Color Green and Brown Azolla as Exposed in Different Environments

INTRODUCTION
Azolla (mosquito fern, duckweed fern, fairy moss, and water fern) is a genus of seven species of aquatic ferns in the family Salviniaceae. They are extremely reduced in form and specialized, looking nothing like conventional ferns but more resembling duckweed or some mosses. Azolla is a free floating aquatic fern, which nowadays can be found in freshwater environments in temperate and tropical regions all over the world. It ranks among the fastest growing plants on Earth and due to its association with the nitrogen fixing cyanobacteria Anabaena azollae, it is independent of extern organic nitrogen. In that way Azolla is not only known to fix huge amounts of carbon, but as well to produce vast amounts of organic nitrogen.

The nutrient, which mainly limits the growth of Azolla, is phosphorous. Azolla is rich in protein, amino acids, vitamins and minerals. It is the only plant used as biofertilizer in agriculture or as a green manure for wetland rice in Northern Vietnam, and Central and Southern China because of its high nitrogen content. In early 1980’s the use of Azolla in South Cotabato, Philippines was widely published. Azolla plants have been described by the Chinese and Vietnamese as being miniature nitrogen fertilizer factories. Azolla is traditionally used throughout Asia and parts of Africa as feed for livestock, poultry and fish such as tilapia, red pacu, carps and other herbivorous and omnivorous species. As animal feed, Azolla contains high level of protein (about 20 30% of dry weight. Some literatures suggest higher protein level of 50 60%) and fat. At times Azolla is also used as human food. Azolla is important to reproduce for it contributes more useful benefits.

Conceptual Framework
REPRODUCTION RATE OF COLOR GREEN AND BROWN AZOLLA AS EXPOSED IN DIFFERENT ENVIRONMENTS OUTPUT
1.) Reproduction rate of green and brown azolla under the sun and on a covered or dim environment PROCESS
Exposition under the sun and on a covered or dim environment INPUT
* Nature
* Characteristics
* color

Statement of the problem
1) What color or type of Azolla (the green or brown) has the fastest reproduction rate in light and dim environments? 2.) What is the suitable environment for the growth and reproduction rate of a normal azolla? Is it under the sun (bright) or a covered dim environment? Hypothesis

Azolla is a plant fern which lives in paddy water and other freshwater environments. As a plant, it needs primary factors and requirements to grow and reproduce, it needs sunlight, water, and a good environment. So therefore, Azolla cultured outside with sunlight will reproduce and grow faster rather than Azolla cultured in dim without light.

Scope and Delimitation of the study
This study was limited and focused to the reproduction rate of the 2 colors (green and brown) of Azolla (Azolla Pinnata) when cultured under indoor and outdoor environment conditions. This research was conducted at GIFTED Learning Centre, Inc. Among the aspects considered were:

1.) Determination of what color of Azolla multiplies faster from indoor environmental condition and outdoor environmental condition. 2.) Determination of what kind of Azolla multiplies the fastest from the over-all samples and considering of what environmental condition ( bright – under the sun and dim – covered environment) it’s being exposed. Importance of the study

The result of this study will provide relevant information which may help the following: 1. Department of Health (DOH). Since azolla is proven to be a mosquito repellant, so by knowing on how and where to reproduce it, this agency can conduct a study in the prevention of dengue here in the Philippines. 2. Agricultural and fish industry owners. Due to its high protein, amino acid, vitamin and nitrogen content, azolla was proven to be substitute food of some herbivore, and omnivore species. It may also be used as biofertilizer in agriculture for some plants. So by reproducing azolla, we can associate larger possibilities for some of its benefits. 3. Squatters area settlers. Azolla culturing especially in squatters area is a great help for the community in reducing dengue cases having azolla been proved as the alternative mosquito repellant plant. 4. Students. Azolla is ensured with significant benefits that will contribute for the betterment of living. Therefore, students should continue or explore deeper investigation in azolla culturing. Definition of terms

1.) Salviniaceae – these are heterosporous floating aquatic plants. The genus Azolla is commonly included within the family. 2.) Duckweed – is the common name for a family of small aquatic herbs known as Lemnaceae that grows in fresh water. They have the unique distinction of being the smallest flowering plants on earth. These plants are also unique in that they do not have any stem or leaf structures. The plant is simply a fleshy ovoid or flattened structure that may or may not bear simple roots. These highly successful plants represent the ultimate in reduction for a vascular plant. Duckweed is ubiquitous to most temperate and tropical regions of the world, making it readily available to most farmers. They are typically found floating in thick mats of homogeneous populations in quiet streams or ponds containing high levels of organic matter.

