Cooperative Education in Agriculture Report

COOPERATIVE EDUCATION IN AGRICULTURE REPORT AT THE AGRONOMIC DEVELOPMENT SOUTHEAST ASIA FIELD RESEARCH AND DEVELOPMENT CENTER OF BAYER CROPSCIENCE IN MASIIT, CALAUAN, LAGUNA, PHILIPPINES

Miss PONSAWAN KHAMPHASAN ID. 543030378-3

INOCULATION METHODS AND MASS PRODUCTION OF DAMPING OFF CAUSED BY Pythium aphanidermatum, Rhizoctonia solani and Sclerotium rolfsii IN LETTUCE AND ONION

THIS REPORT IS PART OF THE COURSE 100495 COOPERATIVE EDUCATION IN AGRICULTURE BACHELOR OF SCIENCE AGRICULTURE (PLANT PATHOLOGY) FACULTY OF AGRICULTURE KHON KEAN UNIVERSITY JULY 2014 COOPERATIVE EDUCATION IN AGRICULTURE REPORT Academic year 2013

Title : Inoculation methods and mass production of damping-off caused by Pythium aphanidermatum, Rhizoctonia solani and Sclerotium rolfsii in lettuce and onion at the Bayer Cropscience (AD-SEA) in the Philippines Name of reporter: Miss Ponsawan Khamphasan Name of company: The Agronomic Development Southeast Asia Field Research and Development Center of Bayer CropScience (AD-SEA) Address: Bayer CropScience, Inc., Agronomic Development Southeast Asia Field Research and Development Center (ADSEA), Barangay Masiit, Calauan, Laguna, Philippines Job supervisors: Mr. Leo Hawod Position: Lead Researcher (Disease Management)

……………………………………………………………………Adviser (Dr. Anan Wongcharone) Board of Cooperative Education …………………………………………………………………… committee (Dr. Anan Wongcharone) …………………………………………………………………… committee (………………………………………………………………..)

COOPERATIVE EDUCATION IN AGRICULTURE REPORT

The Bayer CropScience Agronomic Development Southeast Asia Field Research and Development Center (AD-SED) at Masiit, Calauan, Laguna, Philippines Miss Ponsawan Khamphasan ID. 543030378-3 This report is part of the course 100495 cooperative education in agriculture Bachelor of science agriculture (Plant pathology) Faculty of agriculture Khon kaen university July 2014

Acknowledgements

In performing my internship, it's a successful one I had to take the help and guideline of some respected persons. First of all I am grateful to Khon kaen university who giving a scholarships and opportunity to internship in the Philippines, and also faculty of agriculture who support and help me. I would like to thank you to the Bayer Cropscience and Mr. Pete Davies for accept and giving a chance to internship at AD-SEA, I gain good experience and new knowledge. I also would like to thank you to Mr. Leo Hawod who is my job supervisor for the valuable guidance and advice. I also would like to thank you to Pia, Jennifer, Charish, Macy and everyone at AD-SEA for helps and suggestions I also would like to thank you to my faculty advisor Dr. Anan Wongcharone for support , the valuable guidance and advice. I have to thank you to my lovely family for support me. I also would like to so much thank you to everyone who is a part of helps to successful.

TABLE OF CONTENTS

Detail Page Acknowledgement a Table of contents b Table of figure c Table of table d Chapter 1 Company 1.1 Organization of company 1 1.2 Participation of students 3 Chapter 2 Project 2.1 Abstract 4 2.2 Introduction 5 2.3 Objective 6 2.4 Review of related literature 6 2.5 Material and method 13 2.6 Result 13 2.7 conclusion 26 2.8 references 27 Chapter 3 work achievement recommendation and advantage 28 Chapter 4 Appendix 4.1 company and activities 30 4.2 Project 53 4.3 living in Philippines 57

Table of figure c

Detail Page

Figure.1 Organization Chart of AD-SEA 2 Figure.2 Disease cycle of damping-off and seed decay caused by Pythium sp. 10 Figure.3 Disease cycle of Rhizoctonia solani (Thanatephorus cucumeris) 10 Figure.4: Growth of Pythium aphanidermatum in different media 15 Figure. 5 Growth of Rhizoctonia solani in different media 16 Figure. 6 Growth of Sclerotium rolfsii in different media 18 Figure.7 Rate of germination on lettuce of different pathogenicity methods 19 for damping-off at greenhouse. Figure.8 Pathogenicity testing of damping-off in lettuce 19 Figure. 9 Rate of germination on onion of different pathogenicity methods 20 for damping-off at greenhouse. Figure. 10 Pathogenicity testing of damping-off in onion 20 Figure. 11 Rate of germination on lettuce of different inoculation methods for damping off. 22 Figure.12 Lettuce growth on different inoculum media of damping off 23 Figure. 13 Rate of germination on onion of different inoculation methods 24 for damping-off Figure. 14 Onion growth on different inoculum media of damping off 25

Table of table d

Detail Page

Table 1: Result of diameter mycelia of Pythium aphanidermatum in 14 different culture media within 10 days of incubation Table 2: Result of diameter mycelia of Rhizoctonia solani in 16 different culture media within 10 days of incubation Table 3: Result of diameter mycelia of Sclerotium rolfsii in 17 different culture media within 10 days of incubation

CHAPTER 1 COMPANY

1.1 Organization of company The Agronomic Development Southeast Asia (AD-SEA) Field Research and Development Center is part of Bayer CropScience’s global AD field testing organization. Functions Global: • To evaluate new molecules under local conditions • To profile new or more advanced candidates and products. • To enable the development process Region: • To carry out technical training programs for Regional personnel. • To give technical support to MDT and other Regional forums. • To carry out trial programs as required and where possible. Southeast Asian Countries: • To make information on new candidates available to countries in SEA. • To support Development and Commercial needs of countries in field programs and technical support. Organization • Our team in Philippines consists of 8 Researchers, a Trial Manager and an Administrator. • In Thailand, we have a lean team of 4 in the field. • In Vietnam, we contract up to 20 trials to the local development team 2

Figure1 Organization Chart of AD-SEA Sites • In Philippines we have 2 plantation sites in Davao and a cold weather site in the mountains in Tublay. We have 2 sites with rice paddies and the rest are for upland fruit and vegetables. In Thailand we have an elevated site near Chiang Mai, our other sites are all within 100km of Bangkok. In Vietnam all trials are done in farmers’ fields. • We have one Bayer-owned Field Station in Thailand at Suphanburi. The other four sites in Thailand and all 7 sites in Philippines, including this one are rented.

