Scholarly article on topic 'Response of Prosopis Chilensis to biofertilization under calcareous soil of RasSudr. 2 – Pod production'

Response of Prosopis Chilensis to biofertilization under calcareous soil of RasSudr. 2 – Pod production Academic research paper on "Agriculture, forestry, and fisheries"

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Abstract of research paper on Agriculture, forestry, and fisheries, author of scientific article — Fatma M.K. Faramawy

Abstract An experiment had been done in split design with 4 replicates in RasSudr Research Station, Desert Research Center, at South Sinai Governorate, Egypt, through 2009 on four years old Prosopis Chilensis to evaluate the effect of inoculation with Bradyrhizobium japonicum, Azotobacter chroococcum, Bacillus megatherium (PDB) and +VA mycorrhizae singly or in combination with vegetative (plant height, stem diameter and number of branches) and pod production (pod number/plant and pod weight/plant) and some chemical constituents total flavonoid). Results indicated that different biofertilizer treatments and their interactions significantly increased prosopis yield (pod number and pod weight), chemical analysis of leaves and branches (total carbohydrates, digestive protein, total digestive nutrients and total flavonoids) and chemical constituents of pods (crude protein, ash%, total carbohydrate, total sugar, total flavonoid and macro and micro-elements). Mixed inoculation gave better results than single ones, inoculation treatments can be arranged in descending order as follow, quarto inoculation, triple inoculation, double inoculation double inoculation and finally single inoculation.

Academic research paper on topic "Response of Prosopis Chilensis to biofertilization under calcareous soil of RasSudr. 2 – Pod production"

Annals of Agricultural Science (2014) 59(2), 263-271




Faculty of Agriculture, Ain Shams University Annals of Agricultural Science

Response of Prosopis Chilensis to biofertilization q^a

under calcareous soil of RasSudr. 2 - Pod


Fatma M.K. Faramawy *

Soil Fertility and Microbiology Dept., Desert Research Center, El Matarya, Cairo, Egypt

Received 18 August 2014; accepted 10 September 2014 Available online 12 December 2014


Prosopis Chilensis;


Pod production;

Total carbohydrate; Total sugar;

Total flavonoid and macro and micro-elements

Abstract An experiment had been done in split design with 4 replicates in RasSudr Research Station, Desert Research Center, at South Sinai Governorate, Egypt, through 2009 on four years old Prosopis Chilensis to evaluate the effect of inoculation with Bradyrhizobium japonicum, Azoto-bacter chroococcum, Bacillus megatherium (PDB) and +VA mycorrhizae singly or in combination with vegetative (plant height, stem diameter and number of branches) and pod production (pod number/plant and pod weight/plant) and some chemical constituents total flavonoid). Results indicated that different biofertilizer treatments and their interactions significantly increased prosopis yield (pod number and pod weight), chemical analysis of leaves and branches (total carbohydrates, digestive protein, total digestive nutrients and total flavonoids) and chemical constituents of pods (crude protein, ash%, total carbohydrate, total sugar, total flavonoid and macro and microelements). Mixed inoculation gave better results than single ones, inoculation treatments can be arranged in descending order as follow, quarto inoculation, triple inoculation, double inoculation double inoculation and finally single inoculation.

© 2014 Production and hosting by Elsevier B.V. on behalf of Faculty of Agriculture, Ain Shams



In traditional, cereal-producing farming systems in dry areas, crop failure and famine are expected to become more frequent. Availability of staple foods will decline and food prices are likely to rise, with far-reaching implications. Drought will become the norm rather than the exception and dependence

* Tel.: +20 01121447985. E-mail address: Peer review under responsibility of Faculty of Agriculture, Ain-Shams University.

on annual crops will become increasingly risky. When crops fail, people in dry lands have historically turned to trees for food and fodder. Growing more tree crops is proposed as one of the best coping strategies for reliably and sustainably improving food security. Drought tolerant trees play an essential role in climate change adaptation in these areas though planting them and getting them to survive in desert conditions is still challenging, so it is preferable to make the most of those trees that are already there.

