Field Crops Research 120 (2011) 271–275

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Field Crops Research

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Stalk strength and reaction to infection by Macrophomina phaseolina of brown

midrib maize (Zea mays) and sorghum (Sorghum bicolor)
Tesfaye Tesso a , Gebisa Ejeta b,∗
a
Kansas State University, Department of Agronomy, Manhattan, KS 66506, United States
b
Purdue University, Department of Agronomy, West Lafayette, IN 47907, United States

a r t i c l e i n f o a b s t r a c t

Article history: Reduced lignin concentration in brown mid-rib mutants in both maize and sorghum have resulted in
Received 25 April 2010 improved dry matter digestibility, increased milk yield and higher energy in lactating cows. However,
Received in revised form 11 July 2010 the mutations were not widely deployed due to concern that reduced lignin concentration might increase
Accepted 31 October 2010
vulnerability to lodging and stalk rot incidence. The objective of this study was to determine the effects
of the mutations on stalk strength and stalk rot resistance in both sorghum and maize. Six brown midrib
Keywords:
(bmr) sorghum, four brown midrib (bm) maize, and their normal isolines were evaluated for stalk strength
Brown midrib
and stalk rot disease reaction at two locations in four replications. Three randomly selected plants in
Macrophomina phaseolina
Rind penetration resistance
each plot were inoculated with Macrophomina phaseolina at 14 d after flowering by using the toothpick
Sorghum inoculation technique. On 28 d after inoculation, the plants were rated for disease severity by measuring
Maize the length of necrotic lesions in the stalks. Stalk strength was determined from another three random
Stalk rot disease plants in each plot by using a rind penetrometer. The mutations had no effect on stalk rot disease severity
Stalk strength in either sorghum or maize though stalks strength was markedly affected by the mutations in both species.
While maize bm entries broke easily in response to mild mechanical stress, bmr sorghums did not exhibit
sign of stalk collapse in all backgrounds. The result suggests that the bmr genes in both maize and sorghum
can be deployed without incurring losses to stalk rot disease.
© 2010 Published by Elsevier B.V.

1. Introduction
The evolutionary course that granted plant species the ability
to synthesize and polymerize monolignol residues into complex
lignin has been crucial for development of erect plants and their
adaptation to a terrestrial habitat (Raven et al., 1999; Boudet,
2000). Monolignols are synthesized through the phenyl propanoid
pathway, and enzymes catalyzing their synthesis have been iden-
tified and characterized (Hennion et al., 1992; Baucher et al.,
1998; Whetten et al., 1998). Besides providing mechanical sup-
port and aiding in water and mineral transport by consolidating
the treachery xylem elements (Campbell and Sedroff, 1996; Ye
et al., 2001; Peter and Neale, 2004), lignin confers structural defense
against mechanical stress and enhances fitness (Casler et al., 2002;
Pedersen et al., 2005).
Mutations in lignin biosynthetic pathways have been reported
in several crop species (Porter et al., 1978; Hartley et al., 1992;
Halpin et al., 1998). These are the results of altered activity of
enzymes that polymerize monolignol residues into complex lignin
(Pillonel et al., 1991; Halpin et al., 1998; Ralph et al., 1998). Variants
∗ Corresponding author. Tel.: +1 765 494 4320; fax: +1 765 496 2926.
E-mail address: gejeta@purdue.edu (G. Ejeta).
of sorghum, maize, and pearl millet genotypes with low lignin con-
centration in stems, leaf sheaths, and leaf blades have been reported
(Porter et al., 1978; He et al., 2003; Vogler et al., 2009).
To date five bm loci (bm1, bm2, bm3, bm4 and bm5) have been
identified in maize (Kuc et al., 1968; Haney et al., 2008). More
than two dozens of bmr alleles have been identified in sorghum,
but alleleism tests indicate that most of the mutations are allelic
and only four bmr loci are known (Saballos et al., 2008). Reduc-
tion in the activity of lignin biosynthetic enzymes Caffeic acid
O-Methyltransferase (COMT) and Cinnamyl Alcohol Dehydroge-
nase (CAD) was responsible for reduced lignin concentration and
altered lignin composition in the mutants. The bm3 mutation in
maize and bmr7, bmr12 and bmr26 mutations in sorghum have been
reported to have resulted from reduced activity of COMT (Vignols
et al., 1995; Bout and Vermerris, 2003; Saballos et al., 2008). On
the other hand, the bm1 mutation in maize and bmr6 and bmr28
mutations in sorghum were associated with reduced activity of CAD
(Bucholtz et al., 1980; Pillonel et al., 1991; Halpin et al., 1998). The
bm2 and bm4 in maize and several bmr loci in sorghum have also
been reported to cause reduced lignin concentration. Neverthe-
less, specific pathways leading to these mutations have not been
determined yet. These mutations, which are expressed by a shared
brown midrib phenotype in both maize and sorghum, have impor-
tant biological and economic implications, especially in the feedlot

