Int. Agrophys.

, 2019, 33, 21-30

doi: 10.31545/intagr/103752

Dynamic rheology and microstructure of starch gels affected by triticale genomic

composition and developing stage
Yaeel I. Cornejo-Ramírez1, Francisco J. Cinco-Moroyoqui1*, Elizabeth Carvajal-Millán2,
Francisco Brown-Bojórquez3, Ema C. Rosas-Burgos1, Armando Burgos-Hernández1,
Oliviert Martínez-Cruz1, and Carmen L. Del Toro-Sánchez1
1
Department of Food Research and Graduate Program (DIPA), University of Sonora, Hermosillo, Sonora 83000, Mexico
2
Laboratory of Biopolymers, CTAOA, Research Center for Food and Development, CIAD, A.C. Carretera a La Victoria Km. 0.6,
Hermosillo, Sonora 83304, Mexico
3
Department of Polymers, University of Sonora, Col. Centro, Blvd. Luis Encinas y Rosales S/N, Hermosillo, Sonora, C.P. 83000,
Mexico

Received January 28, 2018; accepted August 9, 2018

A b s t r a c t. Starches of developing triticale grains, differ- The major components of starch are glucose polymers,
ing in genome composition (complete AABBRR or substituted amylose and amylopectin. Amylose is a linear oligosac-
AABBDR), were evaluated in terms of starch granule distribution, charide made of D-glucose units bonded to each other
dynamic rheological behaviour and microstructural characteris-
through α-1,4-glycosidic bonds with a degree of polyme-
tics on several days after anthesis. The starch granules were of
an oblate spheroid shape for A-granules, and of a spherical shape
rization (DP) in the range of 500-6000 glucose residues.
for B-granules. However, those obtained from the complete triti- Amylopectin is a very large and highly branched chain
cale showed a larger diameter size. An X-ray diffraction analysis molecule with a DP ranging from 3 x 105 to 3 x 106 glu-
revealed the common A-type pattern of cereal starches from early cose units, and it consists of α-1,6-linked D-glucose units
development stages. A dynamic rheological analysis showed that attached to α -(1,4)- bonds (Jeon et al., 2010).
the storage and loss moduli reached maximum levels in the tem- Changes in the glucan chain length distribution or the
perature range of 71-86ºC and dropped at around 90ºC. Starches degree of crystallinity can alter starch physico-chemical
from the complete triticale showed lower phase transition tempe-
characteristics (Copeland et al., 2009). In the starch gran-
ratures, compared to those obtained from the substituted genotype
(56.1±0.3 and 60.3±0.8°C, respectively). Scanning electron micro-
ule, amylose and amylopectin are arranged in the alternating
scopy showed that the gels made with the starch of complete triti- amorphous and crystalline regions, respectively, forming
cales were of a less dense sponge-like structure. what is known as the growth rings. Amylose molecules
K e y w o r d s: triticale developing grains, starch, gels, rheo- associated with large branches of amylopectin comprise
logy, microstructure the amorphous region of granules whereas short branches
of amylopectin comprise the crystalline region, as a result
INTRODUCTION
of which a higher proportion of amylopectin in starch gran-
Starch is the major polysaccharide and carbon reserve ules results in a higher crystallinity degree (Cheetham and
of plants (Hizukuri et al., 1981). Starch in the cereal endos- Tao, 1998). Amylose exhibits the most useful functions as
perm is synthesized and accumulated to enhance plant and a hydrocolloid which forms gels and films. In contrast,
grain development. However, it is the grain-filling period amylopectin with long chains interferes with the interaction
that determines the final weight and quality of mature of amylose chains, preventing retrogradation, and leads to
grains (Zhao et al., 2003). viscosity loss (Chung and Liu, 2009).

*Corresponding author e-mail: [email protected] © 2019 Institute of Agrophysics, Polish Academy of Sciences
22 Y.I. CORNEJO-RAMÍREZ et al.

