Minia J. of Agric. Res. & Develop. Vol. ( ) No. 5. (32... · Rehab T. Behairy et al. 324-susceptible cultivars could be as a result of the effect of Orobanche parasitism. Therefore,

FACULTY OF AGRICULTURE

Minia J. of Agric. Res. & Develop.

Vol. (23) No. 5 pp 322 - 348,

3103

EFFECT OF OROBANCHE PARASITISM ON THE BEHAVIOR OF

FOUR FABA BEAN CULTIVARS (Vicia Faba L.)

Rehab T. Behairy*, A. A. M. Ashrie

**

and Mohamed El-Danasoury

***

*Seed technology Research Section, Field Crop Res., Inst., Agric. Res. Cen., Giza

**Food Legumes Research Section, Field Crop Res., Inst., Agric. Res. Cen., Giza ***Biochemistry Dept., Faculty of Agriculture, Al-Azhar University.

Received 33 Sept. 3103 Accepted 34 Dec. 3103

ABSTRACT Four faba bean (Vicia faba L.) cultivars i.e. Giza 438, Misr 0

(tolerant to Orobanche) and Giza 41, Nubaria 0 (susceptible to

Orobanche) were evaluated in Orobanche naturally infested soil

and Orobanche free soil at Giza Research Station, Agricultural

Research Center , in 3101/3100 and 3100/3103 seasons. Results

revealed the superiority of Giza 438 in plant height, number of

pods/plant , number of seeds/plant and seed yield/ plant on the

Orobanche - free soil, mean while Misr 0 was superior number of

pods and seeds/plant and seed yield/plant on the Orobanche-infested

soil. Giza 41 possessed a high level of infestation with the highest

number and dry weight of Orobanche spikes/ m3 .The two tolerant

genotypes Giza 438 and Misr 0 had the low number of Orobanche

infested soil. Results of chemical compositions revealed significant

differences among cultivars under the two soils. The two cultivars

Giza 438 and Misr 0 gave higher values of protein and

carbohydrates, respectively, while Giza 41 gave the lowest values.

The analysis of protein banding pattern is an important tool , where

absence of these bands from total root protein belongs to

Rehab T. Behairy et al.

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susceptible cultivars could be as a result of the effect of Orobanche

parasitism. Therefore, these polymorphic bands can be considered

as a positive marker for tolerant cultivars under Orobanche

infection. In addition, another difference was found in the

expression profile among the studied cultivars. Where, the intensity

of each band was used as an indicator for its level of protein

expression. The tolerant cultivars, Giza-438 and Misr-0, showed

the highest values for protein expression.

ITRODUCTION

Faba bean (Vicia faba L.) is still the backbone of the legume

crops grown in Egypt. It is an important source of high quality and

inexpensive protein supplement for the majority of the Egyptian

population through various popular dishes. Also, it is known as an

efficient atmospheric nitrogen fixer depending on availability of

Rhizobium spp. that colonizes the legume plants which keeping the

Egyptian soil fertile and benefits for the subsequent crop. The average

cultivated area devoted to faba bean during the last three seasons

9008002 – 9020022 represented 238111 feddans with an overage seed

yield equal 8.8 ardab/fed.

Orobanche spp. is root holoparasitic plants causing severe yield

and quality losses in a wide range of dicotyledonous crops. As most

contemporary methods aimed at controlling Orobanche have resulted

in limited success, the search for tolerant genotypes and understanding

the tolerance mechanisms are considered important.

The parasitic weed (Orobanche crenata Forsk) is a major pest

of faba bean (vicia faba L.) in Egypt. For example, in Behera

Governorate (North Delta), Zaitoun et al.(2222) reported that the

percentage of Orobanche infestation amounted to 85.56 of faba bean

cultivated area, with a total yield losses of about 220000 tons.

The food legume breeding program, FCRI, ARC resulted in

releasing three cultivars (Giza 992, Giza 891 and Misr 2) having a

Effect of orobanche parasitism on the behavior of four faba bean

cultivars (vicia faba l.)

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higher level of resistance to Orobanche (Khalil et al 2229, Saber et al

2222 and 9002)

The evaluation of tolerant materials under Orobanche -infested

and Orobanche-free infested fields were investigated by Darwish et al

(2222) Abdalla and Darwish (9009) Morsy and Attia, (9009)El-

Harty.E.H. (9005) Abdalla et al (9008) Darwish et al (9003) Abbes

et al (9008) Abdalla and Darwish (9008) and Ashrie et al (9020).

They concluded that there were significant differences among

genotypes in most traits under study.

