NPDN News Volume 6 Issue 9, September 2011

NPDN Participates in Department of Homeland Security (DHS) Tabletop Exercise (TTX) on Rathayibacter toxicus Marie L. Socha, Contract Support to Planning activities associated with this DHS Science & Tech Directorate, SHRR exercise helped to define informational Consulting, Inc.; Kitty Cardwell, USDA- gaps in sampling and analytical NIFA; Pat Sheil, USDA-APHIS and Karen methodologies. One of several gaps Snover-Clift, Cornell University found includes the lack of an approved method for which to analyze R. toxicus. n September 14 and 15, 2011, the Since R. toxicus is endemic in Australia, National Plant Diagnostic Network, O we requested their recommendations a member of the DHS Integrated for R. toxicus identification testing Consortium of Laboratory Networks procedures and sample collection (ICLN), participated in a two-day methods for field situations and tabletop exercise which focused on harvested products. intentional contamination of fields in Kansas, Colorado and Wyoming with The most significant benefit of this the plant pathogen, Rathayibacter toxicus. exercise was collaboration between The exercise was performed as part of fellow ICLN-members, including the ICLN’s regular practice of evaluating Environmental Protection Agency’s the ICLN Network Coordinating Groups (EPA) Environmental Response (ICLN NCG) Standard Operating Laboratory Network and USDA’s Procedure. During the two day exercise, mock Issue Highlights situational injects were provided to NPDN representatives and they were then evaluated on how they utilized • Daylily leafminer newly the ICLN portal, as well as, how they identified in North America alerted fellow ICLN NCG members • Diagnostic tip: using liquid using either a Preparedness Alert (if an nitrogen for tissue disruption incident has not been confirmed) or a Situation Report (if an incident has been • Pricing commercial image use analytically confirmed), and finally how • IT tip: Facebook security they convened conference calls to define whether they could handle the analytical • First regional workshop for the capacity which resulted from sampling Sentinel Plant Network in the efforts or if they needed to request Northeast assistance from a fellow ICLN network. • In Regional News: new soybean Other activities included utilizing the virus in NYS ICLN portal to upload and merge minimum data element worksheets (a form of data reporting sheet which can National Institute of be merged with other data sheets). Food and Agriculture Volume 6 Issue 9 1 National Animal Health Laboratory view/84826, from 2006, shot in Maine Network (NAHLN). During these on daylily, carries Dr. Lonsdale’s 2008 collaborative efforts, EPA was able to comment to that effect. provide information about sampling Also in 2011, Steck collected and resources which could be used by NPDN identified Florida specimens ofO. for mass sampling assistance while kwansonis. For more details see: Steck, NAHLN was able to inform NPDN G.J. and G.L. Williams, 2011, Florida of their ability to provide analytical DACS, DPI-Pest Alert – “Daylily support for times when NPDN exceeded leafminer, Ophiomyia kwansonis their normal laboratory capacity. This Sasakawa, (Diptera: Agromyzidae), new exercise illustrated how the consortium to North America, including Florida” (in of laboratories can effectively collaborate prep/review). during an animal and plant disease outbreak. Daylily leafminer is most easily detected by the long meandering, serpentine mines caused by larval feeding in the leaf blades (Fig. 1). Mines are usually seen in upper leaf surfaces, but in higher Daylily Leafminer, Ophiomyia populations or reinfestations, may also kwansonis Sasakawa, Newly occur in lower surfaces. One to several Identified in North America larvae may be found in a single leaf. The Gaye L. Williams, Plant Protection & Weed pale yellow larvae (Fig. 2), up to 5mm, Management, Maryland Department of can be detected in plant tissue with a 10x Agriculture and Gary J. Steck, Florida hand lens, usually by spotting the black Department of Agriculture and Consumer mouthparts as they rake back and forth Services, Division of Plant Industry in the mine, feeding on plant tissue Since at least 2008, daylily growers and collectors have been aware of an unknown leafminer damaging leaves of Hemerocallis spp. and varieties (see comments at www.daylilies. org/ahs_dictionary/leafminer. html). In late 2008, at a Maryland daylily garden, Williams noted leafmining damage but found no live larvae. Attempts to rear flies from this site in 2009 and 2010 were unsuccessful. In June 2011, adults reared from Fig. 1. Characteristic larval mining in daylily leaves. puparia collected at the site were Photo courtesy of Sharon Galloway. tentatively identified as Ophiomyia kwansonis Sasakawa. Collected larvae, pupae, adults and reared specimens were sent to Dr. Owen Lonsdale, Canadian National Collections, who confirmed their identity as O. kwansonis, a species previously known only Fig. 2. Larval leafminer. Photo by Gary J. Steck, from Japan and Taiwan. An FDACS. image, www.bugguide.net/node/ 2 NPDN News (Fig. 3). Pupation takes place in the larval specimens in Florida and Maryland, tunnel with spiracles protruding through by the internet image from Maine; the plant’s epidermis (Fig. 4). Pupae diagnosed by larval mining damage (Fig. 5) are tan, about 3-4mm and usually in daylily leaves in Alabama, Georgia, found in the basal half of the leaf. Adult Louisiana, North Carolina, New York, flies are stocky and shining black, often seen on daylily blooms (Fig. 6). This insect may have two or more generations or be continuously present during the growing season, depending on region. Although even severe mining does not appear to Fig. 3. Larva in tunnel. Photo by Gaye Fig. 5. Pupae removed from tunnels. Photo kill plants, mining Williams. by Gaye Williams. damage accumulates and will persist until leaves are removed or replaced by new growth. This means that plants remain disfigured throughout the flowering season, causing major concern for display and tour garden owners. Regulatory and trade Fig. 4. Fly pupa in larval tunnel. Photo by Fig. 6. Adult fly on daylily flower. Photo by impacts are not known Gaye Williams. V.J. Hickey, courtesy of P. Hickey. as yet since this insect has only recently appeared on our radar screens. South Carolina, Texas and Virginia). No formal studies have been conducted With increased awareness, this list will to establish reliable chemical control grow as daylily plants in other states are methods. carefully examined. If you see leafminer Sanitation, including removal and damage in Hemerocallis leaves, please destruction of infested leaves may send images and information to Williams reduce potential numbers of adults. at [email protected]. Unfortunately, control may prove difficult as infested naturalized populations of Hemerocallis fulva, the roadside lily, can serve as sources of reinfestation for nurseries and gardens. At present, distribution of daylily leafminer includes Japan, Taiwan and continental U.S. (confirmed by

