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FNCA (Forum for Nuclear Cooperation in Asia) Biofertilizer Project

Dear Readers,

This 7th issue of the FNCA (Forum for Nuclear Cooperation in Asia) Biofertilizer Newsletter fea-tures the activities pertaining to research and development, industry, use and promotion and impactof Biofertilizers in Philippine agriculture. It's my pleasure to share to you information herein thatyou may find interesting and valuable in line with the same activities in your respective areas.

The need to address the food requirement of the growing population in the Philippines made thefarmers adopt the latest technology in agriculture like the use of high yielding and resistant cropvarieties. These varieties however require high amount of nutrients making the farmers dependenton agricultural input like inorganic fertilizers. It was reported that from 1966 to 1998 fertilizerapplication in the Philippines increased from 20% to 80%. Nitrogen, considered as the most limit-ing factor in crop production, gains the highest mark of all the major nutrients from inorganicsources applied by farmers. The 2001 FAO (Food and Agriculture Organization) Yearbook on fertil-izers stated that from 1987 to 1998, Philippine consumption of nitrogenous fertilizers increasedfrom 371,487 MT to 546,499 MT. Such consumption resulted to high production for some time butadverse soil and other environmental conditions were observed in the long run. This called for theuse of other alternative inputs. The use of biofertilizers has been determined as one of the mainoptions to address the rising concern on agricultural and environmental sustainability.

Biofertilizer industry in the Philippines grown considerably with the promotion of Government ofbioorganic farming and the active involvement of research institutions, private individuals andenterprises. Biofertilizers in various forms have been developed and more researches on rhizobialegume symbiosis, azolla, mychorriza, sesbania, blue green algae and the living microorganism (N-fixing bacteria) were encouraged.

My sincere thanks to the contributors who graciously contributed some highlights of their researchworks and efforts. Without their contributions, this issue would not have materialized.

Best regards,

Richard M. BalogFNCA Biofertilizer Project Leader, Philippines

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Message from the Philippines

Issue No. 7February 2007


The 2005 FNCA Biofertilizer Workshop washeld last 9-13 January 2006 at the PNRI(Philippine Nuclear Research Institute), QuezonCity, Manila jointly hosted by the DOST(Department of Science and Technology) -Philippines and MEXT (Ministry of Education,Culture, Sport, Science and Technology) -Japan. It was attended by 17 participants, pri-marily country project leaders from Indonesia,Japan, Korea, Malaysia, Philippines, Thailandand Viet Nam. Local participants includedPNRI collaborators from Biotech (NationalInstitute of Molecular Biology andBiotechnology) - UPLB (University ofPhilippines Los Banos) and DA (Department ofAgriculture) - BSWM (Bureau of Soils andWater Management). The workshop was high-lighted by the presence and messages of keypersons of the host organizations in person ofDOST Secretary Dr. Estrella Alabastro, PNRIDirector Dr. Alumanda Dela Rosa and Directorfor International Nuclear Cooperation ofMEXT Ms. Takiko Sano. The workshop activi-ties comprise of presentation of country reportsby the respective project leaders, roundtablediscussions of project accomplishments/activi-ties and plans, special lectures by invited lectur-ers and technical visits to biofertilizer facilities.

Dr. Mercedes U. Garcia of the NationalInstitute of Biotechnology-UPLB lectured onthe "Role of Bio-N in Ginintuang MasaganangAni Program for corn in the Philippines". Dr.Gina P. Nilo of DA-BSWM lectured onOrganic-Based Agriculture "Agri-Kalikasan"Program/Tipid-Abono Program in thePhilippines.

The participants visited Biotech-UPLB andBio-N Mixing Plant at Barangay Lawy, Capas,Tarlac. The participants were given overviewabout the Biotech particularly pertaining to itsdeveloped biofertilizers under the program onagriculture and forestry. The participants alsovisited the Bio-N production facility of theinstitute where they were informed about theprocesses of the Bio-N inoculant production.The facility, supervised by the inventor of Bio-N Dr. Mercedes U. Garcia and her staff JulietA. Anarna, was established through the fundsprovided by the DA in 2003 for the develop-ment and application of Bio-N on high valuecrops.

The establishment of mixing plants is one ofthe strategies in the promotion of biofertilizeruse by farmers which was a big help for sus-

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FNCA (Forum for Nuclear Cooperation in Asia) 2005 Biofertilizer Workshop

The workshop participants with Dr. Alumanda Dela Rosa(sitting 2nd from left), Dr. Estrella Alabastro (sitting 3rdfrom left) and Ms. Takiko Sano (sitting 4th from left).

Visit to the Bio-N production facilityat Biotech-UPLB. Supervising thefacility were Dr. Mercedes U.Garcia (standing 4th from right)and her staff Ms. Juliet Anarna(standing center in front). Inset:inside the facility.

The biofertilizer used in the study conductedby the PNRI under the FNCA framework wasBio-N, developed by Dr. Mercedes U. Garciaof the Biotech-UPLB. Though various forms ofbiofertilizer have been developed in the coun-try, Bio-N got the nod from the government asthe most promising technology in crop produc-tion particularly in reducing the use of chemi-cal fertilizers. The government, in line with itsprogram on sustainable agriculture allocatedfunds for the mass production of Bio-N as thiswas foreseen to greatly benefit the farmers.

