Biofertilizer (VITAL N) Techno Demo in Selected Rice Areas

in Western Visayas, Wet Season, 2009

TERMINAL REPORT

Abstract

Improved nutrient management is one of the strategies being adopted by the Philippines’
Department of Agriculture in trying to meet its goal of self-sufficiency in rice. As part of this program, the
DA Regional Field Unit No. 6 conducted a study to verify the effects of micronutrient fertilizer
supplementation, as well as the use of biological nitrogen fixation.
This study was conducted during the 2009 wet season in twelve (12) different sites across
Western Visayas, with two (2) sites per province – one each for irrigated and rainfed condition. There
were four (4) treatments, namely –T1: Full Recommended Inorganic Fertilization (RIF) Rate for NPK, T2:
microbial inoculant fertilizer (VITAL N) plus ½ RIF for N and full RIF for P and K, T3: micronutrient fertilizer
plus full RIF, and T4: microbial inoculant fertilizer (VITAL N) and micronutrient fertilizer plus ½ RIF for NPK.
The nutrient levels were checked using the Minus One Element Technique (MOET) although it
was implemented in only seven (7) sites due to tight schedules as a result of late crop establishment. The
MOET revealed widespread deficiency in Nitrogen and Sulfur and deficiencies of Phosphorous and Zinc in
some areas in Western Visayas. Application of Nitrogen fertilizer, on the other hand, was calibrated based
on Leaf Color Chart (LCC) readings.
The rice harvest data was tabulated and the results the hypothesis that all the treatments was
tested using ANOVA at a level of significance of 0.05. Among the different treatments, Treatment 1 had
the lowest yield, while Treatment 4 had the highest yield under both rainfed and irrigated conditions.
Treatments T2 and T3 had similar yields, although they were both higher than T1. The difference in the
yields were found to be statistically significant.
Based on the result of this study, microbial inoculant fertilizer (VITAL N) supplementation has
been found to be effective should be considered as part of the nutrient management program.

Rationale

The Ginintuang Masaganang Ani Rice Program of the Department of Agriculture is currently
implementing various projects geared towards increased rice production to attain rice sufficiency by year
2013. One strategy is to improve the nutrient management practices of rice farmers. Nutrient
management is one of the most important component in the “Palay Check System” being implemented
in all rice production areas in the Region and nationwide. The higher the production target of a farmer
means higher fertilizer requirement but to an optimum level only. However, there were some reports in
the field that no matter how much they add Nitrogen, Phosphorous and Potassium from any source did
not increase their yield. Intensive cropping has already depleted much of the inherent soil nutrient
especially the micronutrients which are usually neglected by the farmers. There is also a need to
validate the use of biological nitrogen fixing bacteria as it has been used by some farmers and need to
be validated for rice. Moreover, climatic change has indirectly if not directly affected the supply of these
micronutrients necessary for rice growth and development.
Objectives

1. To demonstrate the effectiveness of using biological nitrogen fixing bacteria for irrigated and
rainfed rice culture.

Methodology

Table 1. Demo trial sites and dates of establishment.

Locations and Dates of establishment

The demonstration trials were conducted in all provinces of Region 6 with 2 (two) sites per province, 1 in
irrigated and 1 in rainfed (total of 12 sites) beginning May 2009 to October, 2009. Table 1 shows the
sites of the techno demos. The demo sites were identified by the concerned Provincial and Municipal
Agriculture Offices.

Province Municipality Barangay

Province Municipality Irrigated Date estab. Rainfed Date Estab.

