Teri Micorisa

Thrust areas Major activities/flagship projects Select clients/partners/stakeholders Infrastructure facilities

Technology transfer Patents Services offered

Thrust areas The Centre offers mycorrhizal technology as a biological and eco-friendly means of promoting plant growth at a low cost with a minimum input of chemical fertilizers. It's thrust areas include: Environmental conservation and pollution abatement, including reclamation of fly ash overburdens and other wastelands by integrated mycorrhizal organo-biofertilizer inputs Wastelands are drastically disturbed lands where native vegetation and animal communities have been removed and the top soil has been lost, altered or buried. Examples of wastelands are areas that have been surface-mined for minerals (coal, paoline, bauxite, gravel, sand etc.), mining wastes, or have had the top soil removed for use elsewhere creating 'borrow pits'. Wastelands also include sites which have saline, acidic or calcareous soils, and soils that have been eroded by wind and water. The natural rehabilitation of such lands is a gradual and slow process. The Centre is making efforts to expedite the recovery of such areas using mycorrhizal biofertilizers.

Mass inoculum production The Centre offers the mass production of viable, healthy, genetically pure, and highquality fungal propagules. The technology is commercially viable and has been tested on different crops and forestry plantations in several edapho-climatic regions to prove its wide applicability. Two significant features of this technology are: It permits the standardization of large-scale production technology and distribution of viable inocula, which will assist in the transfer of technological knowledge to the grass roots level; It offers a promosing alternative for the production of AM inocula free of pathogens using Ri T-DNA transformed roots. It has obvious advantages over other systems, permitting observation of fungal morphology and development in vitro.

Establishment of agricultural, horticultural and silvicultural plant species with enhanced yields without using chemical fertilizers Arbuscular mycorrhizal (AM) fungi can utilize phosphorus (P) present at extremely low soil concentrations and even from unavailable sources of P, thereby providing an alternative to costly and polluting phosphate fertilizers. Plants inoculated with AM fungi have increased P concentrations and growth. However, the extent of benefits imparted

by AMF to plants depends upon the plant-fungal combination. The Centre examines hundreds of AM fungi and determine their preference for vegetable and fodder crops, flowers and trees.

Conservation of mycorrhizal germplasm A germplasm bank of mycorrhizal cultures is the largest of its kind in Asia and was established in 1994 at TERI's field station in Gual Pahari. This facility provides researchers in commercial and non-commercial sectors specific cultures of their interest. The facility is well-equipped to preserve the germplasm available in the country and elsewhere. Cultures have been collected from 12 of 20 different agroecological zones in the country. The bank has an impressive collection of over 450 isolates of mycorrhizal fungi.

Major activities/Flagship projects Mass inoculum production The application of mycorrhiza on a large scale is limited by the nature of its proliferation which is strictly biotrophic. TERI has developed technology for the mass production of mycorrhiza by exploiting genetically modified host roots using the bacterium Agrobacterium rhizogenes carrying the Ri T-DNA plasmid. This method allows a huge recovery of inoculum in very short time span and using very little space compared to conventional modes of multiplication in pots. The technology is commercially viable and has been tested with support from the Department of Biotechnology, Government of India. It offers a biological means of assuring plant health in a profitable and ecologically sound manner.

Environmental conservation and pollution abatement The Centre works towards technologies directed at addressing the problematic area of active/abandoned fly ash ponds. The group has demonstrated the reclamation of fly ash using specific microorganisms at Badarpur, Delhi and at Korba, Chattisgarh, without amending good earth and without using chemical fertilizers. (For more information please see the case study)

Sustainable agriculture The Centre is actively involved in activities related to sustainable agriculture. The integrated use of organic manure and mycorrhizal biofertilizers has been found to work wonders in agricultural fields. The Centre has specific mycorrhizal fungi for vegetables (onions, potatoes, etc.) fodder crops (barseem, lucerne, sorghum, etc.), flowers (marigold, gladiolus, aster, etc.), trees (eucalyptus, poplar, etc.). The presence of mycorrhizal fungi has been found to increase yields by 30%-50%.