Volume 6 Issue 9 3 Tip for Using four months with liquid nitrogen. The Liquid Nitrogen purchase of this tank has greatly reduced Diagnostic the amount of liquid nitrogen we waste and a Mini- and has saved us time since we no longer Updates BeadBeater-1 for need to fill a small container several Tissue Disruption times a week. As an added advantage, Jen Olson, Plant Disease the tank can be used for storage of a and Insect Diagnostic Laboratory, culture collection. Oklahoma State When we need to perform DNA or University RNA extractions, we place plant or ur laboratory insect tissue inside a 2.0 ml plastic Oroutinely uses tube filled 25–50% with 2.5 mm glass liquid nitrogen to beads (Fig. 2). It is important to try out disrupt plant and insect different tubes because some plastic tissues in preparation for microcentrifuge tubes will break when nucleic acid extraction. immersed in liquid nitrogen. The beads We do not have access to a may also damage the plastic during liquid nitrogen tank in homogenization so you may need to try our building so we have developed a different size beads. solution to running across campus to The tubes are clipped onto a “stick” get liquid nitrogen for DNA and RNA that consists of three or four cryocanes extractions. It is also much safer than squeezed together to make one long stick having an open vat of liquid nitrogen on (Figs. 3, 4). The lid to the 20L dewar is the lab bench. opened and the stick is placed directly in Several years ago we purchased a 20L the liquid nitrogen (Fig. 5). After 20–30 dewar for liquid nitrogen storage (Fig. 1). seconds, we remove the stick and unclip We fill the dewar approximately every the frozen tube. Although you can wear

1 2

Fig. 1. 20L dewar for liquid nitrogen storage. Fig. 2. Plant tissue placed in 2.0 ml microcentrifuge tube with 2.5 mm glass beads. Fig. 3. Tube is clipped onto stick and ready for 3 immersion in liquid nitrogen. Fig. 4. Four cryocanes are squeezed together to make one 4 long stick.

