Journal of Phytopathology and Pest Management 6(1): 54-65, 2019

pISSN:2356-8577 eISSN: 2356-6507

Journal homepage: http://ppmj.net/

∗

Corresponding author:

Eman F. M. Tolba,

E-mail: dr.mero_83@yahoo.com

54

Evaluation of certain Egyptian wheat cultivars

against rice weevil,

Sitophilus oryzae

(L.)

(Coleoptera: Curculionidae) using biochemical

and biophysical traits

Marwa F. K. Aly

*

, Ali M. Ali

Plant Protection Department, Faculty of Agriculture, Minia University, El-Minya, Egypt

Abstract

Keywords: stored grain pests, wheat cultivars susceptibility, biophysical and biochemical traits, Sitophilus oryzae.

55

1. Introduction

Wheat can be considered the third cereal

crop in terms of world production.

According to FAOSTAT,

wheat

production exceeded 711 million tonnes

in 2013. Also, wheat occupies more than

220 million hectares of land worldwide

(Anonymous, 2015).

Durum wheat

(

Triticum turgidum

subsp.

durum

) is one

of the most important cereal crops. About

half of the total durum wheat-producing

area is located in developing countries,

where it is used for making products such

as pasta and couscous (Herrera-Foessel et

al., 2014). In 2014, the total cultivated

wheat area in Egypt, including bread and

durum wheat, was 8.9 million acres and

the total production was 9.3 million

tonnes (FAO, 2014; CAPMAS, 2014)

which is away less than the consumed

wheat. Cereal grain losses during storage

can reach to 50% of total harvest in some

countries, a worldwide loss quality of

grain is caused by insects (Fornal et al.,

2007). Many pests attack wheat plant in

the field and the stored grain in the

storage and cause a severe damage.

Sitophilus oryzae

(L.) (Coleoptera:

Curculionidae) is a serious cereal pest

that attacks stored wheat grain.The adults

fed on endosperm, hence declining the

carbohydrate contents. The larvae feed on

the germ of the grain and reducing great

amount of protein and vitamins (Belloa et

al., 2000). Rice weevil female chews a

hole in the kernel, lays the egg inside,

and seals the hole with a gelatinous

secretion which protects the eggs

(Arbogast, 1991). Weevil has a life cycle

of 34.8 days in average at 27˚C and 69%

relative humidity (Osman et al., 2012). It

has direct and indirect impact on the

grain. The direct impact comes as a result

of weevils feeding which decreasing the

quantity and the available grain for

human consumption. The indirect impact

is due to affecting the quality of the grain

which may leads to lowering its price or

rejection of grain. In general, the

performance of cereal cultivars against

weevils infestation varied from cultivar

to another (Khan et al., 2014; Shafique &

Ahmad, 2003). In the past few decades

possibility of identifying resistant

cultivars of wheat to stored grain pest

have received attention. Wheat

cultivation in respect to cultivars has

been undergoing a marked change in

recent past. New cultivars are coming up

every year, which resulted a change in

the intensity of insect infestation during

storage. In recent year the laboratory

studies established that many cultivars

have shown some resistances against

S.

oryzae

(Chauhan et al., 2005; Tiwari &

Sharma, 2002; Ram & Singh, 1996).

However, a few studies have been

implemented on evaluating the resistance

of recent growing Egyptian wheat

cultivars against

S. oryzae

. Many factors

can help the development of stored grain

pests and cause losses ranging from 5-

30%, (Khan et al., 2014). High moisture

content of grain, relative humidity and

high environmental temperature during

storage provide suitable condition for

insect’s production (Dars et al., 2001;

Ahmad et al., 1998). The resistant

cultivars, especially in village

cooperative and farmer’s stores, can be

kept for longer time without use of

pesticides (Semple, 1985). Thus, this

study aims to evaluate some Egyptian

spring bread and durum wheat for

S.

oryzae

resistance by studying the effect

of some biochemical and biophysical

traits on the feeding of adult weevils.

56

2. Materials and methods

2.1 Insect rearing

Insects were reared in 1 L capacity glass

gars with a 7 cm diameter which were

covered with muslin cloth and tied by

rubber bands. The gars containing grain

of mixture wheat varieties were kept in a

controlled environment (28 ± 1 °C, and

70% RH). The grain periodically sieved,

and the insects placed in rearing gars

with fresh grain.

