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IJISET - International Journal of Innovative Science, Engineering & Technology, Vol. 6 Issue 2, February 2019

ISSN (Online) 2348 – 7968

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Physiological Responses of Psium sativum L. to Treatment with Plant Growth Promoting

Rhizobacteria, Mycorrhiza and Zinc. Abd El-Monem M.A. Sharaf1, Mahmoud R. Sofy2 and Mahmoud S. Osman3

1-3Botany and Microbiology Department, Faculty of Science, Al-Azhar University,

11884 Nasr City, Cairo, Egypt

Abstract

This study was done to investigate the effect of Azospirillum, Rhizobium, Mycorrhiza and Zinc on growth, productivity and biochemical activity of pea (Psium sativum) as factorial design with three replications. Factors were applied on as fallowing: the first factor was Azospirillum brasilense , the second factor was Rhizobium leguminosarun, the third factor was Mycorrhiza and the final factor was Zinc 100 ppm. Characteristics studied in this research includes growth characters (shoot length, root length and number of leaves/plant), Metabolic activities (Photosynthetic pigment, Carbohydrates, Protein, NPK content and SDS PAGE). The results showed that significant increases in contents of chlorophyll (a), (b) & (a + b), carotenoids, total soluble carbohydrates, soluble proteins, and NPK contents throughout the experimental period . Keywords: Mycorrhiza; Azopirillum; Rhizobium; Psium sativum L; productivity; Zinc; NPK.

1. Introduction

Pea (Psium saivum) is one of the most important Fabaceae vegetable crops grown during winter season in Egypt. It occupies a great figure in the local consumption and export. The green pods and mature seed of pea are rich in protein and vitamins. As pea is short durable crop its cultivation is highly profitable and preferable to the farmers, it can be grown in all types of soil. This crop like many other legumes is cap able of fixing and utilizing atmosprheric nitrogen through symbiotic relationship with Rhizobium (Ahmed et al., 2007). Biofertilization, practicularly those of diazotrophs, as well documented as a great part of plant N demand. Plant growth promoting Rhizobacteria (PGPR) have been used as biofertilization for economically important crops and shown increase crop growth and productivity (Desai et al.,

2012). The nitrogen-fixing interaction between Rhizobia and legumes and the mycorrhizal association between fungi and most of land plants are the most commonly studied symbioses (Talaat and Abdallah, 2008).

Most Agricultural crops are colonized by arbuscula rmycorrhizal fungi (AMF). In this symbiotic association, host plants provide the fungi with carbohydrates and return receive mineral nutrient. AMF can enhance growth of crop plants through increasing nutrient uptake, particularly P (Ryan and Angus, 2003).

2. Material and Methods

2.1. Methods of planting, treatments and collection of samples

Seeds of pea "Psium sativum L." were obtained from Agriculture Research Centre, Ministry of Agriculture, Giza, Egypt.

PGPR strain of Azospirillum brasilense (NO40), Rhizobium leguminosarum bv. Viceae& Rhizobium leguminosarum and Mycorrhizal isolates as spores of three species (i.e. Glomus mosseae, Gigaspora margarita, Acaulospora calaspora) obtained from Microbiology Department, Soil, Water and Environment Research Inst. Agricultural Research Center. Giza, Egypt.

Peat based culture of microorganisms were prepared using the method described by (Vincent, 1970). Cell suspension containing 108cfu/ml of N2 fixing isolates and spores of mycorrhiza were used to impregnate sterilized peat at the rate of 25ml liquid culture/ 100 gm peat and 50 spores of mycorrhiza/100gm peat. Inoculated peat was well mixed and allowed to mature at room temperature for 48 hr. Seeds wetted with 10% arabic gum water solution as an adhesive material (Hamdi, 1982) were inoculated with

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rhizobial, Azospirilum and mycorrhizal peat-based preparation. Seeds were allowed to air dry in the shad for 30 min. and sown immediately. Group of pea seeds are presoaked in (100 PPm Zn) as ZnSo4.

Uniform pea seeds were planted in pots; each pot contained 8 Kg of homogenous loamy- sandy soil. The developed plants were irrigated whenever required with tap water until the complete germination. The plant samples were collected for analysis when the plants were 25(Stage I) and 75 (Stage II) days old. At the end of the growth season analyses of the yielded seeds from the different treatments as well as the control were done.