0378-4290/$ – see front matter © 2010 Published by Elsevier B.V.

doi:10.1016/j.fcr.2010.10.015
272 T. Tesso, G. Ejeta / Field Crops Research 120 (2011) 271–275

Table 1
Analysis of variance for lesion length and resistance to rind penetration among low-lignin and normal sorghum and maize genotypes tested at two locations Purdue
Agricultural Center for Research and Education (ACRE) near West Lafayette and Piney Purdue Agricultural Center (PPAC), Piney, Indiana.

Source dfa Lesion length (mm) Rind penetration resistance

ACRE PPAC Combined ACRE PPAC Combined

Environment (E) 1 – – 1669.86 – – 5.31*

E (Block) 6(3) 420.43 285.14 352.78 6 3.1 4.55
Main Plot (MP) 1 110.02 87.01 3.72 4.58* 0.3 3.62*
MP × E 1 – – 90.61 – – 1.02
Errorb 6(3) 63.31 364.92 514.11 0.59 1.41 1
Genotypes (G) 9 6047.87* 1112.21* 2977.7 5.16* 5.15* 9.82*
Sorghum(S) 5 10,273.48* 1235.46* 4937.37 6.43* 6.10* 11.98*
Maize (C) 3 219.25 346.7 170.49 4.04* 3.96* 7.59
S vs. C 1 648.3 3341.87* 632.76 1.77 1.96 5.5
G × MP 9 119.44 275.4 320.11 3.76* 1.02 3.74
S × MP 5 62.46 126.88 262.8 4.00* 0.66 3.09
C × MP 3 119.81 621.44 485.54 4.32* 1.69* 5.56
S vs. C × MP 1 275.17 62.57 291.39 1.09 0.79 1.87
G×E 9 – – 2968 – – 0.51
S×E 5 – – 3771.62 – – 0.55
C×E 3 – – 388.47 – – 0.58
S vs. C × E 1 – – 3567.61 – – 0.21
G × MP × E 9 – – 80.29 – – 1.04
S × MP × E 5 – – 218.67 – – 1.45
C × MP × E 3 – – 66.93 – – 0.5
S vs. C × E 1 – – 29.75 – – 0.01
Errorc 107(53/54) 432.72 509.62 466.01 0.84 0.64 0.74
*
Statistically significant at P ≤ 0.05 and P ≤ 0.01, respectively.
a
Degrees of freedom in the parentheses are for individual location analysis.

industry, because both maize and sorghum are significant compo-
nents of animal feed. Most of the brown midrib mutations reduce
the level of lignin in plants and remarkably increase intake and
dry matter digestibility (Frenchik et al., 1976; Rook et al., 1977;
Cherney et al., 1991). Brown midrib maize silage alone and mixed
with protein sources has significantly increased milk yield in lac-
tating cows (Keith et al., 1979; Stallings et al., 1982; Oba and Allen,
1999). Similarly, the rates of in vitro dry matter digestibility and cell
wall degradation by rumen bacterium of leaf blades from bmr12
sorghum were shown to be significantly higher than those from
its wild-type isoline (Akin et al., 1986a,b). Besides affecting feed
values, high lignin concentration in pulp woods has also been detri-
mental in the paper industry because lignin removal is a major
step in paper production (Campbell and Sedroff, 1996). Likewise,
in view of the growing interest in bioenergy development, high
lignin concentration in biofuel feedstock sources is obviously unde-
sirable because of its negative effect on conversion of biomass into
fermentable sugars (Grabber, 2005; Vermerris et al., 2007).
Despite the drawbacks of high-lignin, production of low-lignin
cultivars has not been widely adopted, partly because of the con-
cern that altered lignin structure in the mutants may reduce
standability and predispose plants to attack by pests and dis-
eases. However, few studies have been conducted to quantitatively
describe the effect of these mutations on stalk rot disease incidence
in both maize and sorghum. The objectives of this study were to
evaluate the effect of the brown mid-rib mutations on mechanical
strength of stalks and determine reaction of low-lignin maize and
sorghum mutants to the major stalk rotting pathogen, Macrophom-
ina phaseolina.
ficial mutagenesis of a breeding line 954114 while bmr12 was
derived from 954104 in the same way (Porter et al., 1978). Bmr26
and bmr28 are spontaneous mutations discovered in backgrounds
not related to the other mutants. Allelism tests and character-
ization of the mutations revealed that the six mutants occupy
three loci on sorghum genome namely the bmr2 group, the bmr6
group which includes bmr6 and bmr28 and the bmr12 group which
includes bmr7, bmr12 and bmr26. The genotypes were evaluated
under two environments at the Purdue University Agronomy Cen-
ter for Research and Education (ACRE) near West Lafayette, IN,
and at Piney Purdue Agricultural Center (PPAC) in Piney, IN. The
experiment was conducted in a split-plot design with randomized
complete blocks in four replications. The genotypes (six sorghum
and four maize) were assigned to the whole plot unit, and their
brown mid-rib and normal versions were assigned to the subplots.
The whole plot consisted of four 6-m-long rows (two rows each
for the two subplot treatments within each whole plot unit). Seeds
were sown following the standard procedure for both crops, and
optimum agronomic practices were used throughout the grow-
ing period. For treatment application, six plants from each subplot
unit were marked with tagging tapes at half bloom stage. At 14 d
after flowering, three of the marked plants were inoculated on the
basal stalk with M. phaseolina, a causal organism of charcoal rot, by
using a toothpick inoculation technique. Charcoal rot is an aggres-
sive stalk rot pathogen that is particularly devastating when hot,
dry weather overlaps with crop maturation in both sorghum and
maize (Mughogho and Pande, 1984). The remaining three plants
were used to determine stalk strength using digital rind penetrom-
eter, a modified AccuForce Cadet (Ametek, Hatfield, PA) powered
by alkaline battery.