Bio-N is an inoculant in solid powder formthat contains two species of the nitrogen fixingbacteria Azospirillum isolated from he roots of

a local grass talahib (Saccharum spontaneumL.) which have been screened for their effec-tiveness to variety of agricultural crops. It canfix atmospheric nitrogen and transform it intoform usable by plants or crops. Beneficialeffects of Bio-N includes enhancement ofshoot growth and root development, makesplant resistant to drought and pest attack,reduces incidence of rice tungro and corn ear-worm infestation and increases yield andmilling recovery of rice and corn. It wasobserved to provide approximately 30%-50%of the nitrogen required by crops.


tainable crop production in the locality. Themixing plant visited was operated by a farmer'scooperative St. Philip PMPC (Primary Multi-Purpose Cooperative) Inc. in collaboration withthe Biotech-UPLB. The Bio-N mixing plantwas also established in cooperation with theDA-RFU (Regional Field Unit) 3-RegionalSoils Laboratory, Tarlac LGU and PDA-Tarlac.

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Mr. Tolentino, chairman of the St. Philip PMPC talked tothe participants about the cooperative and the mixingplant. (Inset: the participants in discussion about how themixing plant operates).

Bio-N Biofertilizer

Root development of rice as affected byBio-N inoculation

Biofertilizer studies conducted by PNRI under FNCA framework


Method of Bio-N application on crops

Bio-N is generally applied as seed coating forcorn or as suspension for use in watering seedbeds or root dips for rice. The use of 5-6 pack-ets (200g per packet) of Bio-N per hectare isrecommended to inoculate rice or corn.

For corn, seeds are moistened with waterand mixed with sufficient amount of inoculantsuntil seeds are evenly coated. A sticker couldbe use to enhance adsorption of Bio-N to theseeds. Coated seeds are sown immediately andexposure of the inoculated seeds from directsunlight should be avoided.

For direct-seeded rice, germinated seeds(with embryonic roots sprouted) are coatedwith the inoculant and sown directly to the

field or on prepared beds. For transplantedrice, Bio-N is mixed with water to form slurry.Roots of seedlings are pruned into uniformlength and dipped into the slurry for at least 30minutes to an hour before transplanting.

Field experiment and yield trials

Since the development of Bio-N in 1985, fielddemonstration trials have been conducted totest its efficacy to various crops.

The Biotech, as the primary institute for thedevelopment of Bio-N conducted and super-vised several field trials and supervised farmermanaged field trials for assessment on effect ofBio-N to crops. In the study conducted on theeffect of Bio-N and inorganic fertilizer on theyield of corn (Super sweet var.) in aBio-Napplication has higher yield than application ofinorganic fertilizer and was comparable to theyield when application of inorganic fertilizerwas cut into half plus Bio-N. This furthershowed that the use of Bio-N has the leastvalue of input (in peso) per yield (kg) whichindicates that farmers can save big amount offertilizer expenses through the use of Bio-N.

The study conducted by the PNRI on theuse of nuclear techniques in the assessment ofBio-N fertilizer as seed inoculant for corn onlow fertility area in the province of Isabela(around 400km north of Manila) showed thatcorn yield was only 208 kg/ha (kilogram perhectare) when not inoculated with Bio-N andwithout fertilizer application. Corn yield wasincreased tremendously to 2,880kg/ha when

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Bio-N application on rice as root dip

Table 1. Effect of Bio-N and inorganic fertilizer on the yield of corn (Super sweet var.) in farmers field demo, dry sea-son 2001.

Bio-N application on corn as seed coating

Transfers of technology on use of Biofertilizersin line with various initiatives and efforts inpromoting organic agriculture still remains anenormous challenge in Philippine agriculture.Despite of the numerous discoveries and tech-nologies on biofertilizer, bringing it to the endusers particularly the farmers is a difficult task.Convincing farmers is costly as it entailsunlearning and getting rid of their old habits ofusing chemical fertilizers. Different means ofhas been undertaken in the Philippines in its

continuous endeavor of promoting the use ofbiofertilizers. Such means includes conduct ofexhibitions and trade fair, consultations, sym-posia, trainings, field demonstrations, and tap-ping broadcast and print media for informationdissemination.

One of the recent and remarkable efforts bythe government in promoting the use of thebiofertilizer Bio-N was the establishments of


inoculated with Bio-N plus application of Pand K fertilizer. This resulted to an increase inincome from PhP1,456.00 to PhP14,341.50.Bio-N inoculation plus application of half andfull recommended rate of N fertilizer yielded3,460kg/ha and 2,960kg/ha, resulting inincrease in income of PhP16,230 andPhP10,558 respectively. The experiment alsoused N-15 to quantify biological nitrogen fix-ing capacity of Bio-N. N-15 data indicated that19-24% of the nitrogen taken by the plants isnitrogen from the atmosphere that is fixed bythe Bio-N. (Please also see related article byBiotech on biofertilizers)

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A pose with the farmers during"Farmers Field Day" inIsabela province. (Inset:explaining to farmers about theexperiment.)

Field experiment on corn (with yield levels) in low fertilityarea at Isabela province.

