Figure 1. The adult Asian citrus psyllid feeding at the characteristic 45 angle.
|
|
- Easter Woods
- 5 years ago
- Views:
Transcription
1 Worldwide, Huanglongbing (HLB), more commonly known as Citrus Greening, is the single most destructive and economically devastating disease of citrus in the history of agriculture. 1 The specific variety of HLB that is currently wreaking havoc in the state of Florida is caused by the phloem-inhabiting, gram-negative bacterium, Candidatus Liberibacter asiaticus (Las). 2 The bacteria requires a vector to be transmitted from one tree to another. The vector responsible for spreading Las is the Asian citrus psyllid (ACP), Diaphorina citri. It is about 4 mm in length, with a brown mottled body and feeds at a characteristic 45 angle(figure 1). Figure 1. The adult Asian citrus psyllid feeding at the characteristic 45 angle. The psyllid has five nymph stages and the adult stage can jump or fly short distances. The adult female can lay up to 800 eggs during her lifetime. Once a psyllid has acquired the bacteria, it can be transmitted to healthy trees over the course of the psyllids life. The first symptoms usually don t appear for up to two years from infection making early detection difficult and leaving infected trees in the orchard available for continued transmission by psyllids. Typical early symptoms are yellowing of isolated shoots, asymmetrical blotchy yellowing of the leaves, vein corking, yellow veins, and defoliation. Fruit may be small, misshapen, lop-sided, taste medicinal, bitter, or sour, and contain aborted seeds. Once symptoms appear the tree will continue to decline over a period of 3-8 years and may not produce any useable fruit during this period. Death for the tree is certain and unavoidable. Page 1 of 5
2 Figure 2. A citrus leaf displaying the vein corking that is typical of an HLBinfected tree. Figure 3. The orange on the right is typical of those produced by an HLBinfected tree. Notice the blackened, aborted seeds in the smaller than average sized fruit. Believed to have first been discovered in southern China in 1919, HLB has since spread to virtually all of the Asian countries, Africa, the middle east, Brazil, Mexico, Cuba, Costa Rica, Guatemala, the Honduras, Jamaica, the Dominican Republic, and the United States. It was first discovered in Florida in Miami- Dade County in 2005 and is now in all 31 citrus producing counties (Figure 4). More recently it has been discovered in Texas in December of 2011 and finally in California in March of (Figure 4.) Page 2 of 5
3 Florida is the largest producer of citrus in the United States and the second largest producer of orange juice in the world. The estimated economic impact of the citrus industry to the state of Florida during the 2007/08 season was approximately 9 billion dollars and generated over 75,000 jobs. Since the discovery of HLB Florida s growers have suffered 1.3 billion dollars in lost revenue, the state s economy has suffered 3.63 billion dollars in lost revenue and 6,611 jobs have been lost due to a reduction in orange juice production. 3 The potential losses are so great that California s response to the single tree discovered to be HLB-positive in a home-owners backyard in Los Angeles County was to destroy the tree, treat all trees within 800 meters and institute a 93 square mile quarantine. It is currently estimated that 43% of Florida citrus trees have been infected and extrapolation from current data has resulted in predictions that 100% of current citrus plantings in Florida will be affected by HLB within the next decade. 3 Hope, though too late for many, seems to be right around the corner. Erik Mirkov, a scientist at Texas A&M s Texas AgriLife Research and Extension Center has created a genetically modified citrus tree using bacteria-fighting proteins naturally found in spinach. Second, third, and fourth generation trees have now been produced and have proven successful in high psyllid population controlled environments. The trees have been planted in orchards near Lake Okeechobee for field tests in March of Growers are hopeful, but there is no doubt that, even if successful, by the time orchards can replace declining trees with HLB-resistant trees, many more growers will have been put out of business by an invasive bacteria, too small to be seen with the naked eye, that entered our state less than ten years ago. Introduction Prior to this lesson, students should have lab experience with DNA extracton, electrophoresis and PCR (thermocyclers). This lesson could probably be modified to incorporate beginner activities for those students with no exposure, but was not written that way. In this lesson, students learn about some of the job responsibilities of an agricultural technician. These include being able to identify plant diseases, insects, as well as run laboratory equipment and properly prepare samples for Page 3 of 5
4 diagnostic testing. This is much more than the old-fashioned farmer image that most students grew up with. Students will learn how to identify the Asian citrus psyllid and fill out a taxonomic chart identifying its characteristics. Students will learn how to identify symptoms of and HLB-positive tree, leaves, and fruit. Next, students will be given samples to diagnose as HLB-positive or HLB-negative using only photographs, physical samples, an iodine-starch test, and PCR. Once all the diagnoses have been made the students will learn which samples are actually HLB-positive and which are HLB-negative and students will tally the results to statistically analyze which test was most effective for cost benefit purposes. Finally, students will gather all of their information and create a brochure to be handed out at the local farmer s market in our community to bring awareness of HLB to our citizens and assist them in diagnosing their own backyard citrus trees. Objectives At the end of this lesson, students should be able to explain what citrus greening is and why it is of such concern to growers and the citrus industry. Students should be able to list symptoms of an HLB-infected tree and to pick out HLBinfected leaves with a fair amount of accuracy. Students should have a good understanding of why it is important for growers to have a correct diagnosis and which type of test is most likely to provide that. Students should also appreciate another use of DNA extraction, electrophoresis, and PCR as used in the agricultural field. Materials and Supplies Cost (at time of publishing) Item Description Company (if applicable) Catalog Number Liquid Nitrogen Can be obtained from nearest college or university Usually free of cost Absolute EMD Ethanol (500 ml) Fisher Scientific NC $46.30 DNeasy Plant MniKit for 50 reactions Qiagen $ Forward Primer Reverse Primer Integrated DNA Technologies Integrated DNA Technologies $8.03 (plus shipping & handling) $7.70 (plus shipping & handling) Master Mix for PCR (90 units) BioRad EDU $92.50 Page 4 of 5
