**4.2 Laboratories**

*Mycorrhizal Fungi - Utilization in Agriculture and Forestry*

istration, laboratories, storage and packing space.

**4.1 Administration**

the bio-fertilizer production unit must be organized in an H-shaped architecture (**Figure 2**). This comes, according to Alamari [4], This author explained that this structure is based on the fact of its economical aspect and its ability to adapt to the sterilization process with forward walking, in other words from the "soiled", to go towards the "clean" and then towards the "sterile", without possibility of going back and without crossing of flows of "soiled" and "clean". The unit must contain admin-

The administrative team must ensure the respect of directives and guidelines in addition to external relations allowing the best conditions for the scientific team to carry out their work. To achieve these objectives, different tasks must be implemented: (i) Preparation, application and management of the budget and control of its execution; (ii) Establishment of contracts and agreements with different organizations in the same field, while taking care of calls for submissions and the

**86**

**Figure 2.**

*Conventional method for production of mixed AMF bio-fertilizers*

The setting up of a laboratory should meet the criteria approved by the World Health Organization (WHO). A laboratory should be built with walls, ceilings and horizontal surfaces, non-polished, easy to clean, impermeable to liquids and resistant to disinfectants and to antiseptics. Then, to ensure the best work conditions, the laboratory areas must be spacious [42]. In addition, the laboratory must contain:


The production unit laboratory of AMF bio-fertilizer must contain three compartments: (i) Greenhouse for AMF inoculum multiplication; (ii) In vitro multiplication and strains isolation rooms; and (iii) compartment of control, conditioning and storage.

### *4.2.1 Greenhouse AMF multiplication*

The greenhouse for AMF multiplication must be positioned behind the unit occupying a clear space with 204 m2 of approximately area. The greenhouse is used to care for young host plants and to maximize crop productivity by improving the relationship between their growth and AMF biomass. Greenhouse is the most important compartment of the unit, so the geographical location may have to be considered. The attractive location must be related to the adaptability and value of the land, cost of fuel delivered, ample and inexpensive water, in response of number of question: (i) What is the yearly available solar energy? (ii) How much moisture falls, summer and winter? (iii) What are the maximum and minimum temperatures and their duration? (vi) What are the hail and wind belts?(v) Is air pollution a potential problem? (vi) Information on all of the foregoing questions allows the greenhouse operator to determine the degree to which he can maintain near optimum environmental conditions for plant growth and AMF multiplication [43]. Wind is important climatic problem in arid lands, so the wind direction plays an important role in the choice of unit implantation site. So, orientation of the greenhouse is a compromise for wind direction, latitude of location and type of temperature control [44]. After site choice, the greenhouse must follow some recommendations. The greenhouse must be constructed in glass farmed structure on double-sloped with a naturally exposition of natural light for much of the day.

In addition, heat is partly assured by sun rays paired with artificial means, such as circulating steam, hot water, or hot air. Ventilating system is also needed [43, 44]. For low coast, the ventilation must be assured by roof openings and large windows on the side, which can functioned mechanically or automatically. In some conditions, if a financial condition allows automatic ventilation system is installed. For the AMF multiplication, trapping culture must be done in pots or in specialized containers. In this setting, trapping cultures are grown for 3 to 4 months to minimize the accumulation of saprophytes in the medium for excessive growth and senescence of the roots. However, culture maintained for more than 5 months and regulated watering is recommended before areal part cutting and replantation of new seeds as explained above (3.1.1).

#### *4.2.2 In vitro multiplication room*

It represent an aseptic areas separated from the greenhouse and the AMF isolation room in order to avoid any contamination and to control sterility conditions as much as possible.

#### *4.2.3 Drying and conditioning room*

This area is located just before the greenhouse; it is used to dry the contents of the pots and containers for later conditioning. Once the trapping plants are ready to be harvested, they are moved to shelves in this area so that they are not exposed to light. Drying take about 2 to 3 weeks. After this period, the roots of the trapping plants are cut and mixed with a suitable substrate. Conditioning AMF inocula begins by placing the cultures in sealed bags. These bags are provided with codes written both on the surface and on labels affixed to the upper left corner of the crops so that they are easily identified when stacking. In addition, an organization in alphabetical order of cultures is also recommended.

