AFACI Program Workshop on Food Crops Technology Dissemination of Virus-free Seed Potato Production using Hydroponic Production systems in Thailand Orathai Wongmetha Chiang Mai Royal Agricultural Research Center (CMRARC) Horticultural Research Institute (HRI) Department of Agriculture (DOA), Thailand Presentation at Conference room of Angkor Paradise Hotel: 20-24 September 2016, Siem Reap, Cambodia PotatoProductionDOA 19Dec13

(https://www.google.co.th, 2013) Content  Introduction  Potato in Thailand  Main Activities  Results and Discussion  Suggestions for project  Project Impacts & Recommendations  Future Plan (https://www.google.co.th, 2013) PotatoProductionDOA_KOPIA 1Nov13

Thailand Background Flag Emblem Thailand, officially the Kingdom of Thailand, formerly known as Siam. Bangkok is capital of Thailand. Thailand is the world's 51st-largest country. An area of 513,000 km2 (198,000 sq mi). Bhumibol Adulyadej (Rama IX) is the king of Thailand. Flag Deficiency in vitamin A is a major problem in 37 countries, mostly in Southeast Asia and Africa, with 250,000-500,000 people, mostly children, becoming partially or totally blind yearly. More than 2 billion people worldwide are anemic or have insufficient iron intake. Green leafy vegetables are rich in both vitamin A Acedoand iron. Emblem (Wikipedia, 2013) Postharvest_Tomato 19Apr13

Geographically Thailand Thailand is located in the center of peninsular Southeast Asia, 76 provinces. Agricultural regions divided into 5 regions, namely: 1) North 2) Central 3) East 4) Northeast 5) South (Sripinta et al., 2008) 4 TropicalFruitThai_Orathai Nov09

Climate Seasons Months Temperature (oC) %RH Monsoon Hot and dry Feb - May 34 75 - Rainy Jun - Sep 29 87 Southwest Cooler and dry Oct - Jan 20 - 32 Northeast 5 Postharvest_Tomato 19Apr13

Potato in Thailand Potato in Thailand is an important in the north and northeastern part of Thailand because potato chip consumption is an increasing rapidly. The potato farmer income: 2,679-4,464 $US/hec (15,000-25,000 baht/rai). The entrepreneur required more potato production for processing and farmer also required basic seed production (G1) and certified seed production (G2-G3) to support them. PotatoProductionDOA 19Dec13

Potato production in Thailand Table 1. Thailand production of potato in 2011-2016 (Office of Agricultural Economics, 2012). Year Processing Fresh consumption Area (ha) production (tons) Yield (kg/ha) 2011 8,741.6 124,808 14,275 880 14,863 16,887.5 2012 8,447.4 126,044 14,918.8 854.1 13,116 15,356.3 2013 6,589.9 96,096 14,581.3 646.4 9,064 14,025 2014 5,684.6 89,925 15,937.5 644 9,790 15,200 2015 7,158.7 115,141 14,206.3 672.3 10,522 15,650 2016 6,350.7 129,760 20,431.3 660.3 12,543 18,993.8 %Growth -7.72 -4.87 3.09 -7.88 -9.36 -2.14 PotatoProductionDOA 19Dec13

Unit: tones Year Processing Fresh consumption 22 September 2018 Table 2. Quantity of seed potato imported in 2011-2015. Unit: tones Year Processing Fresh consumption Quota Imported 2011 6,062.65 4,659.15 262.7 262.5 2012 8,135.28 3,846.03 237.7 2013 7,967.53 6,898 237.5 2014 10,422.95 6,824.08 237.6 2015 6,124.95 1,421 %Growth 2.72 -16.49 -2.00 -1.99 ศกล.ชม กวก. เทคโนโลยีการผลิตมันฝรั่ง กรมวิชาการเกษตร

Potato Varieties in Thailand 22 September 2018 Spunta for consumption Atlantic for processing ศกล.ชม กวก. เทคโนโลยีการผลิตมันฝรั่ง กรมวิชาการเกษตร

Problem of potato production in Thailand DOA produced pre-basic seed (G0) and basic seed (G1) for farmer but it not adequate. The enterprises of potato chip are imported certified seed >>> high unit cost and difficult for a management system. Farmers are stored low quality (virus, bacteria, scab etc.) certified seed potato for planting in next season. The effect to the quantity and quality of potato, by reduce the total yield and also reduce the marketable quality.