The project, "Integrated nutrient management in poplar-eucalyptus based sustainable agroforestry system" funded by the DBT, Government of India, was carried out by TERI. In this project beneficial microbes including mycorrhizal fungi, were integrated with inorganic fertilizers and manure (INM approach) in a poplar-eucalyptus based agroforestry system with wheat-pulse rotation. Efficient management of fertilizer manure and biofertilizer application for achieving higher productivity - The optimum combination of inorganic and organic manure was identified for achieving higher plant production with attractive cost economics. Field trials were conducted on the wheat-pulse rotation. The application of mycorrhizal fungi resulted in a significant improvement of the overall yield scenario. These findings can now be passed on to farmers in the region. Soil health improvement - The nutrient budgeting was also worked out from the demonstrated model; optimized doses of manure, fertilizers and biofertilizer contributed to improved nutrient gain and productivity. Both these help in achieving sustainable crop production.

The project, "Demonstration of integrated organic farming using VAM fungi in aromatic plants namely Cymbopogon winterianus, C. martinii and Polianthes tuberosa", funded by DBT, carried out at TERI Various types of decomposed organic waste were tested and the microbes enumerated. Some functional properties of microorganisms isolated from mature composts were also tested. AM isolates collected from target species, organically rich and heavy metal polluted areas and Centre for Mycorrhizal Culture Collection (CMCC) were mass multiplied. Compost doses for enhanced crop production of target plant species were optimized and finally, a compost-AM fungus combination was arrived at for the development of an organo-biofertilizer for Cymbopogon spp. and Polianthes tuberosa.

Biodiversity studies The diversity of AMF is studied in different agroclimatic regions of India to identify fungi for potential use in sustainable agriculture. Isolates in one agro-climatic region do not necessarily perform well in other regions. Thus, the study of biodiversity, distribution and diversity pattern of AM fungi in different agro-climatic regions is important.

Ex situ conservation and long-term storage of mycorrhizal germplasm Conservation of mycorrhizae is the first step in the overall conservation process, and begins with the procurement of germplasm and its documentation. The main objectives of the germplasm bank are: To identify and procure promising isolates of both ecto- and endo- mycorrhizal fungi To screen promising native AM fungi To maintain selected AM and EM fungi permanently To provide cultures on request. In the process of developing methodology for effective storage, a technique has been

successfully developed for obtaining a workable protocol for lyophilizing mycorrhizal fungal species for long term storage in the mycorrhizal germplasm bank. (For more information please see Centre for Mycorrhizal Culture Collection)

Select clients/partners/stakeholders Government agencies Department of Biotechnology Department of Science & Technology Ministry of Environment & Forests Fly Ash Mission

Corporate sector National Thermal Power Corporation (NTPC) Associated Alcohols & Breweries Limited (AABL) Tata Chemicals Limited (TCL) Rameshwar Industries KCP Sugar & Industries Corpration Limited Cadila Pharmaceuticals Limited

International Bi-lateral agencies SDC (Swiss agency for development and cooperation) IFCPAR (Indo-french centre for the promotion of advanced research)

Infrastructure facilities Greenhouse We have four greenhouses to meet our requirements: one for maintaining the cultures from different agro-ecological zones, the second for multiplication of the selected promising inoculum and the third and fourth for raising experiments. An autoclave room is situated close to the greenhouse for sterilizing the growth media to be used for raising the cultures. Washing facilities are located nearby. Containers used for pot cultures: 100 ml plastic pots 150 ml containers 300 ml or 500 ml deepots 1500 ml plastic pots 10000 ml cement pots

100ml pots are used only for assessing the Inoculum Potential (IP) of the inoculum and 150 ml pots are used for increasing the inoculum in the initial stages. For producing inocula in bulk, trap plants are raised in pots.