2.2 Source of tested Egyptian wheat

cultivars

Eleven wheat cultivars including 7 durum

wheat (Beni Sweaf-1, Beni Sweaf-3,

Beni Sweaf-4, Beni Sweaf-5, Beni

Sweaf-6, Sohag-3, and Sohag-5) and 4

bread wheat (Misr-1, Misr-2, Giza-171,

and Gimmiza-10) were obtained from

institution of wheat diseases research,

Agriculture research centre, Dokki, Giza

Egypt.

2.3 Experimental procedures

The experiments were conducted in small

glass jar (5cm diam. and 7.5 cm length),

the jars were covered with mesh cloth.

Thirty grams of wheat grain were placed

in each jar. The infestation was

conducted by adding 30 newly emerged

adults to each jar (1 weevil/1gm grain).

Thirty grams of whole grain were

accurately weighted and placed in small

glass jar (5cm diam. and 7.5 cm length),

30 newly emerged adults of each tested

insect species were introduced to glass

jars, which were covered with muslin

cloth and tied by rubber bands. The

adults were left for one month then

removed, and the remaining grain was

used for calculating the % Weight loss

and % damaged grain. As following

formulas:

Weight loss (%) =







Damaged grain (%) =







2.4 Biochemical traits measurements

2.4.1 Starch content

The colorimetric technique was used for

estimation of starch percentage based on

method by (Magel, 1991).

2.4.2 Protein content

The protein content and moisture% (a) of

the flour was determined using Percon

Inframatic 8620 near infrared

spectrometer (Perten Instruments NA,

Inc., Reno, NV) and the data was

processed using Perten’s NetPlus

software.

2.4.3 Gluten estimation

Gluten determination was done as

described by (Imran et al., 2013) which

carried out using a Perten Glutomatic

System, based on International Code

Council (ICC) Standard No. 155, No 158

and American Association for Cereal

Chemist (AACC) method 38-12

(Operational Manual Glucomatic

System). This method uses a glutomatic

gluten washer with a sieve size of 170

mesh (88 microns) and a gluten

centrifuge which provides information on

the quantity and quality of gluten

obtained. 10g of wheat flour was

transferred into the wash chamber and

shaken to obtain a homogenous flour

layer. 4.8 ml of 2% sodium chloride was

added from the dispenser into the

57

chamber at a slight tilt. The chamber was

then agitated to spread the water evenly

over the flour. The remaining mixing and

washing sequence was accomplished

automatically within the washer. Liquid

containing starch was collected in a

beaker placed below the washer and the

gluten mass remained on the sieve. The

gluten mass was centrifuged in a special

sieve cassette in order to force the wet

gluten to pass through the sieve. The

centrifuge allowed for the collection of

both parts of the gluten remaining on the

sieve and that which passed through the

sieve. The total weight of wet gluten was

recorded and expressed as a percentage

of the mass of the original sample. The

following formula was used for the

calculation:

Wet gluten content (%) = [Total gluten (g) x

100/ Weight of wheat flour sample (10g)]

2.5 Biophysical traits determination

The colorimetric technique was used for

estimation of starch percentage based on

method by (Magel, 1991).

2.5.1 Determining the kernel hardness,

weight, diameter, and moisture by

SKCS 4100

Grain moisture, kernel diameter, kernel

weight, and hardness index were

determined using the Perten Single

Kernel Characterization System (model

4100, Perten Instruments, Reno, NV)

(Psotka, 1999; Gaines et al., 1996).

Hardness Measurement by SKCS A

Single Kernel Characterization System

(SKCS) instrument, (model 4100. Perten

Instruments, Reno, NV) was used for

assessment of single wheat kernel

hardness, moisture, and kernel diameter

and weight. Duplicates of each wheat

cultivar were submitted to the SKCS

under normal operating parameters and

conditions, with a selection of 300 seeds

per measurement. During operation, the

instrument isolated individual wheat

kernels, measured, weighed, and crushed

them in a progressively narrower gap

formed by a toothed rotor and a crescent.