How to solve the problem To find technology or protocol are produced high yields, more easy, rapid, flexible, more security for the phytosanitary quality and at lesser cost through research and development. The purpose of this study to find the new technology of propagate potato plantlets by tissue culture technique in solid media or microtubers production by bioreactor and pre-basic seed production under aeroponic system that improve quality and quantity and lower costs.

Main activity Location of experimental site: Chiang Mai Royal Agricultural Research Centre (CMRARC), MaeWang, ChiangMai, Thailand.   The climate condition: At 1,250 m above sea level, a cooler, dry season, lasts from November to February, with an average temperatures ranging from 13.9oC, 11.5 mm rain fall level and 84.3% relative humidity (RH).

เอเฟรม 7 คน 2 หลัง และ เอเฟรม 4 คน 1 หลัง บ้าน VIP 3 หลัง บ้านพักรับรอง หลังใหญ่ ร้านกาแฟ สโมสร/โรงครัว เอเฟรม 7 คน 2 หลัง และ เอเฟรม 4 คน 1 หลัง อาคารสำนักงาน Meeting room. ทางเข้า Chiang Mai Royal Agricultural Research Centre (CMRARC), MaeWang, ChiangMai

Atlantic Variety Characteristics of selected cultivar: Early late maturity (90-120 days after planting) Round shape Shallow eye White flesh Light tan skin Consistently 4 to 9 cm diameter The yield 15.175 tones/hec (2,428 kg/rai) High solid gross >17.5%

Main activity Time schedule Activity and process Year 1: (Jan 2017-Dec 2017) Establishment and research of aseptic techniques (Jan-Apr 17) - Prepare material for micropropagation plantlets.   2. Induction and multiplication of shoots in vitro (May-Aug 17) - To find out the suitable medium for multiplication of potato shoots and roots for produce potato plantlet production form in vitro micropropagation technique. Including monitoring virus infection of plantlet. 3. Mother plant production in net house (Jul-Sep 17) - Tissue plantlets are transplant to a tray, soil media in net-house. The Stem cutting of mother plant is transplant in aeroponic system in 30-45 days after planting. To find out the suitable fertilizer for produce mother plant in soil media. Including monitoring virus infection of mother plant.

Time schedule Activity and process Year 2: (Jan 2018-Dec 2018) 4. Microtubers production in vitro (Jul-Apr 18) - Potato plantlets are induce microtubers by using tissue culture method in solid media or bioreactor system in liquid media under aseptic condition in laboratory. In additional, to find out the suitable medium for microtuber production in both media.   5. Pre-basic seed production (G0) 5.1 G0 from stem cutting in Aeroponic system (Nov17-Apr 18) - Stem cutting of tissue plantlet are inserted to a spongy in holes of form plastic sheets and connected with aeroponic system in net-house. To find out the suitable fertilizer or technique for minituber production in aeroponic system.  Including monitoring virus infection of G0 production. 6. Publication paper or leaflet or book (Apr-Sep 18)

Time schedule Activity and process Year 3: (Jan 2019-Dec 2019) 5.2 G0 from microtuber in vitro in soil media (Nov 18-Apr 19) - Microtubers from in vitro are transplant to a tray, soil media in net house. To find out the suitable fertilizer or technique for G0 production. Including monitoring virus infection. 6. Basic seed production (G1) in the field (Nov 18-Feb 19) - G1 production from G0 minituber in the research field or farmer field (field demonstration). 7. Publication paper or leaflet or book (Jul-Dec 19) 8. Training course for enterprise, student, farmer (Apr-Sep 18)