Aklan Lezo Carugdog May 23, Sta. Cruz June 8,

2009 2009

Capiz Ivisan Pob. Sur May 28, Agustin Navarra May 28,
2009 2009

Murcia and Murcia (Bgy

June 21, May 27,
Negros Occ. Pulupandan Damsite) Pulupandan
2009 2009
Iloilo Dingle Agtatacay July 15, Matangharon May 28,
2009 2009

Guimaras San Lorenzo and Cabano, San May 26, Nazareth,Buenavista May 26,
Buenavista Lorenzo 2009 2009

Antique Sibalom Egana June 13, Bontol June 4,

2009 2009
The treatments were as follows:

Treatment 1 - fertilized with the locally recommended inorganic NPK fertilizers

Treatment 2 - Seeds or roots treated with Vital N inoculants plus half of the recommended
Inorganic nitrogen and full of the recommended phosphorous and potassium fertilizers

Treatment 3 – Micronutrient fertilizer applied at basal plus full recommended inorganic NPK fertilizer

Treatment 4 - Seeds or roots treated with Vital N inoculants plus basal application of
micronutrient fertilizer plus half of the recommended NPK inorganic fertilizers

Each site had a total area of 5000 sqm. The area was divided into four equal plot sizes.
Treatment 1 was applied with the recommended inorganic NPK fertilizer only. In Treatment 2, seeds or
roots were treated with biofertilizer (VITAL N) and fertilized with the full recommendation of basal
Phosphorous and Potassium fertilizers. Only ½ of the recommended Nitrogen was applied in VITAL N
treated plots. Treatment 3 was applied with micronutrient fertilizer in addition to the recommended NPK
rates in the area. Treatment 4 was applied with Micronutrient fertilizer at basal and treated with VITAL N
plus ½ of the recommended inorganic NPK. All other cultural management practices followed the Palay
Check System.

Method of Establishment and Micronutrient Fertilizer Application

The method of rice establishment was either transplanted or direct seeded. Certified seeds of a
recommended variety in the area were used. In Treatments 3 and 4,micronutrient fertilizer was applied
at the rate of 8 kg/ha at basal. Two (2) kg of Micronutrient fertilizer (good for 2 plots- @1,250 sq m/plot)
per site was mixed with 1 kg of Urea. The recommended rate of Nitrogen (Urea) fertilizer was based on
LCC readings beginning at 14 days after seeding (DAS) for direct seeding and 21 days after
transplanting. In Treatments 2 and 4, for direct seeded , 25 grams of VITAL N was dissolved in 1 liter of
water using 10-20 kg pre-germinated seeds treated overnight before seeding (good for 2 plots-1,250 sq
m/plot) In transplanted set-up, the roots were dipped or sprayed with Vital N solution just before
transplanting. Only half of the recommended inorganic Nitrogen fertilizer was applied at basal. The rest
of the recommended Nitrogen was applied as dressing based on Leaf Color Chart (LCC) readings
beginning at 14 DAS. Other care and maintenance practices followed the Palay Check System.

Role of the different Implementers

The Farmer-Partner

The Farmer Partner, with the assistance of the assigned Agricultural Technician in the area,
provided all the labor requirements from land preparation to harvest. All rice produced were owned by
the farmer including the crop cut samples after data was taken.

The DA-RFU6

DA-RFU 6 provided technical support and general supervision and monitoring. The office was
also responsible in consolidating provincial data as well as make general analysis, interpretation and
recommendations.
Provincial LGU

The Office of the Provincial Agriculturist (OPAg) thru its Rice Program Coordinator assisted the
DA-RFU 6 and Municipal LGU (MLGU) in the selection of the municipalities where the demo was
conducted. OPAg also coordinated with the MLGU and DA-RFU in monitoring the demo.

Municipal LGU

The Office of the Municipal Agriculturist in coordination with OPAg and identified the specific
locations/ barangays as well as the Farmer-Partners. The assigned AT was the one responsible for the
establishment and close supervision of the demo and in data gathering and analysis. The OMAg also
initiated the conduct of Field Days in coordination with the OPAg and OMAg as funded by the funding
institutions.