Promotion of biofertilizer use by farmers

One of the Bo-N mixing plantsestablished in province ofCotabato. (inset:: Giving techni-cal assistance to the recipient)

Biofertilizer primarily contains microbial inoc-ulants usually proliferated in a nutrient sourcemedium called as carriers. In the production ofbiofertilizers, carriers undergo a process ofsterilization to create a favorable environmentfor the growth of desired inoculants free fromantagonistic microbes. Conventional method ofsterilization of carriers for biofertilizers is donethrough the use heat-autoclave. This steriliza-

tion process poses some disadvantages such asincomplete sterilization, may result to somechanges in physical and chemical properties ofcarriers, costly and suitable only to small scaletreatments. Alternative means of sterilizationprocess has been found in the utilization of thekilling effect of gamma rays to microorganism.This process has been undergone in thePhilippines in line with its activities inimprovement of the quality of biofertilizers.The PNRI operates a multi purpose Co-60gamma irradiation facility, which is the onlyone in the country. The application of gammairradiation for sterilization of biofertilizer carri-er however is still in its experimental stage.Bio-N for instance, whose carrier is composeof soil and charcoal, gamma irradiation showeddecrease of microbial load at increasing doseand is sterilized at the dose 30kGy. Differentbiofertilizers however have different carriers sofurther experimentation is needed and suffi-cient data should be acquired especially on theadvantages of the gamma irradiation comparedto conventional methods to convince thebiofertilizer producers to adopt the applicationof gamma irradiation for sterilization of biofer-tilizer carriers.


mixing plants. The establishments of mixingplants is one of the projects being undertakenby the DA Ginintuang Masaganang Ani CornProgram, in collaboration with the Biotech-UPLB, LGU's (Local Government Units) andfarmers organization/cooperatives in respond tothe farmers increasing demand of the Bio-N inlate 1996. The mixing plants are consideredsatellite production and sales centers and at the

same time served as distribution channels thatmade Bio-N accessible to the local consumersespecially the farmers who were sourcing itfrom UPLB. There have been 32 mixing plantsstrategically established throughout the countrythat produces and sold Bio-N at price afford-able to farmers. (Please also see related articleby Biotech on promotion and technology trans-fer of Biotech biofertilizers.)

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Co-60 irradiation facility atPNRI (inset: the gammasource)

Application of gamma irradiation for sterilization of biofertilizer carriers


Rationale:

Marginal soils occupy a large area in thePhilippines. The acid upland soil is estimatedto be 17 million hectares or roughly equivalentto 58 % of the total land area of the country(Atienza, 1991). These soils are generally char-acterized as highly weathered soils with lowproductivity (IBSRAM, 1987).

Despite the widespread belief that acidupland soils cannot support intensive and sus-tained agriculture, there are ample evidencethat they can be continuously cultivated andintensely managed for growing annual andperennial crops. To sustain crop production inthese soils, inclusion of legumes in the produc-tion systems had been advocated. Theselegumes can serve as nitrogen sources and theiruse by farmers can reduce the need for nitrogenfertilizers. However, growth and nitrogen fixa-tion of legumes in acid soils are constrained bya number of soil factors which include toxici-ties of Al, Mn and other metal ions (Zn, Cu,Cd), low soil pH, deficiencies of Ca, Mg, P,Mo and other elements including N and K. Tosuccessfully grow legumes in acid soilsrequires soil management strategies such aschoosing legume species that show tolerance tosoil acidity, liming and addition of fertilizersand organic matter.

This study aims to determine the effects oflime, phosphorus and micronutrient applica-tions on the growth, nitrogen fixation and yield

of legumes and to determine the increase inproductivity with the application of theseinputs.

Methodology:

The experiment was established in Kapatalan,Siniloan, Laguna, around 106 kilometers southof Manila. The soil in the experimental areawas taxonomically classified as FineIsohypothermic, Typic, Kanhapludult (Evangelista, pers.com) with a pH of 4.4 and12.6 ppm available P. Peanut (cv. UPL-Pn2)and mungbean (cv. MG-50 L Yellow) wereused as test crops.

The study had the following treatments:a. Lime application

- with lime (5 tons CaCO3/ha)- without lime

b. Micronutrient application- with micronutrient (50 kg fused trace

elements/ha)- without micronutrient

c. Phosphorus application (kg/ha)- 0,40,80,120,160

A split-plot design was used with liming asthe main plot, micronutrient application as thesub-plot and phosphorus levels as the sub-sub-plot. The treatments were replicated threetimes. Nitrogen and potassium fertilizers wereapplied at the rate of 20 kg N/ha and 30 kgK20 /ha. Rhizobium inoculants for peanut andmungbean were inoculated onto the seed priorto planting.

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ENHANCEMENT OF BIOLOGICAL NITROGEN FIXATION IN LEGUMESTO INCREASE PRODUCTIVITY OF ACID UPLAND SOILS

M. J. Palis, A. O.Yambot and J. S. Rojales

Researches on BIOFERTILIZERS in the Bureau of Soils & Water Management,Deapartment of Agriculture, Dilliman, Quezon City, Phillippines





The plants were grown to maturity. At nearblooming stage (7 weeks after planting), 10representative plants from the sampling areawere obtained for determination of dry matteryield, acetylene-reduction activity and noduleweight. The general view of the experiment atmaximum vegetative stage is shown in Figure1. Preparation of samples for acetylene-reduc-tion assay is shown in Figure 2.

Harvesting of the plants was done at physio-logical maturity. All pods from the plants in theharvest area were obtained. The pods were sun-dried, shelled and the weight of seeds wastaken.