Time schedule Activity and process Year 3: (Jan 2019-Dec 2019) 7. G1 and G2 production in the field (Nov18-Apr 19) 7.1 G1 production from G0 microtuber in the research field. To find out the suitable technique for G1 production. Including monitoring virus infection. 7.2 G2 production from G1 minituber in the research field or farmer field (field demonstration). To find out the suitable technique for G2 production. Including monitoring virus infection. Year 4: (Jan 2020-Dec 2020)  8. Potato production for processing (Nov19-March 20) - Potato production for processing from G1 and G2 minituber in the research field or farmer field (field demonstration).

Protocols of Seed Potato Production in Thailand 22 September 2018 Protocols of Seed Potato Production in Thailand Year 1 การผลิตต้นอ่อนปลอดเชื้อ (Pathogen free in vitro plantlets) Year 1 Year 2 การเพาะเลี้ยงเนื้อเยื่อในอาหารแข็ง (Tissue culture) การใช้ระบบไบโอรีแอคเตอร์จมชั่วคราว (Temporary Immersion Bioreactor ; TIB) ห้องปฏิบัติการ (Lab) โรงเรือนกันแมลง (Net house) Year 1 การผลิตต้นแม่พันธุ์ (Mother plants production) วัสดุปลูก (Soil Media) ระบบไฮโดรโปรนิค (Hydroponic) Determine - Virus - bacteria - late blight การขยายต้นปักชำ (Stem cuttings production) โรงเรือนกันแมลง (Net house) โรงเรือนกันแมลง (Net house) Year 1/2 การผลิตหัวพันธุ์หลัก G0 (Pre-basic seed production) ระบบแอโรโปนิค (Aeroponic) โรงเรือนกันแมลง (Net house) การผลิตหัวพันธุ์ขยาย G1 Basic seed production Year 2/3 แปลงผลิตหัวพันธุ์ (Field Seed production) Year 4/5 Year 3/4 ผลิตหัวพันธุ์รับรอง G2 (Certified seed production) การผลิตมันฝรั่งส่งโรงงาน (Potato production for processing๗ ศกล.ชม กวก. เทคโนโลยีการผลิตมันฝรั่ง กรมวิชาการเกษตร

1. Pathogen fee In vitro plantlets

1.1 Tissue culture of CMRARC Year 1 Prepare MS media PotatoProductionDOA 19Dec13

3 weeks–1 month, transplant or sub culture Transfer room Sub Culture 1 week 3 weeks–1 month, transplant or sub culture 2 weeks Culture room PotatoProductionDOA 19Dec13

ศกล.ชม กวก.

1.2 Tissue culture by using Temporary Immersion bioreactor (TIB) 22 September 2018 1.2 Tissue culture by using Temporary Immersion bioreactor (TIB) Year 2 ศกล.ชม กวก. เทคโนโลยีการผลิตมันฝรั่ง กรมวิชาการเกษตร

Prepare bottles 24 oz. and equipments 22 September 2018 Prepare bottles 24 oz. and equipments Prepare 300 ml of MS media Autuclave in 20 min ศกล.ชม กวก. เทคโนโลยีการผลิตมันฝรั่ง กรมวิชาการเกษตร

Prepare TIB equipments in laminar flow clean bench Wait until cool Single node cutting in bottle and connect with TIB system, Feed media 2 times, 2 min/time ศกล.ชม กวก.

1 week 3 weeks 10x10 cm 1.5 months after planting

2. Mother Plants Production 3 weeks–1 month, transplant Year 1 PotatoProductionDOA 19Dec13

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Stem cuttings production 4. 2 leaves and 2-3 nodes stem cutting 1. Mother plant 3. Cut potato stem 5. Soak in Trichoderma + Chitosan 5 min 2. Soak knife in alcohol 50% Year 2 6. Transplant in media or aeroponic PotatoProductionDOA_KOPIA 1Nov13

3.1 G0 production in Aeroponic Year 2 ศกล.ชม กวก.

ศกล.ชม กวก.

