Internship report

COOPERATIVE EDUCATION IN AGRICULTURE 2013At The Agronomic Development Southeast Asia Field Research

and Development Center (AD-SED)of Bayer CropScience at Masiit, Calauan, Laguna, Philippines

Ponsawan Khamphasan 543030378-3Faculty of agriculture(Plant pathology) Khon kaen University

Company

The Agronomic Development Southeast Asia Field Research and Development Center (AD-SED)of Bayer CropScience at Masiit, Calauan, Laguna, Philippines.

Khon Kaen Univercity

Inoculation methods and mass production of

damping off caused by pythium

aphanidermatum, rhizoctonia solani and sclerotium rolfsii in lettuce and onion.

Khon Kaen Univercity

• For vegetable production, Damping-off frequently attacks young seedlings of almost all kinds of vegetables.

• Focus on damping-off in lettuce and onion caused by Pythium aphanidermatum,Rhizoctonia solaniSclerotium rolfsii.

• Damping-off caused by several species of fungi.

INTRODUCTIONKhon Kaen Univercity

• Damping-off is the disease affects seeds, seedlings, and roots of all plants.

• Damage is done during germination either before or after emergence.

Symptoms• Pre-emergence

Seeds become soft and mushy, turn dark brown, and decay.• Post-emergence

Stem tissue near the soil line is decayed and weakened, usually causing plants to topple and die.

Khon Kaen Univercity

About damping-off

OBJECTIVE

To produce mass inoculum for inoculation of damping-offin lettuce and onion caused by

• Pythium aphanidermatum,• Rhizoctonia solani• Sclerotium rolfsii

Khon Kaen Univercity

1.Compare Culture mediaMATERAILAL AND METHOD

1.1 Potatoes Dextrose Agar (PDA) Commercial PDA powder 39 gDistilled water1 L

1.3 Rice Dextrose Agar (RDA)Rice grain 160 gDextrose 15 gAgar 20 g

1.2 Corn Meal Agar (CMA)cornmeal 50 gAgar 15 gDistilled water1 L

RDA PDA CMA

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2. Pathogenicity testing

• Pulpy Agar by blender

• Agar Block

MATERAILAL AND METHOD

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MATERAILAL AND METHOD3. Different Inoculation method of mass production

• Rice hull and Rice grainRice hull 1800 g

Rice grain 300 g

Districted water 2 L

• Sawdust and SorghumSawdust 1800 g

Sorghum 400 g

Distilled water 2 L

Rice hull and Rice grain

Sawdust and Sorghum

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ResultTable : Result of diameter mycelia 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 49.8day 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 +

*PDA : Potato Detrose AgarCMA : Corn Meal AgarRDA : Rice agarDensities of mycelia

* = low

** = Medium*** = high

Khon Kaen Univercity

Result

RDA = Rice Dextrose Agar PDA = Potato Dextrose Agar CMA = Corn Meal Agar Under dark condition, room

temperature Picture 10day

Growth of Pythium aphanidermatum in different media

Khon Kaen Univercity

ResultTable: Result of diameter mycelia of Rhizoctonia solani in different culture

media within 10 days of incubation Date/ media diameter(mm.)

PDA CMA RDAday 1 5.5 8.5 5.2day 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)

*PDA : Potato Detrose AgarCMA : Corn Meal AgarRDA : Rice agarDensities of mycelia

* = low

** = Medium*** = high

( ) = namber of sclerotia

Khon Kaen Univercity

Result

RDA = Rice Dextrose Agar PDA = Potato Dextrose Agar CMA = Corn Meal Agar Under dark condition, room

temperature Picture 10day

PDA CMA

RDA

Growth of Rhizoctonia solani in different media

Khon Kaen Univercity

Date/ media diameter(mm.)PDA CMA RDA

day 1 9 12 10day 2 40.8 42.6 32.6day 3 75 85 56day 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)

ResultTable : : Result of diameter mycelia of Sclerotium rolfsii in different culture media within 10 days of incubation

*PDA : Potato Detrose AgarCMA : Corn Meal AgarRDA : Rice agarDensities of mycelia