The following data were gathered:

1. Soil Analysis /MOET data before planting

2. LCC readings at 14, 21, 28, 35, 42, 49 and 56 DAS/DAP
3. Maturity duration (85% ripening)
4. Grain yield (kg/ha) taken from 2m X 5m crop cut at 14 % m.c.
5. Production expenses and gross income per hectare/Economic Analysis
Results and Discussion

Table 3. Soil Analysis/MOET Results

Province Ecosystem Minus Minus Minus Minus Minus Minus Complete No

N P K Cu Zn S NPK Fertilizer

Aklan Rainfed def suff suff suff Suff suff healthy stunted

Irrigated def def suff suff Suff def healthy No set-up

Antique Rainfed def suff suff suff Suff def healthy “

Irrigated def suff suff suff Def def healthy “

Capiz Rainfed No set-up

Irrigated “

Guimaras Rainfed def def def suff Def def healthy stunted

Irrigated

Iloilo Rainfed def def suff suff Suff def healthy stunted

Irrigated No set-up

Neg.Occ. Rainfed def suff def suff Def def healthy No set-up

Irrigated No set-up

Table 3 shows that among the macro nutrients, Nitrogen was the most deficient element in all provinces.
Phosphorous deficiency was observed in irrigated area in Aklan and rainfed areas in Guimaras and Iloilo. Rainfed
areas of Negros Occidental and Guimaras were deficient in potassium. Among the micro nutrients, sulfur is the
most deficient in all areas except in the rainfed area in Aklan. Zinc is deficient in rainfed areas of Guimaras and
Negros Occidental and in irrigated area of Antique. There was no MOET set-up in Capiz and in all irrigated areas in
Guimaras, Iloilo and Negros Occidental due to late crop establishment. The complete NPK resulted in healthy rice
plants, indicating acceptable pH levels of the soils. On the other hand, the absence of fertilizer treatment resulted
in stunted plants indicating deficiencies in the residual nutrient levels.
Table 4. Leaf Color Chart Reading (Across Treatments)

Province Leaf Color Chart Reading*

7 DAP 14 21 28 35 42 49 56
DAP/DA DAP/DAS DAP/DAS DAP/DAS DAP/DA DAP/DAS DAP/DA
S S S

Rf 4 3 3 4 4 4 4

Aklan Irrig 4 3 2 3 4 4 4

Rf 4 3-4 2-3 4 3-4 3-4

Antique Irrig 4 3-4 2-3 4 3-4 3-4

Rf

Capiz Irrig

Rf 3 3 3 4 3 3 3

Guimaras Irrig

Rf 3 3 2 3 3 2

Iloilo Irrig

Rf 3 4 4 4-5 5

Neg.Occ. Irrig

 LCC reading was taken but not indicated in raw data reports

Table 4 shows the Leaf Color Chart (LCC) readings at 14-56 DAS/DAP across treatments. The leaf color chart
is a convenient and useful tool for checking the relative greenness of the rice leaf. This is an indicator of its N
level and is correlated with the biomass production and photosynthetic rate. The color reflects the N demand of
the rice plant within the growing season. From a maximum chart reading of 5, when the value dips to 2-3, this
indicates the need for subsequent N fertilizer application. Maintaining an optimal foliar N content is vital for
attaining high rice yields and therefore adhered to religiously.
Grain Yield (kg/ha) as affected Micronutrient and biofertilizer treatments, Region6, CY 2009-2010.

Grain Yield (kg/ha)

T4- Micronutrient
Province T3- Micronutrient
T1-Full Rec.rate T2- Vital N+ ½ fertilizer + Vital N
fertilizer + full Rec.
of Inorg. Fertilizer Rec.Nitrogen + ½ of
Inorg.Fertilizer
Rec.Inorg.N,P &K

Aklan Rf 3,200 4,000 3,800 4,100

(Lezo) Irrig 4,290 4,770 5,240 4,950

Antique Rf 4,300 4,000 4,500 5,250

(Sibalom) Irrig 4,100 4,000 4,600 5,550

Capiz Rf 5,000 5,040 4,960 5,400

(Ivisan) Irrig 5,175 6,525 4,950 6,750

Guimaras Rf 3,800 4,800 4,600 5,000

(Buenavista-
Rf/San Irrig 4,900 5,000 5,500 5,750
Lorenzo-Irrig)