5 Tincture of Iodine Drugstore $2.99 Tweezers Razor blades Timers Stereoscopes are helpful, but not required (magnifying glasses will suffice) Thermocycler Electrophoresis Apparatus Mortar and Pestles Centrifuge Vortexer Hot water bath or dry bath HLB-positive and HLB-negative citrus leaves Camera 1 Halbert SE and Manjunath KL, Asian citrus psyllids (Sternorrhyncha: Psyllidae) and greening disease of citrus: a literature review and assessment of risk in Florida Fla Entomol 87: (2004). 2 Pelz-Stelinski K, Stelinski LL and Tiwari S, Effect of Candidatus Liberibacter asaticus infection on susceptibility of Asian citrus psyllid, Daphorina citri, to selected insecticides Pest Manag Sci 67:94-99 (2011). 3 Hodges AW and Spreen TH, Economic impacts of citrus greening (HLB) in Florida, 2006/ /11 1 EDIS document FE903, a publication of the Food and Resource Economics Department, Florida Cooperative Extension Service, Institute of Food and Agricultural Sciences, University of Florida, Gainesville, Fl Published January Stover E, Castle WS, Spyke P, The citrus grove of the future and its implications for Huanglongbing management. Proc. Fla. State Hortic. Soc. 121: (2008) Page 5 of 5
6 Kathy Savage Oviedo High School 601 King Street, Oviedo FL (cell) Title: Citrus Greening is not Green! Abstract: This action plan is designed to allow students to use newly obtained biotechnology lab skills within the context of a real-world problem. This is designed to be a project that encompasses prior skills and knowledge allowing students to discover problems for themselves and apply what they learn to a community awareness project. The heart of this project will be Citrus Greening or Huanglongbing, a disease affecting citrus trees in the state of Florida. In this module students will study how the disease affects citrus trees; how to identify it morphologically and in the lab using an iodine test and PCR, study the cause and factors affecting its spread, track its spread within our state, and design a community awareness brochure. Tests on leaves and psyllids will be performed using kits available from Qiagen. Students will use statistics from government websites to determine how fast HLB is spreading and graphically display their results. Classes will take a field trip to a local citrus grove to observe, learn how to diagnose and identify affected trees, look for psyllids, and collect leaves for testing. Rationale: Students will have the opportunity to study Citrus Greening (Huanglongbing or HLB), a vector-borne bacterial disease that is devastating Florida s citrus industry at an alarming rate. Since it was first detected in Florida in 2005, HLB is now found in all of the states citrus producing counties. Once a tree has become infected, the fruit it produces becomes bitter and salty making it virtually inedible. Death for the tree is inevitable as there is no cure for this disease. The vector that carries and transmits HLB in Florida is a small Asian psyllid that can hop, fly, or be carried longer distances by winds. It is estimated that every citrus grove in the state is within one mile of an infected tree. According to the Florida Department of Citrus, the industry employs approximately 76,000 workers and has an annual economic impact of 9 billion dollars. (1) The United States leads the world in grapefruit production supplying the world with over 30% of its grapefruit, and is the third largest overall citrus producer in the world. The majority of the citrus grown in the United States comes with Florida as the major producer. In fact Florida produces three times as many tons of oranges and four times as many tons of grapefruit as its closest competitor, California. (2) With over ½ million acres of citrus groves and 74 million trees, Florida is second only to Brazil in orange juice production and supplies approximately 80% of the orange juice in the United States during any given growing season. (3) These facts clearly demonstrate that any threat to the Florida Citrus Industry is a threat to the world citrus supply. This module would allow students to reinforce some laboratory skills and techniques they are already familiar with (electrophoresis, PCR) and use some that would be new to them (BLAST). In the past, lab skills have been used in an isolated fashion with various labs designed to teach the skill without a real- Page 1 of 6
7 world frame of reference. This module would allow students to see how these seemingly isolated and unrelated skills work together to identify and diagnose HLB in citrus trees. References Cited: 1. "Florida Grapefruit." Florida Department of Citrus Aug < 2. "Background Statistics: U.S. Citrus Market." United States Department of Agriculture. 22 Jan Aug < 3. "Citrus Facts." Florida Citrus Aug < Lesson Plans: See Below Day One: Students will do a webquest on Citrus Greening (Huanglongbing) Day Two and Three: Discussion and Lecture on the citrus greening disease, it s vector the Asian Citrus Psyllid, and the Economic Impact for Florida using the following PowerPoint Presentations. These are excellent presentations with great graphics and include comments for the presenter. 1. The Asian Citrus Psyllid and the Citrus Disease Huanglongbing by the California Citrus Research Board 2. The Asian Citrus Psyllid & Huanglongbing by Protect U.S. Community Invasive Species Network at the University of Florida 3. Three Exotic Plant Diseases Threatening Florida by the Florida State Agricultural Response Team Day Four: Identify samples of citrus trees with HLB and compare to samples without HLB from a morphological perspective. If possible, taste test citrus that has been harvested from HLB infected trees. Discern whether or not they can tell infected Asian Citrus Psyllid from noninfected Psyllid by visual inspection under a stereoscope. Students will record their observations for each sample of leaf and psyllid and make their predictions as to infected vs. non-infected in a table. Day Five: Students will use the Iodine Starch Test to further solidify or change their initial predictions from the previous day s inspections. This protocol is available from the University of Florida IFAS website Page 2 of 6
8 Day Six: Students will begin preparation for PCR testing of their samples using the protocol provided by Dr. Kirsten Stelinski from UF. Day Seven: Students will work on testing citrus samples and keep a record of results. Day Eight: Continued as above Day Nine: Students will graph results identified by location (city, county, etc.) and compare to actual data obtained from the State of Florida. Day Ten: Students will develop informational flyers/brochures for Seminole County Residents (to be given out at Great Day in the County and local craft fairs) that include morphological detection methods and phone numbers to call for plant inspection and testing. Day Fourteen: Field Trip to Lake Alfred Materials and Supplies: Thermocycler Hot Water Bath Mortar and Pestle Electrophoresis Apparatus 1.5 ml microfuge tubes with caps Microcentrifuge Autoclave Spatulas Stereoscopes Razor Blades Chemicals Required: Item Source for Purchase Catalog No. Cost Liquid Nitrogen University of Central Florida Free Chemistry Department Absolute EMD Ethanol Fisher Scientific NC $ ml DNeasy Blood & Tissue Kit Qiagen $ for 50 reactions DNeasy Plant MiniKit for Qiagen $ reactions Primer IDT (Integrated DNA? Technologies) Loading Dye 1 ml 5x BioRad EDU $50.00 Iodine Tincture or Tincture Drugstore? of Iodine or Lugol s solution Amazon Buses for Field Trip? Page 3 of 6