The Masiit Trial Station • 8.3 hectares • 70% - lowland paddy (rice) • 30% - upland area (vegetable and field crops) • Strategically located near science community of Los Banos. • Serves as the main field research and development center of Bayer Crop Science in Southeast Asia. • It also serves as Training Center for APAC. 1.2 Participation of students Activities All of activities were done at The Bayer CropScience Agronomic Development Southeast Asia Field Research and Development Center at Masiit, Calauan, Laguna, Philippines from March, 24 2014 to July, 11 2014. Mr. Leo Hawod is Job supervisor of Thai student. Working hours started at 8 am to 5 pm (Full time), Monday to Friday. I am plant pathologist assistant. Most of activities were done in the field, green house and in the plant pathology laboratory. 1.2.1 Lecture - Rice production - Vegetable production - Weed of Rice - Important diseases of Rice - Insect pests managements 1.2.2 Field trial Activities - Trials design and lay out - Experimental design and lay-out - Application - Assessment [Insecticide trials in kale (White fly)] - Insecticide trials in kale (White fly) 4

- Seed germination testing (seed treatment) - Data entry 1.2.3 Project (Damping-off)

CHAPTER 2

INOCULATION METHODS AND MASS PRODUCTION OF DAMPING OFF CAUSED BY Pythium aphanidermatum, Rhizoctonia solani and Sclerotium rolfsii IN LETTUCE AND ONION AT THE BAYER CROPSCIENCE (AD-SEA) IN PHILIPPINES

2.1 ABSTRACT This project was conducted at The Bayer CropScience Agronomic Development Southeast Asia Field Research and Development Center (AD-SED) at Masiit, Calauan, Laguna, Philippines. The internship was focus on inoculation method and mass production of damping off caused by Pythium aphanidermatum, Rhizoctonia solani and Sclerotium rolfsii in lettuce and onion. Specific activities included comparison of different culture media for growth of Pythium aphanidermatum, Rhizoctonia solani and Sclerotium rolfsii, growth was compared on three media, Potatoes Dextrose Agar (PDA), Corn Meal Agar (CMA), Rice Dextrose Agar (RDA). Growth of Pythium aphanidermatum, Rhizoctonia solani was best on PDA and Growth of Sclerotium rolfsii was best on CMA. For pathogenicity testing by agar block and pulpy agar. Germination of lettuce and onions seeds were lowest in pots prepared by pulpy agar. For mass production of damping off for the three pathogen were transfer to plastic bag by different substrate, rice hull with rice grain and sawdust with sorghum. Germination of lettuce and onions seeds were lowest in trays prepared by rice hull with rice grain.

2.2 INTRODUCTION With a constantly increasing population and demand for food in the world, there is an urgent need for increased food production and sustainable agricultural development. Damages to crop by pests have been recognized as one of the most important constraints in food production. This is due to the fact the tropical conditions favor the growth and development of not only crops but also plant pest. The plant disease can cause severe yield reduction when left uncontrolled, not only reduce crop yield but also reduce quality of produce and lowering its market value. For vegetable production, Damping-off frequently attacks seeds, young seedlings of almost all kinds of vegetables, It is hard to get older and their stems that is a problem. Losses to damping- off can be severe, especially when cool, wet weather prevails at seeding or seed emergence. Damping-off diseases were found worldwide and can be caused by several species of fungi. Pythium aphanidermatum, Rhizoctonia solani and Sclerotium rolfsii that commonly cause damping-off. Several species of Pythium spp. cause pre- and postemergence damping-off. Pythium sp. Rhizoctonia sp. and Sclerotium sp. produces a white, rapidly growing mycelium. Rhizoctonia solani and Sclerotium rolfsii have sclerotia bodies, easy to notice. This project focus on inoculation methods and mass production of damping-off in lettuce and onion caused by Pythium aphanidermatum, Rhizoctonia solani and Sclerotium rolfsii. Therefore, to disease control and management, growth of pathogen, pathogenicity testing, inoculation method needed to study to solve of damping-off in lettuce and onion.

2.3 OBJECTIVE To produce mass inoculum for inoculation of damping-off caused by Pythium aphanidermatum, Rhizoctonia solani and Sclerotium rolfsii in lettuce and onion.

2.4 REVIEW OF RELATED LITERATURE 2.4.1 Important of damping-off Damping-off is the disease affects seeds, seedlings, and roots of all plants. In all cases, however, the greatest damage is done to the seed and seedling roots during germination either before or after emergence. Losses vary considerably with soil moisture, temperature, and other factors. Quite frequently, seedlings in seedbeds are completely destroyed by damping-off or they die soon after they are transplanted. In many instances, poor germination of seeds or poor emergence of seedlings is the result of damping-off infections in the preemergence stage. Older plants are seldom killed when infected with the damping-off pathogen, but they develop root and stem lesions and root rots, their growth may be retarded considerably, and their yields may be reduced drastically. Some species of the damping-off oomycete also attack the fleshy organs of plants, which rot in the field or in storage. Symptoms When seeds of susceptible plants are planted in infested soils and are attacked by the damping- off fungi, they fail to germinate, become soft and mushy, and then turn brown, shrivel, and finally disintegrate Young seedlings can be attacked before emergence at any point on the plant, from which the infection spreads rapidly, the invaded cells collapse, and the seedling is overrun by the oomycete and dies (preemergence damping-off) (Agrios, 2005). Rhizoctonia, Pythium, Fusarium, Phytophthora, Sclerotinia, Sclerotium, Botrytis, and others that commonly cause damping-off. There are many other species of fungi that occasionally cause this disease (Jones, 2000). In the field, garden, or planter box, seedlings often fail to come up, or die soon after they have emerged from the soil. Seeds may rot before they germinate, shoots may be decayed before they emerge, or stems of seedlings may be attacked near the soil line, causing young plants to collapse. These diseases often are collectively referred to as “damping-off,” and may be caused by a number of soil-inhabiting pathogens.Species of the soil 8

