Notification of amended emergency quarantine measures for Xylella

The following measures apply to plant tissue cultures and nursery stock that are hosts of X. fastidiosa, and are applied in addition to current import requirements:

• nursery stock and plant material coming from countries or regions where X. fastidiosa occurs will need to be tested offshore and certified as being free from X. fastidiosa by the government of the exporting country
• an approved arrangement that ensures the health of plants will need to be in place for off-shore testing and certification of nursery stock from high risk countries/regions.
• material that do not meet the above requirements may be held and tested in an approved post- entry quarantine facility for 12 months, or nursery stock material may be hot water treated, followed by standard post- entry quarantine screening arrangements.

Details of the required measures for tissue culture are outlined in Appendix 1 and for other forms of nursery stock in Appendix 2. Appendices 1 and 2 provide the amended additional declarations required on Phytosanitary Certificates. These emergency measures for tissue cultures and nursery stock apply to all the plant species in the listed regulated families of plants (refer to Appendix 3).

Approved arrangements for the production of nursery stock in high risk countries/regions are outlined in Appendix 4.

List of appendices

Appendix 1	Import conditions for tissue cultures of species from regulated families
Appendix 2	Import conditions for Nursery stock - cuttings, rooted plants, budwood, and some corms and bulbs
Appendix 3	Plant families regulated for Xylella fastidiosa (nursery stock only)
Appendix 4	Offshore approved arrangements for nursery stock from countries/regions with a higher risk of Xylella fastidiosa

Appendix 1. Tissue cultures of species from regulated families

Country category	Off-shore certification	On-shore action if the Phytosanitary Certificate is acceptable (see note 3)	On-shore measures for consignments without an acceptable Phytosanitary Certificate
High risk countries/regions	A Phytosanitary Certificate with the following Additional Declaration or equivalent words: “All tissue cultures in this consignment were derived from mother tissue cultures that were tested by PCR and found free of Xylella fastidiosa as indicated on laboratory test report number ......... (insert number/code here).” (See note 4)	All other current import conditions for the plant species will apply	Tissue cultures must be de-flasked and grown for a minimum of 12 months in a government PEQ (see note 1) before testing by PCR. All plants will be tested. A positive detection of Xylella fastidiosa will result in destruction of the consignment. All other current import conditions for the plant species will apply. OR Export or disposal
All other countries and regions	A Phytosanitary Certificate with the following Additional Declaration or equivalent words: “Tissue cultures in this consignment were derived from plants and tissue cultures that were grown only in ..........(insert country) which is free from Xylella fastidiosa”	All other current import conditions for the plant species will apply.	Tissue cultures must be de-flasked and grown for a minimum of 12 months in PEQ (private or government) (see note 1) before testing by PCR. All plants will be tested. A positive detection of Xylella fastidiosa will result in destruction of the consignment. All other current conditions for the plant species will apply. OR Export or disposal

Appendix 2. Nursery stock - cuttings, rooted plants, budwood, and some corms and bulbs

Country category (see note 5)	Off-shore certification	Action if the Phytosanitary Certificate is acceptable (see note 3)	On-shore measures for consignments that come without an acceptable Phytosanitary Certificate
High risk countries/regions	A Phytosanitary Certificate with the following Additional Declaration or equivalent words: “Plant material in this consignment was produced under an arrangement approved by the National Plant Protection Organisation in accordance with Australian requirements. Plant material in this consignment was tested by PCR and found free of Xylella fastidiosa as indicated on laboratory test report number......... (insert number/code here).” (See note 2)	Current import conditions for the plant species apply.	Plants will be grown for a minimum of 12 months in government PEQ (see note 1) before testing by PCR. All plants will be tested. A positive detection of Xylella fastidiosa will result in destruction of the consignment. All other current conditions for the plant species will apply. OR Plants will be hot-water treated at 50°C for 45 minutes. Following treatment, all other conditions for the plant species will apply. OR Re-export or destroy.
All other countries and regions	A Phytosanitary Certificate with the following Additional Declaration: “Plant material in this consignment and its parent stock were grown only in ..........(insert country) which is free from Xylella fastidiosa”	Current import conditions for the plant species apply.	Plants will be grown for a minimum of 12 months in PEQ (private or government) (see note 1) before testing by PCR. All plants will be tested. A positive detection of Xylella fastidiosa will result in destruction of the consignment. All other current conditions for the plant species will apply. OR Plant will be hot-water treated at 50°C for 45 minutes. Following treatment, all other conditions for the plant species will apply. OR Re-export or destroy.