9 Total Cost $ Bibliography: "Huanglongbing Florida Citrus Greening Information Video." World News, Inc Aug < Protocol: Purification of Total DNA from Plant Tissue (Mini Protocol) Provided by Dr. Kirsten Stelinski Before starting: Buffer AP1 may develop a yellow color upon storage. This does not affect the procedure. All centrifugation steps are carried out at room temperature (15 25 C) in a microcentrifuge. Things to do before starting Buffer AP1 and Buffer AP3/E concentrate may form precipitates upon storage. If necessary, warm to 65 C to redissolve (before adding ethanol to Buffer AP3/E). Do not heat Buffer AP3/E after ethanol has been added. Buffer AW and Buffer AP3/E are supplied as concentrates. Before using for the first time, add the appropriate amount of ethanol (96 100%) as indicated on the bottle to obtain a working solution. Preheat a water bath or heating block to 65 C. Label tubes with sample numbers/codes. You will need: 2 sets of 1.5 ul microcentrifuge tubes, 1 set of QIAshredder spin columns (lilac), and ones set of DNeasy Mini spin columns (white). Autoclave mortars, pestles, and spatulas for use in grinding plant tissue. Procedure 1. Plant tissue (100 mg composite of petioles and midribs) can be ground to a fine powder under liquid nitrogen using a mortar and pestle. Allow the liquid nitrogen to evaporate and transfer the tissue powder to an appropriately sized tube. Do not allow the sample to thaw. 2. Add 400 μl Buffer AP1 and 4 μl RNase A stock solution (100 mg/ml) to a maximum of 100 mg (wet weight) disrupted plant or fungal tissue and vortex vigorously. Spin briefly to remove any sample that has collected on lid of tube. No tissue clumps should be visible. Vortex or pipet further to remove any clumps. Clumps of tissue will not lyse properly and will therefore result in a lower yield of DNA. Note: Do not mix Buffer AP1 and RNase A before use. 3. Incubate the mixture for 10 min- 30 min at 65 C. Mix 2 or 3 times during incubation by inverting tube. This step lyses the cells. 4. Add 130 μl Buffer AP2 to the lysate, mix, and incubate for 5 min on ice. This step precipitates detergent, proteins, and polysaccharides. 5. Centrifuge the lysate for 5 min at 20,000 x g (14,000 rpm). 6. Pour the lysate (avoid transferring perllet) into the QIAshredder Mini spin column (lilac) placed in a 2 ml collection tube, and centrifuge for 2 min at 20,000 x g (14,000 rpm). Page 4 of 6
10 It may be necessary to cut the end off the pipet tip to apply the lysate to the QIAshredder Mini spin column. The QIAshredder Mini spin column removes most precipitates and cell debris, but a small amount will pass through and form a pellet in the collection tube. Be careful not to disturb this pellet in step Add 675 μl Buffer AP3/Eto a new tube (not supplied). Transfer the flow-through fraction from step 6 to the cleared lysate without disturbing the cell-debris pellet, and mix immediately by pipetting or gently vortexing. Ensure that ethanol has been added to Buffer AP3/E. 8. Pipet 650 μl of the mixture from step 7, including any precipitate that may have formed, into the DNeasy Mini spin column placed in a 2 ml collection tube (supplied). Centrifuge for 1 min at _6000 x g (corresponds to _8000 rpm for most microcentrifuges), and discard the flow-through.* Reuse the collection tube in step Repeat step 14 with remaining sample. Discard flow-through* and collection tube. 10. Place the DNeasy Mini spin column into a new 2 ml collection tube (supplied), add 500 μl Buffer AW, and centrifuge for 1 min at 6000 x g (8000 rpm). Discard the flow-through and reuse the collection tube in step 11. Note: Ensure that ethanol is added to Buffer AW. 11. Repeat step 10 by adding 500 μl Buffer AW to the DNeasy Mini spin column, and centrifuge for 2 min at 20,000 x g (14,000 rpm) to dry the membrane. It is important to dry the membrane of the DNeasy Mini spin column since residual ethanol may interfere with subsequent reactions. Discard flow-through and collection tube. Note: Following the centrifugation, remove the DNeasy Mini spin column from the collection tube carefully so the column does not come into contact with the flow through, as this will result in carryover of ethanol. 12. Transfer the DNeasy Mini spin column to a 1.5 ml or 2 ml microcentrifuge tube (not supplied), and pipet 50 μl Buffer AE directly onto the DNeasy membrane. Incubate for 5 min at room temperature (15 25 C), and then centrifuge for 1 min at 6000 x g (8000 rpm) to elute. Store at -20 o C. Page 5 of 6
11 Conventional PCR to Examine the Presence of the HLB-associated Pathogen in Citrus Provided by Dr. Kirsten Stelinski 16S rdna, OI1/OI2 specific to the Ca. Liberibacter asiaticus sp. used for PCR amplification in a 25-μl reaction volume Forward primer: OI1 (5 -GCG CGT ATG CAA TAC GAG CGG CA-3 ) Reverse primer: OI2c (5 -GCC TCG CGA CTT CGC AAC CCA T- 3 ) PCR reaction 25-μl reaction volume consisting of: 1 μl of DNA template 200 nm (each) primer 200 μm (each) dntps 2.5 mm MgCl2 1 PCR buffer dh 2 O 1 unit Platinum Taq polymerase (Invitrogen, Carlsbad, CA) Amplification: 94 C for 2 min 35 cycles of 94 C for 30 s, 62 C for 30 s, and 72 C for 1 min Final extension cycle of 10 min at 72 C Analysis of PCR reactions: Expected product size: 1160 bp 1.0% agarose gel in 1 Tris-acetate-EDTA buffer (40 mm Tris, 20 mm acetic acid, 1 mm EDTA, ph 8.5) Any 1 kb DNA ladder should work e.g. Quick-Load 1 kb DNA Ladder (New England Biolabs) or 1 kb Plus DNA ladder (Invitrogen) Page 6 of 6
huanglongbing Citrus Greening and the Yellow Dragon
huanglongbing Citrus Greening and the Yellow Dragon Coloring and Activity Book Diana C. Schultz, Ronald D. French!!!!!2010 What is Citrus greening? Citrus greening, also known as Huanglongbing (HLB; yellow
More informationDNA Extraction from Radioative Samples Grind plus kit Method
DNA Extraction from Radioative Samples Grind plus kit Method 4 th Edition 2017.5.24 To extract DNA from radioactive sediment samples with low biomass, we are currently not allowed to use chloroform or
More informationDNA extraction method as per QIAamp DNA mini kit (Qiagen, Germany)
APPENDIX 3 (MOLECULAR TECHNIQUES) 3.2.2a) DNA extraction method as per QIAamp DNA mini kit (Qiagen, Germany) Two hundred microliters (200 µl) of the EDTA blood was added to 200 µl of Buffer AL and 20 µl
More informationWorm Collection. Prior to next step, determine volume of worm pellet.