3.) Cyanobacteria – are prokaryotes (single-celled organisms) often referred to as “blue-green algae.” While most algae is eukaryotic (multi-celled), cyanobacteria is the only exception. Cyanobacteria obtain their energy through photosynthesis. They are not strictly unicellular, but can be found in colonial and filamentous forms, some of which differentiate into varying roles. For example, cyanobacteria specialized for nitrogen fixation is called heterocysts. 4.) Paddy water- is a structural form of water like a rice field. 5.) Salinity- the saltiness of the water and can be measured by a refractometer. 6.) Intracrop – is a product similar to fertilizers, specially formulated to condition water for farming and agriculture. 7.) Carbofuran – is one of the most toxic carbamate pesticides. It is marketed under the trade names Furadan, by FMC Corporation and Curater, among several others. It is used to control insects in a wide variety of field crops, including potatoes, corn and soybeans. It is a systemic insecticide, which means that the plant absorbs it through the roots, and from here the plant distributes it throughout its organs where insecticidal concentrations are attained. Carbofuran also has contact activity against pests.

REVIEW OF RELATED LITERATURE

SOURCE: Azolla Utilization Proceedings on the workshops of Azolla use SPONSORED BY: The Fujan Academy of Agricultural Sciences (Fuzhou, Fujian, China) and The International Rice Institute (Los Baños, Laguna ,Philippines) ENVIRONMENTAL REQUIREMENTS

Climate. Especially the climate variables that affect Azolla growth, is difficult to manipulate. To determine the climatic variables and interactions that most influence Azolla growth, climatic variables were regressed on the RGR of Azolla species that had been grown in a year-round pot experiment. The correlation analysis of data recorded during hot humid weather indicated that temperature and humidity had a negative effect on growth rate and solar radiation had a positive influence. Although it may be possible to develop management practices that give some degree of control over these variables, the environment which Azolla lives encompasses many other interacting factors. For example Azolla’s susceptibility to fungal pathogens probably interacts with high temperature and humidity. Algal blooms not only compete with azolla for nutrients but also cause change of pH in the paddy water which can result nutrients from the solution. They also restrict water movement which can result in high water temperatures lethal to azolla. A true picture of Azolla’s environmental requirements cannot be given by describing individual requirements in isolation. Often the interactions or indirect effects of factors are more important than the direct effect. Unfortunately very little research has been conducted on the interactions of environmental factors. ENVIRONMNTAL FACTORS

Water. The lack of good control and availability of water in rice growing areas is the primary constraint to the spread of Azolla use. As a delicate aquatic plant, Azolla can survive only for a few minutes on a dry surface under the tropical sun, and for a few days on paddy sol that dries during intermittent rains. Some varieties can survive indefinitely on moist, shaded mud, but will not multiply to any useful extent without a water surface on which to spread. Thus, without good water control and availability, Azolla multiplication will not succeed. The need for water extends beyond the growing season and remains important to a limited extent throughout the year. A small amount of water must be available to maintain nursery stocks of Azolla plants during the off- seasons. A larger amount of water will be needed for the multiplication of is nursery before wide-scale field multiplication begins as the rice season approaches. Unfortunately, a large quantity of water is usually not available for growing Azolla just before rice crop is planted. The source of water is also important. Like rice, Azolla grows better during dry seasons when irrigation water is available.

If water comes from precipitation, additional problems may occur. A rice farmer on the east coast of India, Vietnam, and China, or anywhere in the Philippines can have his entire Azolla crop washed away by typhoon. Farmers in a monsoon area can suffer the same fate from excessive rainfall, which causes flooding. With the exception of a few areas such as South Cotabato in the Philippines, most farmers will need to invest considerably effort to produce an Azolla crop. This investment can be completely lost due to flooding or a mild drought. Light. The growth rate of Azolla has been reported to saturate at 25-50% of full sunlight, and is not inhibited by full sunlight as long as other factors are not limiting. As an intercrop under rice, the growth rate of Azolla will begin to decline as the developing rice canopy reduces light quantity and quality below that necessary to saturate growth rate. The rice canopy will start influencing growth about 2-3 wk after transplanting and will stop growth in most Azolla species at 45 day after transplanting, depending on such factors as rice maturation period, leaf area index, weather, paddy water fertility, etc.

