
  39. Samayoa, L.F.; Butron, A.; Malvar, R.A. QTL mapping for maize resistance and yield under infes-tation with Sesamia nonagrioides. Mol. Breed. 2014, 34, 1331–1344. [Google Scholar] [CrossRef] [Green Version]
  40. Krakowsky, M.D.; Lee, M.; Woodman-Clikeman, W.L.; Long, M.J.; Sharopova, N. QTL mapping of resistance to stalk tunneling by the european corn borer in RILs of maize population B73 X De811. Crop Sci. 2004, 44, 274–282. [Google Scholar] [CrossRef]
  41. Jiménez-Galindo, J.C.; Ordás, B.; Butrón, A.; Samayoa, L.F.; Malvar, R.A. QTL mapping for yield and resistance against mediterranean corn borer in maize. front. Plant Sci. 2017, 8, 2–11. [Google Scholar]
  42. Krakowsky, M.D.; Brinkman, M.J.; Woodman-Clikeman, W.L.; Lee, M. Genetic components of resistance to stalk tunneling by the European corn borer in maize. Crop Sci. 2002, 42, 1309–1315. [Google Scholar] [CrossRef] [Green Version]
  43. Bohn, M.; Schulz, B.; Kreps, R.; Klein, D.; Melchinger, A.E. QTL mapping for resistance against the European corn borer (Ostrinia nubilalis H.) in early maturing European dent germplasm. Theor. Appl. Genet. 2000, 101, 907–917. [Google Scholar] [CrossRef]
  44. Sandoya, G.; Butrón, A.; Alvarez, A.; Ordás, A.; Malvar, R.A. Direct response of a maize synthetic to recurrent selection for resistance to stem borers. Crop Sci. 2008, 48, 113–118. [Google Scholar] [CrossRef] [Green Version]
  45. Ordás, B.; Revilla, P.; Romay, M.C.; Malvar, R.A.; Butrón, A.; Ordás, A. Eighteen cycles of recur-rent mass selection for early flowering in two maize synthetics. Euphytica 2019, 215, 49. [Google Scholar] [CrossRef]
  46. Velasco, P.; Revilla, P.; Monetti, L.; Butrón, A.; Ordás, A.; Malvar, R.A. Corn borers (Lepidoptera: Noctuidae, Crambidae) in Northwestern Spain: Population dynamics and distribution. Maydica 2007, 52, 195–203. [Google Scholar]
  47. Schön, C.C.; Lee, M.; Woodman, W.L.; Melchinger, A.E.; Guthrie, W.D. Mapping and character-zation of quantitative trait loci affecting resistance against second generation european corn bo-er in maize with the aid of rflps. Heredity 1993, 70, 660. [Google Scholar] [CrossRef] [Green Version]
  48. Santiago, R.; Malvar, R.A. Role of dehydrodiferulates in maize resistance to pests and diseases. Int. J. Mol. Sci. 2010, 11, 691–703. [Google Scholar] [CrossRef] [Green Version]
  49. López-Malvar, A.; Butrón, A.; Samayoa, L.F.; Figueroa-Garrido, D.J.; Malvar, R.A.; Santiago, R. Genome-wide association analysis for maize stem Cell Wall-bound Hydroxycinnamates. BMC Plant Biol. 2019, 19, 1–12. [Google Scholar] [CrossRef] [PubMed]
  50. Barros-Rios, J.; Santiago, R.; Malvar, R.A.; Jung, H.J.G. Chemical composition and cell wall polysaccharide degradability of pith and rind tissues from mature maize internodes. Anim. Feed Sci. Technol. 2012, 172, 226–236. [Google Scholar] [CrossRef]
  51. Santiago, R.; Malvar, R.A.; Barros-Rios, J.; Samayoa, L.F.; Butrón, A. Hydroxycinnamate synthesis and association with mediterranean corn borer resistance. J. Agric. Food Chem. 2016, 64, 539–551. [Google Scholar] [CrossRef]
  52. Gesteiro, N.; Butrón, A.; Estévez, S.; Santiago, R. Unraveling the role of maize (Zea mays L.) cell-wall phenylpropanoids in stem-borer resistance. Phytochemistry 2021, 185, 112683. [Google Scholar] [CrossRef]
  53. Ralph, J.; Hatfield, R.D.; Quideau, S.; Helm, R.F.; Grabber, J.H.; Jung, H.J.G. Pathway of p-coumaric acid incorporation into maize lignin as revealed by NMR. J. Am. Chem. Soc. 1994, 116, 9448–9456. [Google Scholar] [CrossRef]
  54. Zhao, Q.; Wang, H.; Yin, Y.; Xu, Y.; Chen, F.; Dixon, R.A. Syringyl lignin biosynthesis is directly regulated by a secondary cell wall master switch. Proc. Natl. Acad. Sci. USA 2010, 107, 14496–14501. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  55. Ma, Q.H.; Zhu, H.H.; Qiao, M.Y. Contribution of both lignin content and sinapyl monomer to disease resistance in tobacco. Plant Pathol. 2018, 67, 642–650. [Google Scholar] [CrossRef]
Table 1. Contrast analysis for the t large, short and medium tunnel length groups and checks of RILs classified according to resistance to corn borer. Means for agronomic traits and hydroxycinnamic acids with significant differences among groups are included.
Table 1. Contrast analysis for the t large, short and medium tunnel length groups and checks of RILs classified according to resistance to corn borer. Means for agronomic traits and hydroxycinnamic acids with significant differences among groups are included.