6,020
Rf 5,500 5,560 5,300
Iloilo
Infected with
Irrig
tungro

Negros Rf 4,000 4,000 4,000 4,500

Occidental
Irrig 3,100 3,700 4,700 4,900

Mean Yield
4,306 4,672 4,741 5,288
(across locations)
Irrigated Field Results

Based on the graph above, the lowest yield was attained in Treatment 1 or the use of the full
recommended inorganic fertilizer with a mean yield of 4,313 kg/ha. across all provinces in Region VI except Iloilo.
Unfortunately, the irrigated rice trial in Iloilo was infected by Tungro, and hence that particular trial was
discontinued. Treatment 2 or VITAL N with ½ of the inorganic N with full inorganic P and K had the next best yield
with 4,799 kg./ha. followed by Treatment 3 or Micronutrient fertilizer with the full inorganic NPK fertilizer with
4,998 kg/ha. The best yield in the irrigated rice trials was attained in Treatment 4 which was the combination of
VITAL N and Micronutrient fertilizer even if the inorganic N applied was reduced to half, at 5,580 kg./ha.

Irrigated Rice Results Analysis

Rep MS = 27,158,500 d.f. = 4

Treatment MS = 624,810 d.f. = 3
Error MS = 176,402 d.f. = 12
F= 3.541975521 Alpha = 0.05
F(3,12,0.05) = 3.49 p value = 0.048119782
There is a significant difference among treatments!
 These results were tested statistically using the Analysis of Variance (ANOVA) technique with the fertilizer
applied as the treatments and the provinces as the blocks. The p value of the treatments was 0.0481 meaning
that at a level of significance of 0.05, the differences of the yields are significant statistically.

In the rainfed rice trials, the lowest yield was obtained from Treatment 1 or the full inorganic NPK applied
with a yield of 4,300 kg/ha. This was followed by Treatment 3 or full inorganic NPK applied plus Micronutrient
fertilizer at 4,527 kg./ha. then by Treatment 2 (VITAL N + ½ inorganic N, full P & K) at 4,567 kg./ha. The highest
yield was attained with Treatment 4 or the combination of VITAL N and Micronutrient fertilizer even at ½ the
inorganic N with a yield of 5,045 kg./ha.

To reconfim the results, ANOVA was again applied to the data. The resulting p-value was 0.010, meaning
that the differences in the yields were statistically significant at a level of significance of 0.05.

Rainfed Field Results

Rainfed Rice Results Analysis

Rep MS = 24,042,270 d.f. = 5

Treat MS = 320,887 d.f. = 3
Error MS = 59,844 d.f. = 15
F= 5.362079049 Alpha = 0.05
F(3,15,0.05) = 3.29 p-value = 0.010383733
There is a significant difference among treatments!
 The multilocation irrigated and rainfed wet season trials revealed indeed that even if the full
recommended rates of NPK fertilizers are applied, the rice yields did not increase appreciably. Intensive cropping
through the years may have already depleted much of the inherent soil micronutrients in some areas. They were
neglected and neve replenished by the farmers, hence the low yields of 3.28 MT/ha. (as per Bureau of
Agricultural Statistics data for Region VI in 2008). VITAL N is a fortified biofertilizer inoculant, and as such, it
contains macro- and micronutrients. Aside from this, it contains Azospirillum spp., a N fixing bacterium that also
stimulates root and root hair development, solubilizes nutrients in the soil, and provides control against plant
pathogens (Okon, et. Al. 1988; Paul and Clark, 1996, Smith and Collins, 2007). Even at half (1/2) of the
recommended N and the full recommended P and K inorganic fertilizer rates, the VITAL N inoculant biofertilizer
with micronutrients increased the mean yield to 4.67 MT/ha. This resulted in an incremental yield of 366 kg/ha.
In comparison, the Micronutrient fertilizer with the full recommended inorganic NPK fertilizers gave a mean yield
of 4.74 MT/ha. This is an increase of 435 kg/ha. over the treated check. A dramatic increase of mean yield by
982 kg/ha. was attained when VITAL N was complemented by Micronutrient fertilizer, even if the inorganic N was
reduced to half the recommended rate.