Reinke Lab ChIP Protocol (last updated by MK 05/24/13) Worm Collection 1. Collect worms in a 50ml tube. Spin and wait until worms are collected at the bottom. Transfer sample to a 15ml tube and wash with
More informationThe Asian Citrus Psyllid and the Citrus Disease Huanglongbing
Asian Citrus Psyllid and the Citrus Disease Huanglongbing Psyllid M. Rogers Beth Grafton-Cardwell University of California Riverside Huanglongbing Where did Asian citrus psyllid and the HLB disease come
More informationCitrus. Response Program
PATHOLOGY TRAINING Citrus Health Response Program Why is Huanglongbing g g important? In areas where the disease is endemic, citrus trees live 6-8 years, and most never bear usable fruit. So producing
More informationIn Vitro NER Assay. Auble Lab. Reagents:
In Vitro NER Assay Reagents: Water YPD Yeast extraction Buffer (200 ml): 0.2 M Tris-acetate (ph 7.5) (40 ml), 0.39 M (NH 4 ) 2 S0 4 (78 ml), 10 mm MgSO 4 (2 ml), 20% Glycerol (40 ml), 1mM EDTA (ph8.0)
More informationMiniprep - Alkaline Lysis for BACs
Miniprep - Alkaline Lysis for BACs by A. Untergasser (contact address and download at www.untergasser.de/lab) Version: 1.0 - Print Version (.PDF) ATTENTION: This is a low priced protocol. Use it preferably!
More informationEstimating the Greening Effect on Florida Citrus
Estimating the Greening Effect on Florida Citrus Charles B. Moss 1 and Maria Bampasidou 1 1 University of Florida March 26, 2014 1 Citrus Maladies Citrus Greening - The Disease Canker - The Other Citrus
More informationAsian citrus psyllid management and current findings of recent surveys. Xavier Martini
Asian citrus psyllid management and current findings of recent surveys Xavier Martini The Asian citrus psyllid Uninfected - Diaphorina citri, the Asian citrus psyllid. First found in Florida June 1998
More informationCitrus Canker and Citrus Greening. Holly L. Chamberlain Smoak Groves AGRI-DEL, INC. Lake Placid, FL
Citrus Canker and Citrus Greening Holly L. Chamberlain Smoak Groves AGRI-DEL, INC. Lake Placid, FL Hurricanes 2004 and 2005 Challenges Facing FL Citrus Production Citrus Greening Competition Citrus Canker
More informationCitrus Disease Identification for North Florida
Citrus Disease Identification for North Florida Jamie Burrow, Megan Dewdney, and Michael Rogers University of Florida, IFAS, CREC Thursday, January 21, 2016 Citrus Canker Citrus canker is caused by a bacterial
More informationExperiment # Lemna minor (Duckweed) Population Growth
Experiment # Lemna minor (Duckweed) Population Growth Introduction Students will grow duckweed (Lemna minor) over a two to three week period to observe what happens to a population of organisms when allowed
More informationCitrus. Disease Guide. The Quick ID Guide to Emerging Diseases of Texas Citrus. Citrus. Flash Cards. S. McBride, R. French, G. Schuster and K.
E-265 1/12 Citrus Flash Cards S. McBride, R. French, G. Schuster and K. Ong Citrus Disease Guide The Quick ID Guide to Emerging Diseases of Texas Citrus The Quick ID Guide to Emerging Diseases of Texas
More information3.5 Citrus Greening (Huanglongbing) Disease in India : Present Status and Diagnostic Efforts
Page 129 3.5 Citrus Greening (Huanglongbing) Disease in India : Present Status and Diagnostic Efforts Das A. K. National Research Centre for Citrus, Amravati Road, Nagpur 440010, India. Among all diseases
More informationDNA-Miniprep. - Rapid boiling
DNA-Miniprep. - Rapid boiling by A. Untergasser (contact address and download at www.untergasser.de/lab) Version: 1.0 - Print Version (.PDF) ATTENTION: This is a low priced protocol. Use it preferably!
More informationAsian Citrus Psyllid and Huanglongbing (HLB)
Asian Citrus Psyllid and Huanglongbing (HLB) Victoria Hornbaker California Department of Food and Agriculture Feb. 2, 2016 California s iconic citrus is at risk! Approximately 70% of residences have at
More informationScreening Aid for Huanglongbing (HLB) or Citrus Greening Disease Symptoms By Hilda Gomez, Plant Pathologist, USDA, APHIS.
Screening Aid for Huanglongbing (HLB) or Citrus Greening Disease Symptoms By Hilda Gomez, Plant Pathologist, USDA, APHIS. PPQ CHRP Program Huanglongbing (HLB) or citrus greening is a serious disease of
More informationMiniprep - Alkaline Lysis
Miniprep - Alkaline Lysis by A. Untergasser (contact address and download at www.untergasser.de/lab) Version: 1.0 - Print Version (.PDF) ATTENTION: This is a low priced protocol. Use it preferably! 1.