Starch in cereal grains is synthesized in amyloplasts by genotypes on different days after anthesis. The results
four types of enzymes, i.e. ADP-glucose pyrophosphory- might yield further insights into whether immature and
lase, starch synthase, as well as branching and debranching mature, complete and substituted, triticale grains could be
enzymes (Cao et al., 2012). The action of these enzymes is best suited for commercial production.
influenced by cultivar and environmental conditions, which
in turn affects the amount and size of the synthesized starch MATERIALS AND METHODS
granules as well as the chain-length (Ulbrich et al., 2015), Seed samples of a complete triticale (Eronga variety,
the amylose: amylopectin ratio (Altenbach et al., 2003; AABBRR genome) and a substituted triticale (Yoreme
Cornejo-Ramírez et al., 2016; Jiamjariyatam et al., 2015) variety, AABBDR genome) were employed in this study.
and the fine structure of amylopectin (Jane et al., 1999).
Grain samples of the two genotypes of triticale were plant-
All these properties influence the functional properties of
ed during the 2012-2013 planting cycle at the same time
starches, thus contributing to the final quality of food and
and under the same environmental conditions, at the agri-
non-food products. However, there is a continuous demand
culture experiment station of the Agriculture and Livestock
for new cultivars with improved agronomic characteristics,
such as better yield and kernel size, improved nutritional Department of the University of Sonora, Mexico (208 m
properties and optimal performance under extreme envi- above the sea level, with an average annual temperature
ronmental conditions. While the breeding of new and better of 24.2oC and an annual average rainfall of 280 mm).
cultivars through genetic crosses is required to overcome A completely randomized complete block design with three
their demand, it is important to ensure that the expected replications (plots) for each triticale genotype was used.
improved quality has been reached through agronomic and Triticale spike samples were collected on selected days
rheological performance tests. after anthesis (DAA). Eight spikes were cut from plants
Triticale (X Triticosecale Wittmack) is a hybrid cereal within each plot and shelled manually. Only developing
crop created from a cross between durum wheat (Triticum and mature grains from the middle part of the spikes were
durum) and rye (Secale cereale). Triticale varieties have collected and stored at –70 °C until analysis.
been developed as complete (with genomic constitutions Starch of triticale samples collected on different DAA
AABBRR and AABBDDRR) or substituted (with genomic was obtained from single kernels using the method pro-
constitutions AABBDR and AABBDDDR). The first sub- posed by Bettge et al. (1995) with modifications. Briefly
stituted triticale, called Armadillo, was obtained as a result speaking, the embryo and bran of the kernels were manu-
of a spontaneous crossing of triticale with an unknown ally removed with a razor blade; the remaining parts of the
Mexican semi-dwarf bread wheat. Taking advantage of the kernels were lightly crushed between two sheets of wax
highly heritable good agronomic characteristics resulting paper and transferred to a standard 1.5 ml Eppendorf poly-
from the fortuitous crossing, Armadillo materials were dis- propylene microcentrifuge tube. To prevent lumping, the
tributed by CIMMYT among plant breeders from all over solids were steeped twice with 500 µl of petroleum ether
the world, in order to be used for providing high fertility, for 30 min at room temperature with occasional gentle
better hectolitre weights and grain yield, insensitivity to agitation. After centrifugation at 10,000 g for 2 min and
photo-perforation, dwarfism, early maturity and good nutri- removal of the solvent, the precipitate was steeped twice
tional quality to the less promising triticales (Varughese et
in 500 µl of 100 mmol l-1 NaCl at room temperature for
al., 1987). With further genetic improvements and research,
30 min with occasional gentle agitation. Gluten was formed
both types of triticales have shown phenotypes and agro-
by kneading the endosperm inside the microcentrifuge
nomic differences. Complete triticales are more adaptable
tube, with the use of a Teflon micro-spatula, and then man-
and resistant to limiting conditions, showing higher pro-
ductivity, both in fertile areas and in adverse conditions. ually removed. The starch was washed twice with 500 µl of
However, substituted triticales have given rise to more vari- deionised water to remove excess of salt, following which
eties, since they are more stable and of better bakery quality it was centrifuged at 10,000 g for 2 min, re-suspended in
(Mellado et al., 2008). 1 ml of acetone and dried at ambient temperature.
However, few studies have been conducted to compare The size distribution of defatted starch granules
and contrast the differences in certain physicochemical was measured using laser diffraction in a Coulter LS
properties of starches among different triticale genotypes 100Q (Beckman Coulter, Miami, FL, USA). Big starch
during grain development. To our knowledge, no research granules (> 6 µm) are commonly referred to as A-type
has been carried out that would focus on the dynamic rheo- starch granules, whereas the small ones (≤ 6 µm) are
logical properties of starches affected by triticale genomic known as B-type starch granules. For analysis, a small
composition and grain developing stage. The aim of this quantity (20-30 mg) of starch was vortexed in 2 ml of
research was to determine and compare the dynamic rhe- deionised water and pumped through the optical cham-
ological and microstructural gel-forming properties of ber. A GB500 standard, consisting of a glass bead sample
starches isolated from developing grains of two triticale (500 µm nominal mean particle size), was used as a reference.
 RHEOLOGY AND MICROSTRUCTURE OF STARCH GELS FROM DEVELOPING TRITICALE GRAINS 23