Laboratories Microscope laboratory Spores and mycorrhizal roots are studied microscopically. We have three compound microscopes (Olympus BH2 model equipped with UV and fluorescence optics, Leica Galen III and Olympus CH) and two stereomicroscopes (Leica Wild M10 and Leica Zoom 2000). Both the stereo- and compound microscopes have photographic camera attachments. Our image analyser system (Quantimet 500+, Leica) used to produce photovouchers or images consists of a CCD camera which can be positioned on the Olympus microscope BH2 or Leica Wild M10. A colour printer is there to make hard copies of the images. Molecular biology laboratory In the molecular biology laboratory, we have facilities for lyophilization (lyophilizer), shaking (lab shaker with temperature control), gel run (electrophoretic unit), gel drying (gel dryer), observation and photography of the gel (Gel ManagerTM documentation system with the rewuired software and CCD camera, (Biosystematica, UK), PCR cycling (Robocycler 40, Stratagene, USA) and DNA sequencing (Sequencer, Life Technologies).

Soil testing laboratory In the soil testing laboratory, soil samples are analysed by using standardized methods for pH, total soluble salts (as determined by electrical conductivity), organic C, total N, available P and available K. We also have facilities for assessing physical properties of soil (particle density, bulk density, maximum water holding capacity, moisture retention characteristics) as well as analysis of trace and heavy metals (atomic absorption spectrophotometer). The instruments in the soil laboratory include a digital flame photometer (CL 22D), thermocenter (Salvis), cyclomix (BS lab), spectrophotometer (SpectronicR 20 GenesysTM ) and centrifuge (Labofuge, 200).

Clean area We have three laminar flow cabinets, used for the in vitro culture of arbuscular mycorrhizal (AM) fungi, maintenance of ectomycorrhizal (EM) fungal cultures and the culture of other microbes.

Storage of cultured inocula

Storage of AM inocula The pot contents are allowed to dry in situ after harvesting and then placed intact (or broken into segments as large as possible) in plastic ziploc bags and stored in a cold room.

Storage of EM cultures EM cultures are maintained in MMN medium in a petriplate which are incubated in a BOD incubator at 25oC. EM culture discs are also stored in sterile distilled water at 4oC in cold room.

Voucher specimens Voucher specimens are material representations which can be interpreted by present and future biologists. We are in the process of making voucher specimens for AM fungi. We have a collection of hardcopy photographs of AM fungi and have started making slides and digital photographs of different species of AM fungi.

Growth room A growth room has been set up for mass inoculum production of AM fungi in vitro.

Technology transfer In vitro mass production technology of AM fungi to Cadila Pharmaceuticals, Ahmedabad In vitro mass production technology of AM fungi to KCP Sugars, Hyderabad On-farm production technology to Rameshwar Industries Ltd., Raipur.

Patents Lyophilization of EM fungi Lyophilization of AM biofertilizer Mass production AM technology using ROC Improved techniques of mass production of AM fungi

Services offered Physical and microbiological characterization of fly ash and soil samples Macro and micronutrient analyses Heavy metal analyses Tests for suitability of site for agriculture-related activities

Alleviation of metal toxicity by initiating agri-horticultural activities Consultancy services for rehabilitation of fly ash affected sites Consultancy services for setting up laboratory/production facility for mycorrhiza Carrier formulations of biofertilizer micro-organisms Germplasm/starter cultures of mycorrhiza biofertilizer Pilot production facilities of mycorrhiza biofertilizer Efficient inoculants for agri-horticultural and forest plant species Training in mycorrhizal research Identification of mycorrhizal isolates Mycorrhizal culture deposition, maintenance and multiplication Mycorrhiza Network makes available to researchers the growing literature on mycorrhiza and facilitates information-sharing among members. A database is operational for information retrieval and supply to researchers. (For more information please see Mycorrhiza Network web site)

For further information, please contact Dr Alok Adholeya Director, Biotechnology and Management of Bioresources Division