Prosopis is a group of trees that stands out as having huge potential to help feed millions of people, especially the species Prosopis Chilensis. This is now one of the most numerous and

0570-1783 © 2014 Production and hosting by Elsevier B.V. on behalf of Faculty of Agriculture, Ain Shams University.

widespread trees in dry areas of Africa and Asia. It is also one of the most underutilized. Prosopis trees produce masses of nutritious beans even in the driest years. Where they are native in North and South America, prosopis trees of various species have been, and still are, an important staple food for many indigenous peoples. But as a relatively recent arrival in Africa and Asia, people there have not yet learned of its valuable uses and some even think that the beans are poisonous. Prosopis also has the potential to become a source of food for very many poor people in the dry areas of Africa and Asia. The sooner it does, the fewer poor people will go hungry. Prosopis beans as a food Prosopis beans are composed of hard seeds enclosed in a fibrous endocarp, surrounded by a sweet fleshy mesocarp, commonly 15 cm long, but up to 30 cm in some species. There is significant variation in nutritional values between species and even between trees of the same species. However, beans generally contain 10-20 percent crude protein, 30-60 percent carbohydrate including up to 40% sugars in some varieties, acceptable amounts of minerals, and an acceptable amino acids profile. This makes the beans comparable or superior to most cereals, with no anti-nutritional factors detected (Pasiecznik et al., 2001; Felker et al., 2012). In countries such as Argentina, Chile, Peru, Mexico, and the USA where proso-pis is native, the beans were an important staple food in times past (Beresford-Jones et al., 2009). These were traditionally ground into flour using stone mills, and mixed with maize or other flours to make bread, cakes, or a rich gruel. Alternatively, they were boiled into a molasses-like syrup for sweet drinks, and home-made products are still sold in local markets (Felker, 2005; Felker et al., 2012). Prosopis gum is similar to Arabic gum and can be used in cooking and all parts of the tree have traditional medicinal uses (Pasiecznik et al., 2001).

Materials and methods

Soil analysis of RasSudr research station See Tables 1 and 2. Field experiment

Field experiment was carried out on Prosopis Chilensis at four years old trees, which chosen from a previous study during 2006-2007 and given an annual biofertilizers doses till 2010. Experiment was conducted in RasSudr Research Station, Des-

ert Research Center, at South Sinai Governorate; Egypt during 2010. The soil of the location was highly calcareous. Analysis of the experimental soil and irrigated water was presented in Tables 1 and 2.

Production of trees inoculum

Active strains of Azotobacter chroococcum (as nitrogen fixing bacteria), Bacillus megatherium (as phosphate dissolving bacteria), Bradyrhizobium japonicum (as symbiotic nitrogen fixer bacteria) and vasicular arbuscular mycorrhizal fungi (as phosphate solubilizer symbiont) having the capability to withstand the stressed desert conditions were provided by Unit of Soil Microbiology, Desert Research Center, Cairo.

Heavy cell suspension of A. chroococcum or B. megatherium or Bradyrhizobium spp. cells containing about 108 cells ml-1 and VAM spore suspension (50 spore ml-1) were used as.

Mixtures of each strains were prepared just before inoculation by adding equal portion of the culture of each strain to inoculate Prosopis Chilensis seedlings.

Four replicates were chosen to study growth parameters (plant height, stem diameter, number of branches) and yield (number of pods and pods weight/plant) were estimated; also some chemical analyses for leaves and pods were carried out.

The experimental design was split plot in four replicates. The experimental treatments under the investigation were as follows;

1. control

2. B. japonicum

3. A. chroococcum

4. B. megatherium

5. VA mycorrhizae

6. B. japonicum + A. chroococcum

7. B. japonicum + B. megatherium

8. B. japonicum + A. mycorrhizae

9. A. chroococcum + B. megatherium

10. A. chroococcum + A. mycorrhizae

11. VA mycorrhizae + B. megatherium

12. B. japonicum + A. chroococcum + B. megatherium

13. B. japonicum + A. chroococcum + A. mycorrhizae

14. B. japonicum + B. megatherium + A. mycorrhizae

15. A. chroococcum + B. megatherium + A. mycorrhizae

16. B. japonicum + A. chroococcum + B. megatherium + VA mycorrhizae

Table 1 (a) Mechanical properties. (b) Chemical properties.

Depth (cm) Coarse sand % (1-0.5) mm Fine sand % (0.25-0.1) Total sand mm % Silt % (0.05-0.002) mm Clay % mm <(0.002) Class texture

Panel (a) 0-30 30-60 54.51 25.49 25.88 61.12 80.39 86.61 8.46 7.14 11.15 6.25 Sandy loam Sandy loam

Depth (cm) pH EC (ds/m2) CaCO3 O.M% Saturation soluble extract

Soluble anions (meq/L) Soluble cation (meq/L)

CO3 HCO 3 SO3 CI3 Ca+ + Mg+ + Na+ K+

Panel (b) 0-30 30-60 7.7 7.40 4.77 4.16 55.85 51.21 0.60 0.46 0.00 6.00 0.00 3.00 10.50 16.10 31.20 22.50 24.00 16.83 11.00 6.00 10.52 2.18 17.80 1.10

Table 2 Chemical analysis of irrigated water.