2. Materials and methods 2.2. Inoculum preparation and inoculation

2.1. Genetic materials
Six brown midrib sorghum mutants (bmr2, bmr6, bmr7, bmr12,
bmr26, bmr28) and four maize mutants (bm1, bm2, bm3, and bm4)
along with their normal (Nor) isolines were used in this study.
The bmr2, bmr6, bmr7 and bmr12 mutants were derived by arti-
Pure culture of M. phaseolina was provided by Dr. Mitch Tuinstra
from Kansas State University. The strain was originally collected
from infested sorghum fields in Texas in summer 2001 by Dr. Garry
Odvody and shipped to Kansas State in water agar. In 2003, the
strain was grown on fresh water agar and shipped to Purdue Uni-
versity, where it was sub-cultured on potato dextrose agar and
 T. Tesso, G. Ejeta / Field Crops Research 120 (2011) 271–275 273

Table 2 Table 3
Effect of low-lignin mutation on stalk rot disease resistance and stalk strength in Mean lesion length (mm) of brown mid-rib (bmr) and normal (Nor) isolines of
maize and sorghum. sorghum and maize genotypes inoculated with M. phaseolina at Purdue Agricultural
Center for Research and Education (ACRE) near West Lafayette and Piney Purdue
Genotypes Lesion length (mm) Rind penetration resistance Agricultural Center (PPAC), Piney, Indiana.
Sorghum
Genotype Brown mid-rib Normal
bmr 67.1 5.5
Normal 72.2 5.9 ACRE PPAC Mean ACRE PPAC Mean
a
LSD(0.05) ns ns
Maize Sorghum
bm 73.6 5.1b bmr2/Nor2 61.3 43.3 52.3 69.3 44.2 56.8
Normal 72.2 5.7a bmr 6/Nor6 38.3 25.5 31.9 55.5 – 55.5
a
LSD(0.05) ns 0.53 bmr 7/Nor7 76.7 70.6 73.6 84.8 69.8 77.3
bmr12/Nor12 65.7 39.8 52.7 71.2 64.5 67.8
ns, non significant. bmr26/Nor26 134.8 70.3 102.5 141.7 74.8 108.2
a
LSD, least significant difference. bmr28/Nor28 46.3 60.5 53.4 24.2 62.7 43.4
Mean 73.0 59.7 67.1 82.7 58.0 72.2
a
LSD(0.05) 24.36 ns 22.06 49.44 ns 30.89
maintained at room temperature until needed. At 10 d before inoc- Maize
ulation, the pathogen strain was sub-cultured into several fresh bm1/Nor1 65.5 61.0 63.3 75.7 88.8 82.3
bm2/Nor2 72.7 78.0 75.3 64.9 53.5 59.2
potato dextrose agar plates, into which sterile toothpicks were
bm3/Nor3 78.1 65.1 71.6 79.8 76.4 78.1
inserted and incubated at 30 ◦ C until the media was fully covered bm4/Nor4 71.2 89.7 80.4 59.4 75.4 67.4
by the growing sclerotia. At 14 d after flowering, three of the tagged Mean 72.2 75 73.6 72.1 72.3 72.2
a
plants from each plot were inoculated with the infected toothpicks LSD(0.05) ns ns ns ns ns ns
on the basal stalks, approximately 10 cm above the ground. ns, non significant.
a
LSD, least significant difference.
2.3. Data collection and statistical analysis

