bio fertilizers for jatropha curcas l euphorbiaceae grown in different planting media

International Conference on Environmental Research and Technology (ICERT 2008)

Biofertilizers for Jatropha curcas L. (Euphorbiaceae) Grown in Different Planting Media

Ernesto S. Elefan* Life Sciences Department, Central Philippine University Jaro, Iloilo City 5000 Philippines

*Corresponding author, Fax: + 63-33 3203685, Phone: (63-33) 3291971-72 local 1062 Email: [email protected]

ABSTRACT Physic nut (Jatropha curcas L.), commonly known as “Tuba-tuba” in the Philippines, is a small tree of plant tribe oannesieae of Crotonoideae in the Euphorbiaceae family that have approximately 170 known species worldwide (Dehgan and Webster (1979). It grows widely in the Philippines and has recently gained global interest because of its seed oil that can be used for blending with diesel producing an ecology-friendly biofuel as an alternative source of energy. This study compared the early growth response of the Physic nut to the interactions of single inoculation of biofertilizers Azotobacter, Effective Microorganisms (EM), and Bio N with different planting media namely garden soil (Sta. Rita clay loam), humus (decomposed rice hull), river sand, and their mixture on 1:1:1 ratio over the uninoculated control. It was carried out in a 4 x 4 factorial experiment in randomized complete block design with 16 treatment combinations and four replications. Azotobacter x 1:1:1 mixture and EM x humus interactions produced significantly (P<0.05) taller plants. Fresh matter weight per plant was significantly (P<0.05) highest in Azotobacter x Humus and Bio N x Humus interactions. Leaf count, shoot base diameter, and dry matter weight per plant, on the other hand, were not significantly (P>0.05) affected by the biofertilizers and planting media interaction. Among the planting media used, humus showed to be most promising in growing Physic nut as revealed by the significant (P<0.05) results in plant height, leaf count, shoot base diameter, fresh matter weight, and dry matter weight per plant. However, a 1:1:1 mixture can be a good alternative. Using garden soil, either EM or Bio N can be used. However, with sand as a planting medium, EM can be applied as a seed microinoculant.

Keywords: Azotobacter, biofertilizers, bioN, EM (Effective Microorganisms), Jatropha curcas L.

1. INTRODUCTION Physic nut (Jatropha curcas L.), commonly known as “Tuba-tuba” in the Philippines, belongs to plant tribe

Joannesieae of Crotonoideae in the Euphorbiaceae family that have approximately 170 known species worldwide (Dehgan and Webster, 1979). It grows anywhere in the Philippines and has recently gained global interest because the seed oil has both commercial and ecological value. The seed oil, commercially known as curcas oil, has long been used by some Filipinos to illuminate their homes and in making soaps and candles. It can be used for blending with diesel, thus producing ecology-friendly bio- diesel which can be a good alternative source of energy that will be of great help in saving a considerable amount of fossil fuel that destroys the ozone layer (Padilla, 2006). Each inflorescence yields a bunch of approximately 10 or more ovoid fruits which yield about 22 to 37% oil by simple extraction (Quisumbing, 1978; Padilla, 2006 citing Solsoloy & Duldolao, 2004). It is a cure for diarrhea, cough, snake bite, sprains and dislocations, toothache and gum bleeding, skin diseases such as scabies, eczema, ringworm, fever, and rheumatism (Quisumbing,1978). The latex has an alkaloid known as jatrophine, reported to have anti-cancer properties (Sai Petro Chemicals, Pvt. Ltd., 2006). Its bark has tannin, the flowers attract bees and thus the plant has honey production potential. Aqueous extracts of physic nut can be used as bio-control for fungal pathogen and has molluscicidal activity against liver fluke ( Padilla, 2007, citing Garcia & Lawas, 1990; Agaceta, L.M., P.U. Dumag, J. A. Atalos & F.C Bandiola, 1981).