The crushing force and electrical

conductivity between the rotor and

electrically isolated crescent were

recorded. Those data were then

processed by the integrated computer

software to provide the means and

standard deviations for weight, size,

moisture, and hardness index (HI).

2.5.2 Moisture percentage

Moisture was measured with two

different methods first moisture (a)%

which measured by using Percon

Inframatic 8620 near infrared

spectrometer (Perten Instruments NA,

Inc., Reno, NV) and moisture (b)%

which measured by using SKCS A

Single Kernel Characterization System

(SKCS) instrument, (model 4100. Perten

Instruments, Reno, NV).

2.6 Statistical analysis

The experiment was laid out in complete

randomized design (CRD) with three

replications. The obtained data were

analyzed by ANOVA test and significant

means were separated by Duncan

’

s

multiple rang test using a computer

program of SPSS 14.0.

3. Results

Results were evaluated on the basis of

58

percent weight loss, percent grain

damaged of wheat grain because of the

infestation by

S. oryzae

for grain of 11

Egyptian wheat cultivars. Also,

biophysical and biochemical traits for

these cultivars were measured and the

potential correlation between these traits

and

S. oryzae

infestation was calculated.

3.1 Estimation of

S. oryzae

infestation

The percentage of weight loss for

grainand damaged grain were chosen as

an associated parameter to

S. oryzae

infestation. Percentage of Weight loss

and damaged grain weremeasured for11

Egyptian wheat cultivars including (7

durum wheat cultivars and 4 bread wheat

cultivars) (Table 1).

3.1.1 Weight loss percentage

Significant differences were found

among tested cultivars for percentage of

weight loss for grain (ANOVA-One way,

df=10, F=42.86, P<0.01) (Table 1). All

durum wheat cultivars showed less loss

weigh percentages compared with bread

wheat cultivars. Gemmeiza-10 showed

significantly the highest percentage of

weight loss for grain (6.11%) compared

to all tested cultivars. While Beni Sweaf-

3 had the lowest percentage of weight

loss (0.83%) than Beni Sweaf-5, Misr-1,

Misr-2, Giza-171 and Gemmeiza-10 with

a range of 1.96-6.11% of Weight loss.

But no significant differences were found

between Beni Sweaf-3 and other durum

cultivars.

3.1.2 Damaged grain percentage

The infestation by

S. oryzae

had similar

influence on the tested cultivars in

respect to percentage of damaged grain as

that in weight loss. It showed significant

differences among cultivars (ANOVA-

One way, df=10, F=612.20, P<0.01). The

infestation resulted a significantly more

damage for bread wheat cultivars

compared to that in durum. The highest

percentage of damaged grain (22.95%)

was observed in Gemmeiza-10 compared

to all tested cultivars. While, Beni

Sweaf-3 followed by Beni Sweaf 6 had

significantly lowest percentage of

damaged grain (5.28% and 5.68%,

respectively) than other tested cultivars.

3.2 Biophysical parameters

Four biophysical traits were measured

including; moisture (a), moisture (b),

kernel diameter, kernel weight, and

hardness index for the tested 11 Egyptian

wheat cultivars including (7 durum

wheat cultivars and 4 bread wheat

cultivars) (Table 1).

3.2.1 Percentage of moisture (a)

There is no significant differences were

noticed between tested cultivars in

moisture (a) (ANOVA, one-way, df=10,

F= 1.19, P>0.05). Minor differences

were observed among tested

cultivars.Giza-171 showed low

percentage for moisture (a) followed by

Sohag-5 (12.93% and 12.96%,

respectively) than Misr-1and Beni

Sweaf-1 (13.63% and 13.66%). While,

Beni Sweaf-1 had high percentage of

moisture (a) (13.66%) than Giza-171 and

Sohag-5.

3.2.2 Percentage of moisture (b)

The estimation of moisture using SKCS

produced similar results to moisture (a).

