Canadian Journal of Agriculture and Crops

Volume 2, Number 1 (2017) pp 1-10 doi 10.20448/803.2.1.1.10 | Research Articles

 

Response of Chickpea to the Rhizobia Inoculation in Different Region of Morocco

Machehouri Abdelkader 1 Ben Messaoud Btissam 1 Zennouhi Omar 1 Nassiri Laila 1 Ibijbijen Jamal 1 , Siti Nor Ain Mayan 6 Radieah Mohd Nor 6 ,
1 Soil & Environment Microbiology Unit, Faculty of Sciences, Moulay Ismail University, Meknes, Morocco
6 Centre for Global Sustainability Studies (CGSS), Universiti Sains Malaysia, Penang, Malaysia

ABSTRACT

The objective of the present study is to assess the effect of rhizobia inoculations on the growth and nodulation of Chickpea. One rhizobia strains were used as control and soils sampled from eleven sites belonging to six different regions of Morocco. The rhizobia inoculation was applied at the time of the seedling, and the nitrogen fertilizer was performed three times the first application at the seedling moment then two and three weeks after the first application. The biomass and nodulation were evaluated under different treatments. The essay was realized at the greenhouse the Faculty of Sciences, Moulay Ismail University. The Inoculation with these root symbionts, increased the biomass accumulation of the chickpea treated compared to the absolute control. However, the best response result was that of the simple rhizobial inoculation which showed a very good growth as well as an important root infection assisted by the number and weight nodule compared to the rhizobial inoculation combined to the nitrogen fertilization and to the control plants. Finally, the symbiosis with rhizobia is an important biological technology to improve the sustainable production of leguminous plant in different agro-ecological regions.

Keywords: Chickpea, Rhizobia, Inoculation, Growth, Nodulation.

DOI: 10.20448/803.2.1.1.10

Citation: CosticăLupu(2017).Response of Chickpea to the Rhizobia Inoculation in Different Region of Morocco.Canadian Journal of Agriculture and Crops, 2(1): 1-10.

Copyright: This work is licensed under a Creative Commons Attribution 3.0 License.

Funding: This study received no specific financialsupport.

Competing Interests: The authors declare that they have no competing interests.

History: Received: 26 January 2017 / Revised: 18 February 2017 / Accepted: 20 February 2017 / Published: 22 February 2017

Publisher: Online Science Publishing

1.INTRODUCTION

As in many developing countries, Chickpea present an important legume food crop in Morocco. Chickpea as the other grain legumes is considered as main source of proteins in human and animal nutrition and plays a key role in crop rotations. As so, legumes (like chickpea) are cultivated in rotation with other crops with the aim to improve soil fertility and reduce the incidence of weeds, diseases and pests [1]. In Morocco, during the last decade, the chickpea crop has been progressively extended from semi-arid to arid areas where edapho-climatic conditions such as salinity, pH and temperature may have an adverse effect on the establishment of functional nitrogen-fixing symbiosis [2-4].

The present work aims to assess the response of chickpea of winter ILC 195 in different regions of Morocco. The response was tested for nodulation, dry matter yield  in plastic pots carried out in a greenhouse at the Faculty of Science, of Moulay Ismail University.

2.MATERIALS AND METHODS

2.1. The Choice of Soil Sampling Sites

The choice of the sampling sites was based on three criteria. The first type of sapling sites corresponded to the region where the chickpea is usually cultivated, the second type corresponded to the soils having problems according to the chickpea culture and the last one corresponded to the sites where chickpea will be cultivated. For that, the soils were sampled from six regions in Morocco: Sais (Meknes), Chaouia (Settat), Pre-Rif (Tissa), Central Plateau (Maaziz), Middle Atlas (Khénifra) and the Gharb plain (Gharb). The edaphoclimatic characteristics of each region are present in the table 2.

2.2. Soil Sample Collection

In order to preserve the soil in its field  natural conditions, the method “Soil core” described byVincent [5]was used sampling the soil in cylindrical pots (20cm x 16cm) at the rate of 7kg/ha of soil per pot.
Soil samples were collected from 10 sites of 6 different regions in Morocco. From each site 10 kg of soil were randomly sampled. The determination of the different physicochemical characteristics was performed after samples collection.

2.3. Physicochemical Characteristics of Soils

Soil samples taken from representative sites were used for analysis of soil texture and some chemical properties. The analysis were estimated according to the methods described byVan [6].

