Staff Advice Report

June 2021

Advice to the Decision-making Committee on APP204242

Application code: APP204242

Application type and sub-type: Statutory determination

Applicant: idverde Bomendienst B.V.

Date application received: 28 May 2021

Purpose of the Application: Information to support the consideration of the determination of Verticillium nonalfalfae Inderb., H.W.Platt, R.M.Bostock, R.M.Davis & Subbarao (2011)

Executive Summary

Application APP20242 submitted by idverde Bomendienst B.V. seeks a determination on the organism status of Verticillium nonalfalfae in New Zealand.

The applicant provided evidence that the microorganism V. nonalfalfae (formerly known as V. albo-atrum) has been found and collected in various crops across New Zealand since before 1998.

After reviewing the information provided by the applicant and information found in scientific literature, EPA staff recommend that V. nonalfalfae is not a new organism for the purpose of the HSNO Act.

Introduction

1. On 28 May 2021, idverde Bomendienst B.V. applied to the EPA under section 26 of the HSNO Act seeking a determination on the new organism status of Verticillium nonalfalfae.

2. Section 2A(1) of the HSNO Act prescribes that a new organism is, in part, an organism belonging to a species that was not present in New Zealand immediately before 29 July 1998. It is against that criterion that we evaluated the evidence available for the microorganism in the application. Purpose of this document

3. This document has been prepared by the EPA staff to advise the Committee of our assessment of application APP204242 submitted under the Hazardous Substances and New Organisms Act 1996 (the Act). This document discusses information provided in the application and other sources.

4. The decision path for this application can be found in Appendix 3. The application 5. The applicant, idverde Bomendienst B.V., developed a bio-control vaccine called Dutch Trig containing the fungus Verticillium nonalfalfae, an elicitor of the elm tree’s self-defence mechanism against Dutch elm disease. 6. The species previously known as V. albo-atrum was split into three distinct species in 2011 (V. albo-atrum sensu lato, V. alfalfae and V. nonalfalfae) following a phylogenetic analysis based on DNA sequences. The applicant provided evidence that samples of V. albo-atrum isolated in New Zealand before 29 July 1998 were later identified as V. nonalfalfae. 7. Our assessment includes information contained within the application and any other relevant information found in the scientific literature or elsewhere. Organism description

8. Verticillium nonalfalfae (Table 1) is a soil-borne fungus belonging to the order Hypocreales. It is a plant pathogen that causes Verticillium wilt on a wide range of species including hops, kiwifruits, potatoes and eggplants. It infects its hosts through roots and spreads in the plant’s xylem.

9. The species V. nonalfalfae was formerly known as V. albo-atrum (Mellow et al. 2019). Verticillium albo-atrum was split into three species: V. albo-atrum sensu stricto, V. alfalfae and V. nonalfalfae based on molecular systematics work. The three species are difficult to differentiate based on their morphological characters (Inderbitzin et al. 2011). Table 1: Taxonomic description of Verticillium nonalfalfae

Taxonomic Unit Classification

Order Hypocreales

Family Plectosphaerellaceae

Genus Verticillium

Species V. nonalfalfae Inderb., H.W.Platt, R.M.Bostock, R.M.Davis & Subbarao (2011)

Review of information

10. The evidence of V. nonalfalfae’s (formally V. albo-atrum until 2011) continuous presence in New Zealand found in the application form and literature is summarised in Table 2.

Presence of Verticillium albo-atrum 11. The fungus V. albo-atrum was found in tomato and potato crops around the country between 1947 and 1957 (Smith 1965). In 1956, an outbreak of wilt, identified as V. albo-atrum, in Nelson was observed (Christie 1956). 12. The applicant also provided evidence that V. albo-atrum was isolated from diseased plants in March 1973 in Pukekohe, then in 1977 from rockmelon in Hamilton, in 1979 from globe artichoke in Pukekohe and in 1981 from eggplant in the Waikato region (Hill 1982). The fungus was also isolated between 1979 and 1981 from lucerne (Hawthorne 1983).

13. The samples were not analysed after the split of the species in 2011, however, it is reasonable to believe that some of these specimens might have been V. nonalfalfae.

Presence of Verticillium nonalfalfae

Figure 1: Distribution of Verticillium nonalfalfae in New Zealand. Source: GBIF.