2.2. Measurement of growth parameters

Shoot length (cm), root length (cm), number of leaves per / plant, fresh were determined at different growth stages. 2.3. Chemical analysis

Three plant samples were taken during the growing season, at vegetative growth stage I , flowering stage II and yield stage. Photosynthetic pigments were estimated using the method of Vernon and Selly (1966). Contents of soluble carbohydrates were measured according to the method of Umbriet et al.,(1969). Contents of soluble proteins were estimated according to the methods of Lowery et al.,(1951). Nitrogen content (gm/100gm D. wt) was determined in the digested solution by the modified microkjeldahl method as described by Plummer (1971).Phosphorus content (gm/100gm D. wt) was determined colorimetrically according to the method of Jackson (1958).Potassium contents (gm/100gm D. wt) were determined against a standard using flame-photometer (Piper, 1950). 2.4. Statistical analysis The data were subjected to the proper statistical analysis of variance of a randomized complete block design as recommended by Snedecor andCochran (1989). For comparison between treatments means, using LSD, (PASW® Advanced Statistics 20, 2010) at 5%level was used .The values recorded in the values of the biochemical analysis are means of three replicates. 3. Results and Discussion 3.1. Growth responses

Result of the present study figs. 1,2,3 showed that application of Azospirillum brasilense was a stimulatory effects regarding the growth characters of the tested plants.. In pea plants treatment with Azospirillum brasilense increased shoot lengths, weight of pods and 100 seed weight. The resulted stimulatory effects of Azospirillum brasilense regards the growth as well as the yield of the tested plants are in agreement with results

obtained by several other investigators. In this regard Abdel Azeem et al., (2007) found that inoculation of faba bean (Vicia faba) with Azospirillum increase plant height, seed yield, shoot dry mass, no. of pods, root dry weight. Ferreira et al., (2013) found that application of Azospirillum brasilense increase significantly increase shoot , root dry weight 0f maize plants as compared with control. (Abd El Daim, 2007) recorded that inoculation of Hordeum vulgare with Azospirillum brasilense increase shoot, root height, fresh and dry weight of both shoot and root. The same author found that inoculation of Hordeum vulgare with Azospirillum brasilense increase no. of spikes/plants, spike length, spike weight, no of grains/spike, grain weight/spike and weight of 1000 grain.

The positive effect of bacterial inoculation are mainly attributed to the production of phytohormones that improve root development, these developments within the root system are important because they increase absorptive area and volume of soil substrate available to the plant, with subsequent increase in the rate of water and mineral uptake (Okon and Venderleyden, 1997). Plant growth regulators participate in the growth and development of cells, tissues, organs, and in fact the entire plant. Many studies suggest the involvement of indole-3-acetic acid (IAA), produced by PGPR in morphological and physiological changes of the inoculated plant roots (Dobbeleare et al., 2003).

Rhizobium species (PGPR) are widely used in agriculture for legume crops improvement because of their ability to fix atmospheric nitrogen in symbiosis with legumes. The potential for improving faba bean plant performance by inoculation with rhizobia has been investigated in the present study. The obtained results revealed that, most of all investigated growth characteristics (shoot length, pods weight, , number of leaves) of pea plants were markedly increased in response to the application of Rhizobium leguminosarum. The stimulative effects of Rhizobium leguminosarumon plant growth were also obtained by many workers Yadav, Verma (2014) found that in chickpea (Cicer arietinum L.) plant inoculation with Rhizobium leguminosarum increase nodule number , nodule dry weight, shoot, root dry weight and grain yield. Abd-Alla et al.,(2013) showed that Rhizobium leguminosarum inoculation of faba bean increase fresh and dry weight and number of nodule as compared to the control. Wani et al., (2007) reported that pea plants (Psium sativum) inoculated with Rhizobium sp increased dry matter, seed yield. Talaat and Abdallah,( 2008) found that Rhizobia stimulat plant growth by production of plant growth promoting substances, phytohormons, and organic acids. Solimanet al., (2012) recorded that Rhizobium colonization increased plant

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height, root length, number of branches/plant and total dry weight of Acacia saligna plant. Abo El-Soud et al., (2003) found that inoculation of legumes such as faba bean with rhizobium may increase yields and qualities of cultivated crops.