organism Pythium are most often responsible for damping-off, but several other pathogens, including species of Rhizoctonia, Fusarium, Sclerotium and Phytophthora, can also cause decay. (E. J. Perry, 2014) 2.4.2 Symptoms The first evidence of damping-off or seed piece decay (as in potatoes) is the failure of some plants to emerge. If seeds are attacked before they germinate, they become soft and mushy, turn dark brown, and decay. They may have a layer of soil clinging to them when they are dug up because the soil is interwoven with fine, threadlike fungus growth. Germinating seedlings shrivel and may darken. If seedlings are attacked after they emerge, stem tissue near the soil line is decayed and weakened, usually causing plants to topple and die. When only roots are decayed, plants may continue standing but remain stunted, wilt and eventually die. As seedlings get older, they become less susceptible to damping-off pathogens. (E. J. Perry, 2014) 2.4.3 Damage Damping-off is caused by fungus and it usually occurs in small patches at various places in the seedbeds or field. The disease spots often increase from day to day until the seedlings harden. Seedlings are extremely susceptible for about two weeks after emergence. As the stem hardens and increases in size, the injury no longer occurs. Some seedlings are not killed at once, but the roots are severely damaged and the stem is girdled at the ground level. Such plants remain stunted and often do not survive transplanting. The injury from damping-off fungi is of two types - Pre-emergence damping off consists of a decay of the germinating seed or death of the seedling before it can push through the soil. This injury is a common cause of poor stands, which are often attributed to inferior quality of the seed or the untreated seeds. Pythium spp. Sclerotium spp. cause seed decay.

- Post-emergence damping-off which occurs after the seedlings have emerged from the soil but while still small and tender. The roots may be killed, and affected plants 9

show water soaking and shrivelling of the stems at the ground level; they soon fall over and die. Post-emergence damping-off is mostly caused by Rhizoctonia spp.

2.4.4 Disease cycle Most soils contain fungi that can attack seeds and seedlings. When conditions are favorable for grass seed germination and growth, as in the early fall and spring, these organisms usually are of little significance. Warm, wet weather, however, is more favorable for damping- off fungi and less favorable for seedlings. Under these conditions, fungi can severely damage seed and seedlings. Damping-off outbreaks are triggered by high temperatures and humidity, water-logged soils, excessive fertilizer, or an excessive seeding rate. In warm weather, higher-than-normal rates of seed may produce a very dense stand of seedlings that will hinder the escape of moisture from the soil surface. Such wet conditions over long periods are ideal for invasion by damping- off fungi. High rates of fertilizer can produce a succulent turf that is more susceptible to attack by damping-off fungi.With new seeding techniques, such as hydroseeding and hydromulching, seeding is done all year round, and as a result, damping-off has become a more frequent cause of stand failure. Stand loss to seed decay or seedling blight makes successive attempts to reseed the dead areas more difficult. (E. J. Perry, 2014)

Figure 2 Disease cycle of damping-off and seed decay caused by Pythium sp. (Agrios, 2005)

Figure 3 Disease cycle of Rhizoctonia solani (Thanatephorus cucumeris). (Agrios, 2005)

2.4.5 Morphology

The Pathogen: Pythium spp. Several species of Pythium cause pre- and postemergence damping-off. Pythium produces a white, rapidly growing mycelium. The mycelium gives rise to sporangia, which germinate directly by producing one to several germ tubes or by producing a short hypha at the end of which forms a balloon-like secondary sporangium called a vesicle. In the vesicle, 100 or more zoospores are produced, which, when released, swarm about for a few minutes, round off to form a cyst, and then germinate by producing a germ tube. (Agrios, 2005) The Pathogen : Rhizoctonia spp. Represent a large, diverse, and complex group of fungi. All Rhizoctonia fungi exist primarily as sterile mycelium and, sometimes, as small sclerotia that show no internal tissue differentiation. Mycelial cells of the most important species, R. solani, contain several nuclei (multinucleate Rhizoctonia), whereas mycelial cells of several other species. contain two nuclei (binucleate Rhizoctonia). The mycelium, which is colorless when young but turns yellowish or light brown with age, consists of long cells and produces branches that grow at approximately right angles to the main hypha, are slightly constricted at the junction, and have a cross wall near the junction The branching characteristics are usually the only ones available for identification of the fungus as Rhizoctonia. Under certain conditions the fungus produces sclerotia-like tufts of short, broad cells that function as chlamydospores, or eventually the tufts develop into rather small, loosely formed brown to black sclerotia, which are common on some hosts such as potato. (Agrios, 2005)

The pathogen: Sclerotium pp. Sclerotium spp., produces abundant white, fluffy, branched mycelium that forms numerous sclerotia but is usually sterile, i.e., does not produce spores. Sclerotium rolfsii, which causes the symptoms described earlier on most of the hosts, occasionally produces basidiospores at the margins of lesions under humid conditions. Its perfect stage is Aethalium rolfsii. As mentioned earlier, the species S. bataticola, which causes diseases in several different hosts, occasionally produces conidia in pycnidia and is now known as the imperfect fungus Macrophomina phaseolina. (Agrios, 2005)

2.5 MATERAILAL AND METHOD 2.5.1 Compare culture media A disc of mycelium from pure culture of Pythium aphanidermatum growing on CMA, Rhizoctonia solani growing on PDA and Sclerotium rolfsii growing on PDA was cut out and inoculated on to plates with media. Growth was compared on three media, Potatoes Dextrose Agar (PDA), Corn Meal Agar (CMA), Rice Dextrose Agar (RDA). Five replicates were made for each medium and pathogen. The plates were incubated at room temperature , dark condition, incubated 10 days and measurements of the mycelial diameters were recorded daily. 2.5.2 Pathogenicity testing On this trial, mycelial of three pathogen (Pythium aphanidermatum, Rhizoctonia solani and Sclerotium rolfsii) growth straight form the plates were transferred into each pot by 2 methods that is agar block and pulpy agar. then cover agar block and pulpy agar by some soil and sowing the lettuce seeds 50 seeds per pot and onion seeds 25 seeds per pot and cover seeds by some soil, after that water. Lettuce seeds and onion seeds were then allowed to grown for 2 weeks at greenhouse and then checked for damping off symptoms by percent of germination. 2.5.3 Different Inoculation method A disc of mycelium from pure culture of Pythium aphanidermatum growing on PDA, Rhizoctonia solani growing on PDA and Sclerotium rolfsii growing on CMA was cut out and inoculated on to Plastic bag with different inoculum media, rice hull with rice grain and sawdust with sorghum, incubated for 1 week. Mycelial of three pathogen were transferred into each tray. Then cover agar block and pulpy agar by some soil and sowing the lettuce seeds 300 seeds per tray and onion seeds 50 seeds per tray and cover seeds by some soil, after that water. Lettuce seeds and onion seeds were then allowed to grown for 2 weeks and then checked for damping off symptoms by percent of germination.

