International Journal of Agricultural Science and Research (IJASR) ISSN 2250-0057 Vol.

3, Issue 1, Mar 2013, 43-62 TJPRC Pvt. Ltd.

AGRO-MORPHOLOGICAL AND PHYSICAL PADDY SEED ATTRIBUTES OF NERICA AND LOCAL RICE VARIETIES AS AFFECTED BY MYCORRHIZAL INOCULATION AND COMPOST APPLICATION UNDER UPLAND CONDITIONS
NGAKOU A1, MBAIGUINAM M2, NADJILOM Y1 & TOKAM N. M1
1

Department of Biological Sciences, University of Ngaoundere, Ngaoundere, Cameroon

2

Research Laboratory of Natural Substances , Faculty of Applied and Exact Sciences, University of NDjamena, Chad

ABSTRACT
This research was conducted to seek for agro-morphological and physical seed characteristics of local and Nerica rice varieties and determine their behavior under mycorhizal inoculation and organics amendment in the Tandjil regionChad. The experiment was carried out in a split-splot (4x4) x 4, where the varieties were considered as the main factor, and the type of fertilization (mycorhize, compost, fertilizer and control) as the secondary factor or sub-treatment. The results obtained indicate that the biological fertilizers (Mycorrhiza, compost) and fertilizer (NPK) significantly (p < 0.05) improved the growth parameters such as height, number and length of panicles, number of productive tillers on Nerica N6 and NL28 at 60 and 90 days after planting compared to those of the control. Mycorrhiza and fertilizer had a positive impact on yield parameters of Nerica N6 and NL28 varieties, compared to the control. In addition, these fertilizers positively and significantly (0.0001 p < 0.05) affected the physical characteristics of Nerica seeds at harvest, except the bulk density and porosity. In contrast, biological fertilizer did not affect the seed yield of the two local rice varieties Tox728-1 and Madjitolngar, although their yield was greater than that of Nerica varieties. Our results suggest that mycorrhiza inoculum used in this study was not compatible with the local rice varieties. Further research will be focused on isolation and identification of native mycorrhiza associated to the local rice varieties in this region.

KEYWORDS: Mycorrhiza, Compost, Fertilizer , Nerica and Local Rice Varieties, Physical Seed Traits INTRODUCTION
In an effort to reduce growth rate and the high living costs in population of sub-sahara African countries, Chad is undergoing development of projects such as the one on rice production in the south of the country, and particularly in the Tandjile zone to improve and consolidate food security. Rice (Oryza sp, poaceae) is one of the most cultivated cereals in tropical, sub-tropical and temperate regions in the world, and is referred to all plants of the genus Oryza, of which the two cultivated species are O. sativa and O. glaberrima, originated respectively from Asia and Africa (Lizhi et al. 2006). Nerica (New Rice for Africa) variety devives from the crossing between the two species (ADRAO 1992). Rice is an important source of calories, vitamins, mineral elements and rare amino acids (FAOSTAT 2005; Badawi 2004). In Chad, rice is grown either under upland or lowland conditions, with or without irrigation, for its seeds rich in starch, and is consumed as seeds, meal or in soups. However, the plants can only express its optimal potentials if chemical fertilizers are provided for growth to equilibrate their elemental mineral balances, although this practice is subjected to soil nutritional and environmental pollutions (Lachance and Rouleau 2004). Sustainable agronomic researches have revealed that crop production can be better enhanced with more adapted, low costly and safe strategies based on biological agriculture (Thuries et al. 2000; Ngakou 2007). Among these strategies, biofertilizer research is being highly recommended for developing countries to break down their dependency to chemical fertilizer toward to the northern countries (Ngakou et al.

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Ngakou A, Mbaiguinam M, Nadjilom Y & Tokam N. M

2007; Ngonkeu 2009). With the understanding that most of the cultivated plants establish a mutualistic relationship with arbuscular mycorhiza fungi (AMF) (Smith and Read 1997; Khalil et al. 1992), this type of association could be exploited for the benefit of plants (Fresco 2003), since it can further improve the soil structure (Wright and Upadhyaya, 1998; Bethlenfalvay et al., 1998), the resistance of plants to stresses (Bethlenfalvay 1992; Turk et al. 2006)), insect pests (Ngakou et al., 2008a), and pathogens (Hooker et al., 1994; Trotta et al. 1996). Effective nutrient acquisition by mycorrhiza is generally attributed to the extensive hyphal growth beyond the nutrient depletion zone surrounding the root (Tisdale et al., 1995). Previous researches on mycorhizal inoculation and organic amendment in Cameroon have revealed enhancement of plant growth and yield (Ngakou et al., 2006; Ngakou et al., 2008b), and control of plant diseases (Ngakou et al. 2012). Considering the needs of improving rice production, mycorrhiza and soil solarization have been tested as sustainable strategies for Nerica rice production in Cameroon (Natebaye 2010; Signe 2011). However, no research has yet been conducted to assess the effect of mycorrhiza and compost on physical traits of rice. To the best of our knowledge, and unlike Cameroon, no mycorrhizal inoculation research has been investigated in Chad. The Tandjile region-Chad belongs to the same Guinea-savannah agro-ecological zone where most of the researches on mycorrhiza associated to selected crop plants have been carried out in Cameroon. The question derived thereof is: what could be the response of rice to associated introduced mycorrhiza inoculum in this region? In the Tandjile region like other rice growing areas, rough rice (or paddy) enters after harvest a whitening section, during which the hulls and the brownish outer bran layer are removed. By removing the bran particles, the white rice (milled rice) is then produced. Knowledge of physical dimensions plays an important role for designing huller and husker equipments. Moreover, it is an essential issue in marketing and grading, developing new varieties, and drying operations (Ashtiani et al. 2010; Emadzadeh et al. 2010). Therefore, this research was designed to provide rice plants with a biological (mycorrhiza) and organic (compost) fertilizers, to enable better agro-morphological and physical seed traits of Nerica and local rice varieties. The type and quality of information drawn out from this study will allow selection of a sustainable fertilization strategy to be coupled with improvement of rice production in the region, thus to reduce food insecurity within inhabiting population.