Well salinity (ppm) pH E C (ds/m2) Soluble anions (meq/L) Soluble cation (meq/L)

CO3 HCO3 SO4 CI4 Ca+ + Mg+ + Na + K +

45,000 8.60 9.23 0.00 2.50 16.22 81.28 23.65 19.18 57.17 0.51

Densities of plants were 4 plants per each experimental unit (2 x 12 meter).

The experimental site was irrigated by saline water pumped from a well of 4500 ppm salinity. The experiment received 20 m3/feddan organic farm manure during soil preparation before transplanting and 150 kg calcium super phosphate/fed-dan (15.5% P205). And K (100 kg/fed of potassium sulfate 48% k2O), Ammonium sulfate (20.5% N), 60 kg/fed. Four plants were taken at the end of the experiment to evaluate the response the following characteristics:

Microbial analysis

1. Soil samples were collected from rhizosphere region of prosopis plants and analyzed for total microbial count according to Bunt and Rovira (1965). For counting and growing phosphate dissolving bacteria the same medium was used after adding 5 ml of 10% K2HpO4 as a sterile solution followed by adding 10 ml of sterile solution of 10% CaCl2 to each 100 ml of the medium (Abd El-Hafez, 1966), Azotobacter nitrogen deficient medium (Abd El-Malek and Isac, 1968); Azospirilla on Dobereiner's medium (Dobereiner, 1978).

2. Extraction of VA mycorrhizal spores: Spores were collected from rhizosphere and soil samples by wet sieving and decanting technique (Gerdemann and Nicolson, 1963).

Growth characters

The studied growth parameters included (plant height (cm), stem diameter (cm) number of branches/plant.

Chemical nutritive constituents

Samples of the dried leaves, branches and pods were ground for the following chemical analysis:

1. Chlorophyll content: To calculate chlorophyll content as mg/g of tissue the following equation was used (Dhopte and Manuel, 2002).

Total chlorophyll mg/g = 20.2 (OD645) + 8.02(0D663) x(v/1000 x wt)

where OD = Optical Density at certain Wave length (645 or 663 nm). V = Final Volume (10 ml) Wt = Weight of sample (100 mg) Chlorophyll intensity.

The chlorophyll intensity meter (SPAD-520, Minolta, Japan) is a high weight portable diagnostic meter

2. Phosphorus was spectrophotometrically determined as

described by A.O.A.C. (1990).

3. Micronutrients were determined according to Soltanpour and Schwab (1977).

4. Total carbohydrate, according to method described by A.O.A.C. (1990).

5. Ash content, according to method described by A.O.A.C. (1990).

6. Crude fiber (C F) content, according to method described by A.O.A.C. (1990).

7. Crude protein (CP) content, total nitrogen was determined by modified microkjeldahl method described by Jackson (1958) according to Peach and Tracey (1956) and multiplied by 6.25.

8. Extraction, Fresh matter was extracted using 75% methanol (Shan et al., 2005). The mixture was filtered after 24 h of extraction in room temperature through filter paper. The extract was stored in the refrigerator until analysis.

Evaluation of total flavonoid content, the total flavonoid content was measured using a modified colorimetric method (Yoo et al., 2008). The appropriate amount of extract was added to a test-tube together with distilled water. Then was added 5% NaNO2, after 5 min 10% AlCl3 and after another 5 min 1 M NaOH followed by the addition of distilled water. The absorbance was measured against the blank at 510 nm after 15 min. The standard curve was prepared using different concentrations of catechin. The flavonoid content was expressed as g catechin equivalents (CE) per 100 g of dry weight (dw).

9. Estimation of total soluble sugar: The amount of total soluble sugars was estimated by Phenol sulfuric acid reagent method (Dubios et al., 1951).

Statistical analysis

The obtained data were subject to statistical analysis using Michigan Statistical program Version C (MSTSTC) least significance difference (L.S.D) value at 0.05 and for comparison between means of treatments were used as mentioned by Sendecor and Cochran (1981).

Results and discussion

Effect of biofertilizers on total count of soil microorganisms

Results in Table 3 showed the change in count which tends to increase in all treatments compared to the control. Mixed inoculation produced higher increase in the total microbial count. Similarly, Sheraz et al. (2010) reported that microbial inoculants increase the number and biological activities of desired microorganisms and improve the fertility in the root zone.