The crystallinity percent of triticale starches was deter- All data were obtained from three independent experi-
mined using the X-ray diffraction methodology proposed ments and expressed as mean ± standard deviation of
by Song and Jane (2000). Starch samples were equilibrat- triplicates. Data were subjected to one-way analysis of
ed in a 100% relative humidity chamber for 24 h at 27ºC. variance (ANOVA) following general model procedures.
X-ray diffraction patterns (XRD) were obtained with nickel A comparison of sample means was performed by means of
foil-filtered Cu Kα radiation, using a Rigaku RINT-2000 the Tukey´s test with the SAS program (SAS, 2005). In all
diffractometer (Tokyo, Japan), at 27 mA and 50 kV. The cases, the mean values were considered significantly diffe-
scanning region or the two-theta angle (2θ) ranged from rent at p ≤ 0.05.
5 to 50° with a 0.05° step size and a count time of 2 s. RESULTS AND DISCUSSION
The crystalline percent values were calculated according to The granule size distribution of the starch strongly influ-
Hayakawa et al. (1997), by means of the following equa- ences its physicochemical properties and functionality. In
tion: Crystallinity percent = Ac/ (Ac + Aa) 100, where Ac this study, the size distributions of starch granules of deve-
and Aa correspond to the crystallinity and amorphous are- loping triticale grains were measured using laser diffraction.
as, respectively, on the X-ray diffractograms. Table 1 shows the size distributions of starch granules of
Rheology is a convenient tool to study starch gelation complete (Eronga) and substituted (Yoreme) triticale deve-
since a physical gel is formed during starch gelatinization. loping grains. Both triticale genotypes showed a bimodal
The gelatinization process can be analyzed by monitoring starch granule distribution similar to that reported previ-
the rheological properties using a rheometer. The gelatiniza- ously (Cornejo-Ramírez et al., 2015) with oblate spheroid
tion properties of the starches isolated from the developing shape for A-granules and spherical shape for B-granules
grains of the two triticale genotypes were studied using (Bechtel et al., 1990). In general, A- and B-type starch
a stress-controlled Discovery Hybrid Rheometer HR-2 (TA granules of both triticale genotypes increased in size dur-
Instruments, New Castle, DE, USA) equipped with two ing grain development, with differences in size distribution
parallel plates (Peltier Plate Steel-104556) with 40 mm in values as well as in granules volume percent observed at
diameter. A volume of 1.5 ml of 10% starch-water suspen- all stages of development. However, A- and B-type starch
sions was routinely placed on the centre of the bottom plate granules of the complete triticale showed bigger sizes than
while the upper plate was immediately lowered until the those obtained from the substituted genotype throughout
final gap of 1 mm was reached. The edges of the plates were the course of the developing period of grains. The data also
