Insect and Plant Disease Diagnostics at the Hawaii Department of Agriculture Bernarr R. Kumashiro* and Mann P. Ko, Plant Pest Control Branch, Hawaii Department of Agriculture, Honolulu, HI, USA 96814 & Brian Bushe and Scot C. Nelson, College of Tropical Agriculture and Human Resources, University of Hawaii at Manoa, Hilo, HI, USA 96720 ABSTRACT: The Hawaii Department of Agriculture, Plant Pest Control Branch, has a staff of five involved in the diagnostics of insects and plant diseases- three in the Insect Unit and two in the Plant Pathology Unit. Hundreds of identifications are processed through the two diagnostic units. Most of the identifications are done in-house with available resources, but local authorities are consulted within the state for difficult identifications. The Systematic Entomology Laboratory in Beltsville, MD is also often utilized for insect determinations. In events where identifications are of great concern to Hawaii or U.S. agriculture, the NPDN diagnostic protocol has been followed. Thus far this process has been used for the determination and notification of the Asian Citrus Psyllid and the Raspberry Late Leaf . The latter is being used as an example in this poster, describing the multiple steps of a modified NPDN Diagnostic Protocol that led from the initial collection of raspberry rust samples at a farm in the Hilo district, Hawaii, to the eventual confirmation of the rust as americanum by the scientists at the USDA/APHIS/PPQ National Identification Services.

DIAGNOSTIC UNITS AT THE HAWAII DEPARTMENT OF AGRICULTURE (HDOA) IDENTIFICATION OF A NEW UNKNOWN RUST IN HAWAII USING THE NPDN DIAGNOSTIC PROTOCOL

The Insect Taxonomy Unit consists of a Taxonomist, Assistant Taxonomist, and a Technician. This unit The NPDN Standard Operating Procedure for: APHIS-PPQ Pest of Concern Scenario-General SOP ** is responsible for: Revised May 1, 2009 was used for this notification and identification process, with slight modification. • Making authoritative identifications of insects and related arthropods of economic importance The SOP steps are detailed in the document, and only the schematic flow charts are shown here, for the Branch staff, general public, farmers, pest control operators, and other government followed by the respective list of Participants and Agencies. agencies. NPDN Communication Flow Charts • Making identifications of biological control agents introduced and utilized for controlling serious NPDN Notification pests in Hawaii. NPDN Confirmation Results Presumptive Positive • Assisting with identifications of insects and mites from HDOA Plant Quarantine Branch Delayed – backup pathway First Detector Producer APHIS-PPQ NPDN APHIS CDD NIS or SEL Emergency Diagnostician interceptions on imported and exported products. (NIS, CPHST or SEL) incl CPHST Domestic (With SPRO, Program SPHD and First APHIS Detector) Staff First Detector NIS or SEL Headquarter NPDN 1,073 samples were processed by the Insect Taxonomy Unit during Fiscal Year 2009. incl. CPHST and Eastern or Regional Producer Western Director SPRO Region The Taxonomist is a WPDN Principal Investigator and NPDN diagnostician. Program APHIS Eastern or Managers NPDN Western APHIS-PPQ (if agreed by APHIS Emergency and State Dept of Ag) Diagnostician Region ARD (May be LGU SPHD Domestic SPHD Program NPDN Other Other or SDA) National NPDN NPDN labs Program Regional for triage NPDN Leader Directors and surge NPDN National SPRO (NIFA) support Regional Program Director Leader (NIFA) SDA Lab

Bernarr Kumashiro, left, (Taxonomist), Damage on the indigenous naio Myoporum thrips (Klambothrips Janis Garcia (Technician), and Walter (Myoporum sandwicense) caused by myopori) was first discovered in Nagamine (Assistant Taxonomist). Myoporum thrips on Hawaii Island. Hawaii in March 2009.

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Biocontrol agent, Eurytoma erythrinae, Leaves of Erythrina variegata gnarled by Male and female Erythrina gall wasp introduced to control EGW. First released a tiny eulophid wasp. Most of the (EGW) (Quadrastichus erythrinae), in November 2008. Erythrina spp. in Hawaii have been killed. discovered in April 2005. The Plant Pathology Unit consists of a Plant Pathologist and a Technician. The primary function of this unit is to study potential plant pathogens for weed biocontrol and to maintain the Plant Pathology Containment Facility. In the area of diagnostics, this unit is responsible for: • Making authoritative identifications of plant pathogens for the Branch staff, general public, and CHRONOLOGY OF THE SCENARIO (2009) other government agencies. • Assisting with identifications of plant pathogens from the HDOA Plant Quarantine Branch • July 8-14 A raspberry farmer reported an unfamiliar rust disease outbreak on several of his red interceptions. raspberry (Rubus idaeus) cultivars to Brian Bushe (UH-CTAHR). Bushe consulted with Scot Nelson (UH-CTAHR), and together they observed the field, studied the pathogen, and determined that it was 1001 samples were processed by the Plant Pathology Unit during Fiscal Year 2009. probably a new rust to Hawaii. (The farm, totaling 6 acres, is located north of Hilo on the island of The Plant Pathologist is a WPDN participant and NPDN diagnostician. Hawaii. The farmer had been growing 4 varieties of raspberries on 1 acre for 4 years.)

• July 15 Bushe notified Mann Ko (HDOA and NPDN Plant Pathology Diagnostician) of the discovery and asked for assistance in sending samples to a USDA lab for identification, through the NPDN protocol.

• July 23 Bushe hand delivered samples to Ko's lab. Ko consulted with Bernarr Kumashiro (HDOA and NPDN Insect Diagnostician and WPDN representative) and together contacted Carla Thomas (WPDN Deputy Director) for advice on the procedure for sending urgent plant pathology samples for diagnosis. Thomas made arrangements with Mary Palm (USDA National Mycology Lab). Ko and Kumashiro were referred to John McKemy's lab. Unfortunately, Bushe's samples began to deteriorate, so it was agreed that Bushe should send fresh samples directly to McKemy's lab the following Monday.

• July 27 Bushe sent the samples to McKemy via United Parcel Service (UPS). Digital photos of rust Raspberry farm near Hilo, Hawaii where Green and ripe fruits and leaves are infested leaves and microscopic spores taken by Ko and Bushe were sent to Palm, McKemy, and Mann Ko (Plant Pathologist) and Chris a new rust disease was discovered in attacked by the rust. Thomas. Ko and Kumashiro notified Lyle Wong (Hawaii SPRO), Vernon Harrington (Hawaii SPHD), Lao (Technician) in front of Plant 2009. Pathology Containment Facility. Carol Okada (HDOA Plant Quarantine Branch Manager), and others about the suspected new raspberry rust.

• July 29 Samples were received by McKemy's lab. Indira Singh confirmed the as raspberry late leaf rust, Pucciniastrum americanum, on Rubus idaeus in Hawaii. John Bowers (USDA National Survey Coordinator) informed Wong, Harrington, and Joel Floyd (USDA Domestic Diagnostics Coordinator) of the new rust. Yolisa Ishibashi (Pest Survey Coordinator from SPHD office) notified Bushe. Bushe notified Ko, Kumashiro, Nelson, the farmer, and others. Ko and Kumashiro notified Thomas.

• August 10 Nelson and Bushe, after verifying with authorities and literature, concluded that P. Underside of leaf covered with the rust. Ripened fruit destroyed by the rust. Spores of the Raspberry Late Leaf Rust americanum was indeed a new record for Hawaii. There had been some confusion on whether the rust (Pucciniastrum americanum). was previously reported.