2.6 RESULTS 2.6.1 Result of compare culture media PDA was the most suitable media for growth of Pythium aphanidermatum. Based on the results (Table.1), PDA has the highest mycelial growth. Maximum growth of mycelial were observed at day 5 in PDA media. Pictures were also taken at the 10th day, the last day of incubation. (Fig.4) Table 1: Result of diameter mycelial of Pythium aphanidermatum in different culture media within 10 days of incubation. Date/ media diameter(mm.) PDA CMA RDA day 1 65.4 78.6 49.8 day 2 85.0 + 85.0 + 85.0 + day 3 85.0 + 85.0 + 85.0 + day 4 85.0 ++ 85.0 + 85.0 + day 5 85.0 +++ 85.0 + 85.0 + day 6 85.0 +++ 85.0 + 85.0 + day 7 85.0 +++ 85.0 + 85.0 + day 8 85.0 +++ 85.0 ++ 85.0 + day 9 85.0 +++ 85.0 ++ 85.0 + day 10 85.0 +++ 85.0 ++ 85.0 +

Potatoes Dextrose Agar (PDA), Corn Meal Agar (CMA), Rice Dextrose Agar (RDA), densities of mycelia [ * = low, ** = Medium, *** = high]]

Figure 4 Growth of Pythium aphanidermatum in different media: potato dextrose agar (PDA), corn meal agar (CMA), rice dextrose agar (RDA) at 10 days of incubation under dark condition and room temperature. Same as Pythium aphanidermatum, PDA was the most suitable media for growth of Rhizoctonia solani. Based on the results (Table.2), PDA has the highest mycelial growth. Maximum growth of sclerotial bodies were observed at day 5 in PDA media. Pictures were also taken at the 10th day, the last day of incubation. (Fig.5)

Table 2: Result of diameter mycelia of Rhizoctonia solani in different culture media within 10 days of incubation. Date/ media diameter(mm.) PDA CMA RDA

day 1 5.5 8.5 5.2 day 2 85.0 + 85.0 + 85.0 + day 3 85.0 + 85.0 + 85.0 + day 4 85.0 + 85.0 + 85.0 + day 5 85.0 + (68) 85.0 + (4) 85.0 +(9) day 6 85.0 + (68) 85.0 + (4) 85.0 +(9) day 7 85.0 + (68) 85.0 + (4) 85.0 +(9) day 8 85.0 + (68) 85.0 + (4) 85.0 +(9) day 9 85.0 + (68) 85.0 + (4) 85.0 +(9) day 10 85.0 + (68) 85.0 + (4) 85.0 +(9)

Potatoes Dextrose Agar (PDA), Corn Meal Agar (CMA), Rice Dextrose Agar (RDA) Densities of mycelia [ * = low, ** = Medium, *** = high], ( ) number of sclerotia.

Figure 5 Growth of Rhizoctonia solani in different media : potato dextrose agar(PDA), corn meal agar (CMA), rice dextrose agar (RDA) at 10 days of incubation Under dark condition and room temperature.

CMA was the most suitable media for growth of Sclerotium rolfsii. Based on the results (Table.3), sclerotial bodies were observed at day 8 in CMA media. Pictures were also taken at the 10th day, the last day of incubation. (Fig.6) Table 3: Result of diameter mycelia of Sclerotium rolfsii in different culture media within 10 days of incubation. Date/ media diameter(mm.) PDA CMA RDA day 1 9 12 10 day 2 40.8 42.6 32.6 day 3 75 85 56 day 4 85.0 + 85.0 + 85.0 + day 5 85.0 + 85.0 + 85.0 + day 6 85.0 ++ 85.0 + 85.0 + day 7 85.0 ++ 85.0 + 85.0 + day 8 85.0 ++ 85 .0+ (40) 85.0 + day 9 85.0 ++ 86 .0+ (40) 85.0 + day 10 85.0 ++ 87 .0+ (40) 85.0 + (30)

Potatoes Dextrose Agar (PDA), Corn Meal Agar (CMA), Rice Dextrose Agar (RDA) Densities of mycelia [ * = low, ** = Medium, *** = high], ( ) number of sclerotia.

Figure 6 Growth of Sclerotium rolfsii in different media: potato dextrose agar (PDA), corn meal agar (CMA), rice dextrose agar (RDA) at 10 days of incubation Under dark condition and room temperature. 2.6.2 Result of Pathogenicity testing Lettuce seeds were grown in pots and were inoculated with three different pathogens; Pythium aphanidermatum, Rhizoctonia solani, and Sclerotium rolfsii. Germination percentage was done at 14 DAS to assess the incidence of damping-off in lettuce. Germination of lettuce seeds were lowest in pots prepared by pulpy agar method (Fig.7). Lettuce inoculated with Pythium aphanidermatum (3%), Rhizoctonia solani (15%), and Sclerotium rolfsii (12%) have lowest germination rate in pulpy agar inoculum; compared to the untreated with almost 70-90% germination rate for the three pathogens (Fig.8). Pictures were also taken at 14 DAS.

100% 87% 81% 80% 72%

60% 48%

40% 29.00% 16% 15% 20% 12% 3.00% 0% Uninoculated Agar block Pulpy agar pythium Rhizoctonia Sclerotium

Figure 7 Rate of germination on lettuce of different pathogenicity methods for damping-off at greenhouse.

Figure 8 Pathogenicity testing of damping-off in lettuce at 14 DAS under screenhouse condition. a. Pythium aphanidermatum b. Rhizoctonia solani c. Sclerotium rolfsii 20

Onion seeds were grown in pots and were inoculated with three different pathogens; Pythium aphanidermatum, Rhizoctonia solani, and Sclerotium rolfsii. Germination percentage was done at 14 DAS to assess the incidence of damping-off in onion. Germination of onion seeds were lowest in pots prepared by pulpy agar method (Fig.9). onion inoculated with Pythium aphanidermatum (26%), Rhizoctonia solani (24%), and Sclerotium rolfsii (21%) have lowest germination rate in pulpy agar inoculum; compared to the untreated with almost 70-90% germination rate for the three pathogens (Fig. 10). Pictures were also taken at 14 DAS.