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3.2 G0 production in soil media Year 2 PotatoProductionDOA 19Dec13

4. Basic Seed Production (G1-G2) Year 3 PotatoProductionDOA 19Dec13

Important Diseases/ Pests in Thailand Fungus Bacterial Virus & Nematode Pests Late blight (Phytophthora infestans) -Bacterial wilt (Ralstonia solanacearum), Mosaics (PVX, PVY, PYS) and Potato leaf roll virus (PLRV) Aphid (Myzus persicae Sulzer and Ahis gossypii) Early blight (Alternaria solani) Black leg and soft rot (Erwinia carotovora and E. chrysanthemi)   Root-knot nematode (Meloidogyne incognita) Thrips (Thrips palmi, Scirtothrips dorsalis) -Downy mildews (Peronospora destructor) Ring rot (Clavibacter michiganensis) Potato tuber moth (Phthorimaea operculella) Fusarium dry rot and wilt (Fusarium spp.) Common scab (Streptomyces scabies) Leaf miner flies (Liriomyza brassicae) Stem rot (Sclerotium rolfsii) Cutworm (Spodoptera exigua, Spodoptera litura, Agrotis ipsilon Stem canker and black scurf (Rhizoctonia solani)

Late blight detection CIP scale value Blight (%) Symptoms mean limits 1 - No late blight observed. 2 2.5 Traces < 5 - Late blight present. Maximum 10 lesions per plant. 3 10 5 < 15 -Plants look healthy, but lesions are easily seen at closer distance. Maximum foliage area affected by lesions or destroyed corresponds to no more than 20 leaflets. 4 25 15 < 35 - Late blight easily seen on most plants. About 25 % of foliage is covered with lesions or destroyed. 5 50 35 < 65 - Plot looks green; however, all plants are affected. Lower leaves are dead. About half the foliage area is destroyed. 6 75 65 < 85 - Plot looks green with brown flecks. About 75 % of each plant is affected. Leaves of the lower half of plants are destroyed. 7 90 85 < 95 - Plot neither predominantly green nor brown. Only top leaves are green. Many stems have large lesions. 8 97.5 95 < 100 - Plot is brown-colored. A few top leaves still have some green areas. Most stems have lesions or are dead. 9 100 - All leaves and stems dead. (Henfling, 1987)

Virus Glift kit of mother plants

Determined every 1 month

2-3 bands = virus detection

Bacteria Test kit of G0-G1

2 bands = bacteria detection

Project Impacts & Recommendations The high temperature and unsuitable fertilizer ratio are affected to impact crops. Potato virus are seed-borne, reduce mass production of seed potato. The technique of in vitro micropropagation in potato cv. Atlantic will be done. The technique of induce microtuber in solid and liquid media should be develop. The minituber production under aeroponic system should be improves technique, lower costs and increase the farmers’ income. Aeroponics is a soilless method for producing pre basic potato seed. The method can produce higher yields, easier, rapid, flexible and more security for the phytosanitary quality.

Future plan Activities 2017 2018 2019 1-3 4-6 7-9 10-12   1-3 4-6 7-9 10-12 1. Project Activities Preparation 2. Preparation of material 3. Establishment and Research of Aseptic Techniques 4. Tissue plantlets production/ Microtubers production 5. Mother plant production in net house 6. G0 production from stem cutting in aeroponic system and microtuber in vitro culture in soil media under net house

Activities 2017 2018 2019   1-3 4-6 7-9 10-12 7. G1 production in the field 8. Certified seed production (G2) in the field 9. Potato production for processing 10. Virus and disease elimination 11. Publication paper or leaflet or book etc. 12. Training/Visiting scientists/Workshop/Conference meeting (1-2 weeks) 13. Training course for enterprise, student, farmer 14. Report Preparation

Thank You FOR YOUR ATTENTION & SAWASDEE PotatoProductionDOA 19Dec13

Results and Discussion

Suggestions for project