A simple cost and return analysis was alsodone after harvest to determine whether grow-ing legumes in acid soils is profitable for themarginal farmers.

Salient Findings:

1. The application of lime at the rate of 5 tonsCaCO3 /ha gave a marked increase on thedry matter, nitrogenase activity, noduleweight, and pod and seed weight of bothpeanut and mungbean as shown in Figures 3-6.

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Figure 1. General view of the experiment.

Figure 2. Researchers preparing samples for acetylene-reduction assay.

Figures 3 and 4. Effect of liming on the dry matter andacetylene-reduction activity of peanut and mungbeangrown in an Ultisol.


2. Phosphorus application likewise increasedthe growth, nitrogen fixation and yield ofpeanut, but the increase was not as pro-nounced as when compared to the effect oflime (Figures 7 and 8).

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Figures 7 and 8. Effect of rate of phosphorus fertilizationon the growth, nitrogen fixation and yield of peanut andmungbeans grown in an Inceptisol.

Figures 5 and 6. Effect of liming on the nodule weightand pod and seed yield of peanut and mungbean grownin an Ultisol.

3. Based on simple cost and return analysis,growing of mungbean in acid soils could notbe recommended to farmers. A net incomeof P21,563.00/ha (USD 431.26) can beobtained with planting of peanut in acid soilswith the application of 5 tons lime, micronu-trient, and 40 kg of P2O5 per hectare.

4. Peanut seemed to be more tolerant to soilacidity when compared to mungbean.

<References>1. Atienza, R.N. 1991. Research Status of Acid

Soil Management and Utilization in thePhilippines. In. Management and Utilizationof Acid Soils. PCARRD. Los Banos,Laguna, Philippines.

2. IBSRAM (International Board for SoilResearch and Management). Management ofacid tropical soils for sustainable agricul-ture.Bangkok.

3. Evangelista, P.P. . Bureau of Soils & WaterManagement, Diliman, Quezon City,Philippines (pers.com.)


Introduction:

A commonly used technique to improvelegume growth and seed yield is inoculationwith useful Rhizobium strain with a highpotential for nitrogen-fixation. These strainshowever often fail to nodulate because they areunable to overcome environmental stresses.The inoculant strains also have to compete withthe indigenous rhizobia which are often unableto fix N in symbiosis. In order to competeeffectively with indigenous strains, inoculahave to be applied in a titer several times high-er than the native rhizobia in the soils.

Testing the competitive ability of inoculastrain is very important to enhance the effect ofinoculation. There are no easy and accuratemethods for determining competitiveness ofrhizobia in soils and identifying the strainsresponsible for nodulation. However, with thedevelopment of a technique using the resist-ance of rhizobial strains to antibiotics, strainidentification can be carried out without toomuch difficulty. The method therefore wasadopted in this study to select effective andcompetitive strains of rhizobia for mungbeanin marginal soils.

Methodology:

a. Development of antibiotic resistant strains ofrhizobia for mungbean

To investigate the successful establishmentand survival of applied rhizobia in the field, theantibiotic resistance marker was employed.

This method is very sensitive and reliable indetecting and quantifying the inoculated strainsin the nodules of plants.

The intrinsic antibiotic resistance of fourrhizobial strain (MCO, CY 44, TAL 441, TAL209) were developed by exposure of the rhizo-bial strains to different concentrations of theantibiotics like streptomycin, kanamycin,erythromycin and spectinomycin. Retention orloss of the symbiotic effectiveness of the four(4) antibiotic-resistant rhizobial strains wasassayed by using the plant-infection test.

b. Field Experiments (Tanay National Soil andWater Resources Research Center)

Two subsequent field trials were done inTanay which was previously classified as anUltisol. The treatments in the experiments werethe following:

1. Liming:- no lime- with lime (3 tons dolomite)

2. Rhizobium strains- no inoculation- MCO- CY 44- TAL 209- TAL 441- Mixture of the strains

The treatments were replicated 3 times andwere arranged following the RandomizedComplete Block Design (RCBD). In bothexperiments, the mungbean cultivar planted

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SELECTION OF COMPETITIVE STRAINS OF RHIZOBIAFOR MUNGBEANS IN MARGINAL SOILS

M. J. Palis, A. O.Yambot, J. S. Rojales, V. F. Naboa, S. Arai,H. Ito and M. Nakamura


was PSB Mg-6. Fertilization was based on soilanalysis. (29-60-60 kg NPK/ha). At full bloom-ing stage, 10 representative plants from thesampling area in each plot were taken for dryweight determination and nodule occupancyanalysis. The plants were grown to maturity fordetermination of yield and yield components.The general view of the experiment is shown inFigure 1.

Salient Findings:

1. All the strains used in the study developedtheir resistance to antibiotics after three (3)sub-culturing. Result of the confirmatorytrial to test retention or loss of infectivenessof the antibiotic resistant strains showed thatall the strains retained their infectiveness ontheir host as shown by the presence of nod-ules in plants inoculated with the four strains(Fig. 2).

2. For single strain inoculation, strain MCO

was the most competitive among the strainstested during the first field trial (82 % nod-ule occupancy in unlimed soil and 100 % inlimed soil) as shown in Table 1. The otherstrains were not as competitive as strainMCO. However, during the second fieldtrial, recovery of the strains from the noduleswas lower. Strain MCO still occupied mostof the nodules of plants in soils not amendedwith lime (75.3 %) while strain TAL 209occupied 65.4 % of the nodules of plants insoils applied with lime.