The specific mechanisms involved in plant growth enhancement by PGPR have not been well studied (Kloepper, 1993).The observed increase growth parameters of broad bean and pea plant might result from the capacity of these bacteria to form nitrogen-fixing nodules as well as of releasing secondary metabolites, including plant growth regulators; as well as from solubilizing phosphate, so facilitating the uptake of nutrients from the root environment (Aldesuquy,2000, Antoun and Prevost 2005, Sahran and Nehra 2011, Zanatta et al., 2007)

The obtained results revealed that, in pea plants shoot length, number of leaves were mostly significant increased due to mycorrhizal treatment. This effects on the plant growth were also obtained by many workers Abd-Alla et al., (2013) showed that AM fungal inoculation of faba bean increase fresh and dry weight and number of nodule as compared to the control. Soliman et al., (2012) recorded that mycorrhizal colonization increased plant height, root length and total dry weight of Acacia saligna plant. Mosaad (2014) showed that AM fungal inoculation of faba bean increase number of pods and seeds/plant, number of branches/plant, 100 seed weight as well as total yield as compared to the control. Wu, Xia (2006) indicated that, there was a marked increase in the plant hight, shoot, root dry weight leaf number per plant, stem diameter with application of AMF on tangerine (Citrus tangerine).

Results of the present study showed that, application of Zn was of stimulatory effects regarding the growth characters of the tested plants. In pea plants application of zinc increased (shoot length, number of leaves/plant, root lengths. These effects of Zn on plant growth were also obtained by many workers Seilsepour (2006) , Hamdy (2002), El Sayd et al., (2002), Wanas (2002) reported that, treatment of faba bean plants with Zn (50and100 ppm) significantly enhanced the plant height, number of leaves per plant, stem and leaf dry weights per plant and total leaf area per plant. Sofy (2009) found that application of Zn on broad bean plants significantly increased shoot and root length, number of leaves/ plant, fresh and dry weight of shoot, roots, and number of pods/plant, number of seeds / plant and weight of 100-seeds. 3.2. Photosynthetic pigments

Results of the present work figs. 4,5,6&7 showed

that, in pea plants, treatment with Azospirillum brasilense caused in most cases significant increases in the content of chlorophyll a,b total a+b and carotenoids. Increases in photosynthetic pigments observed in the present work in response to Azospirillum brasilense agreed with those observed by Zarea et al., (2012) reported that inoculation of wheat seedling with Azospirillum sp increase chlorophyll content, Abd El Daim (2007) recorded that inoculation of Hordeum vulgare with Azospirillum brasilense increase chlorophyll content.

In the present investigation, results figs. figs. 4,5,6&7 clearly revealed that in pea plants treatment with Rhizobium leguminosarum caused in most cases increase in the contents of photosynthetic pigment a, b, a+b and carotenoids. These findings are supported by the study of Attia (2014) found that inoculation of broad bean (Vicia faba) plant with Rhizobium leguminosarum significantly increase chlorophyll a, b and caretenoids, Ahmad et al., (2013) reported that inoculation of mung bean with Rhizobium leguminosarum increase chlorophyll content, Ahemad and Khan (2011) reported that, inoculation pea plants with Rhizobium sp significantly increase chlorophyll content.

The present study figs. 4,5,6&7 showed that, content of chlorophyll a, b, a+b and carotenoids (stage I)in pea plants were significantly increased in response to the application of mycorrhiza. These results are in agreement with the results obtained by Soliman et al., (2012) found that, chlorophyll content of Acacia saligna were markedly increased by using Mycorrhizal inoculation. Also Abdel Latef and Chaoxing (2011) reported that inoculation of tomato (Lycopersicone sculentum L. cv. Zhongzha105) with AMF(Glomus mosseae) cause highly increase in the content of chlorophylls compared to control.

With respect to treatments of Zn, the obtained results figs. 4,5,6&7 showed different responses as regards the contents of chlorophylls of the tested plants. In pea plants, application of Zn was found to be, mostly effective in increasing contents of chlorophyll a, b ,a+b and carotenoids.. These results are in accordance with other investigators Samreen et al., (2013) found that application of Zn onmung beans plant (Vignaradiata) significantly enhanced chlorophylland crude protein contents. Also, Akay (2011) found that application of Zn on three types of chickpeas (Cicerarietinumcv.Canitez-87, cv. ILC-482 and cv. Gokce) significantly increase leaf chlotophyllcocentration.

Results of the present work figs. 4,5,6&7 showed that, in pea plants results of the present work showed that, Azospirillum was more effective followed by mycorrhiza than other treatments in enhancing chlorophyll a, b and total a+b contents during the first stage of growth. Also

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Rhizobium and mycorrhiza was more effective than other treatments in enhancing chlrorophyll a, a+b content during the second stage. 3.3. Carbohydrates

Results of the present study figs. 8,9&10 showed that, contents of soluble carbohydrates in shoots, roots as well as in the yielded seeds of the pea plants were , mostly increased in response to application of Azospirillum brasilenset treatment.