MATERIAL AND METHODS

Study site: Tandjile N 92325 E 161224

Figure 1: Map of Chad Indicating the Study Site (in Green Colour) Descrption of the Study Site The experimental field was located in the Sategui Deressia valley, south of Chad, in the Tandjile region, between the Logone and Chari rivers, but toward the right border of Logone. The soils are formed on recent alluvions rich in clay

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and limon. The climate is of the sudano type, characterized by two seasons: the rainy season extending from May to October, and the dry season from November to April. Analysis of the Experimental Soil Experimental soils were randomly sampled before sowing and after harvest on the whole study area at between 10 and 15cm depth along the medians and diagonals. After homogenization, three replicates of 1kg soils were sub-sampled from each group. These soils as well as compost samples were physico-chemically analyzed in the Laboratory of Soil and Plant Analysis (LASEP) of ITRAD (Research Institute for Agronomic Development) of NDjamena, Chad, following AFNOR (1982) method. Rice Seeds, Biological and Chemical Fertilizers Mycorrhiza as biological fertilizer was produced at the University of Ngaoundere, and made of a mixture of soil, root fragments and spores of the genus Glomus (10 25 spores/1g of soil). Compost as organic amendment was produced within the experimental sited from 1/1 volume of cow dungs/kitchen manures. The chemical fertilizers NPK (fertilizer ), with the formula 20:10:10 was graciously offered by Malla Farm company of Tandlile-Chad. Rice seeds, also provided by the same company were four varieties, two of which were local (Madjitolngar and Tox 728-1) and the two other Nerica (NL28 and Nerica N6).

(a) Nerica NL28

(b) Nerica N6

(c) Tox 728-1

(d) Madjitolngar

Figure 2: Rice Varieties (a) Nerica NL28, (b) Nerica N6, Local (c) Tox 728-1 and (d) Madjitolngar Experimental Design and Treatments A tract was used to plough a 1000 m2 experimental field at Malla farm in Tandjile region. The work was carried out in a split-plot (4x4) x 4 experimental design, in which the main factor was represented by the four types of fertilization (mycorrhiza, compost, fertilizer , nothing), and the four rice varieties (Tox 728-1; NL28, N6; Madjitolngar) as the subtreatments. The field was then subdivided into 9m2 subplots known as experimental units separated one another by 1m and

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Ngakou A, Mbaiguinam M, Nadjilom Y & Tokam N. M

replicated four times for each treatment. The four treatments were: the control in which no experimental unit receiced neither mycorhiza, nor compost or fertilizer ; biological fertilizer in which experimental units were inoculated with 10g of mycorrhiza in their sowing holes before planting; organic fertilizer (compost) in which experimental units were inoculated with 40g of compost in their sowing holes before planting; fertilizer in which each plant of the experimental unit received 30g of fertilizer (20:10:10) at 30 days after planting (DAP). In each of the 9 m2 experimental plot comprising 11 rows and 14 columns, 3 rice seeds were directly planted per hole on June 10th 2011. Plants were thinned to 1 per hole two weeks after germination to respect the rice planting guide lines. Two plants were separated 25 cm on a row and 20 cm apart. In total, each experimental unit has 154 plants, thus 616 plants per treatment. A partial view of an experimental unit is given by Figure 3 showing planting holes amended with compost on which rice seeds were sown. Nerica and local rice varieties were cultivated up to maturity under upland field during the raining season and without watering condition.

Figure 3 : Partial View of an Experimental Unit Showing Planting Holes Amended with 40 g Compost and Sown with Rice Seeds Measurement of Agromorphological Parameters The number of emerging seeds from planting holes were determined by counting at 14 DAP. The germination rate was then estimated as a ratio of the number of plantlets on the total number of planted seeds. Growth parameters were evaluated at 60 and 90 DAP on 10 randomly selected plants per experimental unit and per rice variety. Thus, the number of productive tillers and panicles/plant were assessed by visual counting, whereas the length of panicle and height of plants were measured using a graduated ruler. The colour of leaves and the symptoms of diseased were also observed. Evaluation of Yield Components At maturity, yield components were assessed on 10 randomly selected plants per experimental unit and per rice variety. Thus, the number of panicles/plant, the number of grain/panicle were determined by counting the grain per panicle per variety and per treatment. Similarly, a 2m ruler was used to measure the length of panicle per plant per rice variety and per treatment. An electronic balance Adam PGW 153i (Max 150 g, 0.001 sensibility) was used to evaluate the weight of 1000 grains. From the weight of grains per experimental unit, the grain yield per hectare (10000 m2) was estimated by extrapolation for each of the rice varieties.