covered with paraffin oil in order to prevent dehydration of show that the A- and B-type starch granules of the com-
the samples by evaporation. Starch samples were examined plete triticale grew faster and reached larger diameters than
in the angular frequency sweep scan range of 0.01-60 Hz the granules of the substituted triticale. A similar behaviour
at a constant strain of 1%. The start temperature was 25°C, was observed in wheat starch granules during development
and a ramp temperature of 5°C min-1 was used until the (Bechtel et al., 1990; Kim et al., 2003).
final temperature of 90°C was reached. After 30 s of releas- Starch molecules arrange themselves in the grain in
crystalline granules. Amylose molecules comprise the
ing the stress, the samples were cooled to 25°C at a cooling
amorphous region of granules, whereas amylopectin mole-
rate of 5ºC min-1. After 300 s of soaking time, the gels were
cules with short branches comprise the crystalline region.
frozen at -20 °C, lyophilized and stored for further analysis.
Crystallinity of starch granules has been directly associated
The rheological parameters elastic modulus (G′) and vis-
with amylopectin concentration (Cheetham and Tao, 1998)
cous modulus (G′′) were obtained at the frequency sweep
and inversely related to onset temperature (T0), peak tem-
range indicated above. The phase transition temperature T0,
perature (Tp) and enthalpy (DH) (Kim et al., 2003). There
i.e. the crossover where the G’ and G’’ values started to are three types of crystalline structures, i.e. A-type found in
increase at the same temperature and representing the sol- cereal starches, B-type found in tubers and C-type present in
to-gel transition, was determined in the dynamic modulus legumes (Singh et al., 2003). In our study, the starches pre-
spectrum (G’ and G’’) during heating (Ulbrich et al., 2015). sented the typical A-type X-ray diffraction pattern (Fig. 1)
Scanning Electron Microscopy (SEM) was used to ana- observed in cereal starches (Kim et al., 2015). The diffrac-
lyze the microstructure of the triticale starch gels using the tograms of triticale starches showed peaks at 15, 17.1, 18
method proposed by Jane et al. (1994). The lyophilized and 23° from the early stage until their maturity.
starch gels were placed on a 13 mm silver tape, metal- The starch samples displayed the same X-ray dif-
shadowed with gold/palladium (60/40) (Sputter coater fraction pattern because their chemical composition and
SPI-Module; West Chester, PA, USA) and mounted on physicochemical properties are similar. Also, intensities of
a brass disk. Starch samples were analyzed using a mag- each peak of the starches isolated from developing grains
nification of 1500 X under low vacuum, using a JEOL on 16, 22 and 31 DAA are similar, whereas the intensi-
JSM-5400LV scanning electron microscope (Peabody, ties of the peaks of these starch samples are different from
MA, USA), at an acceleration voltage of 15 kV. those obtained from mature grains. This suggests that the
24 Y.I. CORNEJO-RAMÍREZ et al.