Biofertilizers are natural fertilizers or microbial inoculants of bacteria, algae, fungi alone or in combination that augment the availability of nutrients to the plants. Their preference to chemical fertilizers offers economic and ecological benefits by way of soil health and fertility to farmers because these can accelerate the decomposition of organic residues releasing plant nutrients through mineralization. These also facilitate the uptake of plant nutrients, increase the nitrogen content of plants through symbiosis and improve plant growth by providing plant growth-promoting substances (Bashan, et al, 2004; Chiu-Chung Young, 1994, Higa, T. and J.F. Parr, 1994). Their beneficial influence plant growth and development can be enhanced by the prevailing hospitable planting media or soil environment. This means there must be enough organic materials, suitable host plants, water, and minimal disturbance of the soil.

Environmental technology & management 308


The host plant provides the microorganisms with soluble carbon sources which, in return, provide the host plant with an increased capacity to absorb water and nutrients from the soil. Adverse conditions are a pervasive feature in both natural and agronomic soils. The soil environment is constantly changing with regard to moisture, temperature and nutrient availability so that soil properties are often manipulated to improve crop yield. In many cases, soils may be contaminated through disposal of chemicals that are toxic to plants and microorganisms (Entry, J.A., P.T. Rygiewicz, L.S. Watrud and P.K. Donnelly, 2002)

The efficacy of biofertilizers applied as seed inoculants to improved plant growth is affected by the quality of planting or rooting media which is determined by soil composition, moisture, temperature, pH, and cation exchange capacity; and also by anthropogenic stressors including soil compaction, metals and pesticides (Gislerod, 1983; Parr, J.F., S.B. Hornick, & D.D. Kaufmann ,1994). These are phenomena of natural soils that can only operate in a range of mineral availability found under natural condition (St. John, T.V., 1985) and is an important consideration in plant propagation.

Physic nut is primarily propagated by seeds, however, scientific studies on the use of biofertilizers in promoting their agronomic characteristics that will ultimately determine its seed production ability under the local condition are limited. Similarly, insufficient data are available on germination and growth performance of this plant as influenced by planting media as basis of recommendation for its mass production under the greenhouse condition. The researcher therefore found it imperative to conduct similar study under Central Philippine University condition utilizing the commercially available biofertilizers as seed inoculants to Jatropha grown in different planting media consisting of garden soil, river sand, and humus used alone, and their mixture on a 1:1:1 ratio.

This study evaluated the early growth performance of Physic nut (Jatropha curcas L) when seed-inoculated with commercially-produced biofertilizers Azotobacter, Effective Microorganism (EM) and Bio N and grown in different planting media using garden soil (Sta. Rita Clay loam), pure river sand, and humus (decomposed rice hull) applied singly, and a mixture on a 1:1:1 ratio under a greenhouse condition. Specifically, this study was done to 1)assess the effects of commercial bio-fertilizers and different planting media on the early growth response of Physic nut under greenhouse conditions; 2) determine if there are significant differences in the early growth response of Physic nut to different commercial bio-fertilizers; 3) determine if there are significant differences in the early growth response of Physic nut to the different planting media, and 4) find out if there are significant interactions between the use of commercial bio-fertilizers and different planting media based on the early growth response of Physic nut to these growth factors.

2. MATERIALS AND METHODS This greenhouse experiment, consisting of two variables namely biofertilizers and different planting media, was

laid-out in a 4 x 4 factorial in a randomized complete block design with four replications. Planting media used were garden soil (P1); humus using decomposed rice hull (P2), sand (P3), and 1:1:1 mixture of garden soil, humus, and river sand (P4). Biofertilizers used to inoculate Physic nut seeds were Azotobacter (B1), EM or Effective Microorganisms (B2), and Bio N (B3) which were compared with the Uninoculated Control (B0) with a total 16 treatment combinations. Biofertilizers were prepared on manufacturers’ recommendation. Azotobacter was prepared by dissolving 100 grams of the material in 1 liter of distilled water. A 1% EM solution was prepared by mixing 1 ml of EM in enough distilled water then raised to 1 liter while Bio N was applied on the recommended rate of 200 grams per 4 kg seeds. All planting media were sterilized by oven-drying for six hours at a temperature of 120 ℃.