4 NPDN News special gloves to handle the tube, we find that nitrile gloves are adequate for 5 the brief amount of time we handle the tubes. The frozen tube is placed in the Mini-BeadBeater-1 (BioSpec Products, Inc.) and processed for 20–30 seconds at 4200–4800 rpm (Fig. 6). We generally repeat the freezing and homogenizing process once to ensure that all tissue is disrupted. If there is a lot of tissue on the lid of the tube, we will briefly centrifuge the tube to concentrate the tissue at the bottom of the tube. The sample is ready for nucleic acid extraction (Fig 7).

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Check out the Summer 2011 edition of the WPDN First Detector News on Invasive Horticultural Plants

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Fig. 5. Stick and tube are immersed in liquid nitrogen directly in the dewar. Fig. 6. Frozen tube is placed in Mini-BeadBeater-1 and ready for homogenization. Fig. 7. Microcentrifuge tube containing homogenized plant tissue.

Volume 6 Issue 9 5 Pricing Commercial Image Use book. For each request, Sam charges How Much Should You Charge? $400. Assuming that GP decided to use Joseph LaForest, Center for Invasive Species the images in all cases, Sam has received & Ecosystem Health, University of Georgia $1,600 for use of the image. Photographers who have posted images Royalty Free in the Bugwood Image Database often Despite the word FREE being in the receive requests to use their images in name, the images do cost money. commercial projects. For the purpose Royalty Free simply means that after of this article, I’m going to assume that paying money for use of the image, the the photographer owns the copyright to buyer has permission to use the images their images…working at a land-grant as much as they want without paying university, federal institution, or other additional royalties. non-profit can throw this into question Royalty Free rates vary widely by (but I’ll cover that in a different article). photographer. Examples I have seen When thinking about charging fees for can range widely from $10 for small commercial use of images, everyone has images (~200 pixels on the long edge) to to answer the question “What should I $800 for very large images (~5,000 pixels charge for use of an image?” Although on the long edge). Depending on the there are many nuances that can possibly uniqueness of the images and prestige apply to commercial licensing of images, of the photographer, higher fees are not there are four main ways that I have seen uncommon. Some photographers also people use to determine a price: place a time limit on the license. 1. Flat-Rate Using the previous example: Gopher’s Publishing (GP) pays Sam Arnold $400 2. Royalty-Free for four year, Royalty Free license to an 3. Rights Managed image that will be used on commercial projects. They still have permission 4. Modified Flat-Rate to use the image in the circular since Flat-Rate the license was Royalty Free. When This is pretty straight forward. For they choose to use the images in the any request to use an image, a set fee textbooks, they find that the license has is charged. Permission is granted for expired. They contact Sam and pay an the image to be used in the project additional $400 for a new license. Over that was specified in the request only. the entire use of the image, Sam has Any other uses would be a separate received $800 for allowing use of the request. A photographer can set their image. price at whatever they like. Common Rights Managed values I’ve seen from photographers This is a far more detailed way to license in the Bugwood Image Database range the use of images. The price charged between $50 and $500. for the images depends on how the Here’s an example: Gopher’s Publishing image will be used. Since there is more (GP) requests use of an image by Sam detail in the licensing, there is more Arnold for a billboard. Sam’s flat rate is work required in figuring out the cost. $400. After paying Sam for the image, Typical considerations for the calculation GP decides to use the image on a circular include: to customers. They make a request and • Usage type — advertising, retail, Sam charges $400. Five years later, GP publishing/editorial, electronic, etc. decides to use it again as the cover of a book and an interior photo of another article continues on page 8 ... 6 NPDN News PROGRAM AREA COMMITTEES PAC Highlights