At 28 d after inoculation, infected plants from each plot were
harvested for disease scoring. Disease severity was rated by longitu-
dinally splitting the stalks and measuring the length of the necrotic
lesion. Stalk strength for sorghum was measured by puncturing
rind penetrometer into the rind at approximately 15 cm above the
ground. For maize, penetrometer readings were taken according to
the procedure described by Flint-Garcia et al. (2003).
For statistical analysis, blocks and locations were considered
random effects, and the entries were considered fixed effect. Before
analysis, data were checked for normality and equality of variance
by using the t-Max test and then subjected to analysis of variance by
using the General Linear Model (GLM) procedure in SAS (Statistical
Analysis Systems, 2004). The RANDOM statement with TEST option
was used to identify random variables.
3. Results
3.1. Effect of brown mid-rib mutation on stalk rot resistance
Analysis of variance for reaction to stalk rot disease and resis-
tance to rind penetration is presented in Table 1. The bmr mutations
did not significantly alter genotypic response to stalk rot infection
in either sorghum or maize at either location. Mean disease score in
bmr sorghum mutants was slightly lower than that for the normal
genotypes 67.1 mm vs. 72.2 mm (Table 2). Similarly, the bm maize
mutants had mean lesion length score (73.6 mm) that was com-
parable to the score in the normal maize genotype (72.2 mm). It
appears that there is no marked relationship between mechanical
strength of stalk conferred by accumulation of lignin and develop-
ment of stalk rot disease induced by M. phaseolina in both sorghum
and maize.
On the other hand, disease severity was significantly different
among sorghum genotypes in both test environments (Table 3).
Mean lesion length ranged from 31.9 to 102.5 mm for the bmr
entries and from 43.4 to 108.2 mm for the wild-type entries. Bmr6
had the lowest score of all bmr sorghum entries at ACRE and PPAC,
and bmr26 had the highest score at both locations. The variation
in disease score did not reflect differences in allelic grouping of
the mutations. Bmr28, a member of the bmr6 family had differ-
ent score from bmr6 at both locations. Similarly members of the
bmr12 group (bmr7, bmr12, bmr18 and bmr26) had markedly dis-
tinct scores. Thus differences in disease reaction are most likely
due to variation in genetic backgrounds than the effect of the bmr
alleles. The wild types exhibited a similar trend, except the normal
counterpart for bmr6 was missing at PPAC (Table 3). Differences
in disease severity among genotypes in the combined analysis,
however, were not significant, although the genotype mean square
was similar to the individual location analysis (Table 1). This was
perhaps a result of the relatively high mean square value of the
G × E source used as the denominator to test the genotype effect.
Furthermore, there was no significant difference in disease sever-
ity between bm and normal maize genotypes at either location
(Table 3). Mean lesion length ranged from 63.3 to 80.4 mm for the
bm genotypes and from 59.2 to 82.3 mm for normal maize geno-
types. The variation between sorghum and maize genotypes was
significant only at PPAC (Table 1). Mean lesion length was 67.1 mm
for bmr sorghum mutants and 73.6 mm for bm maize. There was
no marked difference between normal maize and sorghum except
at PPAC where the score for maize was remarkably lower. Dis-
ease severity was generally higher at ACRE than at PPAC for
low-lignin mutants and normal genotypes of both maize and
sorghum.
3.2. Effect of brown mid-rib mutation on stalk strength
The brown mid-rib mutation remarkably reduced mechanical
strength of stalks. Rind penetration resistance for normal geno-
types was significantly higher than that for bm maize (Table 2).
The trend was similar for sorghum but the difference was not sig-
nificant. About 5.5 kg of force was required to penetrate through
the rinds of bmr sorghum, whereas 5.9 kg was required for similar
penetration of the normal genotypes. Similarly, 5.1 and 5.7 kg were
required to penetrate stalks of bm and normal maize genotypes,
respectively, and this trend was consistent across locations.
Moreover, variation in penetration resistance among sorghum
and maize entries was significantly different. Among bmr
sorghums, bmr26 had the lowest score and bmr2 had the highest.
Among normal lines, Nor26 had the lowest score, and Nor7 had the
highest followed by Nor2 and Nor28. For bm maize entries, bm2 and
bm3 had the lowest and highest resistance to penetration, respec-
tively. Among normal maize lines, the highest and lowest readings
were observed in Nor2 and Nor4, respectively (Table 4).
274 T. Tesso, G. Ejeta / Field Crops Research 120 (2011) 271–275