The increase in carbohydrates contents that were observed in pea plants in response to the treatment with Azospirillum brasilense may be due to the stimulatory effect of that treatment as regards the alteration in phytohormone.

On the other hand, the obtained results as regards the increased content of total soluble carbohydrates that were recorded in pea plants are in agreement with other investigators. In these regard El shanshoury (1995) reported that inoculation with Azospirillum brasilense significantly increased total soluble carbohydrates contents in wheat shoots. Also, Abd El Daim (2007) found that inoculation of Hordeum vulgare with Azospirillum brasilense increase carbohydrates content.

Results of the present work figs. 8,9&10 revealed that, contents of total soluble carbohydrates were significantly increased in shoots, roots and yielded seeds in pea plants in response to the treatment with Rhizobium leguminosarum. The stimulatory effects of Rhizobium as regards contents of total soluble carbohydrates in different plants were recorded by other investigators Ismaielet al., (2014) studied the changes in carbohydrates in broad bean (Vicia faba L.) in response to Rhizobium inoculation (Rhizobium leguminosarum bv. viceae ) . They found that, soluble sugars were increased. Also Soliman et al., (2012) reported that, carbohydrates content of Acacia saligna were significantly increased due to inoculation by Rhizobium.

Result of the present work figs. 8,9&10 revealed that, in pea plants application of mycorrhiza resulted in, mostly, significant increase as regards the content of total soluble carbohydrates in shoots, roots as well as in the yielded seeds. These effects were recorded by other investigators Ismaiel et al., (2014) studied the changes in carbohydrates in broad bean (Vicia faba L.) in response to AMF inoculation. They found that, soluble sugars were increased. Also Abbaspour et al., (2012) recorded that mycorrhizal colonization increased soluble sugars of Pistachio (Pistaci avera L.) plant.

Results of the present work figs. 8,9&10 revealed that, contents of total soluble carbohydrates were significantly increased in shoots, roots and yielded seeds of the pea plants in response to the treatment with Zn. The

stimulatory effect of the micronutrients (Zn) as regards content of total soluble carbohydrates in different plants were recorded by other investigators Khalifa (2005) stated that, application of Zn significantly increased protein contents of peanut plants. Also Sofy (2009) reported that application of Zn on broad bean plants significantly increased total soluble carbohydrates in shoots, roots as well as yielded seeds.

Results of the present work figs. 8, 9&10 showed that, in pea plants the highest increases in carbohydrates contents were observed in plants treated with mycorrhiza followed by Azospirillum in shoot at stage I, Zinc, mycorrhiza in shoot at stage two. While in root rhizobium then zinc as well as in yielded seeds Zinc and mycorrhiza showed the greatest increases. 3.4. Soluble Protein

Results of the present work figs. 11,12&13 revealed that, in pea plants treated with Azospirillum brasilense resulted in, mostly, significant increase in protein contents in shoots, roots and also, in the yielded seeds. These results are in harmony with those obtained by Rahim Naseri et al., (2013) found that inoculation of Rapeseed (Brassica napusL.) with rhizobacterial strain significantly increase seed oil and seed protein. Yolcu et al., (2011) reported that application of PGPR strains on Italian ryegrass (Lolium multiflorum Lam.) significantly increase crude protein yield than control.

In present study figs. 11,12&13, it was found that protein contents in shoots, roots as well as in the yielded seeds of pea plants were significantly increased in response to the treatment with Rhizobium leguminosarum. In accordance with the obtained results, Rugheim, and Abdelgani (2012) revealed that, protein content of faba bean plant were increased in response to Rhizobium inoculation. Ahemad and Khan (2011) recorded that Rhizobium sp inoculation increased seed protein of Psium sativum plant. Also Chaudhary et al., (2011) found that inoculation of Vigna radiate and Vigna angularis L with Rhizobium sp increase total protein.

Results of the present study figs. 11,12&13 revealed that under the treatment of mycorrhiza, protein content in shoots, roots and also, in the yielded seeds of pea plants were, mostly, significantly increased. This is due to phosphorus is an essential element in synthesis of protein in the leguminous plants and mycorrhiza increased phosphorus contents in treated plants. In this regard Mosaad (2014) reported that protein percentage in faba bean (Vicia faba) plants significantly increased in response to AM fungal inoculation. Abbaspour et al., (2012) stated that mycorrhizal infection of (Pistaci avera L.) increased soluble protein more than control plants.

