Chapter 2

REVIEW OF RELATED LITERATURE

Upland Rice Production

Upland rice is grown in areas without bunds or paddies. It is usually aromatic, tall

and late maturing, and is usually grown organically, with lesser cultural practices or

management intervention. It requires lesser amount of water to grow, and growing this

type of rice is closely tied up with the IP's culture and traditions. These plants are very

location-specific and usually thrive in areas with higher elevation. However, these rice

varieties are low-yielding compared to lowland rice varieties.

Upland rice production is an important component of the agricultural economies

of many countries. The yields are generally low, but can be increased by the development

of improved varieties and cultural practices to suit the soil, climatic, and social

conditions. These improvements are entirely possible through research and extension.

Rice (Oryza sativa L.) is probably the most important cereal in the world and

serves as food for about 50% of the world’s population (Ladha et al., 1997). This crop

requires about 10 million tons of N fertilizers every year worldwide (IRRI, 1996). Rice is

usually grown in N-deficient soils, and this element must be supplied to the field by

commercially available N fertilizers. However, a substantial amount of urea-N or NO3-N

applied as fertilizers is lost through different mechanisms, thus causing environmental

pollution problems. Use of biological N2-fixation technology can decrease N fertilizer

 5

application and reduce environmental risks (Raimam et al., 2007). This process can

contribute as much as 75 kg N ha-1 per crop cycle with means of 8 to 30 kg N ha-1

(Irissarri and Reinhold-Hurek, 2001). These N-fixing bacteria may be free-living or

naturally associated to rice plants. Recent studies have shown that several bacteria may

be isolated from sterilized surface roots of rice plants, suggesting endophytic colonization

(Raimam et al., 2007). The most likely candidates for biological N fixation in rice are

species of Alcaligenes, Azospirillium, Bacillus, Enterobacter, Herbaspirillium,

Klebsiella, Pseudomonas, and Rhizobium (Malik et al., 1997; James et al., 1999; Yanni

et al., 1997).

Organic Agriculture

Organic farming has emerged as an important priority area globally in view of the

growing demand for safe and healthy food and long term sustainability and concerns on

environmental pollution associated with indiscriminate use of agrochemicals. Though the

use of chemical inputs in agriculture is inevitable to meet the growing demand for food in

world, there are opportunities in selected crops and niche areas where organic production

can be encouraged to tape the domestic export market.

Bio-fertilizers are being essential component of organic farming are the

preparations containing live or latent cells of efficient strains of nitrogen fixing,

phosphate solubilizing or cellulolytic micro-organisms used for application to seed, soil

or composting areas with the objective of increasing number of such micro-organisms

and accelerate those microbial processes which augment the availability of nutrients that
 6

can be easily assimilated by plants. Bio-fertilizers play a very significant role in

improving soil fertility by fixing atmospheric nitrogen, both, in association with plant

roots and without it, solubilize insoluble soil phosphates and produces plant growth

substances in the soil. They are in fact being promoted to harvest the naturally available,

biological system of nutrient mobilization (Venkatashwarlu, 2008).

Bio-fertilizers

Bio-fertilizers add nutrients through the natural processes of nitrogen fixation,

solubilizing phosphorus, and stimulating plant growth through the synthesis of growth

promoting substances. In fact, bio-fertilizers like Bio-N enhance shoot growth and root

development, makes plant resistant to drought and pest attack reduces incidence of rice

tungro infection, increases yield and milling recovery of rice (FNCA Bio-fertilizer

Newsletter, 2007).