100% 90% 80% 70% 60% 50% 40% 30% 20% 10% 0% Uninoculated Agar block Pulpy agar

pythium Rhizoctonia Sclerotium

Figure 9 Rate of germination on onion of different pathogenicity methods for damping-off at greenhouse. 21

Figure 10 Pathogenicity testing of damping-off in onion at 14 DAS under screenhouse condition. a. Pythium aphanidermatum b. Rhizoctonia solani c. Sclerotium rolfsii

2.6.3 Result of mass production 1) Result of mass production for damping-off in lettuce Lettuce seeds were grown in trays and were inoculated with three different pathogens; Pythium aphanidermatum, Rhizoctonia solani, and Sclerotium rolfsii. Germination percentage was done at 14 DAS to assess the incidence of damping-off in lettuce.Germination of onion seeds were lowest in trays prepared by rice hull with rice grain (Fig.11). Lettuce inoculated with Pythium aphanidermatum (17%), Rhizoctonia solani (71%), and Sclerotium rolfsii (7%) have lowest germination rate in rice hull with rice grain inoculum (Fig.12). Pictures were also taken at 14 DAS.

100% 90% 80% 70% 60% 50% 40% 30% 20% 10% 0% uninoculated Rice hull-grain Sawdust-sorghum

pythium Rhizoctonia Sclerotium

Figure 11 Rate of germination on lettuce of different inoculation methods for damping off.

Figure 12 Lettuce growth on different inoculum media of damping off a. Pythium aphanidermatum b. Rhizoctonia solani c. Sclerotium rolfsii

2) Result of mass production for damping-off in onion Onion seeds were grown in trays and were inoculated with three different pathogens; Pythium aphanidermatum, Rhizoctonia solani, and Sclerotium rolfsii. Germination percentage was done at 14 DAS to assess the incidence of damping-off in lettuce. Germination of onion seeds were lowest in trays prepared by rice hull with rice grain. (Fig.13) Onion inoculated with Pythium aphanidermatum (17%), Rhizoctonia solani (71%), and Sclerotium rolfsii (7%) have lowest germination rate in rice hull with rice grain inoculum. (Fig.14) Pictures were also taken at 14 DAS.

100% 90% 80% 70% 60% 50% 40% 30% 20% 10% 0% uninoculated Rice hull-grain Sawdust-sorghum

pythium Rhizoctonia Sclerotium

Figure 13 Rate of germination on onion of different inoculation methods for damping-off. 25

Figure 14 Onion growth on different inoculum media of damping off a. Pythium aphanidermatum b. Rhizoctonia solani c. Sclerotium rolfsii 26

2.7 Conclusion PDA was the most suitable media for growth of Pythium aphanidermatum. PDA was also found the most suitable media for growth of Rhizoctonia solani and sclerotial bodies were observed on PDA. CMA was the most suitable media for growth of Sclerotium rolfsiiand and sclerotial bodies were observed at day 8 in CMA media.From result of pathogenetic method, Pulpy agar by blender was suitable pathogenetic method for damping-off. From result of inoculation method in mass production of the three pathogen, Rice hull with rice grain was suitable inoculation method in mass production for damping-off.

2.8 References AGIOS, G. N. 2005.Plant pathology 5th ed. Academic press, New York. Peter j. Batt. 2007. The Vegetable Industry in the Philippines. ACIAR GPO Box 1571Canberra ACT 2601, Australia.

CHAPTER 3 work achievement 3.1 work achievement This project is a part of cooperative education in agriculture subject was done at The Bayer CropScience Agronomic Development Southeast Asia Field Research and Development Center (AD-SEA) at Masiit, Calauan, Laguna, Philippines from 24 March – 11 July 2014. The results of this experiment will be useful for management and control of damping-off in other trials. 3.2 Recommendations The Bayer CropScience Agronomic Development Southeast Asia Field Research and Development Center (AD-SEA) at Masiit, Calauan, Laguna, Philippines is a great learning company for interns helps interns to learn and develop their skills and the people in the station so nice. I would recommend, should have schedule of activities for internship period to inform before start internship, that it good for interns to prepare yourself. 3.3 Advantage For student The past 16 weeks of my internship have been very instructive for me. Bayer CropScience has offered me opportunities to learn and develop myself that I can say, I gained a lot of new experience, new knowledge, especially work life.

1. Learned about the process of work, responsibility. 2. Learned to adapt to the work environment. 3. Learned to plan for work and organizing of a project. 4. Learned to increase skills for understanding and working with other people. 5. Learned to acquire additional interpersonal communication and interaction skills. 6. Get experience in working in another country with persons from another culture. 7. Learned about crop and rice production, field trial activities, weed of rice, important diseases of rice, insect pest managements. 8. Learned about life, food, culture, language in Philippines 9. English skills was developed. 29

Chapter 4 Appendix 4.1 Company

The Agronomic Development Southeast Asia Field Research Center of Bayer CropScience in Masiit, Calauan, Laguna, Philippines

Map of The Agronomic Development Southeast Asia Field Research Center of Bayer CropScience in Masiit, Calauan, Laguna, Philippines 30

DESCRIPTION AND SCHEDULE OF ACTIVITIES

 Time and Place of Apprenticeship Place of Apprenticeship : The Bayer CropScience Agronomic Development Southeast Asia Field Research and Development Center at Masiit, Calauan, Laguna, Philippines. Time : March, 24 2014 to July, 11 2014 for 16 weeks.

 Schedule of activities Activities All of activities were done at The Bayer CropScience Agronomic Development Southeast Asia Field Research and Development Center at Masiit, Calauan, Laguna, Philippines from March, 24 2014 to July, 11 2014. Mr. Leo Hawod is Job supervisor of Thai student. Working hours started at 8 am to 5 pm(Full time), Monday to Friday. Most of activities were done in the field, green house and in the plant pathology laboratory.

 Lecture - Rice production - Vegetable production - Weed of Rice - Important diseases of Rice - Insect pests managements

 Field trial Activities - Trials design and lay out - Experimental design and lay-out - Application - Assessment [Insecticide trials in kale (White fly)] 31

- Insecticide trials in kale (White fly). - Seed germination testing (seed treatment). - Data entry.  Project(damping off)

Table : Schedule of activities at the Agronomic Development Southeast Asia Field Research Center of Bayer CropScience in Masiit, Calauan, Laguna, Philippines from March, 24 to July, 11 2014. Activities Date  Arrived to Philippines 23 May  Orientation in the office, field, greenhouse and 24-25 May laboratories  Learning : side dressing, trellising, pruning at field 26-27 May  Introduce my self to everyone in office 28 May  Assignment : weight and select peanut for herbicide trials  Observe : seedling preparation for rice, soaking, 31 March - 4 incubate, wed bed, dapog April  Assignment : pricking, planting, counting rice seeds, cutting - soaking - incubating bitter gourd seeds, pruning and trellising bitter gourd  Lecture : white fly trial, compound, assessment for white 8 April fly trial  Observe : compound preparation, mixing compound, spray compound  Observe : seed treatment trial for rice 10 April  Lecture : AD-SEA, weed and herbicide, Plant diseases 11 April  Lecture : rice production, crop production, trials in AD- 14 - 18 April SEA, insecticide 32

