Potato Nematode Research: with Special Reference to Potato-Early Die, Corky Ringspot and Soil Enzymes
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POTATO NEMATODE RESEARCH: WITH SPECIAL REFERENCE TO POTATO-EARLY DIE, CORKY RINGSPOT AND SOIL ENZYMES By Loren George Wernette A Thesis Submitted to Michigan State University In partial fulfillment of the requirements For the degree of Masters of Science Entomology 2011 Abstract POTATO NEMATODE RESEARCH: WITH SPECIAL REFERENCE TO POTATO-EARLY DIE, CORKY RINGSPOT AND SOIL ENZYMES By Loren George Wernette Potato production faces numerous challenges three of these include Potato-Early Die (PED), Corky Ringspot Disease of Potato (CRSD), and soil quality issues. This thesis consists of three chapters: 1) Impact of Alfalfa on Soil-Borne Enzymes in Potato Systems. 2) Vertical Distribution of Paratrichodorus pachydermus, Pratylenchus penetrans (Nematoda), and Verticillium dahliae (Mycota) in Michigan Potato Systems. 3) Soil Fumigation Guide. Soil quality was evaluated comparing soil enzyme activities associated with fields that had been in alfalfa for ten years in two year potato rotations with corn. We found that the alfalfa rotation had higher enzyme activities for phosphatase and tyrosine aminopeptidase. We also found that the mineralization rate of carbon was significantly higher in the alfalfa system, compared to the potato rotation. The nematode pathogens for CRSD and PED were evaluated to determine the depth at which the highest population density of Pratylenchus penetrans, Paratrichodorus pachydermus, and Verticillium dahliae were found in a potato system. We found that the majority of P. pachydermus was found below, a 30 cm soil depth in September. We also found that P. penetrans was found most commonly in the upper 30 cm of the soil. When targeting the virus vector, P. pachydermus. Fumigant and non-fumigant nematicides were tested. As the amount of active ingredient of the chemicals increased, however, the percent of CRSD symptom expression in the tubers decreased. We also showed that there was no increase in symptom expression during tubers storage. i Dedication I would like to dedicate this thesis to my beautiful fiancé Meghan who has loved and supported me throughout this process. iii Acknowledgements I would like to thank my major professor Dr. George W. Bird for all his time and guidance I could not have completed this research without this help. I would also like to thank my committee members Dr. William Kirk and Dr. Chris Dizonzo for all of the help they have given me during the research and writing process. I would also like to thank Dr. Stuart Grandy, and Kyle Wickings and their lab for assisting with soil enzymes and carbon mineralization analysis. I would not have been able to accomplish what I have without the help of John Davenport, Rob Schafer, and Chris Long. They helped me with everything from chemical applications harvesting and storage sampling as well as just being good friends. Without Walther’s Farms, especially KarI Richie, I would not have had the field sites to do a good portion of my research. I would like to thank Mark Otto for his guidance and understanding as I progressed through this process. I look forward to working with you for many years. iv Table of Contents List of Tables……………………………………………………………………………………………………………………..……vi. List of Figures…………………………………………………………………………………………………………….………….viii. Chapter I. Impact of Alfalfa on Soil-Borne Enzymes in Potato Systems……………………………………….1 1. Introduction……………………………………………………………………………………………………………..2 2. Methods…………………………………………………………………………………………………………………..5 3. Results……………………………………………………………………………………………………………………10 4. Discussion………………………………………………………………………………………………………………12 5. Conclusions…………………………………………………………………………………………………………….19 Chapter II. Vertical Distribution of Paratrichodorus pachydermus, Pratylenchus penetrans (Nematoda), and Verticillium dahliae (Mycota) in Michigan Potato Systems……………………………21 1. Abstract………………………………………………………………………………………………………………….21 2. Introduction……………………………………………………………………………………………………………22 3. Methods…………………………………………………………………………………………………………………26 4. Results……………………………………………………………………………………………………………………29 5. Discussion………………………………………………………………………………………………………………30 6. Conclusions…………………………………………………………………………………………………………….33 Chapter III. Corky Ringspot Disease of Potato Fumigant and Non-fumigant Nematicide control…………………………………………………………………………………………………………………..