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Results of the present work figs. 11,12&13 revealed that in pea plants, treatment with Zn, resulted in, mostly, significant increase in protein contents in shoots, roots and also, in the yielded seeds. These results are in agreement with those obtained by Gamal El-Din (2005) reported that, Zn treatments (100 and 200mg/L) increased crude protein in fenugreek plants. Also Sofy (2009) studied the effect of Zn on broad bean plants they found that, the content of total soluble protein were increased. 3.5. Mineral content

It has been found in the present work fig. 14 that treating pea plants with Azsopirillum resulted in, generally, contents of phosphorus markedly increased in response to the same treatment. This is consistent with pervious findings by El shanshoury (1995) reported that inoculation with Azospirillum brasilense in sterilized soil stimulated plant growth and significantly increased P3+ , N, K+ , in wheat shoots. Deubel et al., (2000) found that Sugars, like phosphate-deficient substrates enhanced the capacity of Azospirillum sp. To solubilize normally insoluble Ca3(PO4)2.Rai and Gaur, (1982) found that inoculation of wheat crop with Azospirillum enhances mineral uptake by increasing uptake of N. Fages (1994) found that plant inoculation with Azospirillum results in enhance uptake of K+, H2PO4 and other microelements.

The increases in the levels of mineral contents N content as a result of Azospirillum treatment may be attributed to the fact that the nitrogen fixation was naturally the first major mechanism of action for the enhancement of plant growth by A. brasilenseas suggested by Warembourg et al., (1987).

Rhizobial inoculation act as an economical importance, it improves N concentration as a result of increasing N2 fixation capacity. Moreover, inoculation with Rhizobium leguminosarum increased the nodulation of broad bean and pea plants and consequently the nutrients (nitrogenous materials) secreted into the soil, it is also increased rhizospheri cmicroflora especially the acid producers and phosphate dissolvers causing more available phosphorus, as the finding of Lipman and Conybeare (1936).

In pea plants, the obtained results fig. 14 revealed that, contents of nitrogen, phosphorus and potassium were markedly increased in response to the treatment with Rhizobium. The obtained results are in agreement with those of other investigators. Abd-Allah and Omar (2001) found that Rhizobium leguminosarum inoculation significantly increased N and P content of faba bean plants.

Results of the present work fig. 14 , revealed that, in pea plants, contents of P and K were increased in

response to the treatment with mycorrhiza, however content of N were markedly increased in pea plants, decreased in broad bean plants. This is consistent with pervious findings by Evelin et al., (2009) found that arbuscular mycorrhizal fungi enhanced nutrient acquisition (P, N, Mg, and Ca), maintenance of the K:Na ratio. Also Cliquet and stewart (1993) observed that N uptake increased in AM plant due to a change in N metabolism brought about by changes in the enzymes associated with N metabolism. Mycorrhizal inoculation can increase P concentration in plants by enhancing its uptake facilitated by the extensive hyphae of the fungus which allows them to explore more soil volume than non-mycorrhizal plants (Ruiz-Lozano and Azco′n, 2000).

The mechanism of increasing plant P concentration by mycorrhiza fungal inoculation is based on the relatively high activity of AM fungal hyphae in absorbing soil P and and then translocate onto the host roots through a specific efficient active translocation as mentioned by Cooper and Tinker (1978). AM fungi may also lower the pH value of the soil, which increase the solubility of phosphorus.

Seguin et al.,(2003) reported that the AM fungi expand their hyphae in soil and plant root, and this hyphael network promote bi-directional nutrient movement where soil nutrients and water move to the plant and plant phostosynthates flow to the fungal network.

Results of the present work fig. 14 revealed that, in pea plants contents of N and P increased in response to the treatment with zinc. In pea plants content of phosphorus increased due to Zn application, while in broad bean P content decreased in response to the same treatment. This results was in agreement with other investigators Abd El-Hady(2007) found that nitrogen, P, and K concentration and their uptake of barley plants were increased by increasing Zn application. Also Rawia et al., (2010) indicated that the treatments of Zn application significantly increased all nutrients in leaves, i.e N, P, K, as compared with the control treatment in (Polianthes tuberosa ) the same author found that content of carbohydrates significantly increased due to application of zinc .