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Determination of Physical Traits of Nerica Rice Seeds

Seed Dimensions and Weight Thirty rice seeds of each of the treatments were randomly selected for the determination of the physical traits according to Baryeh (2001), and Kaptso et al. (2008). The individual dimensions of seeds such as the length (L), the major diameter (D) and the minor diameter (d) were measured from the three main dimensions which are in three mutually perpendicular directions using a micrometer gauge reading at 0.01 mm sensitivity. The length was defined as the distance from one end to the opposite end of the seed, while D and d taken in the two opposite perpendicular directions of seed represented the major and the minor seed diameters. The seed weight was evaluated by an electronic balance (type Sartorius, Prodilab, France), with 0.001 sensitivity. Using the different readings, the geometric mean diameter Dg (mm) was calculated as follow:

The sphericity () was calculated using the equation:

The volume Vg was calculated by the equation:

True and Bulk Seed Density and Porosity The true mean volume of seed was determined according to the water displacement method of Karababa (2006). The true seed density (TSD) expressed in g/ml was calculated as the ratio of weight of seeds to the true volume. Practically, twenty five seeds of given weight, M, were introduced in a graduated flask containing 10 ml distilled water, thus inducing a change in the volume V of water that allows the determination of the true seed density expressed as:

The bulk density (BSD) was determined and expressed in g/ml (Okezie and Bello 1988). In this respect, a 30 ml (V) volumetric glass was filled with seeds. Considering M as the weight of seeds introduced in the glass, the ratio of M on the volume V determines the bulk seed density expressed as:

Following the above, the porosity () of the rice seeds which is the fraction of the space in the bulk seed not occupied by grains, was computed from the values of true seed density and bulk density using the relationship determined by Baryeh (2001): =

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Ngakou A, Mbaiguinam M, Nadjilom Y & Tokam N. M

Statistical Analysis All the data collected were subjected to analysis of variance (ANOVA) using the StatGraphics plus software program. Treatment means were compared using the Least Significance Difference (LSD) at 5%. Correlations between variables were determined using the statistical package for social science (SPSS)

RESULTS AND DISCUSSIONS

Soil Physico-Chemical Attributes Table 1: Chemical Composition of the Experimental Soil before Sowing Soil Composition K (mg/l) N (mg/l) P (mg/l) pH Control 3.0b 0.72b 21.0b 6.51a Compost 2.5a 0.01a 5.5a 8.34b p-value 0.04 < 0.0001 < 0.0001 < 0.0001 For each mineral element values of the same column affected by the same letter are not significantly different at 5% Table 1 displays results on the physico-chemical composition of the experimental soil and compost before this research. Compost and the native soil had different physico-chemical attributes, with compost exhibiting more organic matter (78%), more C/N ratio (41) than that of the native soil. The pH of compost was basic (8.34) compared to that of the native soil (6.52). Compost has been reported to positively influence the soil organic matter that empedes soil acidification (IRAB 2001). In contrast, the native soil contained more nitrogen, potassium and phosphorus contents than compost. This was explained by the history of the native soil that was revealed by the farm technician (personal communication) to have received fertilizer during preceding cultures. According to Bordeleau (1999), compost is very rich in organic matter and colloids that are able to form complexes with soil cations, thereby avoiding leaching of mineral elements. Table 2: Chemical Composition of the Experimental Soil after Harvest Soil Composition K (mg/l) N (mg/l) P (mg/l) pH C (%) MO (%) Control 3.0a 0.72a 21.0a 6.51c na na Compost 19.5b 11.46b 75.0c 6.18b 1.1b 1.88b Fertilizer 22.0c 17.63c 39.1b 6.02a 1.22b 0.71c Mycorrhiza 25.0d 18.13c 23.0a 6.43c 0.18a 0.31a p-value .0001 .0001 .0001 .0001 .0001 .0001 For each type of fertilization values of the same column affected by the same letter are not significantly different at 5% After harvest, soils that received fertilizer and mycorrhiza were enhanced in nitrogen, phosphous and potassium contents more than the control soil (Table 2). One of the mechanisms by which mycorrhiza act has been reported to be the increased elemental mineral zone within the rhizosphere by plant root (Smith and Read 1997; Ngakou et al. 2007). After harvest, all soil pH were acid, but with compost, mycorrhiza and fertilizer significantly (p 0.0001) reducing the pH compared to the control soil. Andrianasolo (2009) has shown that pH values slightly lower than 7 correspond to optimal assimilation conditions of available soil nutrients by plants, as well as development of mycorrhiza, optimal nitrogen fixation and phosphorus solubilization. Under these conditions, there is a possibility for the fungus partner to get access to organic forms of P and N that are translocated to the host plant (Nouaim et al., 1994). Similarly, the use of compost allows the soil to acquire a buffer status against modifications induced by application of agro-chemicals, and that enable availability of nutrients, particularly N and P to the host plant (Amadou et al. 2003). Soil Types Soil Types