Notes

1. There are scheduled fees associated with the growth of nursery stock in an Australian Government post- entry quarantine facility, which must be met by the importer. The importer is responsible for contacting the facility to confirm all arrangements, including space availability and number of plants, prior to the plant material arriving in Australia. Importers must clearly nominate the facility that their material will be sent to on the import permit application.
2. Nursery stock and tissue cultures from high risk countries/regions must be produced through an arrangement approved by the National Plant Protection Organisation of the exporting country to meet Australia’s requirements (Appendix 4). These arrangements have also been distributed to NPPOs through the IPPC notification (https://www.ippc.int/en/)
3. The department will reserve the right to undertake testing to verify that a consignment is free of X. fastidiosa.
4. PCR tests that will detect X. fastidiosa including recognised sub-species. PCR testing will require the following two tests:
   1. the rimM gene sequence real-time PCR test of Harper et al. (2010),¹,
      AND
   2. the conventional PCR of Minsavage et al. (1994)² or an equivalent PCR that detects X. fastidiosa sub-species pear leaf scorch (PLS).
5. The department will temporarily delay implementation of X. fastidiosa emergency conditions for affected host certified bulbs (Narcissus, Hyacinths and Hippeastrum) produced under the Bloembollenkeuringsdienst (BKD) scheme from Netherlands. The department will continue to collaborate with the NPPO, to determine if alternative approved arrangements can be established for both certified and non-certified bulbs.

Appendix 3: Plant families regulated for Xylella fastidiosa (nursery stock only)

Acanthaceae	Boraginaceae	Escalloniaceae	Myrtaceae	Rosaceae
Adoxaceae	Brassicaceae	Euphorbiaceae	Nyctaginaceae	Rubiaceae
Altingiaceae	Bromeliaceae	Fabaceae	Oleaceae	Rutaceae
Amaranthaceae	Cannabaceae	Fagaceae	Onagraceae	Salicaceae
Amaryllidaceae	Cannaceae	Geraniaceae	Orobanchaceae	Sapindaceae
Anacardiaceae	Caprifoliaceae	Gesneriaceae	Oxalidaceae	Scrophulariaceae
Annonaceae	Caryophyllaceae	Ginkgoaceae	Passifloraceae	Simmondsiaceae
Apiaceae	Celastraceae	Haloragaceae	Paulowniaceae	Simaroubaceae
Apocynaceae	Cistaceae	Hamamelidaceae	Phytolaccaceae	Solanaceae
Aquifoliaceae	Clethraceae	Hydrangeaceae	Pinaceae	Strelitziaceae
Araliaceae	Commelinaceae	Hypericaceae	Pittosporaceae	Talinaceae
Araucariaceae	Convolvulaceae	Juglandaceae	Plantaginaceae	Tamaricaceae
Arecaceae	Cornaceae	Lamiaceae	Platanaceae	Theaceae
Argophyllaceae	Corynocarpaceae	Lauraceae	Poaceae	Ulmaceae
Asparagaceae	Cucurbitaceae	Linaceae	Polemoniaceae	Urticaceae
Asphodelaceae (Xanthorrhoeaceae)	Cupressaceae	Lythraceae	Polygalaceae	Verbenaceae
Asteraceae	Cyperaceae	Magnoliaceae	Polygonaceae	Violaceae
Athyriaceae	Dennstaedtiaceae	Malpighiaceae	Portulacaceae	Vitaceae
Balsaminaceae	Ebenaceae	Malvaceae	Proteaceae	Zygophyllaceae
Berberidaceae	Elaeagnaceae	Meliaceae	Ranunculaceae
Betulaceae	Equisetaceae	Montiaceae	Resedaceae
Bignoniaceae	Ericaceae	Moraceae	Rhamnaceae

Appendix 4: Offshore approved arrangements for nursery stock from countries/regions with a higher risk of Xylella fastidiosa (Pierce’s disease)

Note: As at December 2023, there are currently no offshore approved arrangements in place for nursery stock from high risk countries/regions.