 Assignment : transplanting weed in tray for herbicide trial  Planning : Project 21 - 25 April  Assignment : entry data for insecticide trials  Learning : use autoclave(not automatic)  Discuss : project 28 April - 2 May  Inform to my advisor about the title of project  Assignment : entry data for insecticide trials 5 - 7 May  Prepare culture media 19 - 23 May  Transfer pathogen to media 26 - 30 May  Measure diameter of pathogen 31 - 9 June  Transfer pathogen to inoculum media and pathogenicity 10 June testing  Present of progressive report to professor 13 June  Transfer pathogen for Inoculation (mass production) 17 June  Assessment for pathogenicity testing 24 June  Assessment for Inoculation (mass production) 1 July  Gather data for internship report 2July - 8 July  Present internship activities 10 July  Back to Thailand 11 July

Lecture  Rice production  Land Preparation 1. Plowing 1 month before transplanting (1 time) 2. Harrowing (2 times) 3. Final harrowing day before transplanting & Leveling before transplanting

Figure Harrowing of field using hand tractor.  Cultivation Methods 1. Direct Sown Rice (DSR) : Seeds are broadcast sown directly in the field. - Wet land - Dry land 2. Transplanting Rice (TPR) : Seedling are raised in the nursery bed and planting in the field by hand or machine.

Table: Compare Direct Sown Rice and Transplanting Rice Detail DSR TPR Management (harvesting, weed) Hard Easy Cost Low High Yield Low High Planting Easy Hard

 Seed Preparation

Figure Seed preparation for rice  Seedling Preparation Table: Compare seed preparation two method, wet bed and dapog. Detail Wet bed Dapog Age of seedling for transplanting 20-25 days 9-12 days Nursery bed size 300-500 15-25 ( m²/hectare) Kg. seed/hectare 35-45 45-65 Advantage -Seedlings grow rapidly -Less area is needed to -Seedlings easy to uproot raise seedlings -Strong seedlings -Seedlings are raised faster by this method

Figure. Picture showed seed preparation 2 method, wet bed and dapog.

 Transplanting Rice age 20-25 days(wed bed) or 9-12 days(dapog) Remove seedling from the nursery, make leveling before transplanting and planting rice in the field(2-3 seedling/hole).

Figure. Step of rice transplanting.  Water Management Water levels should be around 3 cm initially, and gradually increase to 5−10 cm. and remain there until the field is drained 7−10 days before harvest.  Nutrient Management 1. Apply fertilizer(NPK) 30 kg./ha. 1 day before transplanting or seedling. 2. After 12-15 days apply fertilizer(N) 30 kg./ha. 3. After 20-30 days apply fertilizer(N) 40 kg./ha. 4. After 45-50 days apply fertilizer(N) 30 kg./ha.  Harvesting 1. Hand 2. Machine  Threshing 1. Threshing by foot is a great way to make rice without losing quality. 2. Grains easy to remove from the stalks by machine.

Figure. a. threshing by foot b. machine 36

 Vegetable production  Land Preparation 1. Plowing 1 month before transplanting (1 time) 2. Harrowing (2 times) 3. Final harrowing day before transplanting & Leveling before transplanting  Nursery Management Seedling Media 1. Vermicompost 2. cabonized rice hull 3. coconut coir dust 4. soil garden is optional.

Figure. Ingredient of seedling media planting.

Figure. Step of planting for seedling 37

Hardening In nursery, seedling must be exposed to direct sunlight 3-5 days before transplanting and during this period, watering should be minimized.

Figure. Hardening of seedling Water management 1. Furrow irrigation : furrow irrigation is suitable for many crops, especially row crops. 2. Sprinkler system : automatic sprinkler is effective for watering vegetables planted in a large field the overhead sprinkler but Not really for irrigation for increate humidity to induce disease 3. Perfo - rain system : This system consists of a rubber hose perforated with tiny pores that leak water. You can lay the hose between rows or curve it around plants. weed seed germination will be high. Less labor 4. The watering can : Easy to use the watering can you can focus directly to water plant but slowly and more labor

Figure. Water management

 Weed of Rice  Weed A weed is a plant undesirable. Crops up naturally in plants in the garden. Often damage to economic plants such as competing for food.  Problems caused by weed - Weed increase production costs, higher labor or input costs. - Weed reduce grain quality and price. - Contaminate harvest with undesired weed seed. - Provide refuges for vectors of pests and diseases.  Weed Types of rice Grasses Monocotyledonous - Usually Herbaceous - Narrow leaves growing from the base - Stems are usually cylindrical - Forms alternate leaves

Broadleaves Monocotyledonous - Herbaceous - Superficially resembles grasses - Stems with triangular cross-sections - Leaves are spirally arranged Sedges - Dicots (two-seed leaves) - Herbaceous or woody stems - Leaves are “broad”

Figure. Weed Types of rice a. Grasses b. Broadleaves c. Sedges  Red rice or Wild rice - Rice species producing fewer grain than cultivated rice. - Introduced after TPR shift to DSR. - 3 main types of weedy rice: Jumping rice 80%, Awn rice 10% and Red rice 10%. - Infested areas are estimated at 300,000 in Thailand. - Rice harvesting machines and rice seeds are the main spreading factor of the problem. - Perennial common wild (Oryza rufipogon) – characterized by red percarp and early shattering. - Wild rice is only found in natural habitat and not in man-made ecosystem. - Gene flow between common wild rice and cultivated rice is very high. 40

 Different weed control method 1. Hand-weeding spent 120 hour/ha 2. Mechanical spent 50-70 hour/ha 3. Chemical spent 4 hour/ha  The work of Herbicides - The herbicide molecule inhibits one step within a biochemical pathway which eventually result in plant death for the weed. - Selectivity mechanisms keep the crop safe. - Generally, many processes must occur before the herbicide reaches the site of action. - Understanding the processes help to make sense of what can and does occur in the field.  Herbicide Resistance - Resistance is naturally occurring and inheritable ability to survive a herbicide. - Herbicide resistance naturally exist at very low levels in the overall weed population. - Repeated use of herbicides selects resistant population determines. - The selection pressure and the initial resistant population determines the speed and extent of development of herbicide resistance problems. - Herbicides do not cause resistance.  Important diseases of rice Disease A disease is impairment of the normal physiological functioning of a plant or plant part. It interferes with its normal structure, condition, economic value. Cause Fungi - Rice blast caused by by Pyricularia oryzae - Sheath blight caused by Rhizoctonia solani - Dirty panicle disease caused by multiple fungus species - Narrow brown leaf spot caused byCercospora oryzae 41