In fact, breeding for tolerance and identification of genes

responsible for it is one of the most promising methods to increase the

tolerance of Faba bean towards Orobanche parasitism. Biotechnology

tools such as marker-assisted breeding, tissue culture and genetic

engineering can be useful in achieving this goal. However, only limited

success has been gained. So we need to increase our efforts to get more

progress in this area of research.

The current study was aimed to detect proteins involved in the

tolerance of Faba bean for the parasitism of Orobanche spp. using

SDS-PAGE protein analysis.

MATERIALS AND METHODS

The present investigation was conducted at the farm of Giza

Research Station, ARC, Egypt in 9020022 and 9022029 seasons to

study the performance of four faba bean cultivars (Giza 992, Misr 2

tolerant to Orobanche and Giza90, Nubaria 2 susceptible to

Orobanche ) under both Orobanche-free and Orobanche-naturally

infested soils. The pedigree and reaction of the materials used to

Orobanche are shown in Table (2).


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Table 0. Pedigree and reaction to Orobanche of four faba bean

genotypes under study.

A Randomized Complete Block Design (RCBD) with three

replications was used, in both infested and non infested Orobanche

soils. The experimental plot consisted of three ridges 1 m long, 80 cm

apart , with single seeded hills, 90 cm apart on both sides of ridges.

Cultivation conditions were followed as normally recommendation for

Vicia faba L.. In turn, five random plants were taken from each

replicate every interval time, after 10, 90 and 50 days of sowing date,

and inspected for the presence of Orobanche tubercles. At harvest

time, ten guarded plants were taken at random from each experimental

plot. The following data were recorded :-

0 - Agronomic traits:

Plant height (cm), number of branches/plant, number of

pods/plant, number of seeds/plant, seed yield /plant (g), 200-seed

weight (g) , number of Orobanche spikes and dry weight of

Orobanche spikes ( g/m 9) were recorded at maturity of faba bean

plants for all accession. The susceptible check variety Giza 90 and

Nubaria 2.In Giza 90 and Nubaria 2, Orobanche spikes were

collected just when its plants started to death, and their data were

recorded immediately.

Reaction to Orobanche Pedigree Genotype

Tolerant to Orobanche An individual selected plant

from Giza 909 Giza 438

Tolerant to Orobanche G.1x 291A/95038 Misr 0

Susceptible to Orobanche An individual plant selection

from Rebaya 90 Giza 41

Susceptible to Orobanche An individual selected plant

from Spanish variety Reina

Blanca

Nubaria 0



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3- Chemical composition analysis:

Protein and carbohydrate percentages were determined according

to the procedures outlined by AOAC, (9000). Percentage of protein

was obtained by multiplying nitrogen percentage by 8.95 as stated by

Sadasiuam and Manickam (0886).

2- SDS- PAGE analysis:

Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS

– PAGE), was used to investigate different protein banding profiles

among studied cultivars under both infected and non infected

conditions. Analysis of protein banding patterns was performed

according to methods reported by Laemmli (0881) using the Mini-

Protein Electrophoresis Cell, manufactured by Bio-Rad Corporation.

4-Statistical Analysis

Statistical analyses were performed using the SPSS software

(version 3.5 for windows) and as outlined by Gomez and Gomez

(2289(.

RESULTS AND DISCUSSION

Agronomic traits

The mean performance of four faba bean cultivars under

Orobanche-infested and free soils is presented in Table (9). Results

showed significant differences among cultivars for plant height,

number of branches/plant, number of pods/plant, number of

seeds/plant, seed yield /plant, 200-seed weight. number and dry

weight of Orobanche spikes under infested soil. However, significant

differences were found among cultivars for plant height, number of

branches/plant, number of pods/plant, number of seeds/plant, , and

200-seed weight were observed under Orobanche- free soil, On the

other hand differences between seed yield /plant of both sides were

insignificant.


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Orobanche- free field

The cultivar Giza 992 recorded the highest values of plant

height(208.1), number of pods/plant(95.5), number of

seeds/plant(39.9) seed yield /plant (52.0), Nubaria 2 recorded the

highest number of branches/ plant,(5.5), and200-seed weight (209.9),

under free soil.

Orobanche- infested field

The tolerant cultivar Misr 2 recorded the highest values of

plant height.(202.8), number of pods/plant, (29.0), number of

seeds/plant,(12.5) seed yield /plant, (12.9), followed by Giza

992under infested soil .

These findings are clearly supported by those finding obtained

by Darwish et al (2222), Abdalla and Darwish (9009), Morsy and

Attia, (9009), El-Harty.E.H. (9005) Abdalla et al (9008), Darwish et

al (9003), Abbes et al (9003), Abdalla and Darwish (9008), and

Ashrie et al (9020) who reported that faba bean genotypes differed

from each other in their yield and yield components regarding

Orobanche infestation.