Table 3 Microbial counts of Prosopis Chilensis rhizosphere as influenced by fifteen bio-fertilizer treatments at pod production stage.

Treatments Total microbial N2 fixers PDB counts cfu x VM spores g 1 soil

counts cfu x 107 dry soil Azotobacter Azospirillum 103 dry soil

counts MPN x counts MPN

104 g_1 dry soil x 104 g_1 dry soil

Control 15.1 21 18.1 7 220

Bradyrhizobium 16.6 30 19.3 11 280

Azotobacter 17.2 46 20.0 18 320

PDB 15.9 28 18.8 32 235

VAM 16.5 32 19.7 16 420

Brady + Azotobact 18.2 52 22.0 22 300

Brady + PDB 17.9 33 20.0 38 290

Brady + VAM 17.3 37 20.2 24 440

Azotobact + PDB 17.4 50 21.6 38 280

Azotobact + VAM 18.4 57 22.4 24 465

PDB + VAM 17.5 35 21.0 37 340

Brady + Azoto + PDB 18.7 62 22.6 40 295

Brady + Azoto + VAM 19.5 64 22.8 27 480

Brady + PDB + VAM 19.1 39 22.5 38 380

Azoto + PDB + VAM 19.3 62 22.7 41 390

Brady + Azoto + PDB + VAM 19.8 65 23.0 43 420

Initial counts before biofertilization 70 x 105 17 x 104 15 x 104 15 x 102 3.0

L.S.D at 0.05 0.356 1.88 1.47 1.046 25.5

Effect of biofertilizers on nitrogen fixers

The initial count of N2 fixing azotobacters was 17 x 104 MPN/ g dry soil. Data recorded in Table 3 showed that the change in count which tends to increase in all treatments compared to the control. The counts under A. chroococcum inoculation showed the highest counts all over the mixed biofertilizer treatments while PDB (phosphate dissolving bacteria) inoculation caused the least increase of azotobacters count. Also, mixed applications of A. chroococcum + B. jponicum + B. megatherium + AM reported the highest counts. The obtained results proved that N2 fixers A. chroococcum enrich the soil by nitrogen fixation which increase soil fertility. The promoting effect due to application of A. chroococcum not only due to the nitrogen fixation but also to the production of plant growth promoting substances, production of amino acids, organic acids, vitamins and antimicrobial substances as well, which increase soil fertility, microbial community and plant growth (Revilla et al., 2005). The same trend was recognized in case of Azospir-illum counts, the initial count was 15 x 104 MPN/g dry soil. All biofertilizer applications recorded higher Azospirillum counts compared to control with superiority to Azotobacter applications in both single and mixed inoculations.

Effect of biofertilizer on the counts of Phosphate dissolving bacteria

Table 3 shows that the counts of PDB under inoculation with the same organism showed the highest counts all over the bio-fertilizer treatments. Also, a mixed application of B. megatherium reported higher count compared to single one being 32 x 103. The highest value was recorded in of A. chroococcum + B. jponicum + B. megatherium + AM treatment. It is worthy to notice that the initial count of phosphate dissolving bacteria was 15 x 102 cfu/g of dry soil. B. megatherium inocu-

lation stimulated the organism and increased its density compared to other treatments. A similar trend was recorded by Khan et al. (2006).

Effect of bio fertilization on VM production

It was evident from the present study that all the plants under investigation exhibited colonization by the arbuscular mycor-rhizal fungi were VM spores were present in the rhizosphere regions. The data in Table 3 showed that all biofertilizer treatments significantly increased VM spores numbers compared to control (without inoculation). Treatments inoculated with VM spores were the higher in number of spores in both single and mixed inoculated treatments. It was observed that adding azo-tobacter with AM had the highest positive effect on spore numbers, followed by bradyrhizobium being 380,420 spores/g. This was in agreement with finding of Rabin and Chikkaswamy (2014). It is worthy to notice that inoculation with PDB had the least increase in VM spore numbers. A similar trend was observed by Abou-El-Seoud and Abdel-Megeed, 2012.

Effect of biofertilizer on chlorophyll

It was observed in the present study that Bio-fertilizers used (Fig. 1) induced chlorophyll formation. The percentage increase on over control quantity of total chlorophyll and chlorophyll intensity. Treatments of mixed inoculation when applied were more promotory than single biofertilizer treatments. The significant variation in the level of total chlorophyll content in physiologically active leaves of prosopis plants may be due to variable rate of biosynthesis of chlorophyll and photosynthesis depending up. Significant enhancement in total chlorophyll content in leaves of all the treatments of biofertil-izers with respect to control (0.44) due to increased uptake of

2 1.5 1 0.5 0

¡Chlorophyll content mg/g Chlorophyll intensity

Comparative effect of biofertilizers on chlorophyll content and intensity of Prosopis Chilensis at pod production stage.