Visit the NPDN homepage at www.npdn.org for more information on specific Program Area Committees. Login and password required

Diagnostics Committee Anne Vitoreli, Committee Chair, University of Florida, Department of Plant Pathology ics

t The Diagnostics Committee held a conference call on The next call is tentatively scheduled for October 13,

s September 15, 2011, and the following agenda items 2011. We will cancel if there are no new agenda items

o were discussed: to be discussed.

n • Protocols for NPDN lab accreditation There will be NO November Diagnostics

g • SOP’s in progress Subcommittee conference call. • Proficiency panels

ia • Sentinel Plant Network regional workshops The December Diagnostics Subcommittee conference call is scheduled for December 8, 2011. d Co mmi ttee • Diagnostics Committee poster for Nat’l Meeting

National Database Committee Nancy Gregory, Committee Chair, University of Delaware, Department of Plant and Soil Sciences Following the last newsletter, the National Database • Discussion of change submissions Subcommittee held a conference call on September • Sentinel Plant Network data 14, 2011. The subcommittee continues to work on • Discussion of Insect pest files reviewing the extensive NPDN pest lists. The agenda • Poster for National Meeting included: The next meeting will be held on October 12, 2011. N a t i o na l Da t a b as e

Training and Education Committee Dick Hoenisch, Committee Chair, University of California at Davis, Department of Plant Pathology The Training and Education Committee conducted • National Meeting First Detector awards a conference call on September 19, 2011, and the • Web update following agenda items were discussed: • Protect US update • Sentinel Plant Network (SPN) update • Review and update of modules • National Meeting in Berkeley, CA The next meeting will be held on October 18, 2011. T RAININ G ED UCA T I O N

Website Committee Karen Scott, Committee Chair, Cornell University, Department of Plant Pathology and Plant–Microbe Biology The Website Committee conducted a conference call • Website Committee poster on September 12, 2011 and the following agenda • Access policy of the website items were discussed: The next Website Committee conference call is scheduled for October 3, 2011. • National Meeting WEBSITE Co mmi ttee