Number and dry weight of Orobanche spikes/ m3

Results in Table (1) showed that Nubaria 2 caltivar possessed

the highest level of infestation with high number and dry weight of

Orobanche spikes/ m9 . The two tolerant cultivars Misr 2 and Giza 992

had the lowest number of Orobanche spikes , and the lowest dry

weight of Orobanche spikes, followed by Misr2and Giza 992. The

similar results were obtained by Morsy and Attia, (9009) , Abbes et al

(9003) and Ashrie et al (9020).



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Table 3. Performance of the four faba bean genotypes under

Orobanche –infested and - free soils in 3101/3100 and

3100/3103 seasons. No. of branches /plant Plant height (cm)

Genotype

Free Infested Free Infested

Co

mb

ined

31

00

/03

31

01

/00

Co

mb

ined

31

00

/03

31

01

/00

Co

mb

ined

31

00

/03

31

01

/00

Co

mb

ined

31

00

/03

31

01

/00

9.0 9.1 1.3 9.8 9.0 5.9 208.1 209.1 208.1 202.8 200.0 201.3 Giza 438

9.3 9.3 9.9 9.3 9.8 9.5 209.9 209.3 202.3 202.3 28.1 205.0 Misr 0 1.2 9.2 1.8 1.8 1.5 9.0 22.9 28.1 200.0 32.3 88.1 35.0 Giza 41

5.5 5.3 5.1 5.8 8.0 5.3 29.5 20.0 25.0 32.9 38.3 82.3 Nubaria 0 1664 0610 1634 1686 0633 1688 8653 8621 8624 2654 5624 8628 LSD 1615

Table 3.Cont. No. of seeds / plant No. of pods / plant

Genotype


Co

mb

ined

31

00

/03

31

01

/00

Co

mb

ined

31

00

/03

31

01

/00

Co

mb

ined

31

00

/03

31

01

/00

Co

mb

ined

31

00

/03

31

01

/00

39.9 38.3 32.3 15.8 11.1 18.1 95.5 98.3 99.1 29.5 22.3 21.1 Giza 438

89.0 89.1 82.3 12.5 12.0 90.0 92.8 99.3 92.0 29.0 21.3 29.1 Misr 0

39.0 38.3 32.1 1.3 1.1 9.0 95.1 98.1 99.1 2.5 2.1 2.3 Giza 41

51.1 55.0 52.3 8.9 3.1 5.0 29.8 25.0 29.3 2.3 9.0 2.1 Nubaria 0

0063

8

NS NS 6605 6613 8630 2618 5681 5645 0688 3625 3642 LSD 1615

Ns: non significant


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Table 3.Cont.

011-seed weight (g) Seed yield / plant (g)

Genotype


Co

mb

ined

31

00

/03

31

01

/00

Co

mb

ined

31

00

/03

31

01

/00

Co

mb

ined

31

00

/03

31

01

/00

Co

mb

ined

31

00

/03

31

01

/00

80.3 82.2 80.1 82.0 32.2 89.9 52.0 53.1 59.3 92.9 98.3 12.3 Giza 438

38.1 32.9 33.9 38.8 33.8 32.9 98.5 92.1 93.3 12.9 10.1 19.0 Misr 0

89.0 89.5 81.5 85.0 81.2 88.0 93.1 92.1 95.1 9.9 9.2 9.3 Giza 41

209.9 209.9 209.8 201.2 209.1 201.2 55.3 53.1 59.0 8.1 3.3 5.9 Nubaria 0

0681 3680 3683 4658 5625 5623 NS NS NS 5668 5686 6658 LSD 1615

Ns: non significant

Table 2. Mean number and dry weight of Orobanche spikes

under infested soils during 3101/00 and 3100/03

seasons.

Chemical analysis

The mean performance of the studied faba bean cultivars under

Orobanche - infested and Orobanche - free field conditions is

presented in Table 9. The results showed significant differences among

Orobanche dry weight(g/m3) No. of orobanche /m

3 Genotype

Combine

d

3100/03 3101/00 Combined 3100/03 3101/0

0

289.5 282.3 281.1 90.8 91.1 18.1 Giza 438

238.3 280.0 231.1 13.8 92.0 19.3 Misr 0

999.5 902.3 981.1 218.1 298.3 298.0 Giza 41

930.0 958.3 981.1 218.9 295.3 250.3 Nubaria 0

66683 NS 36665 00603 08621 03624 LSD 1615



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cultivars in protein and carbohydrates. The percentage of protein and

carbohydrates in Giza 992 were higher in the Orobanche - free field

than those in the Orobanche - infested field.