No significant differences were found

59

among tested cultivars (ANOVA, one-

way, df=10, F= 2.10, P>0.05). High

percentage of moisture (b) was recorded

for Giza-171 followed by Gemmeiza-10

(13.32% and 13.37%) compared to Beni

Sweaf-1, Beni Sweaf-3, Beni Sweaf-4,

Beni Sweaf-5 and Misr-1 with a range of

12.61%-12.78%. Otherwise, Misr-1 had

the lowest percentage of moisture (b)

(12.61%).

Table 1: Susceptibility of wheat cultivars to the infestation of S. oryzae and the variation between biophysical traits.

Wheat cultivar

Biophysical traits (Mean± SE)

Weight loss (%)

Damaged grain (%)

Moisture (a) (%)

Moisture (b) (%)

Diameter (mm)

Weight/ kernel (mg)

Hardness index

Beni Sweaf-1

1.79±0.66

7.35±0.12

13.66±0.27

12.78±0.14

3.06±0.06

43.40±0.13

97.61±0.22

Beni Sweaf-3

0.83±0.30

5.28±0.09

13.33±0.22

12.67±0.16

2.89±0.03

42.61±0.15

98.82±0.23

Beni Sweaf-4

1.57±0.40

7.00±0.38

13.56±0.25

12.69±0.15

3.25±0.06

52.51±0.17

93.98±0.24

Beni Sweaf-5

1.96±0.55

8.98±0.55

13.51±0.25

12.73±0.13

2.89±0.03

39.20±0.13

96.99±0.26

Beni Sweaf-6

1.55±0.17

5.68±0.09

13.39±0.24

12.82±0.14

3.18±0.06

48.31±0.16

93.91±0.24

Sohag-3

1.19±0.66

7.40±0.12

13.23±0.21

12.89±0.17

3.45±0.07

56.49±0.14

94.53±0.23

Sohag-5

1.58±0.66

6.40±0.12

12.96±0.20

12.95±0.18

3.29±0.06

51.01±0.15

100.0±0.26

Misr-1

3.42±0.10

13.78±0.09

13.63±0.26

12.61±0.14

3.44±0.07

57.93±0.16

72.95±0.22

Misr-2

2.26±0.51

11.08±0.14

13.15±0.22

12.84±0.17

3.31±0.06

50.42±0.17

77.54±0.23

Giza-171

4.66±0.82

16.10±0.06

12.93±0.20

13.32±0.19

3.03±0.05

43.66±0.15

67.05±0.22

Gemmeiza-10

6.11±0.86

22.95±0.03

13.23±0.23

13.37±0.18

3.12±0.06

45.72±0.16

48.45±0.20

3.2.3 Diameter (mm)

Wheat grain diameter (mm) was

measured for all tested cultivars.

Significant differences were found

among tested cultivars (ANOVA, one-

way, df=10, F= 11.48, P<0.01). Results

revealed that Misr-1 and Sohag-3 had

high and almost similar grain diameter

(3.44 mm and 3.45 mm) compared to

tested cultivars except Sohag-5, Misr-2

and Misr-1 (3.29 mm, 3.31 mm and 3.44

mm, respectively). Low and similar grain

diameter was recorded for Beni Sweaf-3

and Beni Sweaf-5 (2.89 mm) compared

to other cultivars except for Beni Sweaf-5

and Giza-171 (2.89 mm and 3.03 mm).

3.2.4 Weight kernel (mg)

Wheat kernels were weighted for tested

cultivars, and results showed that there is

a significant difference in weight kernel

among wheat cultivars. High kernel

weight was recorded for Misr-1 and

sohag-3 (57.93 mg and 56.49 mg)

compared to the other tested cultivars.

However, Beni Sweaf-5 had significantly

the lowest kernel weight (39.20 mg).

3.2.5 Hardness index

Significant differences were found

between cultivars in hardness index. All

durum wheat showed significantly higher

hardness index than bread wheat. Sohag-

5 showed the highest hardness index

value (100 hardness indexes) compared

to all cultivars except Beni Sweaf-5,

Beni Sweaf-1 and Beni Sweaf-3 (96.99,

97.61 and 98.82 respectively). While,

Gemmeiza-10 showed the lowest

hardness index value (48.45).

3.3 Biochemical parameters measured

for eleven Egyptian wheat cultivars

Biochemical parameters including starch,

protein, gluten index and wet gluten

content were assayed for 11 Egyptian

wheat cultivars (Figure 1).