More informationThreats From Beyond Our Borders: Exotic Diseases and Pests in Citrus
Threats From Beyond Our Borders: Exotic Diseases and Pests in Citrus MaryLou Polek, Plant Pathologist Vice President Science & Technology Citrus Research Board Disease: The Malfunctioning of Cells & Tissues
More informationGreening and Canker Training for Master Gardeners
Greening and Canker Training for Master Gardeners Ryan Atwood, Extension Agent, Lake County & Dr. Tim Spann, Extension Specialist, Citrus Research and Education Center Created: September 2007 Citrus in
More informationYeast nuclei isolation kit. For fast and easy purification of nuclei from yeast cells.
ab206997 Yeast nuclei isolation kit Instructions for use: For fast and easy purification of nuclei from yeast cells. This product is for research use only and is not intended for diagnostic use. Version
More informationAccuID TM _V1. Bone DNA Preparation Protocol. SNP based New Human Identification Technology. Protocol Version
AccuID TM _V1 SNP based New Human Identification Technology Bone DNA Preparation Protocol Protocol Version 1.0 2013.10.02 Copyright 2013 DNA Link, Inc. All rights reserved. AccuID TM Bone Preparation Protocol
More informationHuanglongbing in Belize Current Situation & Activities
Huanglongbing in Belize Current Situation & Activities Stephen Williams & Veronica Manzanero Majil Belize Citrus Growers Association Francisco Gutierrez Belize Agricultural Health Authority Fermin Blanco
More informationEconomic Contributions of the Florida Citrus Industry in and for Reduced Production
Economic Contributions of the Florida Citrus Industry in 2014-15 and for Reduced Production Report to the Florida Department of Citrus Alan W. Hodges, Ph.D., Extension Scientist, and Thomas H. Spreen,
More informationHarvesting Charges for Florida Citrus, 2016/17
Harvesting Charges for Florida Citrus, 2016/17 Ariel Singerman, Marina Burani-Arouca, Stephen H. Futch, Robert Ranieri 1 University of Florida, IFAS, CREC, Lake Alfred, FL This article summarizes the charges
More informationThe Incidence of Greening and Canker Infection in Florida Citrus Groves from September 2007 through August
FE823 The Incidence of Greening and Canker Infection in Florida Citrus Groves from September 2007 through August 2008 1 Robert A. Morris, Candice Erick, and Mark Estes 2 Background In early summer of 2008,
More informationDisease-Carrying Asian Citrus Psyllid. Communication Outreach
Disease-Carrying Asian Citrus Psyllid Communication Outreach Objective Support the California citrus industry in its efforts to keep the disease-carrying Asian citrus psyllid from establishing in California
More informationDooryard Citrus Production: Citrus Greening Disease 1
HS1131 Dooryard Citrus Production: Citrus Greening Disease 1 Timothy M. Spann, Ryan A. Atwood, Jamie D. Yates, Michael E. Rogers and Ronald H. Brlansky 2 Citrus greening disease, also known as Huanglongbing
More informationPsa and Italian Kiwifruit Orchards an observation by Callum Kay, 4 April 2011
Psa and Italian Kiwifruit Orchards, 2011 The Psa-research programme in New Zealand draws on knowledge and experience gained from around the world particularly in Italy, where ZESPRI, Plant & Food Research
More informationChestnut DNA extraction B3 Summer Science Camp 2014
Experiment Type: Experiment Goals: Sample Label: Scientist Name: Date: General Idea: extract the nucleic acid from leaf tissue by grinding it in a reducing medium (the betamercaptoethanol, which smells
More informationFOR PERSONAL USE. Capacity BROWARD COUNTY ELEMENTARY SCIENCE BENCHMARK PLAN ACTIVITY ASSESSMENT OPPORTUNITIES. Grade 3 Quarter 1 Activity 2
activity 2 Capacity BROWARD COUNTY ELEMENTARY SCIENCE BENCHMARK PLAN Grade 3 Quarter 1 Activity 2 SC.A.1.2.1 The student determines that the properties of materials (e.g., density and volume) can be compared
More informationSYSTEMS USED TO COMBAT OTHER VECTOR TRANSMITTED BACTERIA, PIERCE S DISEASE IN GRAPES. Don Hopkins Mid Florida REC, Apopka
SYSTEMS USED TO COMBAT OTHER VECTOR TRANSMITTED BACTERIA, PIERCE S DISEASE IN GRAPES Don Hopkins Mid Florida REC, Apopka Vascular Diseases Caused by Fastidious Prokaryotes Fastidious Phloem-Limited Bacteria
More informationEXPERIMENT 3 - IDENTIFYING FEATURES OF MUTANT SEEDS USING NOMARSKI MICROSCOPY (GENE ONE)
EXPERIMENT 3 - IDENTIFYING FEATURES OF MUTANT SEEDS USING NOMARSKI MICROSCOPY (GENE ONE) STRATEGY I. OBSERVATION OF SEEDS USING LIGHT MICROSCOPY AND FIXING SEEDS FOR OBSERVATION WITH NOMARSKI OPTICS II.
More informationMaxiprep - Alkaline Lysis
Maxiprep - Alkaline Lysis by A. Untergasser (contact address and download at www.untergasser.de/lab) Version: 1.0 - Print Version (.PDF) ATTENTION: This is a low priced protocol. Use it preferably! 1.
More informationARE THE SEEDS OF ALL FRUITS THE SAME?