Fig. 1

magnesium from the soil in the form of Mg + 2 under the influence of VM fungi application and also the beneficial effects of bacterial inoculation on increased chlorophyll content due to higher availability of nitrogen to the growing tissue and organs supplied by aerobic nitrogen fixes. Results also confirm the earlier findings of Kowsar et al. (2014). The total chlorophyll content in leaves of prosopis under VM treatments was found to be stimulatory with respect to control which may be due to the stimulating effect of VM toward intensifying the green color of foliage (Anilkumar and Muraleedharakurup, 2012). The increased level of total chlorophyll concentration in leaves of all the VAM treated plants might be due to the influence of growth retardant on delaying leaf senescence and hence keeping the green pigment from degradation (TirupathiRao et al., 2013). The results are also in agreement with some earlier works (Banerjee et al., 2012).

Effect of biofertilizer on growth parameters of prosopis

Data in Table 4 show that, all tested biofertilizer treatments increased the plant height of Prosopis Chilensis plant as com-

pared with untreated plant (control) with superior for the quarto inoculation (Brady + Azoto + PDB + VM) as it gave 292 cm plant height. The differences between the four triple inoculated treatments of biofertilizers were not significant, as the plants under such applications had nearly close plant height values, while in the double inoculation, the application of Azotobacter + VM showed its superiority in this concern. Also, the plant height of Prosopis Chilensis plant was progressively increased within single inoculation, the highest value was recorded in azotobacter treatment (248.9 cm) followed by AM (247.5), then bradyrhizobium (241.1) and finally PDB (240.4).

Results revealed that all treatments of biofertilizers significantly increased stem diameter of Prosopis Chilensis plants, especially the triple inoculation treatments which scored the highest values, while the inoculation with Brady + Azoto + AM showed its superiority in this concern (15.1 cm). The differences between the single inoculation treatments did not reach level of significance as shown in Table 4. However, the highest value of stem diameter was obtained by using the combined treatment between Brady + Azoto + PDB + AM as it recorded 15.2 cm.

Table 4 Effect of biofertilizer treatments on vegetative growth of Prosopis Chilensis.

Treatments Growth parameters

Plant height (cm) Stem diameter (cm) No of branches

Control 213.4 9.6 57.5

Bradyrhizobium 241.1 12.8 60.5

Azotobacter 248.9 13.0 61.0

PDB 240.4 12.7 60.0

AM 247.5 12.9 60.5

Brady + Azotobact 262.5 13.8 65.0

Brady + PDB 256.2 13.6 64.5

Brady + AM 258.8 14.2 65.5

Azotobact + PDB 260.3 13.9 67.0

Azotobact + AM 265.7 14.8 64.5

PDB + AM 261.0 13.5 66.0

Brady + Azoto + PDB 279.3 14.9 66.5

Brady + Azoto + AM 283.4 15.1 68.5

Brady + PDB + AM 281.6 14.3 66.0

Azoto + PDB + AM 278.2 14.4 65.5

Brady + Azoto + PDB + AM 292.0 15.2 69.0

LSD at 0.05 5.4 0.45 0.8

Table 5 Pod yield of Prosopis Chilensis as influenced with different biofertilizer treatments.

Treatments Pods

Number/tree Weight(kg)/tree

Control 465 9.8

Bradyrhizobium 635 12.6

Azotobacter 697 13.1

PDB 618 12.2

AM 645 12.5

Brady + Azotobact 715 13.4

Brady + PDB 710 13.2

Brady + AM 665 13.8

Azotobact + PDB 728 13.6

Azotobact + AM 790 14.2

PDB + AM 740 13.6

Brady + Azoto + PDB 815 14.8

Brady + Azoto + AM 882 15.1

Brady + PDB + AM 805 14.5

Azoto + PDB + VAM 822 14.6

Brady + Azoto + PDB + sAM 918 15.8

L.S.D at 0.05 60 0.25

Table 4 shows that, all tested bio-treatments led to increase the number of branches/plant, particularly the AM treatments which induced the greatest number of branches/plant. Moreover Brady + Azoto + PDB + AM showed to be the most promising bio-treatment for producing the highest number of branches/plant (69 branch/plant). Besides, an increase in the number of branches was observed when prosopis plant received triple inoculation, so the highest value of branches number was recorded by Brady + Azoto + AM treatment (68.5 branch/plant).