14. The applicant provided evidence of Verticillium samples isolated from New Zealand crops between 1968 and 1978 that were later identified as V. nonalfalfae (MWLR 2021). The same data are used on GBIF to show the wide distribution of V. nonalfalfae across the South Island and the North Island (Figure 1). 15. Evidence of V. nonalfalfae’s presence after 29 July 1998 is supported by its classification as ‘present’ in the New Zealand Organisms Register (NZOR ND), based on the Manaaki Whenua Landcare Research-curated New Zealand Fungi database (MWLR 2021). In 2019, the fungus was considered to have been present in the country for many years in a study on Verticillium spp. on kiwifruit in New Zealand (Mellow et al. 2019). 16. Verticillium nonalfalfae is also listed as present in New Zealand by the United Kingdom Plant Health Risk Register (Department for Environment 2020) and the European Food Safety Authority (Health 2014), however, the original source of the information is unknown.

17. In addition to the specimens found in New Zealand, the applicant noted that the fungus was widely spread around the world including Canada, the USA, China, Cuba, Japan, Middle Asia, Belgium, Slovenia, the Netherlands and the UK on common crop species.

Table 2: Samples containing Verticillium nonalfalfae and V. albo-atrum in New Zealand.

Year Sample Reference

1947 to 1957 V. albo-atrum isolated from tomato and potato crops (Smith 1965) throughout New Zealand

1956 V. albo-atrum was isolated from the diseased hops (Christie 1956) following an outbreak of wilt in a hop garden in the Tadmor district (Nelson)

1968, 1969, Potato (Alexandra, Invercargill), cucumber (MWLR 2021) 1974, 1977, (Whangarei, Auckland), lucerne (Hawkes Bay), 1979, 1984 tomato (Gisborne, Auckland, Nelson), tobacco (Motueka). Subsequently reclassified as V. nonalfalfae after molecular analysis of stored samples.

1973, 1977, V. albo-atrum isolated from diseased plants (Hill 1982) 1979, 1981 (Pukekohe), rockmelon (Hamilton), artichoke (Pukekohe), eggplant (Waikato)

1979 to 1981 V. albo-atrum isolated from lucerne (Taupo, (Hawthorne 1983) Canterbury and Central Otago)

1986 Present in New Zealand (EPPO Global Database ND)

2003 V. albo-atrum isolated from kiwifruit (Te Puke) (Mellow et al. 2019)

ND V. albo-atrum (V. nonalfalfae under the new (Health 2014) taxonomy) confirmed to be present in New Zealand

ND Presence of V. nonalfalfae but not the highly (Mellow et al. 2019) pathogenic strain found in Chile

ND Listed as present in New Zealand (Department for Environment 2020)

ND Listed as present in New Zealand (NZOR ND)

18. Due to the wide distribution of V. nonalfalfae pre-1998 in New Zealand on various crops and its presence post-1998 in the country, we considered that the fungus is highly likely to have had a continuous presence in New Zealand since before the 29 July 1998 when the Act came into force. Evaluation against legislative criteria

19. For an organism to be determined as “not new” under section 26 of the Act, the organism must be shown to lie outside the definition of a new organism as defined in section 2A(1) of the Act: 20. A new organism is- a. an organism belonging to a species that was not present in New Zealand immediately before 29 July 1998: b. an organism belonging to a species, subspecies, infrasubspecies, variety, strain, or cultivar prescribed as a risk species, where that organism was not present in New Zealand at the time of promulgation of the relevant regulation:

c. an organism for which a containment approval has been given under this Act: d. an organism for which a conditional release has been given: e. a qualifying organism approved for release with controls: f. a genetically modified organism: g. an organism that belongs to a species, subspecies, infrasubspecies, variety, strain, or cultivar that has been eradicated from New Zealand. 21. The decision pathway for a section 26 determination is outlined in Appendix 3.

22. Section 2A(1)(a) of the Act states that a new organism must belong to “a species that was not present in New Zealand immediately before 29 July 1998”. We have evaluated the information regarding V. nonalfalfae against this criterion. Comments from Agencies

23. In accordance with section s26(5) of the Act, and the Methodology, the Department of Conservation (DOC) and the Ministry for Primary Industries (MPI) were notified and provided with the opportunity to provide further information on the application.