Table 4. Protein and carbohydrates content of the four studied faba

bean genotypes under Orobanche –infested and –free fields

in 3101/00 and 3100/03 seasons.

Ns: non significant

The susceptible cultivar (Giza 90) gave the lowest values for all

studied traits, while the tolerant genotypes (Giza 992 and Misr 2) gave

the highest values for protein and carbohydrates, respectively. It could

be concluded that seeds from Orobanche – free field were better than

those of Orobanche – infested field and had higher protein content.

These findings were clearly confirmed by those obtained by

Frejnage et al (2223), Megahed (9000), Morsy and Attia (9009) and

Ashrie et al., (9020).

3- Analysis of Total Root Protein:

Total root protein of the four Vicia faba cultivars Giza- 992,

Misr-2, Nubaria-2 and Giza-90 were determined to indicate the

differences due to Orobanche parasitism. The data shown in Table (5)

Carbohydrates (%) Protein (%) Genotype


Co

mb

ined

31

00

/03

31

01

/00

Co

mb

ined

31

00

/03

31

01

/00

Co

mb

ined

31

00

/03

31

01

/00

Co

mb

ined

31

00

/03

31

01

/00

89.8 89.2 89.9 89.8 89.8 89.3 95.1 95.5 95.0 98.2 98.9 95.3 Giza 438

85.8 85.0 88.9 85.1 85.8 89.8 95.5 95.3 95.9 99.2 99.0 99.9 Misr 0

81.9 81.3 81.9 89.5 89.2 89.2 91.3 99.9 91.2 91.8 99.0 99.0 Giza 41

89.1 89.2 89.9 89.8 89.3 89.5 99.1 99.1 99.9 91.8 91.9 91.2 Nubaria 0

16163 NS 16835 16854 NS 16380 16835 06138 0635 16626 16588 06338 LSD 1615


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and Figure (2) revealed that infected samples for tolerant cultivars,

Giza-992 and Misr-2 showed two polymorphic bands at 30 and 89

kDa, whil un-infected plants of the same cultivars were similar to each

other in the protein banding pattern except for Giza-90 which missed a

band at 89 kDa. Absence of these bands from the total root protein

banding patterns is due DNA sequence differences between the studied

cultivars which in turn are transcribed to different peptides which are

involved in certain changes due to the effect Orobanche spp.

parasitism on them (Sakr et al. 9020). Therefore, these polymorphic

bands can be considered as a marker for tolerant cultivars under

Orobanche infection (Abd El-maksoud et al. 9003 and Gadalla et al.

9020).

Non Infected Plants Infected Plants

G41 N 0 E0

G3

48

G43

8 E0 N0

G4

1 M



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021

85

83

55

26

31

Figure 0: SDS PAGE banding pattern for total root protein of

the four studied Vicia faba L. cultivars.


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Also, another difference was found in the expression profiles

among the studied cultivars. Where, the intensity of each band can be

used as an indicator for its level of protein expression. Both tolerant

cultivars, Giza-992 and Misr-2, showed the highest values for protein

expression, which are represented in the values indicated for each band

as shown in figure (9).

Figure 3: Protein expression profiles among infected and un-infected

plants.



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Table 5: SDS-PAGE banding pattern for total root proteins of the

four Vicia faba L. Cultivars under infected and un-

infected conditions of Orobanche spp.

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#

Total

RF

Total

MW

Non Infected Plants Infected Plants

G41 N 0 M 0 G438 G438 M0 N 0 G41

2 0.99 30 + + + + + + - -

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20 0.38 95 + + + + + + + +

22 0.32 91 + + + + + + + +

29 0.89 92 + + + + + + + +

21 0.88 90 + + + + + + + +


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ن تأثير تطفل الهالوك على سلوك أربعة أصناف م

البلديالفول

رحاب تودى بحيرى*

الرازق محمدعشرى , عسام عبد**

محمد مبروك الدناصورى ***

مركز البحوث الزراعية -معهد المحاصيل الحقمية -قسم بحوث تكنولوجيا البذور* مركز البحوث الزراعية -معهد المحاصيل الحقمية -قسم بحوث المحاصيل البقولية **

جامعة األزهر -كمية الزراعة -ةقسم الكيمياء الحيوي***

بحؤوث تحت ظروف أرض موبؤة وأرؤرى ريل ؤن مؤن الكؤيلوك بمحطؤن هذا البحث ىأجر وذلؤؤك 0200/0200و 0202/0200 موسؤؤم رؤؤ ل الزراع ؤؤن مركؤؤز البحؤؤوث – الج ؤؤز أصنيف مؤن الوؤول البمؤدي كؤ كؤ التؤربت نا ومؤد الؤتممت الدراسؤن أربع كدراسن سمو بكدف