Bio-fertilizer is a substance which contains living microorganisms which, when

applied to seed, plant surfaces, or soil, colonizes the rhizosphere or the interior of the

plant and promotes growth by increasing the supply or availability of primary nutrients to

the host plant. Bio-fertilizers can be expected to reduce the use of chemical fertilizers and

pesticides. The microorganisms in bio-fertilizers restore the soil's natural nutrient cycle

and build soil organic matter. Through the use of bio-fertilizers, healthy plants especially

rice can be grown while enhancing the sustainability and the health of soil. Since bio-

fertilizers play several roles, a preferred scientific term for such beneficial bacteria is

plant-growth promoting rhizobacteria (PGPR). Therefore, bio-fertilizers are extremely

 7

advantageous in enriching the soil fertility and fulfilling the plant nutrient requirements

by supplying the organic nutrients through microorganism and their by product. Hence,

biofertilizers do not contain any chemicals which are harmful to the living soil. Bio-

fertilizers are Eco-friendly organic agro-input and more cost effective than chemical

fertilizers (http://en.wikipedia.org/wiki/Biofertilizer).

Components of bio-fertilizers

Effective microorganisms

Effective microorganisms or EM is a mixed culture of beneficial microorganisms

which are primarily photosynthetic, lactic acid bacteria, yeast, actinomycetes, and

fermenting fungi that can be applied as an inoculant to increase the microbial diversity of

soils. This in turn, can improve soil quality and health, which enhances the growth, yield,

and quality of crops. The concept of inoculating soils and plants with beneficial

microorganisms to create a more favorable microbiological environment for plant growth

has been discussed for decades by agricultural scientists. However, the technology behind

the concept of effective microorganisms (EM) and its practical application was developed

by Professor Teruo Higa at the University of the Ryukyu in Okinawa, Japan. He has

found microorganisms that can coexist in mixed cultures and are physiologically

compatible with one another. When these cultures are introduced into the natural

environment, their individual beneficial effects are greatly magnified in a synergistic

fashion (http://en.wikipedia.org/wiki/Bio-fertilizer).
 8

Bio-fertilizers like Rhizobium, Azotobacter, Azospirillium and blue green algae

(BGA) are in use since long time ago. Rhizobium inoculant is used for leguminous crops.

Azotobacter can be used with crops like wheat, maize, mustard, cotton, potato and other

vegetable crops. Azospirilluminoculants are recommended mainly for sorghum, millets,

maize, sugarcane and wheat. Blue green algae belonging to genera Nostoc, Anabaena,

Tolypothrix and Aulosira fix atmospheric nitrogen and are used as inoculants for paddy

crop grown both under upland and low land conditions. Anabaena in association with

water fern azolla contributes nitrogen up to 60 kg/ha per season and also enriches soils

with organic matter (http://en.wikipedia.org.wiki/Biofertilizer).

The concept of Effective Microorganisms (EM), their role and application

The concept of effective microorganisms (EM) was developed by Professor Teruo

Higa, University of the Ryukyu, Okinawa, Japan (Higa, 1991; Higa and Wididana, 1991).

EM consists of mixed cultures of beneficial and naturally-occurring microorganisms that

can be applied as inoculants to increase the microbial diversity of soils and plant.

Research has shown that the inoculation of EM cultures to the soil/plant ecosystem can

improve soil quality, soil health, and the growth, yield, and quality of crops. EM contains

selected species of microorganisms including predominant populations of lactic acid

bacteria and yeasts and smaller numbers of photosynthetic bacteria, actinomycetes and

other types of organisms. All of these are mutually compatible with one another and can

coexist in liquid culture.

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EM is not a substitute for other management practices. It is, however, an added

dimension for optimizing best soil and crop management practices such as crop rotations,

use of organic amendments, conservation tillage, crop residue recycling, and bio-control

of pests. If used properly, EM can significantly enhance the beneficial effects of these

practices (Higa and Wididana, 1991).

Carbonized Rice Hull (CRH)

A hydrophilic material made from the incomplete or partial burning of rice hull,

CRH contains potassium, phosphorus, calcium, magnesium, and other microelements

needed for growing crops like rice. As soil conditioner, CRH replenishes air and retains

water in the soil. Because of the heat it undergoes, it is sterile and thus, free from

pathogens. As such, it makes an excellent host for beneficial microorganisms and an

ingredient for bioorganic fertilizer (http://blog.agriculture.ph/tag/philippines-carbonize-

rice-hull).