Volume 6 Issue 9 7 continued from page 6 ... • Image visibility — front cover, non-profit trade magazine, 10 being magazine insert, postcard, home luxury cars) page, etc. 3. Base price — The most you would • Duration of license — pretty much ever charge for your image (e.g. any length of time selling luxury cars in the Super Bowl). This essentially would be • Print size — ¼ page, full page, etc. your flat rate which will be adjusted • Circulation — how many copies based on Exposure and Profit of the will be distributed? use using the formula below. • Regions of use — worldwide, US Usage Fee = (Exposure Rating x Profit only, regional, etc. Rating) % x Base Price • Target audience — agriculture, Applying this to the example: food, education, etc. • Billboard—local use for Ag Expo As you would expect, all of these (3 x 4)% x $2,500 = 12% x $2,500 = $300 variables have wild implications for the • Circular—regional use for new herbicide price that is charged. Using the example (4 x 6)% x $2,500 = 24% x $2,500 = $600 above and couple of the online stock photo calculators, here is a range what • Textbook—national use for biology GP would be looking at: (5 x 3)% x $2,500 = 15% x $2,500 = $375 • Billboard (up to 1 year, 4’x6’ size, up • Textbook—national use for biology to 5 Locations, minor element, in the (3 x 3)% x $2,500 = 9% x $2,500 = $225 US) – $660–$2,500 Judgments on the profitability and • Circular (6 month campaign, ½ page, exposure will vary from person to 5,000 copies, US only) – $500–$1,300 person but this method does allow for taking that into account. By this method, • Textbook (published for 1–2 years, Sam would have received $1,500 for the full page, front cover, 2000 copies, US use of his image. only) – $360–$800 As you can see, the different pricing • Textbook interior (published for 1–2 methods vary greatly in the revenue years , 1/4 page, 10,000 copies, US that may potentially be generated from only) – $120–$290 commercial use. This does not take In this pricing method, Sam would be into account whether or not the person paid anywhere from $1,640 to $4,890 for requesting use of the images chooses the use of the images. to pay what you ask. It is possible that more people would pay for use of an Modified Flat-Rate image through the royalty free system This model takes into account that not rather than dealing with the details all image uses are the same but not and higher price of a rights managed in anywhere near the detail of Rights license. This also does not take into Managed licensing. Three factors are account any fees that may be incurred if looked at: an individual uses an agency or service 1. Exposure rating — How prominent to process the request and licensing. will the image be from 1–10 (1 for Despite these limits, hopefully this quick thumbnail on a small website, 10 for primer on different options will help in Super Bowl commercials) finding a model that works for you. 2. Profit rating — How much does the user intent to profit from using the image for 1–10 (1 being a small ad in 8 NPDN News Security Tip: of such a tool and have Facebook created a presence on Facebook as well. As is IT News Understanding It So You the case with all software Can Secure It tools available on the Mike Hill and Eileen Luke, internet, good security CERIS, Purdue University practices are important for both the user Keith Watson, a Research Engineer with and the provider. CERIAS (Center for Education and Research in Information Assurance and Security) at Purdue University has been collaborating with the NPDN for over seven years, providing security assessments, reviews, training, presentations and general consulting. His expertise has been invaluable to the network in aiding IT staff in achieving the number one goal of operating and maintaining good security practices. Keith Watson, along with Linda As technology and social media McCarthy and Denise Weldon-Siviy, evolve, new demands arise in securing wrote an article titled, ‘A Guide to information. Facebook, a popular social Facebook Security.’ This guide explains networking service, originally was a how you can protect your Facebook means for individuals to link socially account, avoid the scammer, use online and share their information on advanced security settings, recover a a wider scale. Companies, institutions, hacked Facebook account, and stop and organizations have seen the value imposters. Take the time to read this article; it’s worth it. This document is available for free on Facebook and can be downloaded at www.facebook. com/safety/ attachment/ Guide%20to%20 Facebook%20 Security.pdf