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Influence of the Type of Fertilization on Seed Emergence Whereas the local rice variety Madjitolngar significantly (p = 0.0049) expressed a high rate of germination on soil that have received compost, mycorrhiza or fertilizer , Nerica rice variety poorly germinated on all these soil compared to control (Table 3). These results indicate that the local rice varieties are adapted to their local condition, thus behave well as compared to the introduced Nerica rice varieties. It has been stipulated that growing areas that have maintained a high organic matter content may harbour in their soil a large population of micro-organisms to which they are associated to express their potentials (Dalzell, 1988). Table 3: Seed Emergence Rate in Relation to Types of Fertilization Types of Fertilization Control Compost Fertilizer Mycorrhiza Nerica N6 19.33a 25.33a 25.0a 46.0a Nerica NL28 58.0b 56.66b 57.33b 53.0a Madjitolngar 114.33c 64.0c 69.33c 116.33c TOX728-1 66.0d 54.66d 84.0d 80.66b p-value 0.0049 0.0015 0.0241 0.0090 For each type of fertilization values of the same column affected by the same letter are not significantly different at 5% Influence of the Type of Fertilization on Growth Parameters of Rice Varieties at 90 and 60 DAP At 60 days after planting (DAP) the growing parameters of rice were evaluated as indicated by results in table 4. Application of compost did not affect the height of Nerica N6 variety. In contrast, the fertilizer (N,P,K) and mycorrhiza consistently enhanced the rice plant height compared to compost and control plants for Nerica N6 (p = 0.033) and NL28 (p 0.0001). The local variety Madjitolngar positively reacted to all the type of fertilization by increasing the rice plant height compared to the control. Unlike, the height and tillering of the local variety TOX728-1 was only increased by compost and fertilizer . Globally, the type of fertilizer positively impacted the height and tillering of all the four rice varieties compared to the control, although some increments were not significant. These findings are in agreement with other results obtained in Cameroon on Nerica FN-62 and FN-57 rice varieties after mycorrhizal inoculation and fertilizer application (Natebaye 2010). However, the mycorrhiza did not affect tillering of the local rice varieties Madjitolngar and TOX728-1. The numbers of productive tillers and fertile grains are known as primary yield components, which also play an important role in yield formation (Tiwari et al. 2011; Gevrek et al. 2012). Vigorous seedlings and high yield values were ascribed to plant hormones promoted vegetative growth by active cell division, cell enlargement and cell elongation and thus helped in improving growth (Pareek et al. 2000). These findings were in close agreement with the results of Dunand (1998) who also reported significant increase in plant height, stem elongation and yield of rice in response to gibberellic acid application. There is evidence that mycorrhizal plants contain higher concentration of growth hormones than their non-mycorrhizal equivalents (Skimmer, 1985). The improvement of plant growth parameters following biological or fertilizer application have been attributed to acquisition of mineral nutrients mostly those able to be translocated through mycorhiza (Smith and Read, 1997; Ngakou et al. 2007), enhanced soil fertility or structure by compost (Skat 1996; Ngakou et al. 2008b), and/or fertilizer (Schiltine 2002). At 60 DAP, only Nerica N6 was able to produce panicles, with mycorrhiya significantly and positively influencing their number more than any other treatment. Rice Varieties

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Ngakou A, Mbaiguinam M, Nadjilom Y & Tokam N. M

Table 4: Changes in Growth Responses of Rice Varieties as Influenced by Types of Fertilization at 60 DAP Rice Varieties Control Nerica N6 Nerica NL28 Madjitolngar Tox 728-1 LSD 5% p-value Nerica N6 Nerica NL28 Madjitolngar Tox 728-1 LSD 5% p-value Nerica N6 6.46aa 20.0 ab 30.0ac 28.7ba 8.7 0.0001 0.90abc 0.52aa 0.76ac 0.62ab 0.096 0.0001 0.00a Types of Fertilization Compost Fertilizer Mycorrhiza Tiller Number.Plant-1 ab 7.66 a 10.0b a 6.56aa ab c 25.36 b 50.23 d 28.2bc b b 38.5 c 37.16 c 24.9ac b a 29.9 b 21.06 b 18.8ab 8.6 11.07 6.03 0.0001 0.0001 0.0001 Plant Height (cm) 0.86ad 0.98 bc 0.94abc a b 0.53 a 0.62 a 0.56aa a a 0.79 c 0.81 b 0.78ab a a 0.65 b 0.65 a 0.61aa 0.074 0.158 0.164 0.0001 0.0001 0.0001 Panicle Number.Plant-1 0.14b 0.67c 1.03d LSD 5% 3.43 8.2 7.16 7.63 p-value 0.011 0.0001 0.0003 0.003

0.11 0.05 ns ns

0.033 0.0001 0.366 0.183 0.0001

0.36

0.041

For each rice variety, values of the same row affected by the same letter as superscript for a given parameter are not significantly different at 5%. For each type of fertilization values of the same column affected by the same letter as subscript for a given parameter are not significantly different at 5%; ns = non significant Compost and mycorrhiza did not affect the rice plant height of the two Nerica and local Tox728-1 rice varieties at 90 DAP (Table 5). Unlike, fertilizer significantly (p 0.0001) enhanced the plant height of Nerica variety compared to the control and mycorrhizal plants, then that of the local Madjiltolngar compared to Mycorrhizal plants, but has no effect on the local Tox 728-1 variety. Whereas Nerica N6 and Madjitolngar were significantly (p 0,0001) taller than other varieties, Nerica NL28 plants were the shortest of all the varieties at 90 DAP. At this date, panicles emergence was evident for all the varieties. Fertilizer and mycorrhiza consistently increased the number of panicles in Nerica N6 and NL28 varieties compared to the control plants. However, mycorrhiza did not affect the number of panicle per plants for the two local rice varieties.