Bio-N

BIO-N is a microbial-based fertilizer developed by Professor Emeritus Mercedes

U. Garcia from nitrogen-fixing bacteria isolated from the roots of "talahib" grass

(Sacharum spontaneum). It is a product of the ever inquiring mind of Dr. Garcia who had

observed the pervasiveness of talahib grass even on the hostile soil and environment

conditions. She wanted to find out the reason why the talahib could still grow normally in

conditions where other agricultural crops would easily succumb. Dr. Garcia conducted a
 10

research study at the National Institute of Molecular Biology and Biotechnology

(BIOTECH) to satisfy her curiosity and through years of meticulous work, she was able

to isolate a species of nitrogen-fixing bacteria from talahib roots called Azospirillium.

The bacterium has the capability to convert atmospheric nitrogen (N 2) into a form usable

by the plants. These bacteria, once associated with roots of rice, corn, sugar cane, and

some vegetable plants, can enhance root development, growth, and yield (BIOTECH,

2009).

It is in powder form and prepared in a 200-gram packet, which could either be

used as seed coating or used as a dilute solution for root dipping; or drenching already

established young plants. One packet can be used to inoculate 3 kg of corn seeds or 20 kg

of rice. Five packets of Bio-N can provide the needs of a one-hectare rice or corn

plantation for nitrogenous fertilizer. Fields that have sufficient amounts of other elements

respond dramatically to Bio-N. Bio-N enhances root development, growth and yield of

the rice and corn and other vegetable crops, maintains the soil natural properties and

fertility, and keeps plants healthy and green even during droughts and pest infestation

(BIOTECH, 2009).

Farmers in all regions in the Philippines (Regions 1 to 12) specifically those areas

covered by the Department of Agriculture benefit from the use of BIO-N. Trainings on

the preparation of the carrier of the Bio-N and on the application of Bio-N are provided to

interested clients (BIOTECH, 2009).

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Biogroe

Biogroe is a solid-based microbial plant growth promoter containing plant growth

promoting rhizobacteria (PGPR). PGPR are root-associated bacteria, which influence

root growth by producing plant hormones and provide nutrients in soluble form. PGPR

can also protect plant surfaces from colonization by pathogenic microbes through direct

competitive effects and production of anti-microbial compounds.

Biogroe is easy to use and environment-friendly. The benefits of Biogroe include

enhanced root growth and development, increased productivity of crops, and reduced use

of toxic or environmentally damaging chemical fertilizers and pesticides. The product can

be used in vegetable production, propagation of ornamentals and plantation crops, rice

and sugarcane production and in the growth enhancement of fruit and forest trees.

Biogroe showed promising results on test crops in Bataan, Laguna, Tarlac and Palawan.

Scientists at the National Institute of Molecular Biology and Biotechnology

(BIOTECH) have scored another breakthrough with “Biogroe,” a biological fertilizer

containing plant growth-promoting bacteria (PGPB) which enhances growth and

productivity of plants.

Experiments conducted at BIOTECH showed that plants fertilized with Biogroe

produce high yield - 90% for ampalaya and 40% for lettuce. Biogroe also caused

Dendrobium to flower early, and Mussaenda and Hibiscus to grow heavier and longer

roots. It was also found effective in the rooting of black pepper cuttings and as a

substitute for commercial rooting formulations.

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It is also reported that farmers noted early seedling growth in both crops.

Substituting one-half of chemical fertilizers with Biogroe showed no difference in yield

as when 100% chemical fertilizer was used. Since Biogroe is cheaper than chemical

fertilizers, its use meant savings on the cost of production.