Volume 6 Issue 9 9 Sentinel Plant the NEPDN, and Rich Cowles from the Network Regional Connecticut Agricultural Experiment Training and Station, who in addition to highlighting Workshop at the specific insects and plant diseases, Education Arnold Arboretum focused time on improving scouting by Rachel McCarthy, sharpening one’s observation skills. Department of Plant Pathology and Plant-Microbe Biology, The meeting was well attended Cornell University with 28 people from 18 gardens and professionals from horticulture and IPM The Sentinel Plant Network (SPN) held departments, as well as, education and its first workshop for northeast regional outreach departments. Participating gardens at the Arnold Arboretum in gardens were from Massachusetts, Boston, MA, on September New York, 22–23. This was the first Pennsylvania, workshop in a series Connecticut, intended to engage public Maryland, Maine gardens in the early detection and New Jersey. of regionally significant high- consequence pathogens and The next insect pests. workshop is intended for The Sentinel Plant Network, midwestern a cooperative endeavor gardens and is between the American scheduled for Public Gardens Association October 17–18 (APGA) and the NPDN, at the Morton expects to enhance diagnostic Arboretum in capabilities via improved Lisle, IL. The communication between western regional public gardens and expert workshop is diagnostic facilities. The scheduled for workshop introduced Photo top, George Hudler, and December 5–6 garden professionals to the bottom, Rich Cowles, talking to workshop participants at the Arnold at the Rancho NPDN and its mission with Arboretum. Photos courtesy of Dan Santa Ana presentations and hands-on Stern, American Public Gardens Botanic Garden activities on how to use our Association. in Claremont, online resources, how to CA. Additional collect and submit samples, workshops are and where to find information on their planned in early 2012 for gardens in the state’s diagnostic laboratory so they can southern and Great Plains regions. send in suspect samples. There may be funds available for NPDN The program also featured the unveiling diagnosticians and entomologists to of the first three Train-the-Trainer attend as guests; those interested should modules designed specifically for the contact Rachel McCarthy at rachel. SPN—Module 1: Public Gardens and [email protected]. the Sentinel Plant Network, Module 2: Photography for Diagnosis, and Module 3: Submitting Samples to the National Plant Diagnostic Network. Of course the most talked about sessions were the pest walks led by George Hudler, Director of 10 NPDN News viewed photographs with symptoms from a field in Herkimer County, NY that Regional also appear to be caused News Soybean Vein Necrosis Virus by SVNV; the symptoms were predominantly Confirmed in New York on the upper, youngest Gary Bergstrom, Department of Plant leaves and were being described as a ‘top Pathology and Plant-Microbe Biology, dieback’. Cornell University What is known about SVNV? The The presence of a new soybean disease answer is not very much. It is thought to was confirmed this week in New York. be transmitted from soybean to soybean The pathogen is Soybean vein necrosis by thrips (soybean thrips and perhaps virus (SVNV). It was first described in others). Soybean thrips are Tennessee in 2008, and observed in New York along in Arkansas, Illinois, with a number of other thrips Kansas, Kentucky, and I credit the timely diagnosis of species. The virus has been Missouri in 2009. Dr. placed in the Tospovirus Don Hershman of the SVNV in New group of plant RNA viruses University of Kentucky York to excellent (stands for Tomato spotted commented recently wilt virus) which are that the disease was communication among Northeastern transmitted by thrips. Finding observed in late it this year doesn’t mean we season at low levels Plant Diagnostic will find it next year. We don’t in Kentucky many Network participants. know if the virus can also be years before its viral transmitted through seed or etiology was proven. by any other means. We don’t The symptomatic plants in New York know if it will have an impact on yield. came from a soybean field in Ontario Potentially the virus may be transported County. Symptoms observed in the field long distances were vein-clearing followed by ‘scalded’ by thrips in one reddish areas around the veins and a growing season browning of the veins, especially on the or the virus may lower leaf surface (see the photo). Dr. survive locally Ioannis Tzanetakis of the University of in weed hosts Arkansas, a leading researcher on SVNV, (no one has positively identified RNA of SVNV in demonstrated this samples from Ontario County. I credit yet) and then be the timely diagnosis of SVNV in New transmitted locally York to excellent communication among by thrips when Northeastern Plant Diagnostic Network their population participants. An NEPDN listserve increases communication from Nancy Gregory at within a season. the University of Delaware with photos Tospoviruses of SVN symptoms observed in Delaware are circulative/ arrived at the precise time that I was propagative in trying to diagnosis unusual symptoms their association on soybean in New York. Recently with thrips, so Photo courtesy of Gary C. SVNV was confirmed in Delaware, if thrips survive Bergstrom, Cornell University. Maryland, and Virginia, with pending our winters, diagnoses in Pennsylvania. Yesterday I SVNV might well Volume 6 Issue 9 11 survive in the living thrips. Insecticidal cropping practices just because we found seed treatments may have a role to this new virus. For now, I would classify play in killing thrips on young soybean its presence in NY only as a potential plants and reducing the incidence of problem worth keeping an eye on. We early virus infection. Resistant varieties encourage growers and consultants appear to be the main path to sustainable in New York to inspect any still-green management and several investigators soybean plants for possible SVNV and are assessing varieties in other parts to inform their local Cornell Cooperative of the country. There is no basis to Extension field crops educator if they recommend that farmers do anything find symptoms that are similar to substantially different now in their soybean vein necrosis.

Upcoming Events

National Events

November 6–9, 2011 NPDN National Meeting Berkeley, CA

November 13–16, 2011 ESA 59th Annual Meeting Reno, NV

Rachel McCarthy, Editor NEPDN Cornell University

12 NPDN News