An aspect of shading which is of possible interest but has not been studied is the intracrop shading of Azolla as it becomes crowded. As in other crops, crowding probably results in competition for light, nutrients, etc. It is difficult to take measurements within the Azolla canopy because of the smallness of Azolla, but it is not impossible. Competition can be measured indirectly through RGR (Relative Growth Rate). Further research could help optimize productivity of biomass and accumulated Nitrogen if more information was available about intracrop competition. Day length is another aspect of light. Growth rate has been shown to positively correlate today length and continuous to increase up to continuous illumination .higher latitudes , such as Central China, will have a longer day length during the late spring and early summer Azolla growing season than the tropics which have an almost uniform day length all year round. Azolla growth rate is higher and higher latitudes than in the tropics. Temperature. Temperature is probably the most important limiting environmental factor in Azolla cultivation. It is also very difficult to manipulate.

Its direct effects are not as serious s it indirect influences. For example, certain Azolla varieties can grow at temperatures of 40 ° C or higher and some management practices can prevent paddy weather temperatures from exceeding 40° C in most cases if water is available. The most serious problem with temperature is its stimulating effect on azolla pests, e.g. insects, pathogenic fungi, and free living algae. The optimum temperature for most azolla species is within the range of 20-35 ° C. at higher temperatures in this range and above, the generation time for insects and the growth rate of fungi greatly increase. Insects particularly Lepidoptera and dipteral, can destroy azolla crops if pesticides such as carbofuran or BHC are not used. Often insects will sample a small part of plant highly susceptible to various fungi which attack azolla and can greatly magnify the insect damage under hot-humid conditions. Because high temperatures are not a direct limitation, azolla has an excellent potential for successful cultivation in irrigated deserts where humidity is relatively low and alternate host plants for insects are limited. azolla does very well on the northern border of Senegal in west Africa (Van Hove, pers. Comm.) and can probably do well in the traditional rice areas of Mali with good water control.

Mineral Nutrition. Because Azolla is an aquatic plant, essential elements must be available in the water for Azolla to survive. Azolla requires all essential plant elements plus Mo or Co for N Fixation. However, most paddy water does not contain an adequate balance of essential elements for successful Azolla cultivation. Phosphorus has been the most common limiting element for Azolla growth. The threshold concentration of P in Azolla Tissue is probably about 0.2-0.3% on a dry wt. basis. P stressed plants are usually smaller, pink to red less vigorous, and have a low concentration of total N. Under severe stress, the plants become highly compact and dark red, and often develop very long curled roots. P deficiency can be overcome by applying P fertilizer. In some situations, deficient plants may be able to extract sufficient P from paddy soil if the water level can be reduced to the point the azolla roots can touch the soil. Water quality. Several other aspects of water quality have been studied. Pollution from sewage and herbicides is of little concern to developing countries, but may be important as azolla cultivation is attempted. Other factors are known to limit azolla use. pH is perhaps the most important in developing countries. Optimum growth of azolla in culture solution is in pH range of 4.5-7, but azolla can survive in pH 3.5-10 if all essential elements are available. The most important concern about pH is the availability of essential elements in the paddy water.

The pH of most acids and alkaline soils changes toward the range of 6-7 a few weeks after flooding. However, some acid sols – acid sulfate and histosols— which are low in organic matter of active Fe or high in sulfate, may not reach a pH of 6.0 even after months of submergence. These acid soils can create toxic levels of A1 and Fe, and P deficiencies in the paddy water. Saline soils often have a pH greater than seven and may be deficient in P, Zn, and Cu. Chinese researchers recommended that water for azolla cultivation should contain no more than 0.3% salt. Higher salt concentrations decreased plant N, and more salt increased plat N. Haller et al found that A. caroliniana growth ceased when the concentration reached .3%. Salinity or alkalinity may be a problem to rice fields located along the coast and in poorly drained irrigated desserts, such as in areas of Pakistan. Chapter 3 METHODOLOGY

This chapter presents the methodology used in the research. It describes the method and materials used in gathering, presenting, analyzing and interpreting the data.

1. First, the researchers collected 12 empty gallons. Then they removed the upper part of the gallons.
2. The researchers put the soil (600g) on every gallon using the electric
weighing scale.
3. The researchers put 800 ml of freshwater on every gallon.
4. After putting the exact amount of soil and water, the researchers put the green azolla on 6 gallons and brown azolla for the remaining gallons. Each gallon contains 10 azolla.
5. The researchers investigated and observed them for three (3) weeks by putting three gallons containing green azolla and three (3) gallons containing brown azolla outside under the sun, and by putting the remaining three (3) gallons containing green azolla and three (3) gallons containing brown azolla in our classroom which is free from dust and dirt which was instructed by Ms. Mary Joy Gonzales. The researchers also brought the azolla home every weekend to investigate continuously and consistently and to monitor them. 6. Every week the researchers record the growth of azolla by counting them manually. They also took pictures of them.