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B Figure 7: An Overview of 60 DAP NL28 Nerica (A) and Tox 728-1 Local (B) Rice Varieties both Inoculated with Mycorrhiza For the two local rice varieties, compost was more indicated as the best fertilization type for this parameter. Tox 728-1 significantly (p 0.0001) yielded the highest number of panicles per plant, while that of Nerica N6 was the lowest of all the varieties. The type of fertilization had no significant (p 0.297) effect on panicles length of varieties Nerica N6 and local Madjitolngar. As for Nerica NL28 and local Tox 728-1, the effects of mycorrhiza and control were similar, but significantly greater than that of compost. However, the panicles length of Nerica N6 was significantly (p 0.0001) more important than that of the three other varieties (almost around 24 cm). Once again, the number of productive tiller was improved by mycorrhiza for Nerica NL28 variety at 90 DAP, while compost and fertilizer were the best stimulators in tiller production for the two local varieties. The increase in the number of productive tillers as the results of mycoorhizal inoculation could be attributed to better phosphorus nutrition (Hebert and Shaver 2009). Table 5: Variation of Growth Responses of Rice Varieties as Affected by Type of Fertilization at 90 DAP Rice Varieties Control Nerica N6 Nerica NL28 Madjitolngar Tox 728-1 LSD 5% p-value Nerica N6 Nerica NL28 Madjitolngar Tox 728-1 LSD 5% p-value Nerica N6 Nerica NL28 Madjitolngar Tox 728-1 LSD 5% p-value 6.93aa 34.63b b 29.9bb 32.1bb 22.96 0.0001 1.19ac 0.77aa 1.32bd 1.00ab 0.128 0.0001 6.6aa 34.2bb 29.5bcb 31.8bb 22.9 0.0001 Types of Fertilization Compost Fertilizer Mycorrhiza Tiller Number.Plant-1 a 9.96 a 11.0 ba 9.36aba 21.4 bb 45.76cc 46.56cc c b 33.5 b 28.73 b 21.1ab d a 38.26 c 25.6 b 24.2ab 4.76 14.6 11.73 0.0001 0.0001 0.0001 Plant Height (cm) 1.18ac 1 .26bc 1.21abc a b 0.77 a 0.95 a 0.79aa b b 1.23 c 1.32 c 1.10ab a a 1.04 b 1.04 b 1.03ab 0.195 0.073 0.108 0.0001 0.0001 0.0001 Panicle Number.Plant-1 9.8ba 10.7ba 8.96ba a c 21.26 b 45.0 c 45.66cc c b 32.96 c 28.16 b 20.43ab c a 37.6 d 25.0 b 23.5ab 4.63 34.1 11.46 0.0001 0.0001 0.0001

LSD 5% 3.03 11.13 7.63 6.10

p-value 0.020 0.0001 0.0001 0.0001

0.071 0.160 0.130 ns

0.05 0.0001 0.0001 0.472

3.2 10.8 4.8 5.8

0.041 0.0001 0.0001 0.0001

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Table 5: Variation of Growth Responses of Rice Varieties as Affected by Type of Fertilization at 90 DAP Contd., Rice Varieties Types of Fertilization Panicles Length (cm) Nerica N6 29.21ab 29.53ac 30.31ac 29.1ac ns 0.309 b a b Nerica NL28 24.03 a 22.60 a 25.48 b 25.01bb 1.42 0.0005 a a a a Madjitolngar 25.41 a 24.6 b 23.79 ab 23.28 a ns 0.297 Tox 728-1 24.69b a 24.23abab 22.91aa 24.9bab 1.46 0.049 LSD 5% 3.80 1.99 2.56 1.73 p-value 0.0001 0.0001 0.0001 0.0001 Leaves Length (cm) Nerica N6 29.43ac 32.98bc 34.33bb 31.9bb 3.55 0.002 ab a ab b Nerica NL28 22.84 a 22.44 a 22.55 a 23.36 a 2.42 0.030 Madjitolngar 37.68ad 35.33ac 33.41ab 33.48ab ns 0.302 Tox 728-1 25.43abb 26.29bb 23.04aa 25.13aba 3.25 0.045 LSD 5% 2.99 5.44 10.97 6.77 p-value 0.0001 0.0001 0.0001 0.0001 Leaves width (cm) Nerica N6 1.80 ac 1.97bcc 2.10cd 1.95bb 0.145 0.001 b a b Nerica NL28 1.15 a 1.04 a 1.18 b 1.2bb 0.010 0.010 b a ab a Madjitolngar 1.13 a 0.99 a 1.04 a 0.99 a 0.135 0.014 Tox 728-1 1.28abb 1.31bb 1.30bc 1.16ac 0.136 0.046 LSD 5% 0.13 0.27 0.12 0.17 p-value 0.0001 0.0001 0.0001 0.0001