2.4. Experimentation

2.4.1. Treatments & Experimental Design

The essay was realized according to a design in randomized complete block arranged in split-plot, the chickpea used was the variety ILC 195. A Rhizobia reference strain IAPC1 was used as control to assess the efficiency of the native strains of different soils. Three treatments were executed:
T1: Treatment inoculated with the IAPC1strain;
T2: Treatment without inoculation no nitrogen fertilization;
T3: Treatment fertilized with nitrogen but without inoculation.
Five repetitions per treatment were taken.

2.4.2. Seeds Inoculation

The seeds inoculation of the treatment T1 was done at the sowing moment. The seeds used were coated using a mixture of peat and liquid suspension of Rhizobium strain IAPC1 (5.106 viable cells/seed).

2.4.3. Experimentation Conduct

A/ Sowing:
The chickpea seeds were sown at a depth of 1 to 2 cm at a rate of 4 seed per pot. The inoculated seeds were sown at the last time to ovoid contamination of the non inoculated seeds. After two weeks of growth under greenhouse conditions, one plant was removed in order to leave only three plant per pot.
B/ Fertilization:
After the determination of the field capacity of soils, the macro and micro fertilizers were applied in aqueous solution (table 1) before sowing the seeds.

Table-1. Quantities of macro and microelements used for soil fertilization[7]

The plant of the treatment T3 have received 200mg of nitrogen as urea form per kilogram of dry soil. The nitrogen input was applied three times at the rate of 66.6 mg (N)/kg of dry soil. The first application was at the sowing moment, then after two and three weeks after sowing respectively.

2.4.4. Assessment of the Response of Chickpea to the Rhizobia Inoculation

Six weeks after sowing, the plants were harvested. The response was assessed determining the nodulation by measuring the number and the dry weight of the nodules, the dry matter field of aerial parts of different plants, the nitrogen percentage and the yield nitrogen in plant for each treatment.

3. RESULTS

3.1. Characteristics of the Sampling Sites

The edaphoclimatic characteristics of the six regions sampled in Morocco are represented in the following table.

Table-2. Characteristics of the Soil sampling Sites[8]

3.2. Physico-Chemical Characteristics of Soils

The results of the analysis of soil sampled mainly the texture and some chemical properties were depicted in the following table.

Table-3. Physicochemical characteristics of soil samples
Table-4. Response of Chickpea to inoculation in soils from different sites

3.3. Nodulation

Nodulation is a qualitative assessment of the symbiotic fixation of the nitrogen. Based on the results of the present study, we can divide the sites into three groups. The first group concern the sites, which have shown an increase on the nodules number as well as their weight. The second group belong to the site that have demonstrate an increase on the nodules number without affecting their weight. Finally, the last group that depicted a negative response to inoculation (no effect on the nodules number and weight).

For the first group, the inoculation of chickpea with reference strain IAPC1 have significantly increased the nodules number and their weight in the soils of four sites “Maaziz, SidiBettach, Khenifra and Meknes 2”. The nodules number for these sites was respectively 727, 610, 306 and 125. In addition, the nodules dry weight was respectively 688, 518, 305 and 177mg (Table 4). 

For the site, “Tissa” the inoculation have increased the nodules number however, their weight have not increased. The last group belongs to the sites Meknes 1 and Settat. For this group the inoculation have shown no effect either on the nodules number or on their weight. The nodules number was 344 for Meknes 1 and 775.6 for Settat. The nodules weight of Meknes 1 was 850mg and 2.13mg for Settat (Table 4).
The nitrogen input have affected significantly the nodulation even that this inhibition was partial, the nodules were white, very small and their weight was negligible (Table 5).

Table-5. Inhibition of the nodulation by the nitrogen fertilization in soils sampled from different sites

3.4. Dry Matter Yield

The inoculation have increase the dry matter yield that varied from 9.02 and 16.16 g/pot mainly for the five following sites: Maaziz, SidiBettach, Khenifra, Meknes 2 and Fes. The percentage of the dry matter yield was 74% for Maaziz, 64.5% for Khenifra, 62.6% for SidiBettach, 38.8% for Meknes2 and 20.5% for Fes (Table 4). The inoculation have shown no effect on the yield dry matter especially for the two sites Meknes 1 and Settat. However, the application of nitrogen fertilizer have significantly increase the yield dry matter for all the studied sites.

