EU-Alba - Low-cost and pesticide-free treatment of seeds
to improve yield of sorghum and millet in Africa

The project was a collaboration between University of Copenhagen (the Coordinator) and Partners in Burkina Faso, Tanzania and India.

The project consists of 5 work packages (WP):

WP1 E. alba EFFECT

(WP leader: Associate Prof. Ole Søgaard Lund)

This work package aims to identify under which conditions seed treatment of E. alba is efficient. The parameters tested will be botanical dose, timing of treatment, seed quality and seed health. Potential synergism with the application of fungicide and an Indian developed bio-pesticide will also be adressed. In addition, it will be tested in vitro if seed treatment by E. alba efficiently inhibits seed-to-seedling transmission of major seed transmitted pathogens (Fusarium thapsinum, Curvularia lanata etc.). Yield of Sorghum and Millet in field experiments will be used to evaluate the effect of E. alba seed treatment. Results consolidated from this WP will be indicative of the mechanism involved in E. alba seed treatment.

WP2 E. alba SOURCES

(WP leader: Associate Prof. Ole Søgaard Lund)

This WP will compare biological activity of different sources of E. alba.  Naturally occurring E. alba in Burkina Faso and Tanzania will be compared to both natural and commercial sources of E. alba from India, where the plant is cultivated by medicinal herb industry. Comparison will be made with respect to efficiency in seed treatment (field performance, yield) and with respect to specific anti-fungal activity of extracts. In addition, the availability of natural E. alba as a weed in all three countries will be surveyed and the potential for local cultivation or commercial production of E. alba specifically for seed treatment purposes will be evaluated for all three countries.

WP3 CULTIVATION SYSTEMS

(WP leader: Associate Prof. Ole Søgaard Lund)

This WP aims to analyse the effect of E. alba extract as seed treatment under different farming systems for both Sorghum and Millet. Particularly the influence of, agro-ecological zone, plant density, the use of fertiliser and the application of foliar spray (fungicide) will be assessed dependent on the predominant farming system in each country. Baseline Yield for each farming system (no seed treatment) will also be analysed in relation to the effect of E. alba seed treatment. Seed treatment applying commercial fungicide will be included for comparison to treatment with E. alba extract where relevant. Farmer participation in field experiments, particularly in Burkina Faso, will be coordinated by the national extension service.

WP4 SAFETY

(WP leader: Prof. H.S. Prakash)

This WP will describe safety aspects of applying E. alba seed treatment at farm level based on existing literature for E. alba toxicology in general as a medical herb. Specifically, the record of E. alba as a medicinal herb in the three partner countries, Burkina Faso, Tanzania and India, will be documented and the need for toxicological tests in relation to the use of E. alba for seed treatment will be assessed. Subsequently, applications for deregulation of E. alba as a biopesticide for seed treatment will be filed to the governments of Burkina Faso, Tanzania and India including a comparison of toxicity of fungicides already recommended for seed treatment in the partner countries. A protocol for safe handling of Eclipta alba extract and relevant, approved fungicides for seed treatment at farm level will be made. 

WP5 KNOWLEDGE MANAGEMENT

(WP leader: Associate Prof. Ole Søgaard Lund)

By structuring close interaction between researchers, extension service and farmers, this WP aims to shape research activities during the project to provide results of the highest level of relevance. At the end of the project, results obtained will be consolidated in a “best practice” protocol for applying seed treatment at farm level taking into account the different farming systems within and across countries. Specifically farmers in Burkina Faso acquainted with indigenous E. alba technology for seed treatment will participate in farm-testing of the applicability of the harnessed technology of using an aquaeous extract and soaking of seeds. Three project workshops involving senior extensionists from Burkina Faso, Tanzania and India are held in 2013, 2015 and 2016.

Coordinator: 

University of Copenhagen, DENMARK

Faculty of Science
Department of Plant and Environmental Sciences
Coordinator and Principal Investigator:
Associate Professor Ole Søgaard Lund

Partners:

Institut de l’Environnement et de Recherches Agricoles (INERA), BURKINA FASO 
Centre de Recherche de Kamboinsé

University of Mysore, INDIA 

Department of Studies of Biotechnology
Principal Investigator: Prof. Dr. H.S. Prakash
Email: hasriprakash@gmail.com
Co-Principal Investigator: Prof. S.R. Niranjana
Email: niranjanasr@rediffmail.com