Biofertlizers treatment which consisted of (Bradyrhizobium, Azotobacter, PDB and VM fungi) had a considerable effect on prosopis yield compared with untreated plants (Table 5). It could be noticed that all treatments (single, dual, triple and quarto inoculation) increased prosopis yield. Concerning to

single inoculation of azotobacter treatment gave the highest pods yield (697 pod/tree and 13.1 kg/tree), followed by VAM treatment, then Bradyrhizobium treatment and finally PDB treatment. However among dual inoculation, Azotobacter + AM treatment gave the highest pod yield (790 pod/tree and 14.2 kg/tree) followed by AM + PDB treatment, then Azotobacter + PDB, Bradyrhizobium + azotobacter and Bradyrhizobium + PDB showed no significant results in pod yield. Also Bradyrhizobium + Azotobacter + VAM treatment gave the highest pod yield among triple inoculation treatments. The best result was obtained in case of quarto inoculation, Brady + Azoto + PDB + AM (918 pod/tree and 15.8 kg/tree). These results are in agreement with Ahmed et al. (2013) on guar plants. Ehteshami et al. reported that both qualitative and quantitative characteristics in prosopis were significantly increased by phosphate-solubilizing microorganisms (VAM and PDB) and N2 fixers (Azotobacter and Brady-rhizobium) also increased the growth and resistance of plants.

Seed yield and yield attributes leguminous crops require more phosphorus than other crops to attain optimum growth and productivity (Gitari and Mureithi, 2003). P- solubilizing activity of phosphobacteria associated with the release of organic acids and a drop in the pH of the medium. Different kinds of organic acids, namely citric acid, gluconic acid, lactic acid, succinic acid and propionic acid were produced from the cultures of these isolates. This bacterium helps in increasing crop productivity by way of helping in solubilization of insoluble phosphorus, stimulating growth by providing hormones, vitamins and other growth factors (Bhattacharya and Jain, 2000). Also, VA mycorrhizal fungi form external mycelia extending several centimeters from the roots which can improve phosphate intake when phosphate availability is limited, it also realize acid and alkaline phosphatase which help in phosphate availability (Khatoon et al., 2011). The availability of phosphorus to legume crop is a key constraint to its production. The soil AM is responsible for transfer of the immobilized soil phosphorus into available form through which phosphorus becomes easily available to these legume crops (Singh et al., 2008). The stimulatory effects of biofertilizers used are in accordance with the results obtained by Fatima et al., 2007. In addition, Tran et al. (2006) reported that, the inoculation with certain plant growth-promoting rhizobacteria (PGPR) may enhance crop productivity either by making the other nutrients available or protecting plants from pathogenic microorganism (allelopathic effects). Zodape (2001) also concluded that, the increase in yield productivity with biofertilizer application is due to micro-element and plant growth regulator contained in the fertilizer.

Effect of biofertilizers on chemical constituents of prosopis

The total flavonoids of Prosopis Chilensis were determined in leaves and pods (Table 6). The percentages of total flavonoids increased gradually from single inoculation to quarto inoculation passing through double and tetra inoculation in both leaves and pods. Meanwhile in quarto inoculation, the percentages of total flavonoids were recorded the highest values of 6.13 and 8.35 in leaves and pods respectively. This quantitative variation of the flavonoids must be derived from induction by microbial inoculation, Yan-ping et al. (2004) found that inoculation with Streptomyces and some Bacillus spp. increased the

Table 6 Total flavonoid % of Prosopis Chilensis during the period of investigation (2006 and 2007) as influenced with different biofertilizer treatments.

Treatments Total flavonoid (%)

Leaves Pods

Control 3.78 4.09

Bradyrhizobium 4.09 7.12

Azotobacter 4.38 7.65

PDB 4.06 7.98

AM 4.30 7.45

Brady + Azotobact 5.08 8.05

Brady + PDB 4.97 7.94

Brady + AM 5.18 8.02

Azotobact + PDB 4.84 7.99

Azotobact + AM 4.99 8.18

PDB + AM 4.78 8.12

Brady + Azoto + PDB 5.57 8.13

Brady + Azoto + AM 5.74 8.18

Brady + PDB + AM 5.53 8.26

Azoto + PDB + AM 5.68 8.32

Brady + Azoto + PDB + AM 6.13 8.35

Table 7 Proximate analysis of whole Prosopis Chilensis pods as influenced with different biofertilizer treatments.