The use of microbial inoculant fertilizers seem to be effective in addressing N deficiencies, based on te
results of the study. The diazotrophic Azospirillum in the microbial inoculant get established in the rhizonsphere
and intercellular spaces of root cortex and can provide up to 30% of the total N accumulated by plants (Bottomley
and Myrold, 2007). Moreover, Biro et al 2006 demonstrated that Azospirillum can provide a stress buffer effect
and synergistic interaction for promoting plant growth and development. The optimum root and shoot growth,
and consequently increased grain yield and water use efficiency can be achieved through an alternate wetting
and moderate soil drying (Zhang et al, 2009). This is in consonance with the label recommendation for using
VITAL N biofertilizer to attain sufficiency in rice while saving on water use.

Micronutrients depletion was found to be widespread in the rice growing areas of Western Visayas.
Micronutrients are elements that are essential for plants to complete their life cycles, they are needed, however,
only in small amounts. Deficiencies in micronutrients can reduce dry matter yield and quality of plants, and
unfortunately, micronutrient deficiencies occur in Philippine soils. Micronutrient fertilizer contains a combination
of secondary nutrients: Mg, S, Mn, Zn, Cu, B, Fe, Cl and an exclusive stabilizer.

The micronutrient fertilizer was used as an add-on to the recommended NPK fertilizer requirement and
half of the NPK chemical fertilizer requirement plus VITAL N. In both instances, the micronutrient fertilizer was
able to increase yields by addressing the micronutrient deficiencies, particularly that in Sulfur, which was
predominant among the test sites.

Of particular note is the fact that the dramatic increase in yield when a combination of VITAL N and
Micronutrient fertilizer was employed. There seemed to be a synergistic effect between the enzyme and
micronutrients in Micronutrient fertilizer and the formulation of VITAL N. Improved nutrient absorption,
particularly of Magnesium, Calcium, as well as Potassium, was observed in wheat grains inoculated with
Azospirillum (Creus et al, 2004). A complementation such as this may explain for the increased yield in rice and
can be a subject for further investigation.
Summary and Discussion

The wet season 2009 micronutrient fertilizer and biofertilizer techno demo were established in irrigated
and rainfed areas in Aklan, Antique, Capiz, Guimaras, Iloilo and Negros Occidental. The Minus One Element
Technique, or MOET, soil analysis revealed widespread deficiency in Nitrogen and Sulphur, as well as some
deficiencies in Zinc, Phosphorous and Potassium in some of the selected sites. The leaf color chart (LCC) readings
on rice leaves served as guides for timing the subsequent applications of the nitrogenous Urea fertilizer for
attaining optimal yields.

The yields of the different treatments across the various locations around Western Visayas were collected
and calculated to come up with the rice harvest on a per hectare basis. The data was tabulated and the results
the hypothesis that all the treatments was tested using ANOVA at a level of significance of 0.05. Based on the
result of the statistical analysis, the treatments had statistically significant yields.

Among the different treatments, Treatment 1, or the application of the full recommended inorganic
fertilizer had the lowest yield, regardless of whether the field was rainfed or irrigated. Treatment 2 (½ the
Nitrogen fertilizer, full Phosphorous and full Potassium plus VITAL N ) and Treatment 3 (Full inorganic fertilizer
plus Micronutrient fertilizer) had similar yields, with Treatment 3 (Micronutrient fertilizer plus full NPK) slightly
higher under irrigated conditions while Treatment 4 (VITAL N plus ½ N, full PK) higher under rainfed conditions.

Among both rainfed and irrigated conditions, Treatment 4 (VITAL N + Micronutrient fertilizer + ½ NPK)
was superior, with an increase in yield of around 700 – 1000 kgs. per hectare. This can probably be attributed to
the increased nutrient absorption due to the presence of Azospirillum and made available by the application of
Micronutrient fertilizer and VITAL N. This is an interesting finding that can be subject to further study, as this
effect was observed with wheat.
References

1. Biro, B., K.K. Pechy, M. T. Michael and R. J. Strasser. “Role of beneficial microsymbionts on the plant
performance and plant fitness,” Microbial activity in the rhizosphere. (Eds.) K. G. Mukeryi, C.
Manoharatchary and J. Singh. Springer, Heidelberg:2006, p. 349.