In mixed inoculation, the most competitivewere strains MCO and CY 44, but the indige-nous rhizobia occupied most of the nodulesespecially during the 2nd field trial (Table 2).This phenomenon showed the very formidableadvantage of the indigenous rhizobia in form-ing nodules. The introduced rhizobia were notable to completely overcome the resident rhi-

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Fig. 1. General view of the experiment during the firstfield trial.

Fig.2. Plant-infection test showing infectiveness ofantibiotic-resistant strains for mungbean as indicated bythe presence of nodules on the test crop.

Table 1. Percent nodule occupancy of Rhizobium strains in nodules of mungbean plants grown in limed and unlimedsoil in Tanay (Ultisol) during the 1st and 2nd field trials.


zobia which formed a bulk of the total nodulesanalyzed.3. Response of mungbeans to inoculation of the

different strains showed that on the average,a slight increase on the seed yield of theplant was observed with inoculation. Limingseemed to have a negative effect on the yieldof the plants during the first field trial, how-ever, during the second trial, the applicationof lime increased the seed yield irrespectiveof inoculation treatment (Figure 3.)

<References>1. Somesagaran, P. H. and H. J. Hoben. 1994.

Handbook for Rhizobia. Springer-Vorlag.New York. 450 pp.

2. Trinick M. J. 1990. The ecology ofRhizobium. Interactions between Rhizobiumand other soil organisms. Plant and Soil.108: 137-141.

3. Triplett, E. W. 1992. Genetics of competitionfor nodulation of legumes. Annual Reviewof Microbiology. 46: 399-428.

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Table 2. Competitiveness of Rhizobium strains in mixed inoculation.

Fig. 3. Inoculation and liming effects on the seed yield of mungbean grown in an Ultisol.


The Philsurin (Philippine Sugar ResearchInstitute Foundation, Inc.), a non-profit organi-zation created in 1995 has developed biofertil-izer for sugarcane in line with its functions toundertake sugarcane research, development,and extension. Sugarcane production requireshigh fertilizer inputs, thus development ofbiofertilizer for sugarcane entails reduction ofproduction cost. Sugarcane was found to havethe capability of obtaining nitrogen from air forits development through the association estab-lished with nitrogen fixing bacteria called asdiazotrophic endophytes.

Philsurin microbiologists, Dr. Lucille C.Villegas has conducted a research to developmicrobial inoculant for sugarcane by findingsuperior strains of diazotrophic endophytes todevelop viable nitrogen fixing in associationwith sugarcane. Promising isolates have beenselected and identified and the selected strainscan be successfully introduced to sugarcane invitro. Philsurin reported promising results.Inoculation on micro propagated sugarcaneindicates that four isolates positively affected

the tillering capacity of the plants. There wasno significant difference on the number oftillers between the inoculated plants grown atsuboptimal level of nitrogen and the plantsgrown at optimum level of nitrogen indicatingthat bacterial inoculation can replace needednitrogen to meet the optimum level of growth.

Pot experiment was also conducted to evalu-ate the effects of inoculation at different devel-opmental stage of sugarcane. Seven bacterialisolates were inoculated on four sugarcanevarieties (VMC 86-550, VMC 87-599, PS3,and Phil 80-13). Agronomic parameters of sug-arcane were evaluated as affected by the inocu-lation of different bacterial isolates. No resulthowever was reported for this aspect.

<Source>Dr. Lucille C. Villegas, Ph.D. Development

of Microbial Inoculant for Sugarcane. PhilsurinAnnual Report 2002 - 2003. Pg 39-41.

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Philsurin developed Biofertilizer for sugarcane

Diazotrophic endophytes on different mediaA) thick pellicle in semisolid N free medium, B) orangecolonies on N free agar, C) chocolate brown growth onglucose yeast extract CACO3 agar D) greenish browngrown growth on potato agar.

Inoculated plants growing under natural conditions


Over at the National Institute of MolecularBiology and Biotechnology (BIOTECH) in theUniversity of the Philippines Los Banos, themood is upbeat as scientists work their way inproducing agricultural biotechnology break-throughs that would combat global hunger -biofertilizers which they hope would solve thenever-ending problem on food security.

Biotechnology, a set of scientific tools thatmakes use of living organisms, such asmicroorganisms to generate products and serv-ices, has been doing wonders to agriculture andfood sector. At BIOTECH, the first wave ofproducts paid off in the form of environment-friendly yet affordable microbial-based biofer-tilizers and inoculants that can substantiallydouble the yield of rice, corn and other impor-tant crops and substitute for chemical-basedfarm inputs. These products, generated by theBiotechnology for Agriculture and ForestryProgram, underwent a series of trials - fromgreenhouse to larger field testing - to determineif they are ready for commercialization and fortransfer to private entrepreneurs. These areclassified as: biological nitrogen fixers, myc-

orrhiza, plant growth promoters and microor-ganisms that can degrade organic wastes.

Biological nitrogen fixers

The use of microorganisms to harness elemen-tal nitrogen through nitrogen fixation is one ofthe promising alternative technologies toaddress the high cost of chemical nitrogen fer-tilizers and the increasing environmental prob-lems attributed to their use. These microbes,known as biological nitrogen fixers (BNF), candirectly fix nitrogen gas from the atmosphereto make it usable by the plants.