Biogroe is composed of bacteria which produce plant growth regulators such as

auxins, cytokinins and gibberellins. These bacteria also help plants produce more roots

and protect these roots from diseases by producing antibiotics. Plants with more roots

absorb water and nutrients more easily. Moreover, since they are more tolerant to

diseases, they become healthier and produce higher yield. BIOTECH is now studying

Biogroe ability to promote early germination, growth, and rooting of other hard-to-

cultivate crops (BIOTECH, 2009)

Mykovam

Mykovam is a powdered bio-fertilizer which has effective species of vesicular

arbuscular (VAM) fungi. Basically, its functions are similar to that of mycogroe. The

fungi infect the roots, from a close association with the plant and assist in absorbing

water and nutrients such as phosphorus, zinc and nitrogen. Mykovam can be applied to

almost all crops. Mykovam even fights destructive pests and diseases and increases plants

tolerance to drought and heavy metals. Use of this product reduces 60-85% of the

fertilizer requirements of most crops, improves soil properties and fertility, and promotes

absorption of water and nutrients such as phosphorus, nitrogen and zinc.

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Vital-N

Vital N is a wettable powder formulation that is available in a very convenient

100-gram pack. It acts as a virtual nitrogen fertilizer factory at the roots of the plants. It

protects the plants from root and leaf diseases. It also makes plants drought resistant. Not

only is it convenient and powerful, it is also easy to store. It can be stored at room

temperature for at least 3 years.

Vital-N is an FPA-registered organic bio-fertilizer that replaces as much as half of

the recommended fertilizer. It also increases farmer’s profitability by at least 15%, and in

some cases more than 100%. Tests conducted by independent scientists and farmers in

rice and corn show that, Vital-N increases yields by at least one ton per hectare. Vital-N

is an environmentally-friendly, inexpensive, and easy-to-use bio-fertilizer developed by

Arnichem with the assistance of the Philippine Rice Research Institute. It is a seed

inoculant for rice, corn, bananas, onions, garlic, tomatoes, tobacco, cotton, and other

crops. Vital-N is a special formulation of dried Azospirillium sp. cells, as well as other

vitamins and minerals. This special formula increases yield, reduces the amount of

fertilizer needed, protects plants from disease-causing organisms, resulting in healthier

plants and higher yield.

Vital-N comes in powder form. It has a long shelf life of at least three years in

contrast to the rapid loss of viability of moist preparations of Azospirillium sp. It comes in

a very convenient 100g package of powder, which is enough to treat 20-40 kg of rice

seeds, or 18 kg of corn seeds, avoiding the bulkiness of other biofertilizers. Once opened,

the Vital-N should be used all at once. If this is not possible, it should be protected
 14

from air and water and refrigerated. It turns colored when mixed with water or wet seeds.

It is compatible with fungicides and insecticides.

Azospirillium sp. is a naturally-occurring bacterium found in Philippine soil.

Because it generates plant growth hormones like IAA, it promotes extensive root systems

in plants, cytokinins, and gibberellins-like substances, resulting in healthy and sturdy

plants, higher yield, and more solid grains. It also produces substances that help dissolve

nutrients within the soil, allowing these to be more readily available to the plant. This

results in greater nutrient absorption and increased fertilizer efficiency.

In addition, Azospirillium captures nitrogen from the air, making it available as

plant food and lowering nitrogen fertilizer requirements, and produces a bacteriocin that

destroys disease-causing organisms. Other ingredients have been added to the Vital-N

formulation to protect the Azospirillium and keep it viable, to help it stick to the seeds

and roots of the plant, to extend shelf life, and a dye to serve as in indicator that the bio-

fertilizer has already been applied to the plant.

Vital-N promotes seedling vigor, causing seeds to sprout readily. Vital-N hastens

root growth. More extensive root networks allow for better absorption of soil nutrients,

reduced run-off and water take up. This results in more vigorous and sturdy plants.