- Brow spot caused by Helminthosporium oryzae Bacteria - Bacterial leaf blight caused by Xanthomonas oryzae pv. Oryzae - Bacterial leaf streak caused by Xanthomonas oryzae pv. Oryzicola Virus - Rice tungro virus caused by Rice Tungro Bacilliform Virus (RTBV) Rice Tungro Spherical Virus (RTSV) vector is green rice leafhopper. Symptoms of plant disease - Blight(a rapid death of foliage, blossom, or the whole plant) - Spots (brown, narrow,) and stem canker (localized death of an organ or plants) - Wilt - Galls, tumors and witches’ broom (overgrowth) Management plant disease - Sanitation - Host plant resistance - Crop rotation - Cultural practices - Fungicides - Biological control - Improved plant health and nutrition  Insect pest management Mod of action : Animal poisons - Contract poison - Systemic poison - Attractants Pheromones Baits - Repellants 42

Cause Insecticide resistance Insecticide resistance is a reduction in the ability of an insecticide in achieving the desired control. This is reflected in repeated failure of an insecticide?s expected level of control of insects when used according to the product label recommendations and where causes have been eliminated. There are several ways insects can become resistant to the products: Behavioral resistance Resistant insects may detect or recognize a danger and avoid the toxin. This mechanism of resistance has been reported for several classes of insecticides, including organochlorines, organophosphates, carbamates (none of which we will use), and pyrethroids. Insects may simply stop feeding if they come across certain insecticides, or leave the area where spraying occurred. Maxforce has had to change the formulation several times in the last two decades for this reason. Penetration resistance Resistant insects may absorb the toxin more slowly than susceptible insects. Penetration resistance occurs when the insect?s outer cuticle develops barriers which can slow absorption of the chemicals into their bodies. This can protect insects from a wide range of insecticides. Penetration resistance is frequently present along with other forms of resistance, and reduced penetration intensifies the effects of those other mechanisms. This is currently happening with bed bugs and the use of pyrethroids. Metabolic resistance Resistant insects may detoxify or destroy the toxin faster than susceptible insects, or quickly their bodies of the toxic molecules. Metabolic resistance is the most common mechanism and often presents the greatest challenge. Insects use their internal enzyme systems to break down insecticides. Resistant strains may possess higher levels or more efficient forms of these enzymes. In addition to being more efficient, these enzyme systems also may have a broad spectrum of activity (i.e., they can degrade many different insecticides). Cockroaches are known to be able to metabolize pyrethrin when a synergist is not present. Altered target-site resistance The site where the toxin usually binds in the insect becomes modified to reduce the insecticide's effects. This is the second most common mechanism of resistance. 43

Insect pests Whorl maggot :Hydrellia philippina Ferino Damage to rice - feeding damage causes yellow spots, white or transparent patches and pinholes Symptom of rice - Feeding damage causes yellow spots, white or transparent patches and pinholes - White or transparent patches - Pinholes - Damaged leaves easily break from the wind - Somewhat distorted leaves - Clear or yellow spots on inner margins of emerging leaves - Stunting - Few tillers Green leafhopper: Nephotettix sp. Damage to rice - cause direct damage to the rice plant - feed on rice by sucking the plant sap - plugging the vascular bundles with stylet sheaths - symptoms of various viral diseases Symptom of rice - Transmits virus diseases such as tungro, yellow dwarf, yellow-orange leaf, and transitory yellowing. - Plant stunted and reduced vigor. - Number of productive tillers reduced - Withering or complete plant drying

Rice leaf folder: Cnaphalocrocis medinalis Damage to rice - Larva removes the leaf tissues. - folds a leaf blade together and glues it with silk strandsใ - Feeds inside the folded leaf creating longitudinal white and transparent streaks on the blade. Symptom of rice - Longitudinal and transparent whitish streaks on damaged leaves - Tubular folded leaves - Leaf tips sometimes fastened to the basal part of leaf - Heavily infested fields appear scorched with many folded leaves Brown plant hopper : Nilaparvata lugens Damage to rice - hopperburn - ovipositional marks exposing the plant to fungal and bacterial infections - ragged stunt or grassy stunt virus disease plant may be observed Symptom of rice - Hopperburn or yellowing, browning and drying of plant - Ovipositional marks exposing the plant to fungal and bacterial infections - Presence of honeydew and sooty molds in the bases of areas infected - Ragged stunt or grassy stunt virus disease plant may be observed Black bug : Scotinophara coarctata Damage to rice - sap removal by adults and nymphs - plant stunting - reduced tiller number - weakening of plants and preventing them from producing seeds - formation of whiteheads 45

Symptom of rice - Deadheart - Reddish brown or yellowing of plants - Chlorotic lesions on leaves - Decreased tillering - Bugburn - Stunting of plant - Stunted panicles, no panicles, or incompletely exerted panicles, and unfilled spikelets or whiteheads at booting - Incomplete and unfilled spikelets at crop maturation Rice bug : Leptocorisa sp. Damage to rice - feeding causes empty or small grains during the milking stage - feeding causes deformed or spotty grains at the soft or hard dough stage - grains become dark Symptom of rice - Small or shrivelled grains - Deformed or spotty grains - Empty grains - Erect panicles 1.2.2 Field Trials Activities Trials design and lay out Experimental design - Completely Randomized Design(CRD) - Randomized Completely Block Design(RCBD) ( the most commonly used design in ADSEA trials). - Split plot 46

About trials - Trials normally done : compare the test products with reference products standards. - The comparison is mostly on efficacy, plant compatibility and possibly yield effect. - There are usually mono-factorial trials, sometime use multi-factorial trials. Lay-out - Plot in general are rectangular or square in shape and of the same size in one trial. - Plot can be constructed with barrier or canals in between. - This is done to identify the individual plots, water maintence (herbicide) maintain the pest inside the plots.  Application Compound preparation The steps of compound preparation by Fig.

Figure. The steps of compound preparation

Mixing Preparation of the spray solution in the sprayer tank (prepared in a separate container).