An example of this microorganism is therhizobia, which reside inside the nodules ofleguminous plants. The mixture of rhizobia,sterile soil and charcoal led to the developmentof an inoculant known as NitroPlus. It is abiofertilizer specific for soybean, mungbean,cowpea, peanut, garden pea, pole sitao andother food legumes. NitroPlus comes in pow-der form in 100-gram packet which is used tocoat seeds before sowing. One hectare requires4-5 packets which is a lot cheaper than 2-3

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BIOTECH Fertilizers:the approach to improve crop productivity and quality


bags of ammonium sulfate. Field trials done incollaboration with various agencies showed anincrease in soybean yield by 124%, mungbeanby 29% and peanut by 39%. It was also foundto replace 30-50% of the nitrogen requirementsof plants and has a fertilizing value equal to 30kg N/ha. Presently, NitroPlus is being tested onnon-legumes.

BIOTECH is also successful in producingBIO-N , a biofertilizer that containsAzospirillum, a bacterium isolated from thegrass Saccharum spontaneum or talahib. BIO-N enhances root development, growth andyield of plants (rice, corn and other agriculturalcrops) and provides 30-50% of their nitrogenrequirements. Efficacy testing of Bio-N on ricevariety RC 18 showed improvement in theaverage number of tillers, straw and grainyields. Five packs of Bio-N are needed to inoc-ulate a hectare of rice or corn. It was alsofound to increase yield of vegetables (toma-toes, okra, eggplant, lettuce and ampalaya).BIO-N is available in powder form in 200-gpack, which is meant either for seed inocula-tion, direct broadcasting over seeds or mixedwith water as root dip.

Plant Growth Promoters

Some microorganisms produce plant growthregulators such as auxins, gibberilins andcytokinins which can be used in plant propaga-tion.

BioGroe is a biological fertilizer contain-ing plant growth-promoting bacteria (PGPB)that influence root growth by producing planthormones and provide nutrients in solubleform. It is packaged as a powdered inoculantthat can be used for almost all types of plants -vegetables, ornamentals, flowering plants andherbs. Experiments done at BIOTECH showedthat it increased ampalaya yield by 90% andlettuce by 40%. Dendrobium, on the otherhand, had early flowering while Mussaendaand Hibiscus (gumamela) produced heavier

and longer roots. It was also found effective inthe rooting of blackpepper cuttings and hard-to-root ornamentals Eugenia and Bushida.BioGroe also promoted leaf bud formation andplant survival of coffee stem cuttings andenhanced seed germination of the herbs marjo-ram and thyme. In the efficacy test conductedin Tarlac and Cavite, BioGroe was found toenhance seedling growth and lowered farmers'production cost.

Cocogro is a mixture of plant growth hor-mones derived from coconut water. This natu-ral product can be used as a fertilizer supple-ment to enhance and promote growth and yieldof plants, especially tissue cultured orchids andorchid seedlings. Researchers have proven thatorchid seedlings, when sprayed with Cocogro,increased in growth by 100% within a month.Field trials showed that Cocogro can increaseyield of beans by 64%, peanuts by 15%, sweetpepper by 100% and Chinese cabbage and by20-30%. Cocogro is also good for ornamentals,cereal crops, fruit trees and flowering plants. Itincreases root proliferation, shoot develop-ment, bud formation and early flowering.

Mycorrhiza

Certain microorganisms also provide phospho-rus and other nutrients. Mycorrhiza is a symbi-otic association between the roots of plants anda fungus. The association provides many bene-fits to plants such as increased absorption ofnutrients and water. It likewise serves as a bio-logical control agent against root infection andcan improve soil properties. BIOTECH hasproduced four mycorrhizal products:Mycogroe, Mykovam, VAM Root Inoculantand BrownMagic.

Mycogroe is a biofertilizer in tablet formcontaining spores of ectomycorrhizal fungi.The fungi, when inoculated to reforestationseedlings, will infect the roots and help absorbwater and nutrients particularly phosphorus.They can also prevent root infection by

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pathogens and increase plant tolerance todrought and heavy metals. Mycogroe wasfound effective in promoting the survival andgrowth of trees and can replace 60-85% oftheir fertilizer requirement. Application is doneonly once throughout the entire life of the tree.Mycogroe is applicable only for trees such aspines, dipterocarps, Eucalyptus, Alnus, Acaciaand Casuarina. One tablet is inoculated intoeach seedling during transplanting in the nurs-ery.

On the other hand, Mykovam is a pow-dered biofertilizer which has effective speciesof vesicular arbuscular mycorrhizal (VAM)fungi. Basically, its functions are similar tothat of Mycogroe. The fungi infect the roots,form a close association with the plant andassist in absorbing water and nutrients such asphosphorus, zinc and nitrogen. Mykovam canbe applied to almost all plants: agriculturalcrops (upland rice, corn, tomato, eggplant,onion, garlic, pepper, cassava, and sweet pota-to), fruit trees (guava, rambutan, papaya, citrus,banana, lanzones, coffee, guyabano, coconutand mango), ornamentals and reforestationspecies (acacia, gmelina and mahogany).Mykovam even fights destructive pests and dis-eases and increases plants tolerance to droughtand heavy metals. Use of this product reduces60 -85% of the fertilizer requirements of mostplants. It is economical to use since a kilo ofMykovam can fertilize 400 seedlings.