Sokoine University of Agriculture, TANZANIA

Horticulture & Biotechnology
Dep. Crop Science and Production
Principal Investigator: Dr. Paul M. Kusolwa
Email: kusolwap@gmail.com
Co-Principal Investigator: Dr. Ernest Mbega
Email: mbegaernest@yahoo.co.uk
Co-Principal Investigator: Prof. Dr. Robert D. Mabagala
Email: rmabagala@yahoo.com

Protocol 1: Used for sorghum and pearl millet in Burkina Faso

Collection and processing of E. alba material
● Wild, whole plants of E. alba (root, stem, leaves, flowers) are collected in October-November at the flowering stage (1 plant will be sufficient to treat 1000 seeds).
● Plants are dried in the shade for at least 3 weeks. Brush of remaining soil from roots.
● Powderize the material using a wooden pestle and a large mortar
● Remove large particular matter and store the remaining powder dry and in the dark (up to 3 years) until use.

Preparation of extract (Two days before sowing):
For treatment of 2000 seeds:
●Mix 100 ml water and 2.5 gram dry powder (2.5 gram ~ 11 ml powder)
(Leave the mixture overnight).

Seed treatment (One day before sowing):
● Filter the mixture above through a piece of cloth or use it directly.
● Add 2000 seeds (ca 50 grams) to the mixture
● Allow the seeds to soak for 6 hours (Noon to sunset).
● Spread the seeds on a piece of cloth and allow the seeds to dry over night.

Sowing of seeds:
● Sow the seeds by hand
Sorghum: 4-6 seeds per hill.
Millet: 6-8 seeds per hill
After 3 weeks : thin hills to max three plants per hill.

Protocol 2: Used for sorghum and pearl millet in Tanzania

Collection and processing of E. alba material

● Cultivated, whole plants of E. alba (root, stem, leaves, flowers) are collected at the flowering stage (1 plant will be sufficient to treat 500 seeds).
● Plants are dried in the shade for at least 3 weeks. Brush of remaining soil from roots.
● Powderize the material using a wooden pestle and a large mortar
● Remove large particular matter and store the remaining powder dry and in the dark (up to 3 years) until use.

Preparation of extract (Two days before sowing):
For treatment of 2000 seeds:
● Mix 100 ml water and 5 gram dry powder (5 gram ~ 22 ml powder) (Leave the mixture overnight).

Seed treatment (One day before sowing):
Use the mixture above directly (filtering is not recommended):
● Add 2000 seeds (ca 50 grams) to the mixture
● Allow the seeds to soak for 8 hours.
● Spread the seeds on a piece of cloth and allow the seeds to dry overnight.

Sowing of seeds:
● Sow the seeds by hand
Sorghum: 4-6 seeds per hill.
Millet: 6-8 seeds per hill
After 3 weeks : thin hills to max three plants per hill.

Protocol 3: Treatment of Sorghum and Millet seeds with Balanites aegyptiaca extract.

Tested in Burkina Faso

Collection and processing of B. aegyptiaca material

● Barks of B. aegyptiaca are collected from wild trees. Barks (soft parts) are removed from the trunk of the tree with the help of a pick and air-dried for at least 7 days.  
●Powderize the material using a wooden pestle and a large mortar
●Remove large particular matter and store the remaining powder dry and in the dark (up to 3 years) until use.

Preparation of extract  (Two days before sowing):
For treatment of 2000 seeds: 

• Mix 100 ml water and 2.5  gram dry powder (2.5 gram ~ 11 ml powder)
   (Leave the mixture overnight).

Seed treatment (One day before sowing):
Use the mixture above directly (filtering is not recommended):
●Add 2000 seeds (ca 50 grams) to the mixture
●Allow the seeds to soak for 8 hours.
●Spread the seeds on a piece of cloth and allow the seeds to dry overnight.

Sowing of seeds:
●Sow the seeds by hand
       Sorghum: 4-6 seeds per hill.
       Millet: 6-8 seeds per hill
After 3 weeks : thin hills to max three plants per hill.

Eclipta Alba

Below you find photos to identify Eclipta alba. Download the photo in larger format by clicking it.

Balanites aegyptiaca

Below you find photos to identify Balanites aegyptiaca. Download the photo in larger format by clicking it.

Safe use of plant extracts

Eclipta alba:

 No health risk to humans for using an extract of this plant for seed treatment was found.

Aqueous extract from the plants is currently used in traditional Burkinabe, Indian and Chinese medicin.

Review by Jahan et al 2014: Ethnopharmacological significance of Eclipta alba (L.) Hassk. (Asteracea). International Scholarly Research Notices, 2014, 385969.