Individual farmers all over the country report better seedling vigor, increased yields,

lower fertilization costs, and resistance to disease due to the use of Vital-N. Dr. Edwin

Honrade of the University of Southern Mindanao reports that his preliminary experiments

show that Vital-N protects rice from bacterial leaf blight from soil-borne diseases. Rice

farmers from other parts of the country report similar results. Dr. Romulo Davide in
 15

Bantayan, Cebu obtained more than a 100% increase in yield when Vital-N was applied

as compared to when nothing was applied, and is comparable to results obtained when

complete fertilizer was applied (http:// www. scribd.com).

Studies on the effect of biofertilizers and other soil amendments

Effective microorganisms (EM) technology of nature farming was introduced by

Higa (1991). EM culture consists of co-existing beneficial microorganisms, mainly

species of photosynthetic bacteria (Rhodopseudomonasplastris and

Rhodobactersphacrodes), lactobacilli(Lactobacillus plantarum, L. casei, and

Streptococcus lactis), yeast (Saccharomyces spp.), and actinomycetes (Streptomyces

spp.), which improve crop growth and yield by increasing photosynthesis, producing

bioactive substances, such as hormones and enzymes, controlling soil diseases, and

accelerating decomposition of lignin materials in the soil (Higa, 2000; Hussain et al.,

2002).

Study on the use of rice hulls are evaluated at the International Rice Research

Institute. Yields with rice hulls were similar to yields in inorganic fertilizer with a

recommended rate of 90-28-28 kg NPK/ha. The highest cost of production and lowest

rate of return were with 90-30-30 kg NPK/ha. Rice hulls at 1 t/ha as replacement for 2

bags of 14-14-14 gave the highest net income and rate of return (IRRI Newsletter 1987).

Field trials conducted by BIOTECH to test and demonstrate the effects of BOF on

different crops. At a recommended rate of 10 bags/ha, rice yields are increased by 157%

over the control. Increase in yield with BOF application was also achieved in wetland
 16

rice. BOF application appeared to increase in the number of filled grains, as well as the

number of panicles (Espiritu B.M. 1992)

The study conducted by the PNRI on the use of nuclear techniques in the

assessment of Bio-N fertilizer as seed inoculant for corn on low fertility area in the

province of Isabela (around 400km north of Manila) showed that corn yield was only 208

kg/ha (kilogram per hectare) when not inoculated with Bio-N and without fertilizer

application. Corn yield was increased tremendously to 2,880kg/ha when inoculated with

Bio-N plus application of P and K fertilizer. This resulted to an increase in income from

P 1,456.00 to P 14,341.50. Bio-N inoculation plus application of half and full

recommended rate of N fertilizer yielded 3,460kg/ha and 2,960kg/ha, resulting in

increase in income of P 16,230 and P 10,558 respectively. The experiment also used N-

15 to quantify biological nitrogen fixing capacity of Bio-N. N-15 data indicated that 19-

24% of the nitrogen taken by the plants is nitrogen from the atmosphere that is fixed by

the Bio-N (PNRI, 2007)

Potential role of bio-fertilizers in agriculture nitrogen-fixers (NF) and phosphate

solubilizers (PSBs)

The incorporation of bio-fertilizers (N fixers) plays major role in improving soil

fertility, yield attributing characters and thereby final yield has been reported by many

workers (Subashini et al. 2007a; Kachroo and Razdan, 2006; Son et al. 2007). In

addition, their application in soil improves soil biota and minimizes the sole use of

chemical fertilizers (Subashini et al. 2007a). Under temperate conditions, inoculation of

 17

Rhizobium improved number of pods plant-1, number of seed pod-1 and 1000-seed

weight (g) and thereby yield over the control. The number of pods plant-1, number of

seed pod-1 and 1000-seed weight (g) recorded were 25.5, 17.1 and 4.7 per cent more over

the control, respectively which was statistically significant (Bhat et al. 2009). In rice

under low land conditions, the application of BGA+ Azospirillium proved significantly

beneficial in improving LAI and all yield attributing aspects. Grain yield and harvest

index also exhibit a discernable increase with use of bio-fertilizers (Dar and Bali, 2007).