3.6 Protein profile

A question arises: does PGPR and mycorrhiza influence only the plant growth by mobilizing nutrients from the soil (Sylvia et al.,1999) or they induce some qualitative modification in the protein pattern? In order to asset this problem an SDS-PAGE electrophoresis was performed with the protein extracts. The obtained results table 5, figure 15 revealed that, in pea plants there are no significant qualitative differences between the protein electrophoresis pattern of seeds produced by non-inoculated (control) and rhizobacteria (Az, Rz),

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Mycorrhiza (Mz) inoculated pea plants. In this case the tested rhizobacterial strains and mycorrhiza help protein accumulation only by a mechanism which involves probably only an increased availability and uptake of nutrients from soil. This results are in agreement with other investigators, Ştefan et al., (2009) found that application of PGPR strains resulted in no significant difference between the protein electrophoresis pattern of seeds produced by non-inoculated and inoculated plants. Also Stefan et al., (2013) reported that application of rhizobacteria on runner bean resulted in no significant differences between the control and the inoculated samples.

The present work revealed that treating pea plants with Zn resulted in presence of a unique band with MW of 33 KDa. The stimulating effect of Zn attributes to increasing in crude protein. These results are in agreement with other investigators, Wanas (2002) reported that, spraying faba bean plants with Zn (50 and 100 ppm) significantly enhanced total carbohydrates and crude protein contents.

Figure1. Effect of Azospirillum brasilense (NO40), Rhizobium leguminosarum,,Mycorrhiza and Zinc sulphate on shoot length (cm) of

pea plants.

Figure2. Effect of Azospirillum brasilense (NO40), Rhizobium leguminosarum,,Mycorrhiza and Zinc sulphate on root length (cm) of

pea plants.

Figure3. Effect of Azospirillum brasilense (NO40), Rhizobium leguminosarum,,Mycorrhiza and Zinc sulphate number of leaves/plant of

pea plants.

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Figure4: Effect of Azospirillum brasilense (NO40), Rhizobium leguminosarum,Mycorrhiza and Zinc sulphate on chlorophyll a (mg/g

fresh weight).

Figure5: Effect of Azospirillum brasilense (NO40), Rhizobium leguminosarum,Mycorrhiza and Zinc sulphate on chlorophyll b (mg/g

fresh weight).

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Figure6: Effect of Azospirillum brasilense (NO40), Rhizobium leguminosarum,Mycorrhiza and Zinc sulphate on chlorophylla+ b (mg/g

fresh weight).

Figure7: Effect of Azospirillum brasilense (NO40), Rhizobium leguminosarum,,Mycorrhiza and Zinc sulphate on carotenoids (mg/g

fresh weight)

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Figure8: Effect of Azospirillum brasilense (NO40), Rhizobium leguminosarum,,Mycorrhiza and Zinc sulphate on the total-soluble

carbohydrate contents in shoot (mg/g dry weight) of pea plants


carbohydrate contents in root (mg/g dry weight) of pea plants

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carbohydrate contents in yielded seeds (mg/g dry weight) of pea plants

Figure11: Effect of Azospirillum brasilense (NO40), Rhizobium leguminosarum,Mycorrhiza and Zinc sulphate on the protein contents in

shoot (mg/g dry weight) of pea plants.


root (mg/g dry weight) of pea plants.

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yielded seeds (mg/g dry weight) of pea plants

Figure14: Effect of Azospirillum brasilense (NO40), Rhizobium leguminosarum, Mycorrhiza and Zinc sulphate on Nitrogen, Potassium

and phosphorus content (g/100g. dry weight) of pea plants

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Table (1): Effect of Azospirillum brasilense (NO40), Rhizobium leguminosarum, mycorrhiza and Zinc sulphate on protein profile in the

yielded seeds of Psium sativum .

Figure (15): SDS-PAGE protein patterns for five treatments of pea Plants: S1= control, S2= Azospirillum brasilense, S3= Rhizobium

leguminosarum, S4= mycorrhiza, S5= Zinc.

4. Conclusions

From the outcome of the obtained results, it is worth to mention that the use of biofertilizers and Zinc as micronutrient was of a stimulatory effect on growth and metabolism as well as in the yield of both broad bean and pea plants. The potent effects of these treatments were found to be viā the enhancement the biochemical and metabolic activity throughout the different growth stages. References [1] Abbaspour,H.; Saeidi, S.S. and Abedl-Wahhab,MA.

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[3]Abd El-Azeem, S.A.; Mehana, T.A. and Shabayek A.A.(2007): Response of Faba Bean (Vicia faba L.) to Inoculation with Plant Growth-Promoting

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