3.5. Nitrogen Content and Total Nitrogen

The inoculation have increase significantly the nitrogen content in the shoot of the inoculated plants compared to the control that was not inoculated. The best response was observed for two sites SidiBettach and Maaziz with the following percentage 79% and 56% respectively (Table 4).

The total nitrogen yield of the shoot plants was also increased due to inoculation especially for the sites SidiBettach and Khenifra with the following percentage 108% and 200% respectively (Table 4).

4. DISCUSSION

The degradation of the agricultural land have been aggravated hence the use rehabilitation tools is required to sustain their productivity [9, 10]. This act may be ensured through the use of biological alternative such the rotation[11] in order to improve the physico-chemical properties of the soil [12] in addition to increase its flora and fauna diversity, to increase the yield and the quality of subsequent crops[13-15] .

The present study aims to evaluate the response of chickpea to the inoculation using soils sampled from different region in Morocco. The conduct of the essay in the greenhouse provide the possibility to study the behavior of plant in many and different soils. However, the sampling step must be done carefully in order to preserve as possible the natural state of soils and by the way ovoid the nitrogen mineralization[16, 17]. Once the legumes find two sources of nitrogen, they use with preference the nitrogen combined in the soil not the atmospheric nitrogen[18, 19].

These authors have reported that many other factors affect significantly the response of legumes to the inoculation mainly those environmental and nutritional.  

Rhizobia is another tool known by its importance in the rehabilitation of poor soils especially on nitrogen. These microorganisms are characterized by their ability to tolerate the main environmental factors[20, 21] and improve the growth and biomass of legumes and therefore increased yields. These results are in great agreement with ours, which can be explained by the fact that the rhizobial inoculation guarantees better absorption of nutrients, protection against pests and pathogens and the induction of the systemic resistance of legumes[22-26] . In addition, the rhizobia secrete auxins that promote root growth which in turn stimulates root infection[27, 28]. This could explain our results that showed a good infection assisted by the number and the weight of nodules.

However, it is recommended to identify combinations with native strains promoting the growth of plants to improve the local productivity of chickpea crops[20]. Anterior study revealed that the symbiotic effectiveness is dependent on the particular  combination of the strain and legume species, indicating the selective and specific compatibilities between bacterial and plants[29]. This result is verified by our study through which we noticed a significant improvement in shoot biomass of chickpea inoculated with rhizobia. This is explained by the fact that the growth responses of the inoculated legumes are influenced by the symbionts and the compatibility of the interactions between them (reference strain and the native strains) and those with the host plant[29-31] and also by the application of fertilizers mainly nitrogen ones. Moreover, there was a significant decrease in the number and the fresh weight of the nodule of all the studied sites; this could be explained by the inhibitory effect exerted by the nitrogen fertilizer on the rhizobia that is in perfect agreement with the results proved by Franzini, et al. [32].

Beside the importance of rhizobial inoculation with reference strain, it is better to isolate and identify the most efficient strain for each soil. Therefore their use as biofertilizer in order to have a direct application to improve soil fertility, to reduce chemicals and consequently participate to the protection of the environment supporting a sustainable agricultural system.

5. . CONCLUSION

In conclusion, our results have confirmed the important effects of the inoculation with rhizobia on the growth of chickpea in soils sampled from different region of Morocco. These results show an interesting way of rehabilitation of degraded lands that are in current deficient in nutrients. Symbiosis with rhizobia is a biological alternative that ensure the improvement of the sustainable production of legumes in different agro-ecological regions.

References

[1] J. Mwanamwenge, S. P. Loss, K. H. M. Siddique, and P. S. Cocks, "Growth, seed yield and water use of faba bean (Vicia Faba L.) in a short-season mediterranean-type environment," Australian Journal of Experimental Agriculture, vol. 38, pp. 171–180, 1998.View at Google Scholar | View at Publisher

[2] J. Maatallah, E. B. Berraho, S. Munoz, J. Sanjuan, and C. Lluch, "Phenotypic and molecular characterization of chickpea rhizobia isolated from different areas of Morocco," Journal of Applied Microbiology, vol. 93, pp. 531- 540, 2002.View at Google Scholar | View at Publisher

[3] D. J. Lauter, D. N. Munns, and K. L. Clarkin, "Salt response of chickpea as influenced by N supply," Agronomy Journal, vol. 73, pp. 167–169, 1981.