24. DOC noted that samples collected in New Zealand prior to 1998 were recently identified as V. nonalfalfae therefore this fungus is not a new organism (Appendix 1). 25. MPI supports the applicant’s contention that Verticillium nonalfalfae is not a new organism under the HSNO Act and believes that there is sufficient evidence to conclude that the species was present in New Zealand immediately prior to 29 July 1998 (Appendix 2). Effect on New Zealand’s international obligations

26. It is not considered that any international obligations may be affected by this determination. Recommendation

27. After completing our assessment of the information that was submitted by the applicant, as well as our own findings, we consider that V. nonalfalfae was present in New Zealand immediately before 29 July 1998 and has had an ongoing presence in New Zealand. 28. A new organism is defined in section 2A of the Act, and includes: (a) An organism belonging to a species that was not present in New Zealand immediately before 29 July 1998. 29. The following Act criteria were not applicable to this determination as the species under consideration in this application; • has not been prescribed as a risk species (section 2A(1)(b)); • has not been approved to be held in containment or released with controls (sections 2A(1)(c), (ca) and (cb)); • is not genetically modified organism (section 2A(1)(d)); and • has not been eradicated from New Zealand (section 2A(1)(e)). 30. Therefore, we recommend that Verticillium nonalfalfae should be determined to be not a new organism for the purpose of the Act.

References Christie T 1956. A note on an outbreak of Verticillium wilt in a Nelson Hop garden. New Zealand Journal of Science and Technology, Section A 38(1). Department for Environment FRA, 2020. UK Plant Health Risk Register (Verticillium nonalfalfae) Retrieved 27 May. https://secure.fera.defra.gov.uk/phiw/riskRegister/viewPestRisks.cfm?cslref=27818 EPPO Global Database, ND. Verticillium albo-atrum. Retrieved 4 May. https://gd.eppo.int/taxon/VERTAA/distribution/NZ Hawthorne B 1983. Variation in pathogenicity among isolates of Verticillium albo-atrum from lucerne. New Zealand journal of agricultural research 26(3): 405-408. Health EPoP 2014. Scientific Opinion on the pest categorisation of Verticillium albo‐atrum sensu stricto Reinke and Berthold, V. alfalfae Inderb., HW Platt, RM Bostock, RM Davis & KV Subbarao, sp. nov., and V. nonalfalfae Inderb., HW Platt, RM Bostock, RM Davis & KV Subbarao, sp. nov. EFSA Journal 12(12): 3927. Hill C 1982. New plant disease records in New Zealand, 1974–1982. New Zealand journal of botany 20(4): 355-359. Inderbitzin P, Bostock RM, Davis RM, Usami T, Platt HW, Subbarao KV 2011. Phylogenetics and taxonomy of the fungal vascular wilt pathogen Verticillium, with the descriptions of five new species. PloS one 6(12): e28341. Mellow KD, Tyson JL, Manning MA, Wright PJ 2019. Preliminary pathogenicity screening of Verticillium spp. on kiwifruit in New Zealand. New Zealand Plant Protection 72: 89-94. MWLR, 2021. Systematic Collections Data - Verticillium nonalfalfae. Retrieved 26 May. https://scd.landcareresearch.co.nz/Search NZOR, ND. Verticillium nonalfalfae. Retrieved 27 May. http://www.nzor.org.nz/names/9d7af195-08c3- 420c-bd73-026d3137b601 Smith HC 1965. The morphology of Verticillium albo-atrum, V. dahliae, and V. tricorpus. New Zealand Journal of Agricultural Research 8(3): 450-478.

Appendix 1: Comment from DOC

From: Rod Hitchmough [mailto:[email protected]] Sent: Monday, 31 May 2021 11:10 AM To: ...... @epa.govt.nz>; ...... @epa.govt.nz> Cc: [email protected]; [email protected]; Barry Wards ( MPI ) ; Verity Forbes

Subject: RE: APP204242 Statutory determination for Verticillium nonalfalfae

Dear … and ….

Thank you for the opportunity to comment on APP204242 Statutory determination for Verticillium nonalfalfae.

The situation here seems very straightforward. Verticillium nonalfalfae was first identified and split from its parent species on the basis of genetic differences in 2011, but testing of samples collected in New Zealand prior to 1998 has proved it was present in this country then. It is therefore definitely not a New Organism.

Regards Rod Hitchmough Scientific Officer (Biosecurity) Department of Conservation—Te Papa Atawhai National Office, PO Box 10 420, Wellington 6143 18 Manners St, Wellington 6011

Appendix 2: Comment from MPI

General MPI supports the applicant’s contention that Verticillium nonalfalfae is not a new organism under the HSNO Act and believes that there is sufficient evidence to conclude that the species was present in New Zealand immediately prior to 29 July 1998.