T a b l e 1. Starch granule size distribution and volume (%) of A- and B-type starch granules obtained from complete (Eronga) and
substituted (Yoreme) triticales during grain development on different days after anthesis (DAA)

A-type B-type
DAA
Size (µm) Volume (%) Size (µm) Volume (%)

Complete triticale (Eronga)

16 6.7-23 86.3 ± 0.06aB 0.96-3.7 13.7 ± 0.06bC
22 8.5-30 85.5 ± 0.20aC 0.98-4.5 14.5 ± 0.20bB
31 14-36 87.6 ± 0.20aA 1.5-5.2 12.4 ± 0.20bD
40 18-40 80.4 ± 0.03ªD 2.0-11.0 19.6 ± 0.03bA
Substituted triticale (Yoreme)
16 6.7-20 86.7 ± 0.05aB 0.94-3.4 13.3 ± 0.05bC
22 9.0-24 88.0 ± 0.09aA 0.96-3.6 12.0 ± 0.09bD
31 11.2-33 86.3 ± 0.30aC 1.0-4.2 13.7 ± 0.30bB
40 12-38 74.6 ± 0.05aD 1.2-6.0 25.4 ± 0.05bA
Mean values are expressed in percentage ± standard deviation. Values followed by the same lowercase letter in the same row and capital
letter in the same column are not significantly different (p≤0.05, n=3).
amylopectin shapes of starches from mature grains are The gelatinization process triggers several changes in
different from those of starches isolated from immature the physicochemical properties of starches which be meas-
grains. Based on the results provided by the X-ray diffrac- ured with a rheometer. At the sol-gel transition, the starch
tion analysis of inorganic crystals, carried out by Inoue and solution loses its liquid properties and begins to turn into
Hirasawa (2013), the total area, and the longitude and lati- a solid substance. In this study, the gelatinization mecha-
tude of crystals affect the X-ray peak intensities. As shown nism of starches from complete (Eronga) and substituted
by Cornejo-Ramírez et al. (2016), reduction in the intensity (Yoreme) developing triticale grains was studied by means
of the X-ray diffraction patterns of triticale starches might of a dynamic mode rheological analysis. Oscillatory measu-
be due to reduction in the volume of the A-type starch gran- rements were performed within the linear viscoelasticity
ules at the end of the maturation period. Additionally, the region and the results provided information on the dynamic
crystallinity of those starches increases due to a compaction mechanical properties of the starch gels evaluated as the
of the amylopectin chains to form the crystalline areas. elastic modulus G’ and the viscous modulus G”. Figure 2A,B
A comparison of the crystallinity values observed along shows the evolution of the elastic (G’) and viscous (G’’)
the whole developing period showed that the complete moduli of starch granules from the complete and substi-
and substituted triticale starches were similar, reaching tuted triticales, respectively, as a function of temperature.
final values of 36.1% on 40 DAA (Table 2). Furthermore, The changes in G’ and G’’ of the 16 to 40 DAA starch dis-
an analysis of the crystallinity values of each triticale persions displayed similar behaviour. In general, for both
throughout the developing period showed that the complete triticale genotypes the G’ and G’’ moduli values reached
triticale did not exhibit significant differences in crystallin- maximum levels in the temperature range of 71-86ºC, and
ity from 16 to 31 DAA, but on 40 DAA displayed a value dropped with further heating at around 90ºC. Both moduli
significantly higher than those observed at early stages. showed the same profile; however, G’ displayed higher val-
In contrast, the starch granules of the substituted triticale ues than G’’ in both triticale genotypes, although those of
showed continuous changes in crystallinity until the end of the complete genotype were higher than the substituted gen-
the maturity period. The final values in crystallinity of the otype. Higher values of G’ and G’’ have also been observed
triticale starch granules were similar to those observed in in corn (Kaur et al., 2008) and wheat (Ulbrich et al., 2015)
the starch of wheat (Zhang et al., 2013), as well as barley starch dispersions. The effect of frequency on the elastic
and triticale (Ao and Jane, 2007), though they were higher modulus G’ and viscous modulus G’’ of the starch of devel-
than those reported for waxy wheat (Yoo and Jane, 2002). oping complete (Eronga) and substituted (Yoreme) triticale
Differences in percent crystallinity values of starches from grains are shown in Fig. 2C, D. All samples showed a solid-
several sources have been attributed to the amylose (Yoo like system. The values of G’ and G’’ moduli of all triticale
and Jane, 2002), lipid (Finnie et al., 2009), and amylopec- starches were independent of frequency, which is typical
tin contents and amylopectin chain length, as well as the of ‘true gels’ (Sang et al., 2008). Also, all starch samples
orientation of amylose double helices (Zhang et al., 2013). showed G’ values higher than those of G’’, and were higher
 RHEOLOGY AND MICROSTRUCTURE OF STARCH GELS FROM DEVELOPING TRITICALE GRAINS 25

Fig. 1. X-ray diffraction of the starch of complete (AABBRR) triticale on 16, 22, 31, and 40 DAA (A, B, C and D, respectively) and
substituted (AABBDR) triticale on 16, 22, 31, and 40 DAA (E, F, G and H, respectively).
26 Y.I. CORNEJO-RAMÍREZ et al.