Mature seeds were sun-dried first for one day before use and subjected to germination test in sand medium to determine their viability having obtained 90% germination. Seeds were inoculated with biofertilizers before sowing them. With EM and Azotobacter , inoculation was done by dipping on the solutions for 30 minutes, air-dried for one hour, and then sown. Bio N was mixed with seeds, then moistened to allow sticking of the materials to the seeds, and allowed to stand for one hour before sowing at a depth of one inch from the soil surface. At the end of the experiment, uprooted sample plants were cleansed of soil dirt and then data on plant height, leaf count, shoot base diameter, and fresh weight per plant were taken before these were placed in properly-labeled paper packets for oven-drying. The sample plants were oven-dried in a Memmert oven for three consecutive days at a constant temperature of 80℃ and then weighed again to obtain the dry matter weight.

Data Collection. Three sample plants per pot were used in the data collection done once eight weeks from planting. Data gathered on per plant basis were plant height, which was measured from the base to the growing point, leaf count per plant, shoot base diameter, fresh matter weight, and dry matter weight.



3. RESULTS AND DISCUSSION Plant height

Results revealed a significant (P<0.05) influence of planting media x biofertilizers interactions on plant height. Tallest plants (20.22 cm) were obtained in the 1:1:1 mixture x Azotobacter interaction comparable only to plants in the humus x EM combination (19.96 cm) as shown in Table 1. This can be attributed to the availability of energy supplied by humus in the two planting media which also efficiently retain adequate soil moisture and provided good aeration that help ensure greater microbial activity, population, and diversity (Higa, T. and J.F. Parr, 1994). Among the planting media used, humus produced significantly tallest plants (19.81 cm).

Table 1 Plant Height Taken Eight Weeks after Planting

Biofertilizer Planting Media Uninoculated Azotobacter EM Bio N

------------------------------------------cm------------------------------------------------------ Garden Soil 17.65 f 17.65f 19.32c 19.83 b

Humus 19.46 c 19.90b 19.96ab 19.92 b

Sand 18.47 e 18.75d 19.81b 19.01d 1:1:1 Mixture 19.55 c 20.22a 18.81d 19.42 c

abcdef: Treatment means with the same letter superscript did not differ significantly at 5% level of probability

Leaf count No significant (P>0.05) differences only leaf counts were observed among treatment interactions of biofertilizers and planting media. Significant (P<0.05) differences, however, were noted among the planting media where garden soil and 1:1:1 mixture produced comparable mean leaf counts of 4.56 and 4.48, respectively, but were significantly different from the rest of the treatments (Table 2).

Table 2 Leaf Count per Plant Taken Eight Weeks after Planting

Biofertilizers Planting Media Uninoculated Azotobacter EM Bio N

Planting Media Mean

Garden Soil 4.78 4.58 4.25 4.62 4.56 a Humus 4.17 4.46 4.54 4.33 4.38 b

Sand 4.08 4.21 3.91 4.05 4.06 c 1:1:1 Mixture 4.37 4.56 4.69 4.33 4.48 a Biofertilizer Mean 4.35ns 4.45 4.34 4.33

*Treatment means followed by the same letter superscript are not significantly different at the 5% level of probability by DMRT. * ns: Not significant Shoot base diameter Biofertilizer x planting media interactions had no significant (P>0.05) influence on the shoot base diameter. Significant (P<).05) differences, however, were shown when planting media were compared where humus the largest mean shoot base diameter of 8.91 mm (Table 3). This can be attributed to the physical property of humus being highly porous thus has great aeration ability and water-holding capacity and being high in organic matter that provides nutrients to plants. Fresh matter weight Significantly (P<0.05) fresh matter weight was recorded from the plants grown in humus and inoculated with Azotobacter (P2B1) and in humus but inoculated with BioN (P2B3), having obtained 17.56 and 17.25 grams, respectively (Table 4). The significant fresh matter weight can be attributed to the favorable soil environment provided by humus to both the plant and the beneficial microorganisms. Enhancement of plant growth such as root development allows greater absorption of water and nutrients that eventually improve crop quality. Plants inoculated with EM and grown in humus (P2B2) recorded 15.89 grams per plant. It showed that kasla obtained greatest growth