* = low

** = Medium*** = high

( ) = namber of sclerotia

Khon Kaen Univercity

Result

RDAPDA CMA RDA = Rice Dextrose Agar PDA = Potato Dextrose Agar CMA = Corn Meal Agar Under dark condition, room

temperature Picture 10day

Growth of Sclerotium rolfsii in different media

Khon Kaen Univercity

Result of pathogenicity testing in lettuce

0%

20%

40%

60%

80%

100%

Uninoculated Agar block Pulpy agar

Rate of germination on lettuce of different pathogenicity methods for damping off at greenhouse

pythium Rhizoctonia Sclerotium

Germination

Khon Kaen UnivercityResult of pathogenicity testing in lettuce

UNINOCULATED AGAR BLOCK PULPY AGAR

Pythium aphanidermatum

Rhizoctonia solani

Sclerotium rolfsii

• Result of pathogenicity testingKhon Kaen Univercity

0%10%20%30%40%50%60%70%80%90%

100%

Uninoculated Agar block Pulpy agar

Rate of germination on onion of different pathogenicity methods for damping off at greenhouse

pythium Rhizoctonia Sclerotium

• Result of pathogenicity testing in onion

Germination

Khon Kaen Univercity

• Result of pathogenicity testing

UNINOCULATED AGAR BLOCK PULPY AGAR

Pythium aphanidermatum

Rhizoctonia solani

Sclerotium rolfsii

Khon Kaen Univercity

Result of inoculation method

• Result of inoculation method

0%

20%

40%

60%

80%

100%

uninoculated Rice hull-grain Sawdust-sorghum

Rate of germination on lettuce of different inoculation methods for damping off

pythium Rhizoctonia Sclerotium

Germination

Khon Kaen Univercity

• Result of evaluation of inoculation (mass production)

Pythium aphanidermatum Rhizoctonia solani Sclerotium rolfsii

*SS : Sawdust-sorghum*RGH : Rice hull-grain

RHG SS RHG SS RHG SS

Khon Kaen Univercity

0%

20%

40%

60%

80%

100%

uninoculated Rice hull-grain Sawdust-sorghum

Rate of germination on onion of different inoculation methods for damping off

pythium Rhizoctonia Sclerotium

Germination

• Result of inoculation methodKhon Kaen Univercity

• Result of evaluation of inoculation (mass production)

Pythium aphanidermatum Rhizoctonia solani Sclerotium rolfsii

RHG SS RHG SS RHG SS

*SS : Sawdust-sorghum*RGH : Rice hull-grain

Khon Kaen Univercity

• Conclusion

1. Suitable media• Rhizoctonia solani : PDA • Sclerotium rolfsii : CMA • Pythium aphanidermatum : PDA

3. Suitable inoculation method in mass production of the three pathogen

• Rice hull with rice grain

2. Suitable pathogenetic method for three pathogen in damping-off in lettuce

and onion

• Pulpy agar by blender

Khon Kaen Univercity

Advantage

For company

1.เกิดความร่วมมือทางวิชาการและสร้างความสมัพนัธ์ท่ีดีกบัสถาบนัการศกึษา

2.เป็นการประชาสมัพนัธ์และสร้างภาพพจน์ท่ีดีขององค์กรในด้านการสง่เสริมสนบัสนนุการศกึษาและช่วย

พฒันาบณัฑิตของชาติ

3.มีนกัศกึษาท่ีมีความพร้อมทางด้านวิชาการในระดบัหนึง่ช่วยปฏิบติังาน เป็นระยะเวลานาน 4 เดือน

4.พนกังานประจํา มีผู้ช่วยในการปฏิบติังาน ทําให้มีเวลามากขึน้ท่ีจะปฏิบติัหน้าท่ีอ่ืนท่ีสําคญักวา่

5.มีโอกาสคดัเลือกนกัศกึษาเพ่ือเป็นพนกังานประจําในอนาคต โดยไมจํ่าเป็นต้องมีการทดลองงาน

6.ลดอตัราการเปล่ียนงานและลดต้นทนุการฝึกอบรมพนกังานใหม่

Khon Kaen Univercity

1. Learned about the process of work, responsibility.

2. Learned to adapt to the work environment.

3. Learned to plan for work.

4. Learned to increase skills for understanding and working with people.

5. Learned to interpersonal communication and interaction skills.

6. Learned about real life work.

Advantage

For student

Khon Kaen Univercity

Acknowledgements

Bayer CropScience Southeast Asia Field Research and Development Center Experimental Station at Brgy, Masiit, Calauan, Laguna, Philippines.สหกิจศกึษาคณะเกษตรศาสตร์

มหาวทิยาลยัขอนแก่น

Mr. Leo hawod and everyone at bayerอาจารย์ อนนัต์ วงเจริญ อาจารย์ท่ีปรึกษา

Khon Kaen Univercity

Thank you