For each rice variety, values of the same row affected by the same letter as superscript for a given parameter are not significantly different at 5%. For each type of fertilization values of the same column affected by the same letter as subscript for a given parameter are not significantly different at 5%; ns = non significant Similarly, Abid et al. (2002) has shown that the number of tillers increased significantly with application of phosphorus in wheat trial. Unlike Nerica NL28, mycorrhiza significantly (p 0.002) increased the length and width of Nerica N6 plant leaves compared to the control, with similar effect attributed to compost and fertilizer . Previous reported results vindicating the positive responses of mycorrhiza on inoculated plants were attributed to improved nutrients acquisition by plants (Gianinazzi-Pearson 1982) or increased organic matter (Nouaim et al. 1994). Mycorrhiza had no effect on the length and width of the the local rices varieties. For the local variety Tox 728-1, the enhanced length and width of plant attributed to compost was significantly (p = 0.045) was greater than that of other types of fertilization. Leaves of Nerica rice varieties were significantly (p 0.0001) wider than those of the local varieties. Tillering ability in rice, to some extent, is closely related with panicle number and thus yield. It has been well documented that either excessive or insufficient tillering is unfavorable for high yield (Del Mora and Del Mora, 1995; Lafarge et al. 2002). In the present study, Nerica N6 had less tillers, less panicle/plant an low yield, in the opposite of Nerica NL28. The two local varieties averagely had the same number of panicles per plant, similar to that of Nerica NL28. The present results indicated a positive and significant correlation between tillering ability and panicle number (r = 0.99); p < 0.0001), as well as the panicle length and seed per nanicle (r = 0.47); p< 0.0001) which seems to be important components for the improvement of rice yield potential.

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Influence of the Type of Fertilization on Leave Health of Rice Varieties Between 60 and 90 DAP, the colour of plants leaves and the presence/absence of devastating insects were observed. The devastating insect of rice named African cecidomyie (Oryzae oryzivora) was observed only on the control plots. Plants amended with compost were revealed to have the tendency of being less affected by fungal diseases and insect attacks, as previously revealed by Nakasaki et al. (1998). Yield Responses of Rice Varieties as Influenced by Types of Fertilization at 90 DAP For each of the local or Nerica rice varieties the number of grain per panicle was not affected by mycorrhiza as compared to the control (Figure 4). However, Mycrrhiza significantly enhanced the number of grain per panicle compared to compost for Nerica N6, while for the local variety Madjitolngar, compost better increased (p = 0.03) the number of grain per panicle than mycorrhiza. In contrast, the number of grain per panicle was not affected by the type of fertilization as far as the local variety TOX728-1 is concerned. The grain/panicle ratio of Madjitolngar variety was 0.60, 0.73 and 0.55%, respectively for treatments fertilizer mycorrhiza and control, suggesting that the acquisition of nutrients by the host plant positively impact the grains padding (Adama et al. 2008). It seems like the grain/panicle ratio increase with grain padding. Grain/panicle ratio of between 0.47 and 0.55 has been reported in field poor in nutrients (Chantereau and Kondombo 1994). A positive and significant correlation was observed between the weight of grains per experimental unit and the grain yield expressed in t/ha. Hence, for the two Nerica rice varieties, the grain yield/ha was significantly (p = 0.015) more improved than that of the control and compost applied treatments when plants were inoculated with mycorrhiza at sowing or received thrice the fertilizer during their growth cycle (Figure 5). Thus, the grain yields for treatment control, compost, mycorhiza and fertilizer were respectively 0.93, 0.67, 1.36 1.46t/ha for Nerica N6, and 2.13, 1.92, 3.83, 3.98t/ha for Nerica NL28 varieties. These results are lower than previous results obtained on Nerica FKR62 grown in the guinea savannah zone of Cameroon, indicating 6.8 and 5.8t/ha for mycorrhiza and fertilizer treatments, against 4.8 t/ha for the control treatment (Natebaye 2010). However, the finding confirms the compatibility of mycorrhiza to Nerica rice varieties, but at a different degree in another growing area of the same agro-ecological zone. In contrast, the type of fertilization did not have an effect on the grain yield of the two local rice varieties, with the control treatment exhibiting the highest yield than all the other treatments. For the local variety Madjitolngar the control yielded 5.86 t/ha against 4.53t/ha, 4.16t/ha, 3.78t/ha for fertilizer , compost and mycorrhiza treatments. Similarly, treatment control significantly increased the grain yield of the local variety Tox728-1 (5.83 t/ha) more that treatments mycorhiza (5.05t/ha), compost (4.72 t/ha) and fertilizer (4.49 t/ha).
250 200 150 100 50 0 N6 NL28 Madjitolngar Tox 728-1 Rice varieties Control Com post Fertlizer Mycorrhiza ab a ab b c a bc b

Rice grain/panicle

ab

b ab a

a a a

Figure 4: Variation of Rice Grain per Panicle within Varieties as Affected by Types of Fertilization Bars with the same letter are not significantly different at 5%

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Ngakou A, Mbaiguinam M, Nadjilom Y & Tokam N. M