Improved macro and micro nutrients uptake

The improvement of P nutrition of plants has been the most recognized beneficial

effect of mycorrhiza. The mechanism which is generally accepted for this mycorrhizal

role consists of a wider physical exploration of the soil by mycorrhizal fungi (hyphae)

than by roots. A speculative mechanism to explain P uptake by mycorrhizal fungi

involves the production of glomalin. Glomalin contains very substantial amounts of iron

up to 5% of the glomalin pool (Lovelock et al., 2004).

Because of their small size, AM fungal hyphae are better able than plant roots to

penetrate decomposing organic material and are therefore better competitors for recently

mineralized N (Hodge, 2003). By capturing simple organic nitrogen compounds, AM

fungi can short-circuit the N-cycle. It is also reported that the AM- fungi also increases

the uptake of K, and concentration of K has been found more in mycorrhizal than non-

mycorrhizal plants (Bressan et al., 2001). Apart from this, the AM-fungi also increases

the uptake and efficiency of micronutrients like Zn, Cu, Fe etc. by secreting the enzymes,
 18

organic acids which makes fixed macro and micronutrients mobile and as such are

available for the plant.

Theoretical framework of the study

The theoretical framework of the study is given in Figure 1. The use of mixed

cultures of beneficial microorganisms as soil inoculants is based on the principles of

natural ecosystems which are sustained by their constituents; that is, by the quality and

quantity of their inhabitants and specific ecological parameters that is the greater the

diversity and number of the inhabitants, the higher the order of their interaction and the

more stable the ecosystem. The mixed culture approach is simply an effort to apply these

principles to natural systems such as agricultural soils, and to shift the microbiological

equilibrium in favor of increased plant growth, production and protection (Higa, 1991;

1994; Parr et al., 1994).

To enhance the concept of controlling and utilizing beneficial microorganisms for

crop production and protection, one must harmoniously integrate the essential

components for plant growth and yield including light (intensity, photoperiodicity and

quality), carbon dioxide, water, nutrients (organic-inorganic) soil type, and the soil

microflora. Because of these vital interrelationships, it is possible to envision a new

technology and a more energy-efficient system of biological production.

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BIO FERTILIZER
PRODUCTION

CONVERSION
ENHANCING UPLAND OF
SOIL RICE
FERTILITY/NIT PRODUCTION/FOOD
AGIRCULTUR
ROGEN SECURITY AL WASTE
FIXATION TO BOF

MICROBIAL
INOCULATION

Figure 1. Theoretical framework of the study

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Conceptual framework of the study

The framework of the study is given in Figure 2.0. Upland rice was grown in a

field and treated with locally-made bio-fertilizer (which is made up of CRH, rice bran,

chicken dung, molasses and EM), Bio-N Biogroe, Mykovam and Vital-N were used in

the study. Data on agronomic characteristics of upland rice, enhancement of nutrients in the

soil, physiological Characteristics, soil physical characteristics and yield and yield components of

upland rice was determined.

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UPLAND RICE

COMMERCIAL
LOCALLY-MADE BIOFERTILIZERS
BIOFERTILIZERS

CRH BIO-N
RICE BRAN
EM
BIOGROE
MOLASSES MYKOVAM
CHICKEN DUNG VITAL N

FIELD EXPERIMENT: SIX TREATMENTS AND THREE

REPLICATIONS, 18 PLOTS WITH A MEASUREMENT OF
3 m x 4 m per plot/treatment

1. Agronomic characteristics of upland rice;

2. Physiological Characteristics
3. Soil characteristics.
4. Yield and yield components of upland rice.
5. Cost and Return Analysis

MOST EFFECTIVE AND EFFICIENT BIOFERTILIZER

Figure
2.
Conceptual framework of the study