Application scope The applicant has applied for a determination under section 26 of the HSNO Act as to whether the fungal species V. nonalfalfae is a new organism in accordance with that Act.

Application comments The applicant has provided a number of references supporting the contention that V. albo-atrum was present in New Zealand immediately prior to 29 July 1998. These include isolates from a variety of horticultural species from areas in different parts of the country, as far back as 1973.

The applicant refers to the publication of Inderbitizin et al (2011), which concludes, through phylogenetic analysis, that V. albo-atrum should be split into three species, V. albo-atrum, V. alfalfa and V. nonalfalfae. This new taxonomic reorganisation is supported by Index Fungorum and the Global Biodiversity Information Facility (GBIF).

The applicant refers to a number of V. albo atrum isolates (pre-1998) held by Landcare in the International Collection of Microorganisms from Plants (ICMP) that have been redetermined based on the phylogenetic analysis of Inderbitizin et al (2011). This redetermination concluded that 10 of the isolates are V. nonalfalfae.

Based on this evidence, MPI supports the applicant’s contention that V. nonalfalae was present in New Zealand immediately prior to 29 July 1998.

Appendix 3: Revised s26 pathway

Figure 17: Decision pathway for applications under Section 26 for determination as to whether an organism is a new organism

Context This decision pathway describes the decision-making process for applications under Section 26 for determination as to whether an organism is a new organism.

Introduction The purpose of this decision pathway is to provide the HSNO decision maker1 with guidance so that all relevant matters in the Hazardous Substances and New Organisms Act (1996) (the Act) and the Hazardous Substances and New Organisms (Organisms Not Genetically Modified) Regulations (1998) (the Regulations) have been addressed. It does not attempt to direct the weighting that the HSNO decision maker may decide to make on individual aspects of an application.

The decision pathway has two parts –

• Flowchart (a logic diagram showing the process prescribed in the HSNO Act and the Methodology to be followed in making a decision), and • Explanatory notes (a discussion of each step of the process).

Of necessity the words in the boxes in the flowchart are brief, and key words are used to summarise the activity required. The explanatory notes provide a description of each of the numbered items in the flowchart and describe the processes that should be followed.

For proper interpretation of the decision pathway, it is important to work through the flowchart in conjunction with the explanatory notes.

1 The HSNO decision maker refers to either the EPA Board or any committee or persons with delegated authority from the Board.

Figure 17 Explanatory Notes Section 26 pathway A

Item 1 Review the content of the application and all relevant information Review the application, staff advice and any relevant information held by other Agencies, and advice from experts.

Item 2 Is further information required? Review the information and determine whether or not there is sufficient information available to make a decision.

Item 3 Seek additional information (Section 52 and Section 58) If the HSNO decision maker considers that further information is required, then this may be sought either from the applicant (if there is an external applicant) or from other sources. If the HSNO decision maker considers that the information may not be complete but that no additional information is currently available, then the HSNO decision maker may proceed to make a determination. If the application is not approved on the basis of lack of information (or if the organism is considered new) and further information becomes available at a later time, then the HSNO decision maker may choose to revisit this determination.

Item 4 Is it an organism (i.e. fits the “organism” definition in Section 2)? An organism (a) does not include a human being: (ab) includes a human cell: (b) includes a micro-organism: (c) includes a genetic structure, other than a human cell, that is capable of replicating itself, whether that structure comprises all or only part of an entity, and whether it comprises all or only part of the total genetic structure of an entity: (d) includes an entity (other than a human being) declared to be an organism for the purposes of the Biosecurity Act 1993: (e) includes a reproductive cell or developmental stage of an organism If yes, go to item 5. If no, as this is not an organism, it is not regulated under the new organism provisions of the HSNO Act.

Item 5 Is the determination about a potential GMO (Section 2A(1)(d))? If the determination relates to whether an organism is a potential GMO, go to pathway B. If the organism is not a GMO, go to item 6.