2. Bottomley, P. J. and D. R. Myrold. “Biological inputs,” Soil microbiology, ecology and biochemistry, 3 rd ed.
(Eds.) E. A. Paul. Elsevier, NY, USA:2007.

3. Creus, Cecilia M., RJ Sueldo, and Barassi, C.A., “Water relations and yield in Azospirillum-inoculated
wheat exposed to drought in the field,” Can. J. Bot. 82(2): 273–281 (2004)

4. Okon, Y., Y. Kapulnik and S. Sarig. “Field inoculation studies with Azospirillum in Israel,” Biological
nitrogen fixation – Recent Developments. (Eds.) N. S. Subba Rao, Gordo and Breach. N.Y, USA:1988. pp.
337.

5. Paul, E. A. and F. E. Clark. Soil microbiology and biochemistry, 2 nd ed. Academic Press, NY, USA:1996, p.
340.

6. Reza, H. A., Design of experiments for agriculture and the natural sciences, Boca Raton, FL : CRC Press,
2006, pp. 51-118.

7. Smith, J. L. and H. P. Collins. “Management of organisms and their processes in soils, “ Soil microbiology,
ecology and biochemistry, 3rd ed. (Ed.) E. A. Paul. Elsevier, NY, USA:2007, p. 532.

8. Zhang, H. Y., Z. Wang, J. Yong and J. Zhang. “An alternate wetting and moderate soil drying improves root
and shoot growth in rice,” Crop Science 49:2246-2260.
Economic Analysis, Micronutrient and Biofertilizer Trial, Region 6, CY: 2009-2010:
Production cost and net income per hectare (Php)
T3- Micronutrient T4- Micronutrient
Province
T1-Full Rec.rate of Inorg. T2- Vital N+ ½ fertilizer + full Rec. fertilizer + Vital N + ½ of
Fertilizer Rec.Nitrogen and full PK Inorg.Fertilizer Rec.Inorg.N,P &K
  Prod’n Net Gross Prod’n Net Gross Prod’n Net Gross Prod’n Net
Gross Sales cost Income Sales cost Income Sales cost Income Sales cost Income
Rf 54,400 23,442 30,958 68,000 24,857 43,143 64,600 25,592 39,008 69,700 25,732 43,968
Aklan
Irrig 72,930 27,026 45,904 81,090 27,566 53,524 89,080 29,960 59,120 84,321 28,654 55,667
Rf 47,300 25,341 21,959 44,000 24,280 19,720 49,503 26,518 22,985 57,500 27,418 30,082
Antique
Irrig 43,050 26,384 16,666 42,000 25,290 16,710 48,300 27,101 21,199 58,275 28,523 29,752
Rf 46,200 27,123 19,077 46,989 26,039 20,950 45,830 28,316 17,514 49,896 27,775 22,121
Capiz
Irrig 46,284 29,545 16,739 58,359 32,634 25,725 44,273 29,576 14,697 60,372 28,844 31,528
Rf 45,600 23,918 21,682 57,600 24,778 32,822 55,200 25,826 29,374 60,000 23,120 36,880
Guimaras
Irrig 0 0 0 0 0 0 0 0 0 0 0 0
Rf 60,500 22,470 38,030 61,160 21,070 40,090 58,300 22,470 35,830 66,220 21,070 45,150
Iloilo Infected with   0 0 0 0 0 0 0 0 0 0
Irrig
tungro
Rf 36,000 15,140 20,860 36,000 14,620 21,380 36,010 15,760 20,250 40,580 16,290 24,290
Neg. Occ.
Irrig 40,300 24,160 16,140 48,100 22,796 25,304 61,100 25,452 35,648 63,700 24,088 39,612
Ave. Rf 48,333 22,906 25,428 52,292 22,607 29,684 51,574 24,080 27,494 57,316 23,568 33,749
Income
(Across
Irrig 40,513 21,423 23,862 45,910 21,657 30,316 48,551 22,418 32,666 53,334 22,022 39,140
Location)