These requirements are for offshore arrangements approved under Australia’s emergency measures for Xylella fastidiosa. The requirements apply to nursery stock of plants exported from high risk countries/regions. Offshore approved arrangements are currently not available for imports of true botanical seeds that can transmit Xylella.

The National Plant Protection Organisation (NPPO) of the exporting country will approve the arrangements and ensure that Australia’s requirements are met. Several other parties, including the grower and testing laboratory will need to work with the NPPO to do this. It is anticipated that the supplier will contact the NPPO of the exporting country to establish the arrangements. Australian importers should contact their overseas suppliers to ensure that the work is initiated for the arrangements.

This document describes the overarching systems and processes to ensure that nursery stock that is produced for export to Australia is grown, tested and confirmed free from infection by X. fastidiosa. The roles and responsibilities of the key parties involved in the arrangements are also described, as are the requirements for sampling and testing. The exporter and the testing laboratory should verify Australia’s requirements before testing is commenced. The Australian Department of Agriculture, Fisheries and Forestry retains the right to monitor the arrangements by auditing and by sampling and testing consignments after they arrive in Australia.

The arrangements

The arrangements require the following elements:

1. Propagation, growth, testing, certification and export under the authority of the NPPO of the exporting country
2. The facility where plants are grown is insect-proof so that it excludes all insects of the suborder Auchenorrhyncha (leafhoppers, froghoppers, sharpshooters, spittlebugs and treehoppers)
3. Plants for export to Australia are grown for their entire life in the facility regardless of the propagation technique used (e.g. plants grown from seed, grown vegetatively or grown in tissue culture)
4. All the mother plants are tested by the approved protocol (Appendix 5);
   • nursery stock – the mother plants that are the immediate source from which the nursery stock plant lot was propagated are tested
   • tissue cultures –the mother tissue cultures that are the immediate source from which the nursery stock plant lot was propagated are tested
   • The mother plants have been grown under the arrangement, in the facility, for 12 months before they are tested (Diagram 1)
     • nursery stock – the mother plants are protected in the facility for 12 months before testing
     • tissue cultures – the mother tissue culture line is protected and propagated in the facility for 12 months before testing
   • Prior to export, an official sample is drawn from the plant lot and tested according to the approved protocol for X. fastidiosa in Appendix 5. Appendix 5 contains details of sampling and sample size
   • Plant consignments are packed and packaged to prevent infection by X. fastidiosa
   • Phytosanitary certificates issued by the NPPO with additional declarations including information that enables tracing of plant lines for export to Australia to test results and the facility in which the lines were grown.

Roles and responsibilities

The NPPO

The NPPO will provide oversight of the arrangements to ensure that they meet Australia’s requirements. The NPPO is responsible for approving facilities or authorising any independent entities that approve facilities on its behalf. The NPPO may be requested by the Australian Government Department of Agriculture, Fisheries and Forestry to provide records relevant to the approval and management of the arrangements.

The NPPO or an entity acting under the authority of the NPPO will:

• Inspect the production facility to confirm that:
  • phytosanitary requirements are met
  • the facility is insect proof
  • the facility is constructed and equipped to achieve requirements
  • records are maintained by the producer or the facility
• Approve the facility under the arrangement
  • audit the facility and records
• Approve sampling of mother plants by the grower for testing or take samples of mother plants
• Take official samples of the plant lot for testing
• Maintain records of:
  • approved facilities
  • audits of facilities
  • phytosanitary practices within facilities
  • laboratory tests for X. fastidiosa
• Certify that plants exported to Australia are free from X. fastidiosa. This will be based on evidence of systems that confirm:
  • plants were kept in the registered facility throughout their life, from propagation to export
  • testing has been undertaken by a competent laboratory and laboratory reports have been sighted
• Phytosanitary Certificates must include:
  • additional declarations that indicate the status of the plant lot
  • the facility approval code/number
  • the laboratory report code/number

The grower

The grower will ensure that:

• phytosanitary conditions for Australia are met
• plants are appropriately tested by an approved testing laboratory
• provide samples of mother plants for testing, if approved by NPPO
• the facility is free from X. fastidiosa
• the facility is insect-proof.