Treatments Analysis of prosopis pods (%)

Dry matter Crude protein Crude fiber Ash

Control 82.0 7.1 12.6 3.0

Bradyrhizobium 85.0 9.2 13.6 3.4

Azotobacter 86.1 9.8 14.2 3.9

PDB 84.8 8.8 13.5 3.5

AM 85.3 9.0 14.0 3.6

Brady + Azotobact 87.7 12.6 18.7 4.7

Brady + PDB 87.2 11.9 17.6 4.2

Brady + AM 88.3 12.1 18.5 4.5

Azotobact + PDB 87.9 12.4 18.2 4.4

Azotobact + AM 88.5 12.5 18.9 4.6

PDB + AM 87.5 12.2 18.0 4.4

Brady + Azoto + PDB 89.4 14.2 22.8 5.8

Brady + Azoto + AM 90.2 14.7 25.5 5.5

Brady + PDB + AM 89.2 14.2 24.7 5.3

Azoto + PDB + AM 89.6 14.3 25.4 5.4

Brady + Azoto + PDB + AM 93.6 16.2 28.0 6.2

L.S.D. at 0.05 0.3 0.2 0.2 0.1

Table 8 Macro and micro mineral contents in whole Prosopis Chilensis pods as influenced with different biofertilizer treatments.

Treatments Ca% P% Mg% Na% K% Cu (ppm) Zn (ppm) Mn (ppm) Fe (ppm)

Control 0.21 0.17 0.13 0.01 0.32 25.0 48.1 48.8 255.3

Bradyrhizobium 0.41 0.22 0.22 0.02 0.38 28.0 49.6 50.1 262.0

Azotobacter 0.43 0.24 0.28 0.03 0.40 28.0 49.8 50.4 265.0

PDB 0.38 0.27 0.20 0.02 0.36 27.0 49.1 49.4 259.4

AM 0.42 0.30 0.25 0.04 0.50 30.0 55.0 55.1 270.5

Brady + Azotobact 0.48 0.26 0.32 0.03 0.42 29.0 50.3 52.2 267.1

Brady + PDB 0.45 0.28 0.30 0.02 0.40 30.0 50.0 51.7 266.2

Brady + AM 0.46 0.37 0.32 0.04 0.54 35.0 50.8 55.6 265.7

Azotobact + PDB 0.45 0.33 0.35 0.03 0.42 30.0 49.9 51.8 277.1

Azotobact + AM 0.51 0.40 0.37 0.05 0.58 37.0 52.2 55.8 268.5

PDB + AM 0.45 0.48 0.35 0.04 0.55 36.0 50.6 55.5 279.3

Brady + Azoto + PDB 0.58 0.45 0.41 0.03 0.46 31.0 50.2 52.6 275.4

Brady + Azoto + AM 0.61 0.48 0.44 0.06 0.60 38.0 52.8 52.4 280.1

Brady + PDB + AM 0.56 0.55 0.40 0.06 0.62 37.0 52.6 55.7 279.6

Azoto + PDB + AM 0.59 0.57 0.48 0.07 0.60 38.0 52.8 55.9 278.7

Brady + Azoto + PDB + AM 0.71 0.60 0.51 0.09 0.66 40.0 53.3 56.2 280.8

L.S.D at 0.05 0.02 0.03 0.02 0.01 0.01 2.00 1.10 3.10 3.30

flavonoid content and grain yield of Tartary buck wheat. The mixtures of biofertilizers showing the greatest effect are in accordance with the results obtained by Patra et al., 2012, on soybean plant.

Further analysis of Prosopis Chilensis pods is shown in Table 7. All biofertilizer treatments increased the production of the dry matter, crude protein, crude fiber and ash of prosopis pods. It is observed that inoculation with azotobacter alone or in combination with other microorganisms due to its role in N2 fixation and nitrogen is one of the major plant nutrients, which are referred to as the master key elements in crop production (Mohamed et al., 2014). Also inoculation with VAM fungi increased all pod parameters due to its beneficial role in improving all nutrients uptake especially P.