( وصؤؤنو ن حسيسؤؤ ن لإلصؤؤيبن 0، مصؤؤر 904 ن لإلصؤؤيبن بيلكؤؤيلوك ةج ؤؤز صؤؤنو ن متحممؤؤ، كؤ صؤويت 904(ا مد أوضحت النتيئج تووق صنف ج ؤز 0، نوبير ن 92بيلكيلوك ةج ز

األرض كؤؤؤ طؤؤؤول النبؤؤؤيت، عؤؤؤدد ال رون/نبؤؤؤيت وعؤؤؤدد البؤؤؤذور/نبيت ومحصؤؤؤول بؤؤؤذور النبؤؤؤيت ويت عؤؤدد ال رون/نبؤؤيت، عؤؤدد كؤؤ صؤؤ 0الريل ؤؤن مؤؤن الكؤؤيلوك، كمؤؤي توؤؤوق الصؤؤنف مصؤؤر

بيلكؤؤيلوكا كمؤؤي انروؤؤض عؤؤدد اإلصؤؤيبنالبؤؤذور/نبيت، ومحصؤؤول بؤؤذور النبؤؤيت تحؤؤت ظؤؤروف م يرنؤن بيلصؤنو ن ج ؤز 0، مصؤر 904ج ؤز صؤنو حيلؤن كؤ 0ووزن لمير خ الكيلوك /م

ا0ونوبير ن 92ن بؤؤ ن وأظكؤؤرت النتؤؤيئج وجؤؤود ارت كؤؤيت مصنو ؤؤن لمصؤؤويت الك م يئ ؤؤن و الب وك م يئ ؤؤ

األصؤؤنيف تحؤؤت الدراسؤؤن كؤؤ كؤؤ التؤؤربت ن ةالريل ؤؤن والموبؤؤة (، ومؤؤد سؤؤجمت عمؤؤ الترت ؤؤ أعمؤؤ مؤؤ م لنسؤؤبن البؤؤروت ن، الكربوه ؤؤدرات ، كؤؤ حؤؤ ن 0و مصؤؤر 904األصؤؤنيف ج ؤؤز

بإسؤتردام التور ؤؤد حؤزم البؤروت ن نمؤط أمؤل ال ؤ م ا ومؤؤد تؤم تحم ؤل 92سؤجل الصؤنف ج ؤز لؤكل الكؤيلوك وتو ؤد مؤن التطوؤل تجؤي الوؤول البمؤدي أصؤنيف بؤ ن الوؤروق لوصؤف الككربؤ


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والنوبير ؤؤن0مصؤؤر و 904الج ؤؤز الوؤؤول البمؤؤدي مؤؤن مؤؤن فالبؤؤروت ن الكمؤؤ لجؤؤذور أصؤؤني

الص نؤيت مؤن وأظكؤرت النتؤيئج .الكؤيلوك تطوؤل النيتجن بسب االرت كيت لوحص 92 والج ز 0

مم ؤزت ن كؤ الؤوزن حؤزمت ن ظكؤور 0، ومصر904 المتحممن من الج ز لألصنيف المصيبنلؤم لؤنوس األصؤنيف المصؤيبن غ ؤر أن النبيتؤيت كؤ حؤ ن ك مؤو دالتؤون، 49 و 02 الجز ئؤ األصؤنيف الحسيسؤن مؤن البؤروت ن الكمؤ كؤ الحؤزم هؤذ عؤدم وجؤود .ارت كؤيت أي تظكؤر

هؤذ الحؤزم اعتبؤيرولؤذلك، مكؤن .عم كي الكيلوك تطول لتأث ر نت جن مكن أن صتبر لإلصيبن

.الكيلوك عدوى تحت المتحممن لإلصيبن ا جيب ن لألصنيف كمصمميت جز ئ ن

نتيج الج ن التصب ر عم مستوى نمط أررى ارت كيت كمي أظكرت النتيئج وجود وا كمةلر تستردم حزمن مكن أن كل ح ث أن لد وضوح المدروسن األصنيف البروت ن ب ن

النتيئج أن أظكرت الحزم وضوح لد ك االرت كيتب ن يلم يرننوب .البروت ن تصب ر لمستوى

التصب ر لمبروت نا ك أعم م م ( لكي0ومصر 904الج ز ةالمتحممن األصنيف

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