ACTIVITY 1 ARE THE SEEDS OF ALL FRUITS THE SAME? EXPERIMENT OBJECTIVES AND CONTENT In this activity, students gain a better understanding of the seeds of edible fruits. ESSENTIAL KNOWLEDGE Matter: Characteristics
More informationActivity Sheet Chapter 6, Lesson 6 Using Chemical Change to Identify an Unknown
Activity Sheet Chapter 6, Lesson 6 Using Chemical Change to Identify an Unknown Name Date DEMONSTRATION 1. Your teacher poured iodine solution on top of two white powders. How do you know that these two
More informationPECTINASE Product Code: P129
PECTINASE Product Code: P129 Enzyme for sample clarification prior to patulin analysis. For in vitro use only. P129/V1/02.06.16 www.r-biopharm.com Contents Page Test Principle... 3 Kit Components... 3
More informationAsian Citrus Psyllid threat to Santa Barbara County Author: Surendra Dara
Asian Citrus Psyllid threat to Santa Barbara County Author: Surendra Dara February 2, 2011 In light of spotting a couple of Asian citrus psyllids (ACP) in Ventura County about a month ago, it is important
More informationHuanglongbing (HLB), also known as citrus greening,
Plant Disease September 2017 PD-112 Citrus Huanglongbing Michael J. Melzer, Diane M. Sether, John S. Hu, and Anne M. Alvarez. Department of Plant and Environmental Protection Sciences Huanglongbing (HLB),
More informationEXTRACTION OF SEDIMENTS FOR AROMATIC AND CHLORINATED HYDROCARBONS
EXTRACTION OF SEDIMENTS FOR AROMATIC AND CHLORINATED HYDROCARBONS Juan. A. Ramirez, Bo Wang, Donell S. Frank, Thomas. J. McDonald, Rebecca Price, Susanne J. McDonald and James M. Brooks TDI-Brooks International./B&B
More informationDetermination of Melamine Residue in Milk Powder and Egg Using Agilent SampliQ Polymer SCX Solid Phase Extraction and the Agilent 1200 Series HPLC/UV
Determination of Melamine Residue in Milk Powder and Egg Using Agilent SampliQ Polymer SCX Solid Phase Extraction and the Agilent 1200 Series HPLC/UV Application Note Food Safety Authors Chen-Hao Zhai
More informationMONITORING WALNUT TWIG BEETLE ACTIVITY IN THE SOUTHERN SAN JOAQUIN VALLEY: OCTOBER 2011-OCTOBER 2012
MONITORING WALNUT TWIG BEETLE ACTIVITY IN THE SOUTHERN SAN JOAQUIN VALLEY: OCTOBER 11-OCTOBER 12 Elizabeth J. Fichtner ABSTRACT Walnut twig beetle, Pityophthorus juglandis, is the vector of thousand cankers
More informationLoving our Citrus. How to care for our trees. Maggie O Neill Lemon Blossom Heritage Farm Claremont Garden Club Sept 13, 2017
Loving our Citrus How to care for our trees Maggie O Neill Lemon Blossom Heritage Farm Claremont Garden Club Sept 13, 2017 Lemon Blossom Heritage Farm Keeping history alive for future generations Educate
More informationFlorida Citrus Production, Greening & Horticultural Practices
Florida Citrus Production, Greening & Horticultural Practices Stephen H. Futch Extension Agent, Multi County University of Florida Citrus Research & Education Center 700 Experiment Station Road, Lake Alfred,
More informationThe Economic Impact of Bird Damage to Select Fruit Crops in New York
The Economic Impact of Bird Damage to Select Fruit Crops in New York Funding provided by USDA s Specialty Crop Research Initiative Summer 2014 Bird damage to blueberries. Economic Impact of Bird Damage
More informationGrowing divergence between Arabica and Robusta exports
Growing divergence between Arabica and Robusta exports In April 218, the ICO composite indicator decreased by.4% to an average of 112.56, with the daily price ranging between 11.49 and 114.73. Prices for
More informationWALNUT BLIGHT CONTROL USING XANTHOMONAS JUGLANDIS BUD POPULATION SAMPLING
WALNUT BLIGHT CONTROL USING XANTHOMONAS JUGLANDIS BUD POPULATION SAMPLING Richard P. Buchner, Steven E. Lindow, James E. Adaskaveg, Parm Randhawa, Cyndi K. Gilles, and Renee Koutsoukis ABSTRACT Years and
More informationBEEF Effect of processing conditions on nutrient disappearance of cold-pressed and hexane-extracted camelina and carinata meals in vitro 1
BEEF 2015-05 Effect of processing conditions on nutrient disappearance of cold-pressed and hexane-extracted camelina and carinata meals in vitro 1 A. Sackey 2, E. E. Grings 2, D. W. Brake 2 and K. Muthukumarappan
More informationTable of Contents. Syngenta Citrus Scout Summer
Syngenta Crop Protection Citrus Scout Internship Morgan McKenna East Coast Intern Vero Beach, FL May 20-August 16, 2013 Table of Contents Internship Overview 2 Scouting 3 Citrus Soil Assay 6 Florida Citrus
More informationION FORCE DNA EXTRACTOR FAST Cat. N. EXD001
ION FORCE DNA EXTRACTOR FAST Cat. N. EXD001 User Manual Via San Geminiano, 4 41030 San Prospero (MO) Italy : +39 059 8637161 : +39 059 7353024 : laboratorio@generon.it : www.generon.it [1] User Manual
More informationTHE THREAT: The disease leads to dieback in shoots and fruiting buds and an overall decline in walnut tree health.
Taking Control of Botryosphaeria in California Walnut Orchards Summary THE ISSUES: Botryosphaeria, or Bot, is a fungal disease that spreads by spores that germinate and enter the tree through existing
More informationAWRI Refrigeration Demand Calculator
AWRI Refrigeration Demand Calculator Resources and expertise are readily available to wine producers to manage efficient refrigeration supply and plant capacity. However, efficient management of winery
More informationIdentification and Classification of Pink Menoreh Durian (Durio Zibetinus Murr.) Based on Morphology and Molecular Markers
RESEARCH Identification and Classification of Pink Durian (Durio Zibetinus Murr.) Based on Morphology and Molecular Markers Nandariyah a,b * adepartment of Agronomy, Faculty of Agriculture, Sebelas Maret
More informationTomato Potato Psyllid
Tomato Potato Psyllid Bactericera cockerelli CHECK YOUR CROPS REPORT SUSPECTED SIGHTINGS Monitoring Guide What does the psyllid do? Tomato Potato Psyllid (TPP) is an exotic pest with a wide host range,
More information3. Aspirin Analysis. Prelaboratory Assignment. 3.1 Introduction
In this experiment, you will analyze the purity of your crude and recrystallized aspirin products using a method called thin layer chromatography (TLC). You will also determine the percent yield of your
More informationMastering Measurements
Food Explorations Lab I: Mastering Measurements STUDENT LAB INVESTIGATIONS Name: Lab Overview During this investigation, you will be asked to measure substances using household measurement tools and scientific
More informationACP and HLB: The California Situation. Victoria Hornbaker California Department of Food and Agriculture
ACP and HLB: The California Situation Victoria Hornbaker California Department of Food and Agriculture Dangerous Pest Threatens California Citrus California's $2.5 billion citrus industry is at risk. Citrus
More informationDry Ice Rainbow of Colors Weak Acids and Bases
Dry Ice Rainbow of Colors Weak Acids and Bases SCIENTIFIC Introduction Add a small piece of solid carbon dioxide to a colored indicator solution and watch as the solution immediately begins to boil and
More informationLAB: One Tube Reaction Part 1
AP Chemistry LAB: One Tube Reaction Part 1 Objective: To monitor and document the chemical changes occurring in a single test tube containing a predetermined mixture of chemicals. Materials: test tube,
More informationHow Much Sugar Is in Your Favorite Drinks?