Tunnel Length (cm)LargeMediumShortChecksLSD a
Under Control Condition (Insecticide Protected)
Days to Anthesis67.0 a65.8 b63.2 c67.7 a0.8
Days to Silking68.5 ab67.4 b64.1 c69.3 a1.4
Grain Yield (t ha−1)5.48 a5.24 a4.01 b5.37 a0.88
Grain Moisture (%)19.3 a17.8 ab16.4 b18.6 ab2.4
Plant Height (cm)181 a170 b157 c181 d5
Lodging (%)13.23 ab20.67 a22.01 a9.19 b10.69
PCA (mg/g)6.43 a8.07 b10.0 c9.71 c1.58
FA (mg/g)1.68 a1.69 a1.60 a2.42 b0.51
DFA 5-5 (mg/g)0.058 a0.062 a0.067 a0.087 b0.017
DFA 8-O-4 (mg/g)0.090 a0.963 a0.097 a0.134 b0.029
DFA 8-5-l (mg/g)0.064 a0.068 ab0.065 a0.084 b0.011
DFA 8-5-b (mg/g)0.083 a0.090 a0.089 a0.127 b0.017
DFA 8-5 (mg/g)0.153 a0.158 a0.148 a0.211 b0.026
DFAT (mg/g)0.301 a0.317 a0.312 a0.435 b0.079
Under Infestation Condition
Days to Anthesis67.6 a66.0 ab63.2 b66 ab4.3
Days to Silking69.3 a67.3 b64.5 c69.9 a1.2
Grain Yield (t ha−1)5.69 a5.59 a4.20 a5.52 a2.12
Grain Moisture (%)16.7 a15.5 a14.8 a15.8 a5.1
Plant Height (cm)176 a162 ab146 b165 ab26
Lodging (%)40.03 a43.26 a50.40 a43.03 a30.21
a. Least significant differences between means at the 0.05 significant level, according to Fisher protected LSD method. Means showing the same letter do not differ significantly. PCA: p-coumaric acid; FA: Ferulic acid; DFA5-5: diferulic acid 5-5; DFA 8-o-4: Diferulic acid 8-O-4, DFA85l: Diferulic acid 8-5-Linear; DFA85b: Diferulic acid 8-5-Benzofuran; DFA8-5: Diferulic acid 8-5; DFAT: Total diferulic acids content.
Table 2. Genotypic (above diagonal) and phenotypic (below diagonal).
Table 2. Genotypic (above diagonal) and phenotypic (below diagonal).
Tunnel LengthPlant HeightDays to AnthesisDays to SilkingGrain Yield LodgingGrain Moisture
Under Control Condition (Insecticide Protected)
Tunnel Length 1 0.57 *0.47 *0.43 *0.56 *−0.120.54 *
Plant Height0.32 * 0.39 *0.42 *0.63 *0.240.39 *
Days to Anthesis0.27 *0.18 0.96 * *
Days to Silking0.23 *0.190.87 * − *
Grain Yield0.130.20 *−0.07−0.13 0.10 *0.32
Lodging−0.090.10−0.04−0.030.18 0.32
Grain Moisture0.30 *0.150.25 *0.24 *0.16 *0.13
Under Infestation Condition
Tunnel Length 0.60 *0.43 *0.48 *0.68 *−0.041 *
Plant Height0.43 * 0.320.350.56 *−0.110.40
Days to Anthesis0.130.07 0.97 *0.10−0.100.71 *
Days to Silking0.130.060.93 * 0.04−0.090.55 *
Grain Yield0.22 *0.38 *0.05−0.10 0.040.44
Lodging0.020.08−0.01−0.060.17 0.41
Grain Moisture0. *0.160.15
* Significant correlation coefficient because it exceeded twice its standard error. 1 Tunnel length data comes from trial under infestation condition. Correlations above 0.50 (in absolute value) are highlighted in bold
Table 3. Multiple linear regression models (using stepwise selection) of tunnel length on hydroxycinnamic acids.
Table 3. Multiple linear regression models (using stepwise selection) of tunnel length on hydroxycinnamic acids.
Step Wise Selection
Tunnel Length (cm)
StepVariable introduced in the ModelR2 PartialR2
1PCA (mg/g)0.150.19
2FA (mg/g)0.280.44
3DFA 5-5 (mg/g)0.060.49
4DFAT (mg/g)0.040.53
ModelTUNNEL LENGTH: 42.13−3.08 × PCA + 5.87 × FA + 132.1 × DFA-8-5-l
R2 partial: percentage of the variance explained by each independent variable; R2: Total percentage of the variance explained by the model. PCA: p-coumaric acid; FA: Ferulic acid; DFA 5-5: Diferulic acid 5-5; DFAT: total diferulates content.
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Share and Cite

MDPI and ACS Style

López-Malvar, A.; Reséndiz, Z.; Santiago, R.; Jiménez-Galindo, J.C.; Malvar, R.A. Relationships between Stalk Resistance and Corn Borers, Agronomic Traits, and Cell Wall Hydroxycinnamates in a Set of Recombinant Inbred Lines from a Maize MAGIC Population. Agronomy 2021, 11, 1132.

AMA Style

López-Malvar A, Reséndiz Z, Santiago R, Jiménez-Galindo JC, Malvar RA. Relationships between Stalk Resistance and Corn Borers, Agronomic Traits, and Cell Wall Hydroxycinnamates in a Set of Recombinant Inbred Lines from a Maize MAGIC Population. Agronomy. 2021; 11(6):1132.

Chicago/Turabian Style

López-Malvar, Ana, Zoila Reséndiz, Rogelio Santiago, José Cruz Jiménez-Galindo, and Rosa Ana Malvar. 2021. "Relationships between Stalk Resistance and Corn Borers, Agronomic Traits, and Cell Wall Hydroxycinnamates in a Set of Recombinant Inbred Lines from a Maize MAGIC Population" Agronomy 11, no. 6: 1132.

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop