Biotechnology and sustainable

agriculture: biofertilizers and

biopesticides

Dr. Nicolás Medina Basso Dr. Ondina Le ón Díaz

National Institute of National Center for
Agricultural Sciences Animal and Plant Health
medina@inca.edu.cu ondina@censa.edu.cu

2004 Pugwash Workshop

 SOME
INTRODUCTORY
REMARKS

2004 Pugwash Workshop

 IMMEDIATE ANTECEDENTS

GREEN REVOLUTION (early 50´s XX century)

- Introduction of new, more attractive varieties (higher yields)

- Intensive use of agrochemicals (mineral fertilizers and pesticides)

- Progressive mechanization

- Increased monoculture

- Significant increases in crops yields

It seemed to be a suitable agricultural technology for highly

developed countries.

But....

2004 Pugwash Workshop

 MAIN CONSEQUENCES OF THE USE OF
TRADITIONAL TECHNOLOGICAL MODELS FOR
AGRICULTURE

- Depletion of natural resources.

- Incresed erosion and loss of natural fertility of soils.

- Sudden and frequently apparition of new pest and diseases.

- Colapse of hydric supply systems.

- Reduction of biomass production and biological diversity.

2004 Pugwash Workshop

ENVIRONMENTAL IMPACT OF HIGH RATES OF
NITROGEN FERTILIZATION
Increased rates of soil organic matter mineralization

Decrease of soil Modification of soil microbial

organic matter pool composition and balance

Decrease in soil fertility Increased incidence of

Increase soil compactation pests and diseases

A decrease in crops yields and increased needs of

agrochemicals to rise soil fertility
and control pest and diseases

2004 Pugwash Workshop

 AN INTEGRATED CONCEPT OF
SUSTAINABILITY IN AGRICULTURE

A new paradigma for agriculture, based on agroecological

concepts, being the agroecosystem the foundation stone,
above which men with their experience and knowledge , as
results of the continuous advances in science and technology,
manage soils, plants, animals and weather factors, in order
to satisfy the always growing and changing food needs of
mankind, without deteriorating the environment

2004 Pugwash Workshop

 Characterization of a sustainable agroecosystem
The one who allows conservation of renovable resources, crops
adaptation to environment and obtention of a high level of
productivity. In this kind of agriculture, emphasis should be put
more on the long-term ecological sustainability than in short-
term productivity, and must fulfil the following objectives:

- Utilization of production methods that restore the natural

mechanisms of community stability.

- Optimized recycling of organic matter and nutrients.

- Maximal utilization of system´s multiuse capacity.

- Reduction of energy and inputs comsumption.

- Decreasing of global costs.

2004 Pugwash Workshop
BIOFERTILIZERS

2004 Pugwash Workshop

Evolution of mineral fertilizers consumption in Cuba
in the last 15 years
1000
890.4
900

800

700

600 11 times less

Mt

500

400

300 240.7
224.1 207.9
200 164.4
107.9 118
100
81.1

0
1989 1997 1998 1999 2000 2001 2002 Est 2003

2004 Pugwash Workshop

 SOME ALTERNATIVES TO THE USE OF
MINERAL FERTILIZERS

✹ GREEN MANURES
✹ MANURES, COMPOSTS AND BIOEARTHS
✹ EARTH WORM MANURE
✹ CROP AND AGROINDUSTRIAL RESIDUES
✹ BIOFERTILIZERS

2004 Pugwash Workshop

 POPULATION DENSITY AND BIOMASS OF THE
MAIN GROUPS OF MICROOORGANISMS FOUND IN
SOILS (Average values)

Microorganism Population density Biomass

N. g-1 N. m-2 N. ha-1 kg. ha-1 % weight

Bacteria 108 1012 1024 2 000 0,100

Actinomycetes 106 108 1016 2 000 0,100

Fungi 105 105 1015 2 500 0,100

Algae 104 105 1015 500 0,005

Protozoa 104 108 1014 150 0.005

2004 Pugwash Workshop
 SOIL MICROORGANISMS MAIN EFFECTS
ON AGRICULTURAL PRODUCTIVITY

. Decay of organic residues with nutrients and CO2 release.

. Direct or indirect supply of nutrients to plants.

. Production of plant growth promoting substances.

. Enhancement of soil chemical and physical properties.

. Enaction of mutualistic symbiotic relationships with plants.

. Biological control of soil pest and diseases.

. Degradation of xenobiotic toxic compounds, mainly pesticides.

2004 Pugwash Workshop

 ANNUAL ATMOSPHERIC NITROGEN FIXATION
Fixation source 106 t N.year-1
Industrial (fertilizer production) 49
Atmospheric (electrochemical processes) 10
Other chemical processes 35
Total biological fixation 175
- Oceans 36
- Earth ecosystems 139
. Leguminous plants (140 kg N.ha-1.year-1) 35
. Rice ( 30 “ ) 4
. Pastures ( 15 “ ) 45
. Other crops ( 5 “ ) 5
. Forest ecosystems ( 10 “ ) 40
. Other ecosystems ( 2 “ ) 10

2004 Pugwash Workshop

 A CONCEPT OF BIOFERTILIZERS

Biopreparates based on living soil-borne and endophitic

microorganisms at rates of population several times higher
than those normally found in nature, which are able, by
mean of their biological activity, to supply to plants,
directly or indirectly, most of the nutrients they need for
development, as well as plant growth promoting substances.

2004 Pugwash Workshop

 General methodology for obtaining and using biofertilizers

Prospection, isolation and characterization of strains

Production of certified inoculant

Production of commercial inoculant

Inoculation

Seed coating technology Field application

(Direct sowing crops) (Nursery crops, seed-
beds, “in vitro”
Single Combined plantlets, perennial
inoculation inoculation crops, etc.
(Coinoculation)

2004 Pugwash Workshop

 Biofertilizers and bioestimulators developed in Cuba
• DIMARGON (Azotobacter chroococcum)………………………………INIFAT
• DIMAZOS (A. chroococcum)…………………………………………………….INIFAT
• FOSFOSOL-P (A. chroococcum & other genera).…………………INIFAT
• FOSFOSOL-B (A chroococcum & other genera).………………….INIFAT
• FOSFORINA (Pseudomonas sp. & other genera).…………………I. SUELOS
• FOSFORINA plus A (Pseudomonas sp. & other genera)……..I. SUELOS
• FOSFORINA plus R (Pseudomonas sp. & other genera).…….I. SUELOS
• BIOFER (Bradyrhizobium sp.)…………………………………………………..I. SUELOS
• BRADYRIZOBIUM (Bradyrhizobium sp.)……………………………….I. PASTOS
• AZOSPIRILLUM (Azospirillum brasilense)……………………………ICIDCA
• AZOFERT-G (A. brasilense)………………………………………………………INCA
• AZOFERT-L (Bradyrhizobium sp.)……………………………………………INCA
• ECOMIC (Arbuscular mycorrhizal fungi)………………………………..INCA
• MICOFER (Arbuscular mycorrhizal fungi)……………………………..IES
• INDOLACETIC ACID………………………..……………………………………….ICIDCA
• GIBBERELIC ACID……………………………………………………………………..ICIDCA
• JASMONIC ACID……………………………………………………………………….ICIDCA
• BIOBRAS (Brassinosteroids) …………………………………………………..UH/INCA
• RIZOBAC (Metabolites from Bulkholderia cepacia)……………INCA
2004 Pugwash Workshop
 PRODUCTION AND APPLICATION OF
BIOFERTILIZERS IN CUBA
(YEARLY AVERAGE FOR THE LAST DECADE)

Biofertilizer Production Surface (ha)

Rhizobium 10 t 15 000

Azotobacter 800 ML 15 000

Fosforina 80 ML 3 000

Arbuscular mycorrhiza 25 t 3 000

2004 Pugwash Workshop

 AVERAGE DOSE FOR
SOLID BIOFERTILIZERS

1-5 kg/ha

AVERAGE DOSE FOR

LIQUID BIOFERTILIZERS

20 L/ha

2004 Pugwash Workshop

BIOPESTICIDES

2004 Pugwash Workshop

 Philosophy
Philosophythat
thatuses
usesthe
thebest
best
combinations
combinationsof oftactics
tacticsof
of
management
managementfor forpest
pestlimiting
limiting
IPM with
withthe
theminimum
minimumimpact
impactinin
environment
environmentandandeconomy.
economy.
••Dynamic
Dynamic
••Flexible
Flexible
 Cultural
Cultural Chemical
Chemical

Physical
Physical Etology
IPM Etology

Genetic
Genetic Biological
Biological
Legal
Legal
I Main bioproducts developed and
commonly used in Cuban agriculture
BIOPESTICIDES:

Bt (Bacillus thuringiensis) formulations to control mainly lepidopterae
species.

Trichoderma sp mainly to control soil fungi

Verticillium lechanii to control whiteflies, aphids, thrips and other
insects

Bauveria bassiana to control thrips and other insects

Paecelomices fumosoroseus controlling different insects, mainly
beetles, fire ants and nematodes

Corynebacterium paurometabolum to control nematodes and other
pests.

Metarhizium anisopliae to control termites, various coleoptera insects,
leafhoppers and aphids .
Biological Control Target Crop
Bacillus thurigiensis Lepidoptera, Mites Vegetables, roots and
strains LBT-1, LBT-13, tuber, citrus, tobacco,
LBT-21, LBT-24 potato, plantain, grass
lands.
Beauveria bassiana Coleoptera (weevils), Sugarcane,
strain LBB-1 ants, Trips palmi plantain,citrus, rice,
potato,beans
Verticillium lecanii Bemisia tabaci Vegetables, roots and
strain Y-57 Myzus persicae fruits.
Metarhizium anisopliae Lepidoptera and grass lands, rice and
strain LBM-11 Coleoptera plantain

Trichoderma harzianum Phytophthora, Tobacco, vegetables,

Trichoderma spp. Rhizoctonia, Phytium, ornamentals, grains
Sclerotium
 Biological Control Pest Crops
Trichogramma pretiosum, Lepidoptera Sugarcane,
T. pintoi tobacco, Cassava,
Trichogramma spp cabbage,
cucumber
Phytoseiulus macropilis Mites Cassava, banana,
plantain
Telenomus spp. Spodoptera frugiperda Corn, sugarcane

Pheidole megacephala Cylas formicarius Sweet potato

Encarcia spp. Bemisia spp Bean

 For 1 Million of Hectares
Biological agents Production: 1000 – 3000
T/year

CREE:
- M I N A G : 132 working and 11 in
construction; 3 industrial plants
(one almost finished).
- M I N A Z : 53 working.

Indicators of use:
-4,5 kg/ha biopests.
-30,0 th/ha entomophagous
CREE: General Characteristics

 Location
 Workers: Woman
 Materials
 Strains and Insects populations.
 Facilities
 Diversification
 Technologies
 Distribution
 Popularization and Capacity building
 Quality Control.
 Thousand
of Ha.

1200
1500
1800

300
600
900

0
19
88
19
89
19
90
19
91
19
92
19
93
19
94
19
95
19
96
19
97
19
98
19
99
20
00
20
01
20
02
20
03
 25

21,100

20

11.1 Times Less

15
Ton

10
7,900

4,200
5

1,900

0
1990 1998 2002 2003
Urban Agriculture: 28 Sub-programs
Vegetables and Fresh Condiments

3345.0 3500.0

2370.2

U.M:Mt 1681.2

872.1
478.5
24.5 57.3 140.0

1995 1996 1997 1998 1999 2000 2001 2002 Plan

2003

2003: 3.5 MMt

Plan 2004: 3.7 MMt
 RKN Pc

Losses per 70 millions of USD/Year in the World

High losses in Vegetable Crops under Tunnel in Cuba
 • Identity
• Biodiversity
RFLP
• Mass Production (Control Quality)
• Post Release Monitor

New challenger: Real Time PCR

Specific Primer
Different forms of presentation of the
ended product and application in the
field.
Toxicological and Eco-
toxicological studies