Table 4 study, results from previous evaluations that involved these same
Differences in rind penetration resistance (lb) among brown midrib (bmr) and nor-
mal (Nor) isolines of sorghum and maize genotypes at Purdue Agricultural Center for
genotypes showed that the brown mid-rib mutants have a low
Research and Education (ACRE) near West Lafayette and Piney Purdue Agricultural lignin concentration in stems, leaf sheaths, and leaf blades (Barriere
Center (PPAC), Piney, Indiana. et al., 1994; Gentinetta et al., 1990; Vogler et al., 2009). However,
the variation in stalk strength between the bm and normal maize
Genotype Brown midrib Normal
appears to be larger than the differences revealed by rind pene-
ACRE PPAC Mean ACRE PPAC Mean tration resistance. The bm maize mutants were highly brittle, and
Sorghum the slight stress commonly encountered under normal field condi-
bmr2/Nor2 6.5 6.3 6.4 6.7 6.5 6.6 tions caused complete breakage. Though no comparison was made
bmr 6/Nor6 5.1 5.2 5.1 5.0 5.2 5.1
between the mutations, large number of the maize plants in the
bmr 7/Nor7 4.4 6.02 5.2 7.3 6.6 6.98
bmr12/Nor12 5.5 6.1 5.9 5.8 6.2 6.0 bm rows were broken, often at the crown, at the time of data
bmr26/Nor26 3.9 4.7 4.3 4.2 5.0 4.6 collection.
bmr28/Nor28 5.7 6.3 6.0 6.3 6.8 6.6 Despite the high digestibility of forages and silages from bm
Mean 5.2 5.8 5.5 5.9 6.1 5.9 maize (Rook et al., 1977; Cherney et al., 1991) and the small
a
LSD(0.05) 1.50 ns 1.12 1.32 1.29 0.89
Maize
effect of the mutation on M. phaseolina-induced stalk rot sever-
bm1/Nor1 5.1 5.8 5.4 5.3 5.4 5.3 ity, the mutants’ vulnerability to mechanical damage under normal
bm2/Nor2 4.1 4.5 4.3 7.0 6.2 6.6 field conditions may pose a challenge to widespread cultiva-
bm3/Nor3 6.0 6.4 6.2 5.8 6.4 6.1 tion. However, a favorable mutant by background interaction that
bm4/Nor4 4.2 4.6 4.4 5.1 4.8 5.0
would minimize stalk delicacy without affecting the low lignin
Mean 4.9 5.3 5.1 5.9 5.7 5.7
a
LSD(0.05) 0.95 1.18 ns 1.46 0.64 ns content could be exploited if suitable backgrounds are identi-
fied. In contrast, none of the sorghum mutants were particularly
ns, non significant.
a prone to damage by wind or mechanical stresses imposed dur-
LSD, least significant difference.
ing routine field operation. Considered along with the small effect
of the mutation on stalk rot severity, this result suggests that
4. Discussion bmr sorghum genotypes can be grown under normal sorghum
production environments without fear of stalk-rot-induced crop
Absence of a noticeable difference in disease score between lodging.
normal and low-lignin maize and sorghum lines may partly allevi- Although the differences in penetrometer scores among maize
ate concerns of forage and biofuel sorghum producers. This result mutants can be fully attributed to the different type of mutations,
suggests that degree of lignification is not associated with stalk rot- the differences noted among normal genotypes at individual loca-
induced lodging in either sorghum or maize, although resistance to tions are difficult to explain because these genotypes are of the
mechanical lodging is expected to be higher in normal genotypes. same genetic background. The small sample size (three plants) and
Our result is in agreement with previous findings in sorghum. An possible variation in microenvironments might be partly respon-
earlier report by Esechie et al. (1977) refuted the presumed rela- sible for the observed variation. However, those differences were
tionship between lignin deficiency and stalk-rot-induced lodging not significant in the combined analysis.
in sorghum. Recent studies that involved a diverse set of sorghum Some of the bmr mutations included in this study are allelic
genotypes showed no observable difference in stalk rot resistance but found in genetically distinct backgrounds. Hence, variation
between stiff (perhaps highly lignified) and weak-stalked sorghum in rind penetration resistance observed among mutants might be
lines Tesso et al. (2004, 2005). However, the situation may be dif- the result of interaction between the mutations and background.