However, suitable AM fungal inoculations for wetland rice varieties was reported to benefit the plants in terms of yield (grain and shoot), corresponding to half of the normal dose of phosphatic fertilizer (Gupta and Ali 1993), in agreement with acceleration of N and P transfer from shoots and/or from soils to grains under flooded conditions by AMF (Solaiman and Hirata 1995). Other results on the positive responses of upland rice plants (Gangopadhyay and Das 1984; Iqbal et al. 1978), and many cereals including pear millet (Rao et al. 1983), maize (Lu and Miller 1989) to AMF inoculation have been reported.
7 6
Rice grain yield (t/ha)

c b a

c a a b

5 4 3 a 2 1 0 N6
Cnntrol

a b

b b a a

NL28
Compost

Madjitolngar
Fertilizer

Tox 728-1
Mycorrhiza

Rice varieties

Figure 5: Variation of Rice Yield within Varieties As Affected By Types of Fertilization Bars with the same letter are not significantly different at 5% Table 6 : Changes in1000 Seed Weight (g) of Rice Varieties as Affected by Types of Fertilization Types of Fertilization Control Compost Fertilizer Mycorrhiza Nerica N6 30.14d 30.14c 30.14d 30.14d Nerica NL28 24.22b 24.22b 24.22b 24.22b Madjitolngar 23.32a 23.32a 23.32a 23.32a TOX728-1 29.73c 29.73c 29.73c 29.73c p-value < 0.0001 < 0.0001 < 0.0001 < 0.0001 For each type of fertilization values of the same column affected by the same letter are not significantly different at 5% Results in table 6 indicate that the 1000 grain yield was not influenced by the type of fertilization (compost, mycorrhiza, fertilizer ), in agreement with the constancy of 1000 grain weight of rice, irrespective to treatments and varieties previously reported (Emadzadeh et al. 2010). According to the same author, the main factor involved in grain feeding is the sun light ray that acts in the same manner on all experimental units of our homogenous flat experimental field. Conversely, Nerica rice varieties significantly (p 0.0001) had the greatest 1000 grain yield compared to that of the local varieties, with Nerica N6 and the local variety Madjitolngar exhibiting respectively the highest (30.14 g), and the lowest (23.32 g) weights. This 1000-grain weight values of Nerica N6 was close to values of between 31.03- 33.76 g (Gevrek et al. 2012), and 29. 48-31.89g (Wang et al. 2007) recently reported. However, the 1000 weight of the local variety TOX728-1 (29.73g) and Nerica NL28 (24.22 g) were respectively closer to those of Nerica N6 and local Madjitolngar. The improvement of rice (Natebaye 2010) and cowpea seed weights (Ngakou 2007) by mycorrhiza was justified by their positive influence on growth parameters of the crops. Rice Varieties

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Influence of the Type of Fertilization on Rice Grains Physical Traits The responses of local Tox 728-1 and Madjitolngar rice varieties to mycorrhiza inoculation were poor. Therefore, only the Nerica rice varieties were considered for characterization of physical seed traits. As indicated in table 7, mycorrhiza and compost significantly improved the geometric diameter of the two Nerica rice varieties compared to the negative control treatment. Table 7: Differences in Nerica Rice Grains Physical Traits as Affected by Types of Fertilization Nerica Varieties Control Nerica N6 Nerica NL28 LSD 5% p-value Nerica N6 Nerica NL28 LSD 5% p-value Nerica N6 Nerica NL28 LSD 5% p-value Nerica N6 Nerica NL28 LSD 5% p-value Nerica N6 Nerica NL28 LSD 5% p-value Nerica N6 Nerica NL28 LSD 5% p-value Nerica N6 Nerica NL28 LSD 5% p-value Nerica N6 Nerica NL28 LSD 5% p-value Nerica N6 Nerica NL28 LSD 5% p-value 0.91aa 0.93aba ns 0.367 28.24ab 23.69aa 4.55 0.0001 0.32ab 0.216aa 0.109 0.0001 0.69bb 0.60aa 0.095 <0.0001 0.079abb 0.061aa 0.018 0.0002 0.184ab 0.165aa 0.019 0.006 0.816aa 1.24cb 0.425 < 0.0001 0.76 bb 0.65aa 0.12 0.0001 0.007aa 0.008ab 0.001 0.002 Compost Types of Fertilization Fertilizer Mycorrhiza LSD 5% ns 0.043 p-value 0.721 0.0006

Seed Length (mm) 0.92aa 0.90 aa 0.92 aa c b 1.00 b 0.96 b 0.92aa 0.074 0.058 ns 0.0008 0.0008 0.984 100 Seed Weight (g) 29.02ab 29.00ab 30.15bb b b 25.25 a 25.48 a 24.67ba 3.76 3.51 5.47 0.0001 0.0001 0.0001 Minor Diameter (mm) 0.35bb 0.31ab 0.33abb 0.23b a 0.236ba 0.22aba 0.12 0.079 0.11 0.0001 0.0001 0.0001 Geometric Diameter (mm) 0.68aba 0.64aa 0.66abb b b 0.64 a 0.63 a 0.60aa ns ns 1.73 0.080 0.425 0.0001 Geometric Volume 0.075aa 0.072aa 0.085ba b a 0.128 a 0.068 a 0.095aba ns ns ns 0.864 0.700 0.0001 Bulk Density (g/ml) 0.188aa 0.185aa 0.192aa a 0.188 a 0.200a 0.192aa ns 0.014 ns 0.161 0.026 0.656 True Density (g/ml) 0.972 bb 0.898ab a 0.994ba a a 0.82 a 0.93 a 1.07ba 0.144 ns ns 0.033 0.405 0.271 Sphericity 0.75bb 0.69ab 0.71ab a a 0.64 a 0.65 a 0.65aa 0.11 0.04 0.06 0.0001 0.008 0.0005 Porosity 0.007aa 0.00aa 0.007aa a a 0.007 a 0.007 a 0.010aa ns ns ns 0.104 0.976 0.263