Figure. Step of compound mixing for spray  Assessment [Insecticide trials in kale (White fly)] - Count a number of dead white fly by next day after application. - Count white fly is dead in front of leaf, the sixth leaf of kale

Notice White fly is alive will be not spread wings but when it is dead will be spread wings and fall on in front of another leaf, We estimated when we count not exactly amount of white fly.

Figure. White fly is alive and dead.

 Insecticide trials in kale (White fly) * Big plots whiteflies 6 day after application

Figure. Show Phytotoxicity in kale on 5 treatment (IA14FPIGUKOE1D) * Small plots whiteflies 6 day after application

Figure. Show Phytotoxicity in kale on 5 treatment (IA14FPIGUKOE1D) 49

Result From Insecticide trial in kale found treatment 2,3,4 and 5 is Richard appear Phytoxicity but treatment 6 is Ivy result is no Phytoxicity. So should use Ivy because no affect to kale.  Seed germination testing (seed treatment) Methodology 1. Prepare materials (soil, rice seed, pots, trays, compound, water) 2. Mix seed treatment in rice seed. 3. Planting rice to pots (5 row/pot) and then cover soil and not cover soil. 4. Keep rice pots in tray and then add water to trays. Assessments done by check of seedling growth (height, crop stand). Result Table: Result of Seed germination testing(seed treatment) Datail Don’t cover soil Cover soil Phytotoxicity Treatments 4, 5, 6 Treatments 4, 5, 6 Advantage -seedlings grow fast -no falling seedlings -seedlings no process delay -strong seedlings Disadvantage -falling seedlings -seedlings process delay

Recommendation For in the future should be cover soil because the one with cover soil that not fall and seedling stronger, they will not dry up easily lose moisture.

Figure. Rice growth of seed germination testing (seed treatment) with not caver soil and cover soil. 50

Figure. Phytotoxicity of Rice (not cover soil)

Figure. Phytotoxicity of Rice (cover soil)  Data entry - Entry data in excel.

4.2 Project  Preparation Culture Media 1. Corn Meal Agar (CMA) Ingredient: Distilled water 1 L Agar 15 g Cornmeal Infusion 250 ml Cornmeal Infusion: Distilled water 1L Yellow cornmeal 50g Preparation of Corn Meal Infusion: Add corn meal to distilled water and bring volume to 1L. Gently heat and bring to a boil. Simmer for 10 minutes. Filter through cheesecloth. Return volume to 1L.

Figure. Step of Corn Meal Infusion preparation Preparation of CMA media: Add Agar to 250ml cornmeal infusion and bring volume to 1L with distilled water. Dissolve thoroughly. Gently heat and bring to a boil. Simmer approximately 2 minutes. Transfer to autoclavable flask and autoclave for 15 minutes at 121° C at 15 psi.

Figure. Step of Corn Meal preparation 52

2. Potatoes Dextrose Agar (PDA) Ingredient: Distilled water 1 L Commercial media powder 39 g Preparation: Add PDA powder into the 1L distilled water. Mix thoroughly to dissolve. Transfer to a pan and bring to a boil with continuous stirring. Simmer until clear (approximately 2 minutes). Transfer to autoclavable flask and autoclave for 15 minutes at 121° C at 15 psi.

Figure. Step of Potatoes Dextrose Agar (PDA) preparation. 3. Rice Dextrose Agar (RDA) Ingredient: Rice grain 160 g Dextrose 15 g Agar 20 g Distilled water 1 L

Preparation Add Rice grain to distilled water and bring volume to 1 L. Gently heat and bring to a boil. Simmer for 20 minutes. Filter through cheesecloth. Return volume to 1L. Volume then add Dextrose and Agar.

Figure. Step of Rice Dextrose Agar (RDA) preparation.  Pathogenicity testing Agar Block - Used ¼ piece per one pot or 1 plate per 4 pots.

Figure. Step of pathogenicity testing by Agar Block

Pulpy Agar by blender - Used agar 1 plate per 300 ml for 4 pots.

Figure. Step of pathogenicity testing by Pulpy Agar

 Preparation of inoculum media(mass production)  Rice hull and Rice grain Rice hull 1800 g Rice grain 300 g Districted water 2 L

Figure. Step of transfer inoculum to trays (Rice hull and Rice grain )  Sawdust and Sorghum Sawdust 1800 g Sorghum 400 g Distilled water 2 L

Figure. Step of transfer inoculum to trays (Sawdust and Sorghum)

4.3 Living in Philippines This is my experience in Philippines for 16 weeks, that it is so long time enough for make me can sing a Philippines song. I am Thai student, I have to apprentice at Bayer CropScience (AD- SEA) in Philippines. I wanted to do my internship abroad to experience the international company and to learning, improve and develop skills. One of my main goals is to improve my English. Personally, I know Philippines not much that make me excited to go to Philippines. This is my first time to stay other place without family. In Philippines, my daily life not hard but not easy because I can’t to go everywhere that I want to go, no car on motorcycle that I have to ride the jeepney every day to go to work. I can eat Philipino food. I went to travel with company and friends, It’s was so much fun. So I will describe about Philippines during my internship period based on my experience.

Languages Filipino is based on Tagalog, English is generally used for educational, governmental and commercial purposes. It is common to hear Filipinos use a mixture English and Filipino words in their everyday conversations. Meeting customs - A handshake, with a welcoming smile, is the standard greeting. - Close female friends may hug and kiss when they meet and say goodbye. - Pagmamano, the practice of respect by asking for an elder’s hand and touch one’s forehead while bowing.

Food For Philipino food, personally I think too salty, too sweet, too sour. They love cheese and love fast food, Jollibee is the most famous. My favorite Philipino food is sinigang, sisig with eeg, lechon, chrispy pata, squid, chicken in maung inasal resterant, longanisa, barbecue and my favorite Philipino dessert is halo-halo, turon, malunggay pandesal. Transportation - Tricycles (motorcycles with a sidecar attached) are especially for short trips)The prices depend on the distance. 56

- Jeepneys are used for short and long distances from town to town. - Busses are classified into not air-conditioned and air-conditioned busses. - Pumpboats(bancas) Most of the time Filipinos will take the boat to travel from one island to another island in the Philippines. The traditional type is the bancas. Pumpboats are motorized bancas, outrigger boats. The pump-boats are normally used for the shorter distances. Traveling - Baguio - Escudero village - Enchanced kingdom - Sn.pablo city - Puerto galera