ferent under natural conditions, in which the parasite normally On the other hand, some of the mutations, though allelic, may
progresses through different steps of the infection process to estab- differ in degree of alteration from the wild-type gene such that
lish in the host tissue (e.g., overcoming mechanical resistance to variants of mutant alleles can occur at a given locus and thus
penetration of the host tissue). The effect of lignin content may be a have a different effect on lignin deposition and differently affect
factor in the initial phase of natural infection and subsequent steps stalk strength. In a previous study, we noted significant varia-
of disease development. In this experiment, inoculum was deliv- tion in lignin accumulation between sorghum mutant bmr26 and
ered directly into the pith of the host tissue such that the role lignin bmr12 in all tissue types (Vogler et al., 2009), but these muta-
could have played as mechanical barrier to pathogen penetration tions were allelic (Bittinger et al., 1981). On the other hand, bmr18
was bypassed. Further research should take this into account and and bmr26 have also shown to be allelic (Vogler et al., 2009),
compare bmr lines and their wild-type counterparts under both suggesting that that bmr12, bmr18, and bmr26 mutations were
natural and artificial infection. Nevertheless, the absence of a sig- either the same gene or variants of the same gene. Furthermore,
nificant difference in disease severity between normal and bmr penetrometer score might also be affected by internode length,
genotypes has important implication in forage and biofuel sorghum which is influenced by genetic factors other than those involved in
production. Because disease scores in the bmr mutants were not dif- lignin accumulation. Genotypes that have relatively shorter intern-
ferent from, and in some cases were even lower than, those of their odes tend to require higher force to penetrate their rinds than
respective wild-type counterparts, bmr sorghums with improved those with longer internodes. In this study, the bmr26 background
dry matter digestibility likely can be grown in much same way as that carries the mutation that is allelic to bmr12 is significantly
normal sorghums with no predicted loss of dry matter yield due to taller than the rest of the entries and required the least force to
stalk rot diseases. penetrate its rinds. This suggests a general hypothesis that stalk
Unlike stalk rot resistance, low lignin concentration in both strength is affected by a combined effect of alleles that deter-
bm maize and bmr sorghum (Chabbert et al., 1994a,b; Vogler mine content and composition of lignin molecules and height
et al., 2009) appeared to have noticeably altered stalk strength. genes that affect internode length. Therefore, to develop low-lignin
Rind penetration resistance was higher in normal genotypes than varieties, one may need to consider how the possible interaction
in low-lignin mutants in both species. This finding concurs with between genes at other loci and the bmr mutations affect agro-
the common knowledge that tissue lignin concentration affects nomically and nutritionally important plant characteristics such as
mechanical strength of plants (Raven et al., 1999). Though no standability, dry matter digestibility, and ethanol conversion effi-
attempt was made to assay lignin content of genotypes in this ciency.
 T. Tesso, G. Ejeta / Field Crops Research 120 (2011) 271–275
5. Conclusion
Brown mid-rib mutations due to their effect on lignin content in
both sorghum and maize resulted in reduced resistance to mechan-
ical penetration. The mutations, particularly in maize, appeared to
remarkably alter the structural integrity of the stalk tissue that the
mutants were brittle and broke easily in response to mild mechani-
cal stress. No lodging or stalk damage associated with the mutations
was observed in sorghum. On the other hand, there was no dif-
ference among both maize and sorghum genotypes in response
to infection by M. phaseolina. Absence of a functional relationship