Lesson 3 How Much Sugar Is in Your Favorite Drinks? Objectives Students will: identify important nutrition information on beverages labels* perform calculations using nutrition information on beverages
More informationDry Ice Color Show Dry Ice Demonstrations
Dry Ice Color Show Dry Ice Demonstrations SCIENTIFIC Introduction Add a small piece of solid carbon dioxide to a colored indicator solution and watch as the solution immediately begins to boil and change
More informationFood Allergen and Adulteration Test Kits
Food Allergen and Adulteration Test Kits Overview Neogen offers food allergen test kits to detect almond, egg, gliadin, hazelnut, milk, mustard, peanut, sesame, shellfish, soy and walnut residues (see
More informationDR. RENEE THRELFALL RESEARCH SCIENTIST INSTITUTE OF FOOD SCIENCE & ENGINEERING UNIVERSITY OF ARKANSAS
Challenges in Muscadine Juice and Wine Production DR. RENEE THRELFALL RESEARCH SCIENTIST INSTITUTE OF FOOD SCIENCE & ENGINEERING UNIVERSITY OF ARKANSAS RTHRELF@UARK.EDU Muscadine juice and wine production
More informationESTIMATING ANIMAL POPULATIONS ACTIVITY
ESTIMATING ANIMAL POPULATIONS ACTIVITY VOCABULARY mark capture/recapture ecologist percent error ecosystem population species census MATERIALS Two medium-size plastic or paper cups for each pair of students
More informationTitle: Genetic Variation of Crabapples ( Malus spp.) found on Governors Island and NYC Area
Title: Genetic Variation of Crabapples ( Malus spp.) found on Governors Island and NYC Area Team Members: Jianri Chen, Zinan Ma, Iulius Sergiu Moldovan and Xuanzhi Zhao Sponsoring Teacher: Alfred Lwin
More informationSequential Separation of Lysozyme, Ovomucin, Ovotransferrin and Ovalbumin from Egg White
AS 662 ASL R3104 2016 Sequential Separation of Lysozyme, Ovomucin, Ovotransferrin and Ovalbumin from Egg White Sandun Abeyrathne Iowa State University Hyunyong Lee Iowa State University, hdragon@iastate.edu
More informationEnzymes in Industry Time: Grade Level Objectives: Achievement Standards: Materials:
Enzymes in Industry Time: 50 minutes Grade Level: 7-12 Objectives: Understand that through biotechnology, altered enzymes are used in industry to produce optimal efficiency and economical benefits. Recognize
More information- NSES-C, NSES-F, NSES-G, NHES-1
Lesson 1.3 The Good Samaritans Estimated time: One 50 min period Instructional overview Lesson 1.3 will show students the importance of beneficial microorganisms in food production and increase awareness
More informationRationale or Purpose: This lesson introduces students to the process of prehistoric hot rock cooking in earth ovens on the Edwards Plateau of Texas.
Lesson Title: What s Cookin at Honey Creek? Subject: Texas History Grade level: 4th (includes 4 th grade TEKS, but can easily be adapted to 7th grade) Rationale or Purpose: This lesson introduces students
More informationAsian Citrus Psyllid and the Citrus Disease Huanglongbing
Asian Citrus Psyllid and the Citrus Disease Huanglongbing Psyllid Huanglongbing Beth Grafton-Cardwell Department of Entomology UC Riverside Photography: M. Rogers, S. Halbert and E. Grafton- Cardwell Adult
More informationHLB Causal Agent HLB is caused by a highly fastidious, phloem- limited bacterium. As yet, has not been cultured singularly. Dr. Mike Davis has managed
HUANGLONGBING AKA YELLOW SHOOT DISEASE YELLOW DRAGON DISEASE CITRUS GREENING HLB HLB Causal Agent HLB is caused by a highly fastidious, phloem- limited bacterium. As yet, has not been cultured singularly.
More informationAi Arizona Citrus Trends. Scott Halver Appraiser Ganado Group
Ai Arizona Citrus Trends Scott Halver Appraiser Ganado Group 25,000 Yuma Mesa 20,000000 Price/Acre e 15,000 10,000 5,000 0 2000 2001 2002 2003 2004 2005 2006 2007 2008 2009 2010 2011 2012 2013 2014 Lemons
More informationAGRABLAST and AGRABURST TREATMENT OF COFFEE FUNGUS AND BLACK SIGATOKA ON BANANAS
AGRABLAST and AGRABURST TREATMENT OF COFFEE FUNGUS AND BLACK SIGATOKA ON BANANAS Coffee Leaf Rust is a major problem facing commercial coffee producers mainly in Africa, India, Southeast Asia, South America,
More informationTeacher s Manual. Rebecca W. Keller, PhD
Teacher s Manual Rebecca W. Keller, PhD Cover design: David Keller Opening page: David Keller, Rebecca W. Keller, PhD Illustrations: Rebecca W. Keller, PhD Copyright 2013 Gravitas Publications, Inc. All
More informationJuly marks another month of continuous low prices
July marks another month of continuous low prices In July 2018, the ICO composite indicator price decreased by 2.9% to an average of 107.20 US cents/lb, which is the lowest monthly average for July since
More informationProgress Report Submitted Feb 10, 2013 Second Quarterly Report
Progress Report Submitted Feb 10, 2013 Second Quarterly Report A. Title: New Project: Spotted wing drosophila in Virginia vineyards: Distribution, varietal susceptibility, monitoring and control B. Investigators:
More informationTitle: Western New York Sweet Corn Pheromone Trap Network Survey
Title: Western New York Sweet Corn Pheromone Trap Network Survey Project leader(s): Marion Zuefle Cooperator(s): Abstract: The New York sweet corn pheromone trap network (SCPTN) is an affiliation of extension
More informationSeparating the Components of a Mixture
Separating the Components of a Mixture Introduction: Mixtures are not unique to chemistry; we encounter them on a daily basis. The food and drinks we consume, the fuel we use in our vehicles, building
More informationThe GOODELL laboratory
The GOODELL laboratory Author Title Introduction Materials Protocol Shannon L.McKinney, KathyJo Jackson, Corinne Sonnet, Margaret A. Goodell 1996 Isolation of a heterogenous muscle-derived cell population
More informationVineAlert An Economic Impact Analysis