Item 6 Does the organism belong to a species that was known to be present in NZ immediately before 29 July 1998 (Section 2A(1)(a))? Determine on the basis of the available information whether on balance of probabilities the organism is known to belong to a species that was present in New Zealand immediately prior to 29 July 1998. For the purposes of making a Section 26 determination an organism is considered to be present in New Zealand if it can be established that the organism was in New Zealand: (a) immediately before 29 July 1998; and (b) not in contravention of the Animals Act 1967 or the Plants Act 1970 (excluding rabbit haemorrhagic disease virus, or rabbit calicivirus). If yes, go to item 7 to test the organism against the next criterion. If no, go to item 12.

Item 7 Is the organism prescribed as a risk species and was not present in New Zealand at the time of promulgation of the relevant regulation (Section 2A(1)(b))? Determine whether the organism belongs to a species, subspecies, infrasubspecies, variety, strain, or cultivar that has been prescribed as a risk species by regulation established under Section 140(1)(h) of the Act. If the organism is prescribed as a risk species, determine whether it was present in New Zealand when it was

prescribed. The organism is a new organism if it was not present in New Zealand at the time of the promulgation of the relevant regulation. Note: at this point it may become apparent that the organism is an unwanted organism under the Biosecurity Act. If this is the case, then MPI and DOC may be advised (they may already have been consulted under items 1, 2 and 3). If yes, go 12. If no, go to item 8 to test the organism against the next criterion.

Item 8 Has a containment approval been given for the organism under the Act (Section 2A(1)(c))? For the purposes of making a Section 26 determination, this will also include the following organisms which are “deemed” to be new organisms with containment approvals under the HSNO Act: (a) animals lawfully imported under the Animals Act 1967 before 29 July 1998 pursuant to Section 254 of the HSNO Act; (b) animals lawfully present in New Zealand in a place that was registered as a zoo or circus under the Zoological Garden Regulations 1977 pursuant to Section 255 of the HSNO Act (except where other organisms of the same taxonomic classification were lawfully present outside of a zoo or circus – see section 2A(2)(c)); (c) hamsters lawfully imported under the Hamster Importation and Control Regulations 1972 pursuant to Section 256 of the HSNO Act; or (d) plants lawfully imported under the Plants Act 1970 before 29 July 1998 pursuant to Section 258 of the HSNO Act. If yes, go to item 12. If no, go to item 9 to test the organism against the next criterion.

Item 9 Has a conditional release approval been given for the organism (Section 2A(1)(ca))? If yes, go to item 12. If no, go to item 10 to test the organism against the next criterion.

Item 10 Has a qualifying organism with controls approval been given for the organism (Section 2A(1)(cb))? A “qualifying organism” is an organism that is or is contained in a “qualifying medicine” or “qualifying veterinary medicine”. These terms are defined in Section 2 of the HSNO Act. If yes, go to item 12. If no, go to item 11 to test the organism against the next criterion.

Item 11 Is the organism known to have been previously eradicated (Section 2A(1)(e))? Determine whether the organism belongs to a species, subspecies, infrasubspecies, variety, strain, or cultivar that is known to have been previously eradicated. Eradication does not include extinction by natural means but is considered to be the result of a deliberate act. If yes, go to item 12. If no, then the organism is not a new organism.

Item 12 Has HSNO release approval without controls been given for an organism of the same taxonomic classification under Sections 35, 38 or 38I of the Act or has an organism of the same taxonomic classification been prescribed as a not new organism (Section 2A(2)(a))? If a release approval has been given for an organism of the same taxonomic classification under Section 35 or 38 of the Act then the organism is not a new organism. If a release approval has been given for an organism of the same taxonomic classification under Section 38I of the Act without controls then the organism is not a new organism, however, if this approval has been given with controls then it is a new organism. If an organism of the same taxonomic classification has been prescribed by regulations as not a new organism2 then it is not a new organism.

2 http://www.legislation.govt.nz/regulation/public/2009/0143/latest/whole.html#DLM2011201

If yes, the organism is not a new organism. If no, the organism is a new organism. Section 26 pathway B

Item 1 Have the genes or other genetic material been modified by in vitro techniques or inherited from genes or other genetic material that has been modified by in vitro techniques?

If yes, go to item 2.

If no, the organism is not a genetically modified organism. However, you must check whether it meets the other new organism criteria so go to Pathway A item 6 onwards.

Item 2 Does the organism result solely from selection or natural regeneration, hand pollination, or other managed, controlled pollination (Regulation 3(1)(a) of the Regulations)?

Is the organisms solely the result of selection or natural regeneration, hand pollination, or other managed, controlled pollination?