In the present project extract from dried leaves and aqueous extract of whole plants (roots, stem and leaves) were tested for toxicity according to OECD protocols of acute toxicity following the animals (rabbits and rats) every day up to 14 days after exposure to the extract.

OECD guideline test 423 (Toxicity when eaten)

OECD guideline test 404 (Toxicity to the skin)

OECD guideline test 405 (Toxicity to the eye)

Testing was carried out by accredited laboratory M/s Vipragen Biosciences Private Limited, Mysore, India.

• Leaves

No toxicity of the extract was found

Publication Udayashankar et al., 2015.

• Whole plant

No toxicity of the extract was found.

Report conclusions 2018

Balanites aegyptiaca

The extract from bark (Protocol-3) was not tested for toxicity in the present project.

A previous study did not find indications of acute toxicity in rabbits:

Mohamed et al 1999: Some pharmacological and toxicological studies on Balanites aegyptiaca bark

Phytotherapy Research, Volume13, Issue5, Pages 439-441

[10] Zida PE, Néya JB, Soalla RW, Paco Sérémé P, Lund OS (2018) Seed treatment with Eclipta alba plant extract and application of fertilizer confers a combined effect on the yield of sorghum. International Journal of Agronomy and Agricultural Research, 13:96-102.

[9] Zida PE, Néya JB, Soalla RW, Paco Sérémé P, Lund OS (2018) Seed-priming of sorghum with antifungal extracts from Balanites aegyptiaca and Eclipta alba in different agro-ecological zones of Burkina Faso. African J Agricultural Research 13: 2516-2525.

[8] Nandhini M, Rajini SB, Udayashankar AC, Niranjana SR, Lund OS, Shetty HS, Prakash HS (2018) Diversity, plant growth promoting and downy mildew disease suppression potential of cultivable endophytic fungal communities associated with pearl millet. Biological Control 127:127-138.

[7] Zida PE, Néya BJ, Stokholm MS, Jensen SM, Soalla WR, Sérémé P, and Lund OS (2018) Increasing sorghum yields by seed treatment with an aqueous extract of the plant Eclipta alba may involve a dual mechanism of hydropriming and suppression of fungal pathogens. Crop protection 107: 48-55 https://doi.org/10.1016/j.cropro.2018.01.001

[6] Zida PE, Néya BJ, Soalla WR, Sérémé P, Lund OS (2017) Hydropriming of pearl millet (Pennisetum glaucum L.) in Northern and Central Burkina Faso applying six hours of soaking and overnight drying of seeds. African J. Agricultural Res. 12:3441-3446.

[5] Udayashankar AC, Rajini SB, Nandhini M, Suhas YS, Niranjana SR, Lund OS, Prakash HS (2016) Acute oral toxicity, dermal irritation and eye irritation study of Eclipta alba aqueous extract in Sprague Dawley rats and New Zealand white rabbits. International Research Journal of Pharmacy 7 (6) 103-109.

[4] Stokholm MS, Wulff EG, Zida EP, Thio IG, Néya JB, Soalla RW, Głazowska SE, Andresen M, Topbjerg HB, Boelt B, Lund OS (2016). DNA barcoding and isolation of vertically transmitted ascomycetes in sorghum from Burkina Faso: Epicoccum sorghinum is dominant in seedlings and appears as a common root pathogen. Microbiological Res. 191: 38–50.

[3] Zida PE, Néya BJ, W. Soalla WR, Jensen SM, Stokholm MS, Andresen M, Kabir MH, Sérémé P,  Lund OS (2016) Effect of sorghum seed treatment in Burkina Faso varies with baseline crop performance and geographical location. African Crop Science Journal 24(2):109-125.

[2] Zida PE , Néya BJ, Soalla WR, Prakash HS, Niranjana SR, Udayashankar AC, Nandini M, Rajini SB, Barrocas EN, Andresen M, Sérémé P, and Lund OS (2015) Increased feasibility of treating sorghum seeds with Eclipta alba extract by lowering concentration of plant extract and soaking time of seeds. Int. J. Tropical Agriculture. ISSN: 0254-8755, 33:2391-2400.

[1] Andresen M, Wulff EG, Mbega ER, Stokholm MS, Glazowska SE, Zida EP, Mabagala RB, and Lund OS (2014) Seed treatment with an aqueous extract of Agave sisalana improves seed health and seedling growth of sorghum. Eur J Plant Pathol (2015) 141:119–132. DOI 10.1007/s10658-014-0530-6