3.3.1 Starch percentage

Starch analysis showed significant

differences between tested cultivars.

60

Gemmeiza-10 and Misr-2 recorded the

highest values (78.41% and 78.13 %)

than all tested cultivars. While, Beni

Sweaf-3 had the lowest value for starch

(54.16 %) compared to tested cultivars.

Almost similar starch values were

recorded for Beni Sweaf-5, Sohag-3 and

Sohag-5 (58.61%, 58.83% and 59.85 %

respectively).

3.3.2 Protein percentage

Measurement of protein in the tested

wheat cultivars resulted a significant

difference among the tested cultivars.

Results indicated that Misr-1 showed the

highest protein percentage (17.19%)

compared to all tested cultivars. While,

Beni Sweaf-1 had the lowest protein

percentage (11.7%). Almost similar

percentage of protein was recorded for

Beni Sweaf-6, Giza-171, Sohag-3 and

Beni Sweaf-5 (13.19%, 13.35%, 13.55%

and 13.93% respectively). Similar results

for Beni Sweaf-4, Sohag-5 and

Gemmeiza-10, where percentages of

protein were 14.23%, 14.81% and 14.82

respectively. The same trend was

recorded for Beni Sweaf-3 and Misr-2,

where percentages of protein were 15.28

% and 15.70%.

3.3.3 Percentage of wet gluten content

The content of wet gluten was measured

for the tested cultivars, and the results

revealed that there is a significant

difference among tested cultivars. Misr-1

and Misr-2 showed a high value for wet

gluten content (37.3% and 35.6%)

compared to all tested cultivars. While

Beni Sweaf-1 had the lowest value for

wet gluten content (20.2%). Similar wet

gluten content values were recorded for

Beni Sweaf-3 (22.5%) and Beni Sweaf-5

(22%).

Figure (1): A comparison between biochemical traits of eleven Egyptian wheat cultivars.

0

20

40

60

80

100

% Starch

Egyptian wheat cultivars

% Starch

0

10

20

30

40

% Wet gluten content

Egyptian wheat cultivars

% Wet gluten content

0

5

10

15

20

% Protein

Egyptian wheat cultivars

%Protein

61

3.4 Correlation between percentages of

damaged grain for eleven Egyptian

wheat cultivars and their biophysical

and biochemical parameters

Highly significant positive correlation

was observed between percentage of

damaged grain and percentage of weight

loss (r = 0.968**, P<0.01). So, the

percentage of damaged grain was chosen

to correlate with the biophysical and

biochemical parameters of the eleven

Egyptian wheat cultivars as showed in

Table (2). In respect to the biochemical

traits, a significant positive correlation

was noticed between % of damaged grain

and starch (r = 0.713**) and wet gluten

(r = 0.551**), while no significant

correlation was found with the protein

content. On the other hand, high

significant negative correlation was

noticed between hardness index and

percentage of damaged grain (r=-

0.973**). However, no significant

correlation was found between moisture

(a), moisture (b), diameter (mm), and

weight kernel (mg) and percentage of

damaged grain.

Table (2): Correlation between percentages of damaged grain for

eleven Egyptian wheat cultivars and their biophysical and

biochemical traits.

Biophysical traits

r value

P value

Weight loss (%)

0.968**

P<0.01

Moisture (a) (%)

-0.136 ns

P>0.05

Moisture (b) (%)

0.286 ns

P>0.05

Diameter (mm)

0.016 ns

P>0.05

Weight/ kerenl (mg)

-0.049 ns

P>0.05

Hardness index

-0.973**

P<0.01

Biochemical traits

Starch (%)

0.713**

P<0.01

Protein (%)

0.262 ns

P>0.05

Wet gluten content (%)

0.551**

P<0.01

*Significant at 0.05 Probability level; ** Significant at 0.01

Probability level; ns is not significant.