between the bmr mutation and disease resistance indicates that
low-lignin genotypes of both sorghum and maize could be pro-
duced without incurring losses to stalk rot disease.
Acknowledgements
The authors thank the Purdue University Department of Agron-
omy for providing field and laboratory facilities for this study. We
also thank Terry Lemming for his assistance in data collection. This
work is a contribution by Kansas agricultural Experiment Station
(Contribution No. 10-251-J).
References
Akin, A.E., Hanna, W.W., Rigsby, L.L., 1986a. Normal-12 and brown midrib-12
sorghum I. Variations in tissue digestibility. Agron. J. 78, 827–832.
Akin, A.E., Hanna, W.W., Snook, M.E., Himmelsbach, D.S., Barton, F.E., Windham,
W.R., 1986b. Normal-12 and brown midrib-12 sorghum II. Chemical variations
and digestibility. Agron. J. 78, 832–837.
Barriere, Y., Argiller, O., Chabbert, B., Tollier, M.T., Monties, B., 1994. Breeding silage
maize with brown midrib genes. Feeding value and biochemical characteristics.
Agronomie 14, 15–25.
Baucher, M., Monties, B., Van Montagu, M., Boerjan, W., 1998. Biosynthesis and
genetic engineering of lignin. Crit. Rev. Plant Sci. 17, 125–197.
Bittinger, T.S., Cantrell, R.P., Axtell, J.D., 1981. Allelism tests of the brown midrib
mutants of sorghum. J. Hered. 72, 147–148.
Boudet, A., 2000. Lignins and lignification: selected issues. Plant Physiol. Biochem.
38, 81–96.
Bout, S., Vermerris, W., 2003. A candidate gene approach to clone the sorghum Brown
mid-rib gene encoding caffeic acid O-methyltransferase. Mol. Genet. Genomics
269, 205–214.
Bucholtz, D.L., Cantrell, R.P., Axtell, J.D., Lechtenberg, V.L., 1980. Lignin biochem-
istry of normal and brown midrib mutant sorghum. J. Agric. Food Chem. 28,
1239–1245.
Campbell, M.M., Sedroff, R.R., 1996. Variation in lignin content and their composi-
tion. Plant Physiol. 110, 3–13.
Casler, M.D., Buxton, D.R., Vogel, K.P., 2002. Genetic modification of lignin concen-
tration affects fitness of perennial herbaceous plants. Theor. Appl. Genet. 104,
127–131.
Chabbert, T., Monties, B., Barrière, Y., Argillier, O., 1994a. Biological variability in
lignification of maize: expression of the brown midrib bm3 mutation in three
different maize cultivars. J. Sci. Food Agric. 64, 349–355.
Chabbert, T., Monties, B., Barrière, Y., Argillier, O., 1994b. Biological variability in
lignification of maize: expression of the brown midrib bm2 mutation. J. Sci. Food
Agric. 64, 455–460.
Cherney, J.H., Cherney, J.R., Akin, D.E., Axtell, J.D., 1991. Potential of brown mid-
rib, low lignin mutants for improving forage quality. Adv. Agron. 46, 157–
198.
Esechie, H.E., Maranville, J.W., Ross, W.M., 1977. Relationship of stalk morphol-
ogy and chemical composition to lodging resistance in sorghum. Crop Sci. 17,
609–612.
Flint-Garcia, S.A., Mcmullen, M.D., Darrah, L.L., 2003. Genetic relationship of stalk
strength and ear height in maize. Crop Sci. 43, 23–31.
Frenchik, G.E., Jhonson, D.G., Murphy, J.M., Otterby, D.E., 1976. Brown midrib maize
silage in dairy cattle rations. J. Dairy Sci. 59, 2126–2129.
275
Gentinetta, E., Bertolini, M., Rossi, I., Lorenzoni, C., Motto, M., 1990. Effect of brown
midrib 3 mutant on forage quality and yield in maize. J. Genet. Breed. 44, 21–26.
Grabber, J.H., 2005. How do lignin composition, structure, and cross-linking affect
degradability? A review of cell wall model studies. Crop Sci. 45, 820–831.
Halpin, C.K.H., Chojecki, J., Oliver, D., Chabbert, B., Monties, B., Edwards, K., Barakate,
A., Foxon, G.A., 1998. Brown mid-rib maize (bm1) – a mutation affecting cynamyl
alchol dehydroginase gene. Plant J. 14, 545–553.
Haney, L.L.H., Hake, S.S.H., Scott, M.P.S., 2008. Allelism testing of maize coop stock
center lines containing unknown brown midrib alleles. Maize Newslett. 82, 4–5.
Hartley, R.D., Morisson, W.H., Borneman, W.S., Rigsby, L., O’neill, M., Hanna, W.W.,
Akin, D.E., 1992. Phenolic constituents of cellwall types of normal and brown
midrib mutants of pearl millet (Pennisetum glaucum (L) R Br) in relation to wall
biodegradability. J. Food Sci. Agric. 59, 211–216.
