Effect of Dual Inoculation with Rhizobium Bacteria, A- Mycorrhizal Fungi and Micronutrients on Productivity of Egyptian clover

INTRODUCTION

Egyptian clover (Triflouim alexandrinum, L.) is the most important forage crops in Egypt, it is cultivated in about 2.5 million feddan and used as animal feed and soil improvement. Forage quality is the most important character of feed staff producing and feeding the highest quality forage possibly increases animal performances reduces feeding costs and ultimately results in an increased return on time and money invested in forage production (Abdel- Sattar et al., 1996, Abdel-Halim et al., 1993 and Abdel – Gawad, 2003).

The foliar spray is more essential than soil application due to higher utilization which makes the nutrients more efficient. It can, also, be used to satisfy a cut- need of macro, micronutrients. Moreover, some soil fertilization problem can early to solve by foliar spray application. It acts as micronutrient on one hand and environmental toxic factor on the other hand and is known to affect nodulation and nitrogen fixation (Gaure et al., 2012), successful development of nodules by rhizobial species at many different stages of development (Brewin, 1991).

Rhizobial surface component play an important role in deciding the host compatibility and abringing about the infection leading to nodulation and nitrogen fixation (Swamynthan and Singh, 1995).

Mycorrhizal are multifunction organisms in agro ecosystems that improve soil physical, chemical and biological properties by developing mycelium, increasing nutrients absorption and soil nutrients (Cardoso and Kuyper, 2006). Chaicki et al. (2015), reported that dry matter of berseen clover (triflouim alexandrinum, L.) inoculation with mycorrhizal was significantly more than control treatment. According to the same report co- inoculation by Rhizobium bacteria and Mycorrhizal fungi increased clove shoot dry weight and leaf area index compared by control. Therefore, the objective of the research is to study the effect of dual inoculation with rhizobium & A- mycorrhizal fungi and micronutrients on productivity of Egyptian clover.

MATERIALS AND METHODS

Two field experiments were carried out at the Experimental Farm of Faculty of Agriculture (Saba Basha), Alexandria. University, at Abees region Alexandria, Egypt, during the two growing seasons of 2014/2015 and 2015/2016 to study the effect of dual inoculation with Rhizobium & A- mycorrhizal fungi and micronutrients on productivity and quantitative traits of Egyptian clover.

The experiment was designed in as split plot with three replicates. The main plots were allocated to the calcium and micronutrient B at 100 g/fed for foliar spray (untreated, one spray and two spray), the four dual inoculation i.e. (uninoculation, Rhizobium, A-mycorrhizal and mixture Rhizobium + A-mycorrhizal) were allocated randomly to the sub plots. Analysis of chemical and physical properties of the experimental soil (0 to 30 cm) is shown in Table (1) according to methods reported by Page et al. (1982).

Table (1).The physical and chemical properties of the experimental soil during 2014/2015 and 2015/2016 seasons

The plot area was 10.5 m² (1/400 feddan) and seed were broadcasted at the rate of 20 kg/fed. (Variety Giza 6). All plots received 30 kg P₂O₅/fed, prior to planting date (Oct. 10^th and Oct 12^th first and second seasons).Berseem seed was inoculated prior to sowing with Rhizobium legumonsarumbiovartrifolii. The rhizobia strains were provided by the biofertilizer production unit. Soil. water and Enviromental Research institute. ARC. Vie ARC 101 (RE1) isolated from nodulated between (Triflouim alexandrinum, L.) root plants isolation and purification were done according to the method described by Vincent (1970). Apeat – based inoculum containing ≥ 10⁸ cell/g was used in seed inoculation.

A-mycorrhizal fungi (Glomusm acrocarpuim) strain was obtained from Department of Plant Production, Faculty of Agriculture (Saba Basha), Alexandria. University, at the rate of 2550 spores was mixed with seeds and decating technique as described by Radwan (1996) three cuts were taken through the growing period of both seasons. Cutting was done when the stand of plots was about 40- 50 cm height and the stubble height was about 6 cm from the surface. Plant samples were taken on 45 days after sowing to determine the following parameters:

A) Growth and yield characters:

1. Plant height at three cutting.
2. Number of nodules
3. Dry weight of nodules (g/10 plants).
4. Fresh and dry yield (ton/fed).

B) Quantitative traits

Plant samples were collected from each plot at each cutting weighted dried and ground in a grinding mill to pass through a 1mm seive. Samples of each cut were analyzed for forage quality properties. Crude protein (CP), crude fiber (CF),water soluble carbohydrate (WSC%), Ether extract (EE), Ash and digetative dry matter (DOD) according to A. O. A. C. (1990) and natural detergent fiber (NDF%) collected according to Mcdonald et al. (1978)

Statistical analysis

The obtained data were statistically analyzed for ANOVA and LSD values were calculated to test the differences between the mean values of the studied treatments according to Gomez and Gomez (1984).

RESULTS AND DISCUSSION

A- Growth characters and yield:

The obtained results given in Tables (2, 3, 4 and 5) clearly showed that the application foliar micronutrients at two times exhibited a significant effect on growth characters and yield i.e. plant height, number of nodules, dry weight of nodules/10 plants, fresh and dry yield forage (ton)/fed in both seasons. Application foliar at two sprays of some micronutrients significantly increased the growth characters at all sampling and fresh, dry yield (ton)/fed during both seasons. These results may be due to the effect of calcium and Boron on stimulation physiological processes plant photosynthetic carbohydrate and protein accumulation, as well as sugar translocation in plant. Similar results were reported by Mohamed and Helal (1999), Nadian (2004) and Dheri et al. (2007).

Data in the same Tables show the effect of dual inoculation Rhizobium & A- mycorrhizal on growth characters (plant height, number of nodules and dry weight of nodules/10 plant) at all sampling and fresh and dry yield (ton)/fed, in both seasons. Growth characters and yield were improved by the dual inoculation (Rhizobium & A-mycorrhizal) which caused significant increase in growth character and yield in both seasons.

Rhizobium + A-mycorrhizal increase the ability of host plant to uptake soluble nutrient, particulary phosphorus and some micronutrients (Shabani et al., 2011). Also, Nadian at al. (1998) reported that dry matter of berseem clover (Triflouim alexandrinum, L.) inoculated with mycorrhizalwas significantly more than control treatment.

The interaction between application of some micronutrients and dual inoculation was significant for growth characters (plant height, number of nodules and dry weight of nodules/10 plant) at all sampling and fresh and dry yields (ton)/fed in both seasons. Tables (2, 3, 4 and 5).

Table (2). Plant height (cm) as affected by micronutrients and dual inoculation (Rhizobuim, Mycorrhizal) at three cuts of T. alexandrinum during 2014/2015 and 2015/2016 seasons

Mean values in the same column marked with the same letter are not significantly differed at 0.05 levels of probability

* significant at 0.05level of probability