VineAlert An Economic Impact Analysis Goodman School of Business Consulting Group Goodman School of Business Cairns Complex Room 209A 500 Glenridge Ave. St. Cathararines, ON L2S-3A1 905-688-5550 Ext. 5104
More informationHERZLIA MIDDLE SCHOOL
NAME TEACHER S COMMENT TEACHER CLASS PARENT S COMMENT MARK PERCENTAGE PARENT S SIGNATURE HERZLIA MIDDLE SCHOOL GRADE 7 ECONOMIC & MANAGEMENT SCIENCES 27 AUGUST 2015 TIME: 50 minutes MARKS: 70 o This paper
More informationSetting up your fermentation
Science in School Issue 24: Autumn 2012 1 Setting up your fermentation To carry out all the activities, each team of students will need about 200 ml of fermentation must, 200 ml of grape juice and about
More informationTHIN LAYER CHROMATOGRAPHY AND MELTING POINT DETERMINATION: DETECTION OF CAFFEINE IN VARIOUS SAMPLES
EXPERIMENT 8 THIN LAYER CHROMATOGRAPHY AND MELTING POINT DETERMINATION: DETECTION OF CAFFEINE IN VARIOUS SAMPLES Additional Resources http://orgchem.colorado.edu/hndbksupport/tlc/tlc.html http://coffeefaq.com/caffaq.html
More informationPrices for all coffee groups increased in May
Prices for all coffee groups increased in May In May 2018, the ICO composite indicator increased by 0.7% to an average of 113.34 US cents/lb, following three months of declines. Prices for all coffee groups
More information2015/16 Harvesting Charges for Florida Citrus: Picking, Roadsiding and Hauling
2015/16 Harvesting Charges for Florida Citrus: Picking, Roadsiding and Hauling Ariel Singerman, 1 Marina Burani-Arouca, 2 and Stephen H. Futch 3 University of Florida, IFAS, CREC, Lake Alfred, FL A survey
More informationAPPENDIX Thirty Trees Sampling Method for CBB Monitoring
APPENDIX Thirty Trees Sampling Method for CBB Monitoring Based on a Presentation by Luis F. Aristizábal (University of Florida) Revised: January 2014 by CBB Summit Committee I t is not enough to know that
More informationParsnip Pancakes Revised By Mikaela Taylor, FoodCorps
Parsnip Pancakes Revised By Mikaela Taylor, FoodCorps Theme: Science, Cooking, Math Grade Level: 4th - 5th Subject Area : Science, Math, ELA Summary: Students learn about solids, liquids, and gases using
More informationCHAPTER I BACKGROUND
CHAPTER I BACKGROUND 1.1. Problem Definition Indonesia is one of the developing countries that already officially open its economy market into global. This could be seen as a challenge for Indonesian local
More information1. Continuing the development and validation of mobile sensors. 3. Identifying and establishing variable rate management field trials
Project Overview The overall goal of this project is to deliver the tools, techniques, and information for spatial data driven variable rate management in commercial vineyards. Identified 2016 Needs: 1.
More informationPeach and Nectarine Cork Spot: A Review of the 1998 Season
Peach and Nectarine Cork Spot: A Review of the 1998 Season Kevin R. Day Tree Fruit Farm Advisor Tulare County University of California Cooperative Extension Along with many other problems, fruit corking
More informationScientific curiosity as an emerging threat The P. kernoviae story. Dr Mike Ormsby, Senior Adviser, Biosecurity New Zealand
Scientific curiosity as an emerging threat The P. kernoviae story Dr Mike Ormsby, Senior Adviser, Biosecurity New Zealand Hypothesis: That Scientific Research posses a Biosecurity Risk 2 Evidence of Risk
More informationECONOMIC IMPACTS OF THE FLORIDA CITRUS INDUSTRY IN
ECONOMIC IMPACTS OF THE FLORIDA CITRUS INDUSTRY IN 2007- Mohammad Rahmani and Alan W. Hodges Food and Resource Economics Department Institute of Food and Agricultural Sciences FLORIDA CITRUS INDUSTRY HIGHLIGHTS
More informationSmall scale fisheries Big contribution
Small scale fisheries Big contribution Why developing fish health capacity is the insurance to this industry Seminar for OIE National Focal Points for Aquatic Animals Ghana, 20 22 March 2012 Qurban Rouhani
More informationNEW YORK CITY COLLEGE OF TECHNOLOGY, CUNY DEPARTMENT OF HOSPITALITY MANAGEMENT COURSE OUTLINE COURSE #: HMGT 4961 COURSE TITLE: CONTEMPORARY CUISINE
NEW YORK CITY COLLEGE OF TECHNOLOGY, CUNY DEPARTMENT OF HOSPITALITY MANAGEMENT COURSE OUTLINE COURSE #: HMGT 4961 COURSE TITLE: CONTEMPORARY CUISINE CLASS HOURS: 1.5 LAB HOURS: 4.5 CREDITS: 3 1. COURSE
More informationSCENARIO Propose a scenario (the hypothesis) for bacterial succession in each type of milk:
Prokaryotic Diversity! and Ecological Succession in Milk Name INTRODUCTION Milk is a highly nutritious food containing carbohydrates (lactose), proteins (casein or curd), and lipids (butterfat). is high
More informationFlorida's Citrus Canker Eradication Program (CCEP): Summary of Annual Costs and Benefits 1
FE532 Florida's Citrus Canker Eradication Program (CCEP): Summary of Annual Costs and Benefits 1 Marisa L. Zansler, Thomas H. Spreen and Ronald P. Muraro 2 Rapid expansion and integration of international
More informationExtraction of Acrylamide from Coffee Using ISOLUTE. SLE+ Prior to LC-MS/MS Analysis
Application Note AN796 Extraction of Acrylamide from Coffee using ISOLUTE SLE+ Page 1 Extraction of Acrylamide from Coffee Using ISOLUTE SLE+ Prior to LC-MS/MS Analysis This application note describes
More informationGrade: Kindergarten Nutrition Lesson 4: My Favorite Fruits
Grade: Kindergarten Nutrition Lesson 4: My Favorite Fruits Objectives: Students will identify fruits as part of a healthy diet. Students will sample fruits. Students will select favorite fruits. Students
More informationPaper Reference IT Principal Learning Information Technology. Level 3 Unit 2: Understanding Organisations
Centre No. Candidate No. Surname Signature Paper Reference(s) IT302/01 Edexcel Principal Learning Information Technology Level 3 Unit 2: Understanding Organisations Wednesday 3 June 2009 Morning Time:
More information