Brown Magic consists of finely choppedand powdered fruiting bodies of endomycor-rhizal fungi. It is especially effective for in-vitro cultured orchids and provides the nutri-ents necessary for their growth. It also servesas a biological control agent of root infectionand other harmful pests and diseases.Experiments done at BIOTECH showedhealthier, greener and more vigorousDendrobium, Vanda and Cattleya seedlings.Plants had more suckers and spikes and pro-duced flowers earlier and at a longer period oftime.

VAM Root Inoculant is composed ofdried chopped roots infected with VAM fungi.These fungi assist plant roots in absorbingwater and nutrients and thus reduces chemicalfertilizer requirement of crops. VAM RootInoculant also acts as biocontrol agent againstroot pathogens and produces certain substancesthat promote plant growth and improve soilstructure. Experiments done at BIOTECHshowed improved growth and fruiting capacityof papaya, increased yield of peanut andenhanced phosphorus uptake of bitter gourd. Itis also an effective growth promoter of agricul-tural (banana, sugarcane, corn, root crops) andhorticultural crops, vegetables (eggplant, okra,tomatoes, onion), fruit trees and ornamentals.

Microorganisms that decompose agricultural residues

BIOTECH has produced an organic fertilizertechnology through composting which can sig-nificantly enhance crop growth and yield.Chicken, swine or cattle manure can be com-bined with farm wastes such as rice straw, cof-fee hulls and corn stover or with agro-industrialwastes such as sugarcane bagasse and mud-press and processed into bio-organic fertilizer.The technology also involves the use of twoinoculants: Bio-Quick which containsTrichoderma sp. that can degrade organicwaste materials in only 3 to 4 weeks instead ofalmost 6 weeks, and Bio-Fix, the enrichmentinocula containing Azotobacter sp. which isadded to the compost heap to increase thenitrogen content of the final product. The end-product called Bio-Green can be applied to allkinds of plants and can increase yield by 20-30%. It also serves as soil conditioner,enhances the nutrient uptake of plants in soiland thus makes plant healthier. Bio-Greencan be applied basally or directly depending onthe crop. If applied at 10 to 20 bags/ha, it canequal the effects of chemical fertilizer at 60-30-30.

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Promotion and Technology Transfer of BIOTECH Biofertilizers

Research does not end with the discovery of atechnology or process. More important is theutilization of the technology or process byintended clients. Thus, at BIOTECH,researchers are not just keen on producing highquality fertilizers but also in reaching the farm-ers, even those in marginal areas of the country.They go beyond the confines of laboratories togenerate and sustain a high level of awarenessand acceptance of biofertilizers. They do thesein tandem with the extension and communica-tion specialists of the Institute. One promotion-al approach used is the production of an inte-grated information package through print andmultimedia materials. Biofertilizer fact sheetsor leaflets are given to farmers and extensionworkers as supplementary materials duringseminars, forums or trainings, whereresearchers serve as resource speakers. Anotherapproach is to facilitate institutional collabora-tion and establish links with different agencies,including local government units, in the trans-fer or commercialization of BIOTECH biofer-tilizers. They also collaborate with setting-upfield demonstration areas to see the efficacy ofour biofertilizers.

Case in point: to create awareness andencourage extensive support and patronage offarmers, the Bio-N staff work hand in handwith the Department of Agriculture in imple-menting the transfer of the technology in threedifferent phases. Training workshops on Bio-Napplication were held in different regions of thecountry complemented by presentations,posters, handouts and product samples foractual testing. Bio-N mixing plants were like-wise established to address the problem on theunavailability of supply and/or accessibility todistribution centers. BIOTECH also partnerswith the Technology and Livelihood ResourceCenter in marketing Bio-N.

However, Bio-N is just one of the biofertil-izers which is being aggressively promoted bythe Institute. Media exposure for NitroPlus,Mykovam, Mycogroe, BrownMagic andBioGroe were also sought to intensify ourinformation campaign on biofertilizers. As aresult, researchers were interviewed in variousradio and television programs while featurearticles of their technologies were published innewspapers, magazines and newsletters.Demonstration trials done both at BIOTECHand in farmers' field validated the efficacy ofthe biofertilizers, much to the satisfaction offarmers.

To facilitate the efficient technology transferfrom the laboratory to target industries and/orbeneficiaries, BIOTECH prepares pre-feasibili-ty studies, industry analyses and businessplans. Applications for experimental use per-mit, product registration, technology licensing,joint venture are likewise done. Bio-N for riceand NitroPlus for soybeans are now registeredwith the Fertilizer and Pesticide Authority, as aresult of our techno-demo farms utilizing theidle lands around BIOTECH.

Judging from the increasing demand fromfarmers and plantation owners, BIOTECH isindeed fulfilling its mission of developing fer-tilizers that do more than just provide morebountiful yield. The plethora of interest gener-ated by our technologies inspires ourresearchers to continue the research, develop-ment and extension programs they have started.Concomitantly, they hope to further widen thepartnership with different public and privateagencies, biotech firms in particular, to facili-tate the commercialization process ofBIOTECH biofertilizers to make them readilyavailable to farmers in different regions of thePhilippines and in other countries as well.(Imelda V. Garcia , Program Leader,Communication and Technology UtilizationProgram of Biotech)

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A relatively youngbut revolutionarybiotech productsuch as VITAL N R

is now getting a lotof attention. Thedemand for cost-effective and inno-vative VITAL N R biofertilizer is rapidly grow-ing. In fact, VITAL N R now has a fast-growinglist of distributors/dealers all over the country.