Ta b l e 2. Crystallinity percent of starch granules from complete in the complete triticale than in the substituted one, sug-
(AABBRR) and substituted (AABBDR) developing triticale gesting a gel-like behaviour similar to that observed in the
grains on different days after anthesis (DAA) starch of corn (Kaur et al., 2008), sorghum (Sang et al.,
2008) and wheat (Ulbrich et al., 2015). Furthermore, the
Crystallinity (%) starch of both triticale genotypes exhibited very similar G’’
DAA Complete triticale Substituted triticale values, suggesting that its viscous contribution to the gel
(Eronga) (Yoreme) structure was similar throughout its synthesis in the devel-
oping grains.
16 26.5 ± 0.7aB 27.0 ± 0.6aC The sol-gel transition temperature T0 was determined
22 27.3 ± 0.4aB 28.1 ± 0.3aBC at the crossover point of G’ and G’’. Table 3 shows the T0
values of starches from complete (Eronga) and substituted
31 28.6 ± 0.7aB 29.2 ± 0.4aB (Yoreme) developing triticale grains. The starches of the
40 36.1 ± 0.1aA 36.8 ± 0.3aA
developing complete triticale grains showed significant-
ly lower T0 values than those of the substituted triticale
Explanations as in Table 1. throughout the developing period. The T0 values of com-
plete and substituted triticale were the highest on 16 DAA

A B

Temperature (°C) Temperature (°C)

C D

Fig. 2. Changes in elastic G’ and viscous G’’ modulus during heating (the angular frequency sweep scan range of 0.01-60 Hz; constant
strain 1%; ramp temperature 5°C min-1; start temperature 25°C; final temperature 90°C) of 10% starch suspensions of the complete
triticale (AABBRR) and the substituted triticale (AABBDR) by the effect of temperature (A and B) and frequency (C and D) using
a stress-controlled Discovery Hybrid Rheometer HR-2 equipped with two parallel plates with 40 mm in diameter.
 RHEOLOGY AND MICROSTRUCTURE OF STARCH GELS FROM DEVELOPING TRITICALE GRAINS 27

Ta b l e 3. Sol-to-gel phase transition temperature (T0), degree of polymerization of amylose and apparent amylose content of starches
extracted from complete (AABBRR) and substituted (AABBDR) developing triticale grains on different days after anthesis (DAA)

Transition temperature Degree of polymerization

DAA Apparent amylose (%)*
(T0, oC) of amylose*

Complete triticale
16 63.9±0.4aB 375±49a 13.5±0.2b
22 63.6±0.5aB 405±36a 11.7±0.7a
31 59.5±0.7bB 811±42a 16.5±0.8a
40 56.1±0.3cB 1549±42a 23.5±0.8a
Substituted triticale
16 66.3±0.3aA 254±38b 14.1±0.2a
22 65.2±0.2aA 287±43b 11.3±0.7a
31 62.7±0.3bA 756±41b 15.9±0.6b
40 60.3±0.8cA 1313±62b 22.1±0.7b
Values are expressed as mean ± standard deviation. Values followed by the same: lowercase letter and capital letter in the same column
are not significantly different for similar DAA (p≤0.5). *Data taken from Cornejo-Ramírez et al. (2016).