[4] P. M. Singleton and B. B. Bohlool, "Effect of salinity on nodule formation by soybean," Plant and Physiology, vol. 74, pp. 72–76, 1984.View at Google Scholar | View at Publisher

[5] J. M. Vincent, A manual for the practical study of root nodule bacterie. IBP Handbook n° 15 Brackwell 11 Oxford 1-164, 1970.

[6] R. Van, Procedures for soil analysis. Wegningen: International Soil Reference and Information Center, 2002.

[7] D. P. Beck, L. A. Materon, and F. Afandi, "Practical Rhizobum- Legume technology manual," International Center for Agricultural Research in the Dry Areas,Technical Manual No.19, 1993.

[8] H. Ghanem, "Contribution à l'étude des sols du Maroc. Génèse,classification et répartition  des sols des régions des Zaers et de ses bordures," Thèse de Docteur es Sciences, Université de Gand Belgique, 1978.

[9] C. M. C. Costa, U. M. T. Cavalcante, B. T. Goto, V. F. Santos, and L. C. Maia, "Fungos micorrízicos arbusculares e adubação fosfatada emmudas de mangabeira," Pesquisa Agropecuária Brasileira, vol. 40, pp. 225-232, 2005.View at Google Scholar | View at Publisher

[10] A. Ulrich and I. Zaspel, "Phylogenetic diversity of rhizobial strains nodulating Robinia pseudo acacia L," Microbiology, vol. 146, pp. 2997–3005, 2000.View at Google Scholar | View at Publisher

[11] L. C. Snook, "Tagasaste (Tree Lucernes): Chamaecitysus palmensis: Abrowse shrub which will increase production from grazing," Aust. Anim. Prod. Aust., vol. 15, pp. 589–592, 1982.View at Google Scholar 

[12] K. M. Saghir, A. Zaidi, and J. Musarrat, Microbes for legume improvement. Germany: Springer-Verlag / Wien, 2010.

[13] K. Recep, S. Fikrettin, D. Erkol, and E. Cafer, "Biological control of the potato dry rot caused byFusarium species using PGPR strains," Biological Control, vol. 50, pp. 194–198, 2009.View at Google Scholar | View at Publisher

[14] K. B. S. Dileep, "Fusarial wilt suppression and crop improvement through two rhizobacterial strains in chick pea growing in soils infested with fusarium oxysporum f. spciceris," Biology and Fertility of Soils, vol. 29, pp. 87–91, 1999.View at Google Scholar | View at Publisher

[15] B. J. Duijff, J. W. Meijer, P. Bakker, and B. Schippers, "Siderophore mediated competition for iron and induced disease resistance in the suppression of fusarium wilt of carnation by fluorescent Pseudomonas spp," European Journal of Plant Pathology, vol. 99, pp. 277–289, 1993.View at Google Scholar | View at Publisher

[16] M. B. Ben, L. Nassiri, and J. Ibijbijen, "Effects of rhizobia and mycorrhizae inoculations on the growth and nodulation of chamaecytisus proliferus," International Journal of Agricultural Sciences and Natural Resources, vol. 2, pp. 28-35, 2015.

[17] B. Soudi, A. Sbai, and N. C. Claude, "Nitrogen mineralization in semi arid soils of Morocco: Rate constant variation with depth," Soil Science Society of America Journal, vol. 54, pp. 756-761, 1990.

[18] Y. Obaton, "Generalités sur la symbiose fixatrice d’azote rhizobium légumineuse," presented at the I Biol 1 in Fiche Tech de la fixation Symbiotique de l’azote Via la Symbioose Legumineuses-Rhizobium Editée Par la FAO, 1983.

[19] J. J. Drevon, J. Hockman, F. Soussana, and L. Salsac, "Inhibition of nitrogen fixation by nitrate assimilation in legumes-rhizobium symbiosis," Plant Physiology and Biochemistry, vol. 26, pp. 197-203, 1988.View at Google Scholar 

[20] M. Srinivasan, D. J. Petersen, and F. B. Holl, "Influence of indoleaceticacid-producing bacillus isolates on the nodulation of phaseolus vulgaris by rhizobium etli under genotobiotic conditions," Canadian Journal of Microbiology, vol. 42, pp. 1006–1014, 1996.View at Google Scholar | View at Publisher

[21] J. K. Vessey and T. J. Buss, "Bacillus cereus UW85 inoculation effects on growth, nodulation, and N accumulation in grain legumes controlled - environment studies," Canadian Journal of Plant Science, vol. 82, pp. 282–290, 2002.View at Google Scholar | View at Publisher