…………………35 a. Introduction………………………………………………………………………………………………..35 b. Methods……………………………………………………………………………………………………..36 c. Results………………………………………………………………………………………………………..39 d. Discussion……………………………………………………………………………………………………40 e. Conclusions…………………………………………………………………………………………………42 APPENDIX. Fumigation Field guide…………………………………………………………………………………………64 References……………………………………………………………………………………………………………………………….88 v List of Tables Table 1. Soil nutrient and cropping management data for four fields used The first column indicated which field was sampled one or two and the letter indicates if the cropping system was continuous alfalfa (A) or potato seed corn (B). The second column indicated the zone number that was sampled……………………………………………………………………………………………………….43 Table 2. Activity of four soil enzymes: Phosphatase, Tyrosine aninopeptidase, N-acetyl glucosaminidase and β-1,4-glucosidase (nmol/h/g) associated with two potato management systems in Mecosta County, MI in March 2009………………………………………………………………………..45 Table 3. Activity of four soil enzymes: Phosphatase, Tyrosine aninopeptidase, N-acetyl glucosaminidase and β-1,4-glucosidase (nmol/h/g) associated with two soil types in Mecosta County, MI in March 2009……………………………………………………………………………………………………….46 Table 4. Total and mean daily carbon mineralization (μg C/g soil) associated with two management systems and two soil types in Mecosta County MI, in March 2009…………………….47 Table 5. Absolute nematode population density (nematodes/100 cm3 soil) associated with two management systems and two soil types in Mecosta County MI, in March 2009………………48 Table 6. Absolute nematode population density (nematodes/100 cm3 soil) of two management systems and five trophic groups……………………………………………………………………….49 Table 7. Absolute nematode population density (nematodes/100 cm3 soil) associated with two soil types and five tropic groups in Mecosta County MI in March 2009……………………………50 vi Table 8. Soil characteristics of four fields sampled to determine the vertical distribution of Pratylenchus penetrans, Paratrichodorus pachydermus and Verticillium dahliae under Michigan potato systems…………………………………………………………………………………………………………………………51 Table 9. Absolute and relative population densities associated with Pratylenchus penetrans at multiple soil depths in four Michigan fields……………………………………………………………………………..52 Table 10. Absolute and relative population densities of the vertical distribution of Paratrichodorus pachydermus at three soil depths associated in four potato fields in Michigan………………………………………………………………………………………………………………………………….53 Table 11. Verticillium dahliae propagules per gram of soil associated with two soil depths in two potato fields in MI…………………………………………………………………………………………………………….54 Table 12. Vertical distribution of Paratrichodorus. pachydermus in two fields infected with CRSD in Michigan……………………………………………………………………………………………………………………55 Table 13. 16 Chemical treatments for Corky Ringspot Disease of potato control research in White pigeon MI………………………………………………………………………………………………………………………………………………………….56 Table 14. 7 Chemical treatments for Corky Ringspot Disease of potato control research in White pigeon MI………………………………………………………………………………………………………………………………………………………….57 vii List of Figures Figure 1. Four sites, Fields 1A, 1B, 2A, 2B and their associated management zones used for long-term alfalfa vs. potato-seed corn evaluation of soil enzymes, carbon mineralization and nematode community structure in relation to soil quality. Management zones are numbered arbitrarily for organizational purposes……………………………………………………………………………………..56 Figure 2. Potato tuber yields associated with 16 soil fumigant and non-fumigant nematicide treatments at White Pigeon MI in 2008……………………………………………………………………………………57 Figure 3. CRSD potato tuber symptoms associated with soil fumigant and non-fumigant nematicides treatments in White Pigeon MI in 2008……………………………………………………………….59 Figure 4. 2009 Grade A potato tuber yields associated with nematode control for seven treatments……………………………………………………………………………………………………………………………….61 Figure 5. 2009 TRV symptom expressions associated with seven chemical treatments……………………………………………………………………………………………………………………………….62 Figure 6. A-D. Possible tobacco rattle virus crystal structures seen in symptomatic tubers photos taken under Transmission Electron Microscopy…..……………………………………………………...63 viii ix Chapter I. Impact of Alfalfa on Soil-Borne Enzymes in Potato Systems Abstract Potatoes are commonly grown in two-year rotations with corn, wheat or other annual crops in Michigan. Due to the expense and lack of marketability, long-term cover crops like alfalfa are not usually included in the rotation. Four fields were selected in Mecosta County, MI to evaluate the impact of long-term alfalfa on soil enzymes (β-1,4-glucosidase, N-acetyl glucosaminidase, phosphatase, tyrosine aminopeptidase, and phenol oxidase), carbon mineralization potential and nematode community structure; compared to potato-seed corn rotation. The sites have been used for potato production since the 1970’s. Two of these fields, however, were taken out of potato production and used to grow alfalfa hay for ten years. These fields were returned to potato production in 2009 and 2010. The sites were sampled in 2009, for