#### *4.2.4 AMF strain isolation room*

Isolation room is completely isolated from all plant growing areas and the use of unsterilized soil is strictly prohibited. Isolation is practiced as follows: The contents of the dried pots are installed on grounds. The isolation of the spores from the sample is done by wet sieving method [28]. This technic of isolation is practiced in order to produce bio-fertilizer containing AMF spores. In addition isolated AMF strains are conserved in order to develop further research. During the AMF isolation process, a series of precautions must be observed, especially disinfection of surface area of isolation, tables and shelves with draying after cleaning. Asepsis is main condition for the success of this crucial stage of the AMF bio-fertilizer production.

#### *4.2.5 Control room*

This space is used for carrying a series of bio-fertilizer control tests. These tests include AMF spores count with microscopic examination, evaluation of AMF root colonization rate and elaboration of must probable number test. AMF spores number must vary between 10 and 15 pots per day. Once extracted, the spores are transported in glass Petri dishes and stored in the laboratory refrigerator. Indeed, the examination is carried out by a stereo microscope on the day of the extraction. The information thus retrieved is stored in a database and all written notes are

**89**

*Native Arbuscular Mycorrhizal Fungi and Agro-Industries in Arid Lands…*

temperature is not detrimental to the viability of AMF propagules.

**5.1 Why produce native AMF bio-fertilizers for arid lands?**

**5. AMF bio-fertilizers challenges of production and application on** 

of the land areas of the world. These areas are subjected to several desertification phenomena [3]. To counteract this problem, applications of new agricultural technics are required including application of bio-fertilizers. Nevertheless, the use of AMF for the restoration of degraded ecosystems has received poor attention, requiring a different approach [25]. In addition, loss of AMF propagules is usually recorded following soil and cover plant degradation, which could further inhibit natural and/or artificial revegetation processes [3, 25]. Taking into account all the previously cited aspects and the necessity of restoration in these areas, the ecotechnology proposed by some studies [13–15, 18, 20, 22, 23, 25] represents a good alternative. They propose the restoration of degraded areas by re-introduction of

Production of native AMF bio-fertilizers unit require appropriate funding with adoption of a good financing strategy, based on various technical-economic parameters including description of the income elements and those of the expenses (**Table 3**). Indeed, the elements of income include the sale of bio-fertilizers, remunerations, publicity and assurance. **Table 4** describes all the expenses and revenues

Production of native AMF production is an agro-industrial investment for each arid region, in some case the product may be exported to similar edapho-climatic areas. Designed production and marketing chain must support each unit in order to guarantee the success of the project. In **Table 4** we have tried to establish an approximate economic study which will allow the investors to have an idea on the economic situation of the project, but these figures may vary from one country

**5.3 Economical challenge of native AMF bio-fertilizer production?**

Arid lands constitute the most widespread terrestrial biome in earth, with 35%

archived as a physical backup. Systematically, all the files having processed the culture collection are stored centrally on the unit's web server, saved on a separate hard drive on the same computer, and stored on another computer in the laboratory.

The storage is done at an atmosphere of 4°C at the level of the shelves of metal racks. These are characterized by mesh surfaces to optimize air circulation and facilitating their cleaning. The racks are placed in the center of the room and equipped with wheels to facilitate their movement. The storage period can reach a maximum of 3 years [4]. Bio-fertilizer storage process requires certain recommendations mainly: bag surface and their labels must be cleaned and disinfected before they are placed in this room. Floors and shelves are regularly disinfected. AMF bio-fertilizer product should be stored in a corner of the laboratory where the air

*DOI: http://dx.doi.org/10.5772/intechopen.94084*

**agricultural projects of arid lands**

native AMF and plant species [25].

**5.2 How produce native AMF bio-fertilizer unit?**

provided by the bio-fertilizer production unit.

*4.2.6 Storage room*

archived as a physical backup. Systematically, all the files having processed the culture collection are stored centrally on the unit's web server, saved on a separate hard drive on the same computer, and stored on another computer in the laboratory.