The grower will also ensure that

• complete records are maintained of:
• plant lot identifying numbers or codes
• the parentage of the exported plant lots
• dates that plants are introduced to the facility
• mother plants, i.e. plants that are the immediate propagation source from which plant lots were propagated
• pathogens detected in plants in the facility
• arthropods found in the facility
• any plant material destroyed and the reason for the destruction
• all plants destined for export to Australia are transported in insect-proof closed containers or packaging.

The testing laboratory

The testing laboratory is approved by the NPPO as competent to undertake the testing required by Australia, using the prescribed testing methods.

It will:

• use the approved test protocol
• record the plant lots or mother plants that are tested and the number of samples tested
• provide the evidence of tests, results and operating processes to the NPPO, as required.

Diagram 1

The following measures apply to seeds that can transmit X. fastidiosa, and are applied in addition to current import requirements:

• must be grown and held for 12 months at an approved post-entry quarantine facility in Australia
• during the post-entry quarantine period, the resulting plants must be tested to confirm freedom from Xylella.

The above requirements apply regardless of the country of origin or export.

As at May 2022, Xylella regulations in seed is limited to Carya (pecans, hickory) species.

PCR testing will require the following two tests:

1. the rimM gene sequence real-time PCR test from Harper et al. (2010)¹,
   AND
2. the conventional PCR from Minsavage et al. (1994)² or an equivalent PCR that detects X. fastidiosa sub-species pear leaf scorch (PLS).

• all the mother plants must be tested for X. fastidiosa, i.e. the mother tissue cultures or the mother plants that are the immediate source from which the nursery stock plant lot was propagated

AND

• prior to export (up to 8 weeks before), an official sample will be drawn from the plant lot and will be tested for X. fastidiosa
• mother plants or mother tissue cultures will have been grown under the arrangement, within the facility, for 12 months before testing.

• when testing the sample from the plant lot, the sample size (number of units) will be set according to Table 1 in ISPM 31³ and will be sufficient to detect, with a 95% confidence level, that X. fastidiosa is not present in more than 0.5% (level of detection) of each lot, with the units defined as individual plants.
• two tissue samples per unit will be tested from tissue cultures, bulbs and corms
• three tissue samples per unit will be tested from nursery stock plants and cuttings
• samples must include mid-ribs of leaves, if the plant has leaves
• if the material lacks leaves, then living tissue with vascular structures will be sampled

Note: a unit is a tissue culture plantlet, nursery stock plant, a bulb or a corm.

• DNA extracted from up to 10 samples may be tested in a single PCR as a pool or batch, where a sample is defined as a single piece of tissue
• samples from different species should not be pooled

• house-keeping gene positive controls must be run for each batch of tests to confirm that the DNA was extracted successfully
• house-keeping positive controls must be run for each different plant species

• the laboratory must record the plant lots and mother plants that are tested and the number of samples tested
• If the testing is conducted prior to export (nursery stock only):
  • The exporting NPPO must verify the laboratory report and retain a copy
  • The identifying code or number of the laboratory report must be provided on the Phytosanitary Certificate

• Harper, S. J., Ward, L. I., and Clover, G. R. G. (2010). Development of LAMP and real-time PCR methods for the rapid detection of Xylella fastidiosa for quarantine and field applications. Phytopathology 100:1282-1288.
• Minsavage, G.V., Thompson, C.M., Hopkins, D.L., Leite, M.V.B.C. and Stall, R.E. (1994) Development of a Polymerase Chain Reaction protocol for detection of Xylella fastidiosa in plant tissue. Phytopathology 84: 456-461.
• ISPM 31 in International Standards for Phytosanitary Methods, No. 1 to 32 (2009), the Secretariat of the International Plant Protection Convention, Food and Agriculture Organization of the United Nations, Rome. pp. 401–420. https://www.ippc.int/en/publications/588/