(63.9 ± 0.4 and 66.2 ± 0.3ºC, respectively) and 22 DAA tures (T0) and were related to higher amylose contents and
(63.6 ± 0.5 and 65.2 ± 0.2ºC, respectively), and decreased higher amylose DP, in contrast to those of the substituted
with time, reaching the lowest values on 40 DAA (56.1 ± triticale Yoreme (Table 3).
0.3 and 60.3 ± 0.8°C, respectively). The highest T0 values The gels made with the starch of triticales developed
were observed in starches of the substituted triticale from on 16 DAA showed large air cells of sponge-like struc-
the 16 DAA and to 22 DAA with no significant diffe- tures (Fig. 3), which has also been observed in waxy corn
rences. However, the transition T0 values determined in this (Kaur et al., 2008) and corn starch (Alishahi et al., 2015).
However, both gels displayed some differences. The starch
research for both triticale starches were lower than those
gel of complete triticale (Eronga) showed an internal struc-
reported for acid-modified wheat (69.4°C), potato (80°C)
ture more similar to the gels of corn starch, while the starch
and pea (80°C) (Ulbrich et al., 2015). Yet, they fell with- gel of the substituted triticale (Yoreme) displayed an inter-
in the range of values reported for normal corn (65.7°C) nal structure showing small air cells within large ones.
(Kaur et al., 2008) and normal wheat (46-57.1°C) starches The gels made with the triticale starch samples on 22
(Sasaki et al., 2000). The differences in T0 of starches from DAA formed a closed network, creating small cells of
diverse cereal cultivars might be due to variations in the sponge-like structures, similar to the internal structure of
granular structures, degree of polymerization and molecu- the gels made with normal corn starch (Kaur et al., 2008).
lar architecture of amylopectin (Singh et al., 2003). The internal structure displayed by all these gels might be
The higher percent crystallinity of the starch makes the due to higher DP of amylose (Table 3) that could make
granules more resistant to gelatinization because the longer a thick matrix, capable of filling the space between the
chains of amylopectin require higher temperatures to dis- remnants, resulting in a porous structure due to the forma-
sociate completely, while the opposite occurs for amylose tion of voids, similar to a honeycomb-like structure. The
(Yamin et al., 1999). Also, a starch with high amylose con- differences in the gel microstructures might be due to the
tent has a more amorphous region and, therefore, a less differences in the physicochemical characteristics of the
developing starch granules to make the gels. The starches
crystalline region, as a result of which low temperatures
of complete and substituted triticales differed in terms of
are needed to achieve gelatinization (Sasaki et al., 2000).
the DP of amylose (Cornejo-Ramírez et al., 2016), size and
In our study, the starches from the substituted triticale volume percent of A- and B-starch granules (Table 1) at this
genotypes showed, in general, higher crystallinity values stage of development.
throughout the developing period, although no significant The gels made with the starch samples of the triticale
statistical differences were observed. collected on 31 DAA formed a network with larger cells of
The T0 values were strongly influenced by genotype, sponge-like structures, though not as large as those of the
considering that the starches of the developing grains of the gel network made with the starch formed on 16 DAA. The
complete triticale Eronga showed lower transition tempera- internal structures of the gels were similar to those made
28 Y.I. CORNEJO-RAMÍREZ et al.

Fig. 3. Scanning electron micrographs of starch gel samples prepared from starches of developing grains of a complete triticale
(AABBRR) on 16, 22, 31, and 40 DAA (A, B, C and D, respectively), and of a substituted triticale (AABBDR) on 16, 22, 31, and 40
DAA (E, F, G and H, respectively) using a magnification of 350x.
with waxy corn starch (Kaur et al., 2008); however, the sweeps, low stability of pastes at low temperatures and low
starch gel of the complete triticale showed similar areas to resistant to retrogradation. The gels made with substituted
the gels made with wheat starch-wheat fibre (Sun et al., triticale showed the same rheological behaviour as the corn
2015) and corn starch (Alishahi et al., 2015). starch gels, and they could have low cohesiveness and high
The gels made with the triticale starch samples at matu- values of springiness, hardness, gumminess and viscosity
rity (40 DAA) showed differences among them in their (Sun et al., 2015).
internal structures. The gel made with starch of complete CONCLUSIONS
triticale displayed an internal structure with a minor dense
honeycomb structure, with reduced internal pores, and 1. The results showed that the starches of the complete
a large lamellar structure similar to the gels made with genotype displayed different gel properties than those
wheat starch (Sun et al., 2015). Therefore, large starch gels obtained from the substituted genotype, due to differences
have the advantage in competing for water molecules form- in the molecular structure of their amylose and amylopec-
ing a strong network structure that enhances the mechanical tin, synthesized on different DAA during grain developing.
properties of noodles, vermicelli and sheet jelly (Alishahi 2. The complete triticale developed bigger and more
et al., 2015). In contrast, the gel made with the starch of crystalline A- and B-type starch granules than the sub-
substituted triticale displayed more similarities to the gels stituted triticale. Although no significant differences in
prepared with wheat starch (Sun et al., 2015) and normal crystallinity were observed between the starches collected
corn starch (Kaur et al., 2008). The corn starch gels showed n different DAA, the substituted triticale showed higher
low peak viscosity, G’, G’’ and Tp and less pronounced crystallinity values, which coincided with the low amylose
changes in the rheological parameters within the frequency content reported in a previous study.
 RHEOLOGY AND MICROSTRUCTURE OF STARCH GELS FROM DEVELOPING TRITICALE GRAINS 29