1.13 0.986

0.001 0.0005

0.025 0.019

0.001 0.01

0.053 0.029

0.031 < 0.0001

0.01 0.059

0.05 0.034

ns ns

0.172 0.172

0.155 0.136

0.022 < 0.0001

0.037 ns

0.0005 0.683

ns ns

0.513 0.280

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Ngakou A, Mbaiguinam M, Nadjilom Y & Tokam N. M

Nerica N6 exhibited a more greater diameter (0.68 and 0.69 cm) than Nerica NL28 (0.64 and 0.63 cm), respectively in response to mycorrhiza and compost applications. Moreover, compost amendment consistently enhanced the seed weight of N6 variety than the negative control, whereas for NL28 veriety, fertilizer and mycorrhiza more improved the seed weight over the control. Like geometric diameter and seed weight, sphericity of Nerica N6 variety was improved by biological fertilizers (Mycorrhiza and compost), although these fertilizers did not affect the sphericity of Nerica NL28. For the aforementioned parameters, Nerica N6 best responded to biological fertilizers and this differential response in the two varieties might be attributed to their particular adaptation specificities. The enhancement of geometric diameter and sphericity of Nerica N6 variety by compost and mycorrhiza could be allocated to their positive effect on the length, minor and major diameters of seeds, with the understanding that these two seed traits are closely dependent to seed length and diameters. This finding is in agreement with other results obtained on cowpea seeds after dual inoculation with mycorrhiza and rhizobia in three agro-ecological zones of Cameroon (Ngakou et al. 2011). It should be mentioned that the mean values of sphericity for Nerica N6 (0.73) and Nerica NL28 (0.65) fall within the range of 0.32-1 reported for most agricultural products, including rice (Lewis 1999; Mohsenin 1986; Ashtiani et al. 2010). The improved rice seed weight attributed to compost and mycorrhiza could be explained by the performances of these two treatments on rice growth parameters. Compost amendment positively affected the geometric volume of Nerica N6 seeds, while that of Nerica N28 was instead improved by mycorrhiza compared to the negative control. Our results line with those of Koocheti et al. (2007), who demonstrated the significant enhancement of geometric volume and densities of water melon seeds after compost amendment. The true density of cereal grains is important to separate different impurities from them as the true density of cereal crops and most impurities widely differ. The analysis of variance showed that whereas the type of fertilization had no effect on the true density of Nerica N6, the true density of Nerica NL28 seed variety was significantly (p < 0.0001) enhanced by mycorrhiza, compost and fertilizer (positive control) compared to negative control. Bulk density values are necessary in sizing hoppers and storage equipments of grains. They are also useful to determine the product mass or volume in the tank of combine harvester which itself is an appropriate parameter for yield monitoring in precision farming. Bulk density was consistently increased by mycorrhiza and compost for N6, and by fertilizer for NL28 rice variety, thus varies with the rice variety. This finding is supported by the results presented for Brazilian rice (Correa et al. 2007). A significant difference has been revealed between true and bulk densities of two tested rice cultivars (Ashtiani et al. 2010) with one of the cultivar exhibiting lower porosity as the consequence of higher bulk density. The porosity of rice was not affected neither by the type of fertilization, nor by the seed variety. The mean values of porosity 0.8% of our Nerica rice varieties was very low compared to 47.41 to 48.85% for Sorkheh and Sazandegi, rice respectively (Ashtiani et al. 2010), and (64 to 66% for Brazilian varieties (Correa et al. 2007). This difference could be inherent to characteristics of varieties. It has been reported that excessive tillers, in particular initiated at later stage could result in increased difference in grain maturity within a plant (Counce et al. 1996). Therefore, the more the tillers for a plant, the greater the variation of grain quality within a plant, because the plant with more tillers has more chance of being subjected to different temperatures during grain filling. Hence, it is possible that rice quality will be deteriorated due to excessive tillers within a plant.

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CONCLUSIONS
The overall results obtained from this study indicate that although vulgarization of Nerica rice varieties has been going in the south of Chad, its growth adaptation still need to be taken into consideration if high yield are expected. From our investigations, it appears that local rice varieties Madjitolngar and TOX728-1 are more adapted to their indigenous soils for the expression of their agro-morphological potentials. They may have established a mutualistic relationship with their native mycorrhiza fungi partners. Since mycorrhiza are not well distributed in soils of a given region or agroecological zone, improved yield of local rice varieties could be obtained if these native AMF are isolated, produced and coupled to a large scale production. Nevertheless, the biological fertilization with compost and mycocrhiza could be one of the best alternative approaches to be coupled to Nerica rice production, in order to achieve higher seed yield, better physical seeds qualities, with maximum economic return to population of this region.

ACKNOWLEDGEMENTS
The authors are thankful to the Malla Farm company of Tandjile-Chad from where the rice varieties and the chemical fertilizer were obtained for free.

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