The best result obtained among dual inoculation was that inoculated with Azotobacter + AM which gave 88.5 DM%, 12.5CP%, 18.9 CF% and 4.6 ash%. The triple inoculation

behaves the same trend, Brady + Azoto + VAM treatment gave the most significant results, followed by Azoto + PDB + AM. However Brady + Azoto + PDB treatment and Azoto + PDB + AM treatment showed no significance in their results. The best results were obtained in case of quarto inoculations due to improvement of nutrients uptake, Selvakumar et al. (2012) reported that, there was a significant enhancement in pod protein content owing to the application N and P biofertilizer treatments. A significant effect of N and P biofertilizer application on seed protein content has been reported for various leguminous crops, viz. chickpea (Eslam, 2010), groundnut, soybean (Tewari and Pal, 2005), and black gram (Selvakumar et al., 2012). Accordingly, the interaction of N and P biofertilizer treatments resulted in the highest content of seed protein and carbohydrate, with the optimum interaction being the one that also proved optimum in the case of yield parameters. In addition

Table 9 Total carbohydrate and total sugar content in whole Prosopis Chilensis pods as influenced with different biofertilizer treatments.

Treatments Total carbohydrate Total sugar (g)/100g (g)/100 g

Control 50.1 9.5

Bradyrhizobium 53.5 11.2

Azotobacter 59.7 13.5

PDB 61.8 10.2

VAM 54.5 12.5

Brady + Azotobact 51.5 14.4

Brady + PDB 60.5 13.5

Brady + VAM 66.5 13.8

Azotobact + PDB 62.8 13.6

Azotobact + VAM 69.0 14.2

PDB + VAM 64.0 13.6

Brady + Azoto + PDB 71.5 14.8

Brady + Azoto + VAM 68.2 15.1

Brady + PDB + VAM 70.5 15.5

Azoto + PDB + VAM 72.2 15.6

Brady + Azoto + PDB + VAM 71.8 16.2

L.S.D at 0.05 2.80 0.90

a combined application of N and P biofertilizers (Bradyrhizo-bium, Azotobacter, AM and PDB) was generally better than the single application of either of these biofertilizers. The interaction of the P and N biofertilizer treatments was significant for most of the parameters studied., which was also in the regarding N uptake, pod yield, and most of the yield parameters, proved to be the most advantageous and cost-effective interaction for yield parameters, N uptake, and seed quality parameters.

Moreover, significant increase in CP content may be due to the fact that legumes contribute to the total pool of nitrogen in the soil as observed by Ahmad et al., 2001. Higher P content may be due to inoculation and availability of P nutrients in soil by microbes. Soil pH, organic C, total N, P and K, available N, P and K content were significantly increased by the application of biofertilizer application. Concerning micro-nutrients i.e., iron, zinc, sodium, copper, potassium and manganese. The mineral concentration of Prosopis Chilensis was not sufficiently available in control and single inoculation treatments, but mixed treatments especially quarto one contain sufficient amounts of minerals especially calcium and phosphorus (Sharma, 1997), ie, mixed biofertilized treatments induced higher values in comparison with the mineral ones for control or single alone.

From Table 8, it is clear that application of bio-fertilizers led to more plant contents of macro and micro nutrients compare to non biofertilizers application. The maximum values were observed as a result of treating with mixture of Brady-rhizobium + azotobacter + PDB + AM treatment.

Biofertilizers treatments showed that, Ca% increased gradually from single to quarto inoculation passing through dual and triple once. The other elements have the same trend except P which showed dramatic increase in treatments inoculated with VAM and PDB due to their roles in phosphorus availability, this is in agreement of Jakobsen et al. (2002) and El-Quesni et al. (2013).

The bio-chemical parameters such as pod protein, carbohydrate and total sugar, were increased in treated with combined inoculation of biofertilizers (Brady, Azoto, PDB and AM) of Prosopis Chilensis (Table 9). This was well correlated with earlier studies on Vignamungo L. by Selvakumar et al. (2012).

This investigation clearly showed that the potential value of Prosopis Chilensis pods is a significant source of flavonoids, sugar and carbohydrates, therefore prosopis could be considered a good source of natural untraditional source of food. Since commercial prosopis species do not exist, these results could be important to use these species as breeding materials in future.


The dry lands, in which tropical Africa is no exception contain a rich wealth of indigenous fodder tree and shrub species which are regarded as an important source of fodder for livestock. However, only little is known about the nutritive value for most of these species identified. The present findings showed chemical composition as well as the mineral concentrations of Prosopis Chilensis fruits were within the ranges reported elsewhere with the same browse species. The rich content of Prosopis Chilensis fruits (pods) with protein, energy and mineral concentration might give a strong indication that Prosopis Chilensis is potentially a suitable fodder tree that can meet the grazing requirements of livestock for the sustain-ability of animal production. Our current results likely encourage conducting further experimental work on Prosopis Chilensis to enrich knowledge on the nutritional value of this important forest tree species.


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