in terms of fresh matter weight when it is grown in humus medium and inoculated with either Azotobacter and Bio N. The better aeration, excellent water-holding capacity, and the organic matter content of could have enhanced the beneficial microbial activity of Azotobacter and Bio N and consequently improved plant’s immediate environment. Table 3 Shoot Base Diameter per Plant Taken Eight Weeks after Planting


Planting Media Mean

-------------------------------mm------------------------------------ Garden Soil 7.69 6.85 7.68 7.27 7.37 d Humus 8.83 9.00 8.80 9.00 8.91 a

Sand 8.20 8.36 7.59 7.08 7.81 c 1:1:1 Mixture 8.71 8.20 7.46 8.00 8.09 b Biofertilizer Mean 8.36 ns 8.10 7.88 7.83

*abcd: Treatment mean followed by the same letter superscript are not significantly different at the 5% level of probability by DMRT. *ns: Not significant Dry matter weight. Planting media and biofertilizer interactions had no significant (P >0.05) influence on plant dry matter weight. Significant (P<0.05) results were observed only when comparing the different planting media with humus-grown plants having significantly highest dry matter weight per plant (9.73 gm). Table 4 Fresh Matter Weight per Plant Taken Eight Weeks after Planting


----------------------------------------grams----------------------------------------------------- Garden Soil 11.80d 11.66d 14.53bcd 12.81cd

Humus 15.02bcd 17.56a 15.89b 17.25 a

Sand 13.78bcd 14.39bcd 15.19bc 12.76bcd 1:1:1 Mixture 15.20bc 15.22bc 13.99bcd 14.57bcd

*abcd: Treatment means followed by the same letter superscript are not significantly different at the 5% level of probability by DMRT. Table 5 Dry matter Weight per Plant Taken Eight Weeks after Planting


Planting Media Mean

---------------------------------grams--------------------------------- Garden Soil 7.80 7.48 8.34 . 7. 99 7.90 d Humus 10.29 9.15 9.65 9.83 9.73 a Sand 9.18 9.06 10.46 9.09 9.44 b 1:1:1 Mixture 9.39 9.05 8.05 8.91 8.85 c Biofertilizer Mean 9.16ns 8.68 9.12 8.96

*abcd: Treatment means followed by the same letter superscript are not significantly different at the 5% level of probability by DMRT. 4. CONCLUSION

Plant height, leaf count, shoot base diameter, fresh matter weight (roots and shoot), and dry matter weight (root and shoot) did not differ significantly among the biofertilizers used. Plants were significantly tallest when grown in humus. Leaf count on garden soil and 1:1:1 mixture was comparable but significantly higher than in humus and



sand. Shoot base diameter was significantly largest in humus-grown plants. Both the fresh and dry matter weights per plant were significantly highest in humus-grown plants and lowest in garden soil. Plant height and fresh matter weight were significantly influenced by the interactions of biofertilizers and planting media. Azotobacter X 1:1:1 mixture interaction has produced significantly tallest plants comparable only to EM X humus interaction. Significant fresh matter weights were recorded in Azotobacter X Humus and Bio N X humus interactions. Leaf count, shoot base diameter, and dry matter weight were not significantly influenced by the interaction of biofertilizers and planting media eight weeks after planting. 5. REFERENCES Bashan, Yoav, Gina Holguin, & Luz E. de-Bashan (2004). Azospirillum-plant relationships: physiological,

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http://www.niir.org/books/book/zb

http://www.agent.org./library/articlehtml

bio fertilizers for jatropha curcas l euphorbiaceae grown in different planting media

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