He, X., Hall, M.B., Gallo-Meagher, M., Smith, R.L., 2003. Improvement of forage quality
by downregulation of maize O-methyltransferase. Crop Sci. 43, 2240–2251.
Hennion, S., Little, C.H.A., Hartmann, C., 1992. Activities of enzymes involved in lig-
nification during the postharvest storage of etiolated asparagus spears. Physiol.
Plantarum 86, 474–478.
Keith, E.A., Colenbrander, F., Vlechtenberg, V.L., Bauman, L.F., 1979. Nutritional value
of brown midrib maize silage for lactating dairy cows. J. Dairy Sci. 62, 788–792.
Kuc, J., Nelson, O., Flanagan, E.P., 1968. Degradation of abnormal lignins in the brown-
midrib mutants and double mutants of maize. Phytochemistry 7, 1435–1436.
Mughogho, L.K., Pande, S., 1984. Charcoal rot of sorghum. In: Mughogho, L.K. (Ed.),
Sorghum roots and stalk rots – a critical review. Proceedings of the consultative
group discussion on research needs and strategies for control of sorghum root
and stalk rot diseases. Bellagio, Italy, 27 Nov–2 Dec, 1983. ICRISAT, Patancheru,
A.P. India.
Oba, M., Allen, M.S., 1999. Effects of brown midrib-3 mutation in maize silage on
dry matter intake and productivity of high yielding dairy cows. J. Dairy Sci. 82,
135–142.
Pedersen, J.F., Vogel, K.P., Funnell, D.L., 2005. Impact of reduced lignin on plant
fitness. Crop Sci. 45, 812–819.
Peter, G., Neale, D., 2004. Molecular basis for the evolution of xylem lignifications.
Curr. Opin. Plant Biol. 7, 737–742.
Pillonel, C., Mulder, M.M., Boon, J.J., Forster, B., Binder, A., 1991. Involvement of
cynamyl-alchol dehydroginase in the control of lignin formation in Sorghum
bicolor L Moench. Planta 185, 538–544.
Porter, K.S., Axtell, J.D., Lechtenberg, V.L., Colenbrander, V.F., 1978. Phenotype, fiber
composition, and in vitro dry matter disappearance of chemically induced
brown midrib (bmr) mutants of sorghum. Crop Sci. 18, 205–209.
Ralph, J., Hattfield, D., Rpiquemal, J., Yahiaoui, N., Pean, N., Lapierre, C., Boudet, A.M.,
1998. NMR characterization of altered lignins extracted from tobacco plants
down regulated for lignification enzymes CAD and CCR. Proc. Natl. Acad. Sci.
U.S.A. 95, 12803–12808.
Raven, P.H., Evert, R.F., EichhorN, S.E., 1999. Biology of plants, 6th ed. W. H. Freeman
and Company, New York.
Rook, J.A., Muller, L.D., Shank, D.B., 1977. Intake and digestibility of brown midrib
maize silage by lactating dairy cows. J. Dairy Sci. 60, 1894–1904.
Saballos, A., Vermerris, W., Rivera, L., Ejeta, G., 2008. Allelic association, chemical
characterization and saccharification properties of brown midrib mutants of
sorghum (Sorghum bicolor L Moench). Bioenerg. Res. 1, 193–204.
Stallings, C.C., Donaldson, B.M., Thomas, J.W., Rosman, E.C., 1982. In vivo evaluation
of brown midrib maize silage by sheep and lactating dairy cows. J. Dairy Sci. 65,
1945–1949.
Statistical Analysis Systems, 2004. SAS/STAT User’s Guide Version 9.1. SAS Institute,
Cary, North Carolina.
Tesso, T., Claflin, L., Tuinstra, M., 2004. Estimation of combining ability for resistance
to fusarium stalk rot in sorghum (Sorghum bicolor). Crop Sci. 44, 1195–1200.
Tesso, T., Claflin, L., Tuinstra, M., 2005. Analysis of resistance to stalk rot diseases
and genetic diversity among drought tolerant sorghum (Sorghum bicolor). Crop
Sci. 45, 645–652.
Vignols, F., Rigau, J., Torres, M.A., Capellades, M., Puigdomenech, P., 1995. The brown
midrib3 (bm3) mutation in maize occurs in the gene encoding caffeic acid O-
methyltransferase. Plant Cell 7, 407–416.
Vermerris, W., Saballos, A., Ejeta, G., Mosier, N.S., Ladisch, M.R., Carpita, N.C., 2007.
Molecular breeding to enhance ethanol production from corn and sorghum
stover. Crop Sci. 47, 142–153.
Vogler, R.K., Tesso, T., Johnson, K.D., Ejeta, G., 2009. The effect of allelic variation on
forage quality of sorghum mutants with reduced COMT activity. Afr. J. Biochem.
Res. 3, 70–76.
Whetten, R.W., Mackay, J.J., Seferoff, R.R., 1998. Recent advances in the understand-
ing of lignin biosynthesis. Annu. Rev. Plant Physiol. 49, 585–609.
Ye, Z.H., Zhong, R., Morisson III, W.H., Himmelsbach, D.S., 2001. Caffeoyl coenzyme
A O-methyltransferase and lignin biosynthesis. Phytochemistry 57, 1177–1185.