[22] J. M. Ruiz-Lozano and R. Azcon, "Specificity and functional compatibility of VA mycorrhizal endophytes in association with bradyrhizobium strains in cicer arietinum," Symbiosis, vol. 15, pp. 217–226, 1993.View at Google Scholar 

[23] R. Azcon and J. A. Ocampo, "Factors affecting the vesiculararbuscular infection and mycorrhizal dependency of thirteen wheat cultivars," New Phytologist, vol. 87, pp. 677–685, 1981.View at Google Scholar | View at Publisher

[24] R. Azcon, R. Rubio, and J. M. Barea, "Selective interactions between different species of mycorrhizal fungi and rhizobium meliloti strains, and their effects on growth, N2 fixation (N15) in medicago sativa at four salinity levels," New Phytologist, vol. 117, pp. 399–404, 1991.View at Google Scholar | View at Publisher

[25] D. Redecker, P. V. Berswordt-Wallrabe, D. P. Beck, and D. Werner, "Influence of inoculation with arbuscular mycorrhizal fungi on stable isotopes of nitrogen in phaseolus vulgaris," Biology and Fertility of Soils, vol. 24, pp. 344–346, 1997.View at Google Scholar | View at Publisher

[26] P. E. Mortimer, M. A. Pérez-Fernandez, and A. J. Valentine, "The role of arbuscular mycorrhizal colonization in the carbon and nutrient economy of the tripartite symbiosis with nodulated phaseolus vulgaris," Soil Biology and Biochemistry, vol. 40, pp. 1019–1027, 2008.View at Google Scholar | View at Publisher

[27] P. E. A. Asea, R. M. N. Kucey, and J. W. B. Stewart, "Inorganic phosphate solubilization by two penicillium species in solution culture and soil," Soil Biology and Biochemistry, vol. 20, pp. 459-464, 1988.View at Google Scholar | View at Publisher

[28] H. O. Halvorson, A. Keynan, and H. L. Komberg, "Utilization of calcium phosphate for microbial growth at alkaline Ph," Soil Biology and Biochemistry, vol. 22, pp. 887 - 890, 1990.View at Google Scholar | View at Publisher

[29] B. A. McKenzie, G. D. Hill, V. Ganeshan, E. Yamoah, M. E. Andrews, and D. R. Humphry, "The role of legumes in improving nitrogen availability, soil fertility and growth in the following crop," Aspects of Applied Biology, vol. 63, pp. 53–60, 2001.View at Google Scholar 

[30] A. Alfaro, F. López, A. Pérez, J. C. García, and A. Rodríguez, "Integral valorization of tagasaste (Chamaecytisus Proliferus) under hydrothermal and pulp processing," Bioresource Technology, vol. 101, pp. 7635–7640, 2010.View at Google Scholar | View at Publisher

[31] M. Monjardino, D. Revell, and D. J. Pannell, "The potential contribution of forage shrubs to economic returns and environmental management in Australian dry land agricultural systems," Agricultural Systems, vol. 103, pp. 187–197, 2010.View at Google Scholar | View at Publisher

[32] V. I. Franzini, R. Azcon, M. F. Latanze, and R. Aroca, "Interaction between glomus species and rhizobium strains affect the nutritional physiology of drought stressed legume hosts," Journal of Plant Physiology, vol. 167, pp. 614–619, 2010. View at Google Scholar | View at Publisher

About the Authors

Machehouri Abdelkader
Soil & Environment Microbiology Unit, Faculty of Sciences, Moulay Ismail University, Meknes, Morocco
Ben Messaoud Btissam
Soil & Environment Microbiology Unit, Faculty of Sciences, Moulay Ismail University, Meknes, Morocco
Zennouhi Omar
Soil & Environment Microbiology Unit, Faculty of Sciences, Moulay Ismail University, Meknes, Morocco
Nassiri Laila
Soil & Environment Microbiology Unit, Faculty of Sciences, Moulay Ismail University, Meknes, Morocco
Ibijbijen Jamal
Soil & Environment Microbiology Unit, Faculty of Sciences, Moulay Ismail University, Meknes, Morocco
Siti Nor Ain Mayan
Centre for Global Sustainability Studies (CGSS), Universiti Sains Malaysia, Penang, Malaysia
Radieah Mohd Nor
Centre for Global Sustainability Studies (CGSS), Universiti Sains Malaysia, Penang, Malaysia

Corresponding Authors

Ibijbijen Jamal
Radieah Mohd Nor