3. Dynamic rheology determinations revealed that the Cornejo-Ramírez Y.I., Cinco-Moroyoqui F.J., Ramírez-Reyes
complete triticale displayed higher values of elastic G’ and F., Rosas-Burgos E.C., Osuna-Amarillas P.S., Wong-
Corral F.J., Borboa-Flores J., and Cota-Gastélum A.G.,
viscous G’ moduli than the substituted triticale. 2015. Physicochemical characterization of starch from
4. Starches of the complete triticale collected on several hexaploid triticale (X Triticosecale Wittmack) genotypes.
DAA showed lower phase transition temperature values CyTA J. Food, 13(3), 420-426.
than those obtained from the substituted one, suggesting Cornejo-Ramírez Y.I., Ramírez-Reyes F., Cinco-Moroyoqui
F.J., Rosas-Burgos E.C., Martínez-Cruz O., Carvajal-
lower gelatinization temperatures.
Millán E., Cárdenas-López J.L., and Wong-Corral F.,
5. The microstructure of gels prepared from starches 2016. Starch debranching enzyme activity and its effects on
extracted from the complete triticale grains displayed a den- some starch physicochemical characteristics in developing
ser internal structure. substituted and complete triticales (X Triticosecale
Wittmack). Cereal Chem., 93(1), 64-70.
Conflict of interest: The authors have no conflict of Finnie S.M., Jeannotte R., and Faubion J.M., 2009. Quantitative
characterization of polar lipids from wheat whole meal,
interest to declare.
flour, and starch. Cereal Chem., 86(6), 637-645.
Hayakawa K., Tanaka K., Nakamura T., Endo S., and Hoshino
ACKNOWLEDGEMENTS T., 1997. Quality characteristics of waxy hexaploid wheat
(Triticum aestivum L.): properties of starch gelatinization
The authors wish to thank the Wheat Collection and retrogradation. Cereal Chem., 74, 576-580.
Hizukuri S., Takeda Y., Yasuda M., and Suzuki A., 1981.
Wellhausen-Anderson Plant Genetic Resource Center of
Multi-branched nature of amylose and the action of
the International Maize and Wheat Improvement Center, debranching enzymes. Carbohyd. Res., 94(2), 205-213.
CIMMYT-México for kindly donating the triticale seeds Inoue M. and Hirasawa I., 2013. The relationship between crys-
used in this study. Author Yaeel I. Cornejo-Ramírez tal morphology and XRD peak intensity on CaSO4 × 2H2O.
acknowledges the National Council for Science and J. Cryst. Growth, 380, 169-175.
Jane J., Chen Y.Y., Lee L.F., McPherson A.E., Wong K.S.,
Technology (CONACyT, Mexico) for providing a doctoral Radosavljevic M., and Kasemsuwan T., 1999. Effects of
fellowship. amylopectin branch chain length and amylose content on
the gelatinization and pasting properties of starch 1. Cereal
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