4. Discussion

This study was conducted on 11 recent

Egyptian wheat cultivars including (7

durum wheat and 4 bread wheat). The

main aim is to provide more information

about the susceptibility of the tested

cultivars to

S. oryzae

based on two

measured parameters percentage of

damaged grain and weight loss of

infested grain. On the other hand, the

study indicated a correlation between

some biophysical and biochemical traits

of the tested wheat grain and the

infestation by

S. orzyae.

Overall, high

percentage of weight loss and damaged

grain was observed in bread wheat as

compared with the durum wheat. Similar

results was reported by Mahmoud et al.

(2011), who found high susceptibility of

bread wheat compared to durum wheat

against the infestation by

Sitophilus

granarius

(L).The same authors also

reported Gemmeiza-7 as the most

susceptible cultivar and Beni Sweaf-4 as

the most resistance one, which partially

in harmony with this study finding.

According to the obtained data there is a

62

significant difference among wheat

cultivars in respect to the grain content of

starch, protein, and wet gluten as

biochemical traits and in their

biophysical traits (hardness index, kernel

diameter, and kernel weight). There is no

significant variation in grain content of

moisture among all the tested cultivars.

The obtained data indicated no actual

association between grain content of

protein and the percentage of damaged

grain. This finding is supported with Rao

and Sharma (2003) who reported that

grain protein content could only had a

slight role in wheat resistance to the

weevils. In contrary, (Ram & Singh,

1996) and (Tiwari & Sharma, 2002)

stated that

S. oryzae

weevil correlated

negatively with protein. Wet gluten had a

weak correlation to weevil infestation,

that in harmony with a study by (Tiwari

& Sharma, 2002) who stated that no

impact of gluten content on the

resistance and susceptibility to insect

species. The obtained data revealed that

wheat grain content of starch positively

correlated with weevil infestation that

could be due to the enzymology of

digestive such as amylase for some

Sitophilus

weevils required initial stages

of large food polymers such as starch

(Franco et al., 2002; Baker, 1986). Also,

our results are in harmony with Singh

and McCain (1963) who noticed positive

correlation between sugar and starch

content of the kernels and weevil

reproduction. Highly significant negative

correlation between grain hardness index

and percentage of damaged grain was

observed. This observation tremens

dously clear when comparing the

hardness index of durum wheat cultivars

as a hard red spring wheat and the bread

wheat as a soft red spring wheat

cultivars, and the percentage of damaged

grain of these cultivars. Many studies

have also reported hardness index as

negatively correlated with

S. oryzae

infestation (Tiwari & Sharma, 2002;

Ram & Singh, 1996; Sinha et al., 1988;

Rout et al., 1976). However a study by

Rao and Sharma (2003), who screened

different wheat germplasm against the

infestation by

S. oryzae

and reported no

clear relationship between kernel

hardness and weevil resistance. There is

no significant correlation was found

between percentage of damaged grain

and grain size of tested cultivars.

However, a study by Ram and Singh

(1996), Tiwari and Sharma (2002)

indicated positive correlation between

grain size and resistance to

S. oryzae

weevils. The conflict in the results could

be due to the variation in tested cultivars

since none of the previous studies

included durum wheat in their studies.

Some of durum wheat had almost the

same size as bread wheat, but

tremendously varied in other traits that

widely reported as a correlated

parameter to weevil infestation such as

hardness index. Grain weight also

showed no impact on damaged grain.

There is no clear evidence that grain

weight had an impact on host resistance

to insects (Tiwari & Sharma, 2002).

Grain content of moisture has reported

one of the factors that may affecting the

susceptibility to

S. oryzae

(Hussain &

Nasr, 2015; Rashad et al., 2005; Hameed

et al., 1984). However, no correlation

was detected between grain content of

moisture, that measured by two different

63

instruments, and percentage of damaged

grain that could be due to the minor

differences in moisture content among

the tested cultivars. Further investigation

should be done to confirm the

importance of moisture content. It could

be concluded that none of the tested

cultivars were completely resistant to

infestation by the granary weevil but

their susceptibility to the infestation

varied considerably and the tested

cultivars significantly varied in their

biophysical and biochemical traits.

Acknowledgments

We thank professors Drs. David

Marshall and Christina Cowger

(Department of Entomology & Plant

Pathology, North Carolina State

University) foroffering the required

facilities and the instruments for this

study. We also thank Mr. Gabe Supino

for his excellent technical assistance

during the experiment.

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