VITAL N R is a wettable powder formulationthat is available in a very convenient 100-grampack. This small package packs a lot. It actsas a virtual nitrogen fertilizer factory at theroots of the plants. It protects plants from rootand leaf diseases. It also makes plantsdrought-resistant. Not only is it convenient andpowerful, it is also easy to store. It can bestored at room temperature for at least 3 years.

VITAL N R is a breakthrough and sustainableagriculture technology for increasing farmers'incomes and food sufficiency. Furthermore, itis proudly 100% Filipino! The product is theresult of my years of research together with myscientist wife, Dr. Saturnina C. Halos.Together with our children and in-laws, weformed a progressive biotech company,Arnichem Corporation, to make the productavailable to the average Filipino farmer. Withthis product, we hope to re-VITALize thenation.

VITAL N R is an FPA-registered organicbiofertilizer that replaces as much as half of therecommended fertilizer. This translates to asavings of 4 bags of fertilizer per hectare (ha)or about P3,000 for rice and corn and 6-8 bagsof fertilizer/ha (about P5-6,000/ha savings) forvegetables. It also increases farmer's profitabil-

ity by at least 15%, and in some cases morethan 100%. Tests conducted by independentscientists and farmers in rice and corn showthat, VITAL N R increases yields by at least oneton per hectare. In onion, additional yields of15 tons per hectare can be realized by using theproduct. With its efficacy and the low cost ofthe product, it is well suited for the majority ofsmall farmers who can not afford the full costof recommended fertilizer requirement andwho badly need a harvest booster.

The following tables (Tables 1-3) were partof the technical data obtained by Dr. Nora

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VITAL N R : A Revolutionary Biofertilizer Technology with a Fast-growing Market

Dr. Ponciano M. HalosChairman of the Board / Arnichem Corporation

Pleasant Village College, Laguna 4031 Philippineswww.vitaln.com

Table 1. The effect of VITAL N R inoculation inOPV corn (white corn) yields

Table 2. The effect of VITAL N R on yellow hybrid corn (DK818Y) under late drought conditions.


Inciong, Consultant, Bureau of Soils and WaterManagement, Dept. of Agriculture submitted tothe Fertilizer and Pesticide Authority in supportof the registration of VITAL N R as a biofertiliz-er. (RIF-Recommended Inorganic Fertilizationrate).

The following are results obtained by under-graduate thesis students of Dr. Betty Malab,Professor of Soil Science, College ofAgriculture, Mariano Marcos University,Batac, Ilocos Norte. The results show that 1/2RIF plus VITAL NR give similar yields asthose of RIF.

Farmer scientists participating in the pro-gram of Dr. Romulo Davide in Bantayan, Cebuobtained more than 100% increase in yieldswhen they applied VITAL N R (3.86 MT/ha)compared to their practice of applying nothing(1.76 MT/ha). This yield was comparable towhat they obtained when they applied com-plete fertilizer (14-14-14) also at 3.86MT/ha.In Toledo City, using IPB 929, an increase of62.5% of corn treated with VITAL N R (3.77MT/ha) was obtained over that without anytreatment (2.32MT/ha).

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Table 3. The effect of VITAL N R on rice yields

rice

Corn

Kinchay

Table 4. The yield of pepper (Capsicum anuum L) as affected by varying levels of fertilizer and VITAL N R

without VITAL N R

with VITAL N R


Project leader of The Philippines

Richard M. BalogScience Research Specialist ISoils UnitAgricultural Research GroupAtomic Research DivisionPhilippine Nuclear Research Institute (PNRI)Commonwealth Ave., Diliman, Quezon City, 1100 PhilippinesTel. No.: (+632) 929-6011 to 19 local 237Fax No.: (+632) 920-8765E.Mail: [email protected]

Research Interest□　Use of nuclear techniques in agricultural research

●　Use of 15N isotope dilution technique in fertilizer use efficiency and biological nitrogenfixation studies

●　Use of fallout radionuclide 137Cs in soil erosion and sedimentation studies

Positions held□　Project Leader, Forum for Nuclear Cooperation in Asia Biofertilizer Project □　Project Leader, Department of Agriculture-Bureau of Agricultural Research funded project on

the Use of Cs-137 in Estimating Soil Erosion and Sedimentation□　Member of the National Project Team for International Atomic Energy Agency RAS/5/043

project on Sustainable Land Use Management and Strategies for Controlling Soil Erosion andImproving Soil and Water Quality

Check FNCA Web Site!http://www.fnca.jp/english/

FNCA Biofertilizer News Letter No. 7Editor: Mr. Richard M. Balog

Leader of FNCA Biofertilizer Project of the PhilippinesSecretariat: Ms. Yuko Wada

Asia Cooperation Center (ACC)Japan Atomic Industrial Forum, Inc. (JAIF)Tel:+81-3-6812-7104 Fax:+81-3-6812-7110 E-mail: [email protected]

* This Biofertilizer Newsletter is issued by Japan Atomic Industrial Forum, Inc. (JAIF) under the contract of the Ministry of Education, Culture, Sport, Science and Technology (MEXT)

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