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APP202274 S67A Amendment Proposal Sept 2018.Pdf PROPOSAL FORM AMENDMENT Proposal to amend a new organism approval under the Hazardous Substances and New Organisms Act 1996 Send by post to: Environmental Protection Authority, Private Bag 63002, Wellington 6140 OR email to: [email protected] Applicant Damien Fleetwood Key contact [email protected] www.epa.govt.nz 2 Proposal to amend a new organism approval Important This form is used to request amendment(s) to a new organism approval. This is not a formal application. The EPA is not under any statutory obligation to process this request. If you need help to complete this form, please look at our website (www.epa.govt.nz) or email us at [email protected]. This form may be made publicly available so any confidential information must be collated in a separate labelled appendix. The fee for this application can be found on our website at www.epa.govt.nz. This form was approved on 1 May 2012. May 2012 EPA0168 3 Proposal to amend a new organism approval 1. Which approval(s) do you wish to amend? APP202274 The organism that is the subject of this application is also the subject of: a. an innovative medicine application as defined in section 23A of the Medicines Act 1981. Yes ☒ No b. an innovative agricultural compound application as defined in Part 6 of the Agricultural Compounds and Veterinary Medicines Act 1997. Yes ☒ No 2. Which specific amendment(s) do you propose? Addition of following fungal species to those listed in APP202274: Aureobasidium pullulans, Fusarium verticillioides, Kluyveromyces species, Sarocladium zeae, Serendipita indica, Umbelopsis isabellina, Ustilago maydis Aureobasidium pullulans Domain: Fungi Phylum: Ascomycota Class: Dothideomycetes Order: Dothideales Family: Dothioraceae Genus: Aureobasidium Species: Aureobasidium pullulans (de Bary) G. Arnaud, Annales de l'École Nationale d'Agriculture de Montpellier 16 (1-4): 39 (1918) Category 2 (spore forming) Description: The “black yeast” Aureobasidium pullulans is an oligotrophic fungus found worldwide. It has been reported growing in a range of habitats from the phyllosphere (Andrews and Kenerley, 1978) to rocks and monuments (Urzì et al., 1999) to hypersaline waters in salterns (Gunde-Cimerman et al., 2000). It holds potential biotechnological importance due to its ability to produce the biodegradable extracellular polysaccharide pullulan (poly-α-1,6-maltotriose) (Zalar et al., 2008), which has been considered a promising biological packaging material (Rekha, M.R. and Sharma, C.P., n.d.; Singh et al., 2008), and also due to its ability to produce a variety of enzymes (Li et al., 2007; Liu et al., 2008; Ma et al., 2007). Genetic modification of A. pullulans is proposed with the goal of producing bioactive compounds of interest at an industrial scale. Fusarium verticillioides Domain: Fungi May 2012 EPA0168 4 Proposal to amend a new organism approval Phylum: Ascomycota Class: Sordariomycetes Order: Hypocreales Family: Nectriaceae Genus: Fusarium Species: Fusarium verticillioides (Sacc.) Nirenberg, Mitteilungen der Biologischen Bundesanstalt für Land- und Forstwirtschaft 169: 26 (1976) Category 2 (spore forming) Description: Fusarium verticillioides (synonym F. moniliforme), “one of the most common maize-associated fungi”, plays a dual role as a pathogen and an endophyte of its host (Brown et al., 2012; Yates et al., 2005). As an important pathogen of the major cereal food crop maize (Zea mays) worldwide, it causes Fusarium seedling disease, stalk rot, and ear rot (Ridenour and Bluhm, 2017), and contaminates infected kernels with a range of mycotoxins such as fumonisins, fusaric acid, and fusarins (Brown et al., 2012; Desjardins, 2006; Munkvold, 2003). However, as a frequent endophyte of maize (Pamphile and Azevedo, 2002; Ridenour and Bluhm, 2017; Wicklow, 1988), F. verticillioides has been reported to deter a number of pathogenic fungi including Fusarium graminearum (Rheeder, 1990; Wyk et al., 1988) and Stenocarpella maydis (Rheeder, 1990), and reduce the disease severity of corn smut caused by Ustilago maydis (Lee et al., 2009; Rodriguez Estrada et al., 2012). We seek to genetically modify F. verticillioides to eliminate pathogenic features, including toxin production, with the ultimate aim of generating a non-pathogenic bioprotective endophyte of maize via gene editing. Kluyveromyces spp. Domain: Fungi Phylum: Ascomycota Class: Saccharomycetes Order: Saccharomycetales Family: Saccharomycetaceae Genus: Kluyveromyces Species: Kluyveromyces marxianus, Kluyveromyces lactis, Kluyveromyces aestuarii, Kluyveromyces nonfermentans, Kluyveromyces dobzhanskii, Kluyveromyces wickerhamii Category 2 (spore forming) Description: Kluyveromyces, part of the Saccharomyces complex, is a yeast genus with 6 species (Kurtzman, 2003). Best known members of this genus are Kluyveromyces marxianus (type species of Kluyveromyces) and K. lactis, both of which have the ability to assimilate and use lactose as a carbon source, and are considered GRAS (Generally Regarded As Safe) in the United States, and QPS (Qualified Presumption of Safety) in the European Union (Lane and Morrissey, 2010). Both species are used for the production of an array of enzymes (Fonseca et al., 2008; van Ooyen et al., 2006) and have been used in molecular genetic research (Dujon et al., 2004; Lane and Morrissey, May 2012 EPA0168 5 Proposal to amend a new organism approval 2010). Two other Kluyveromyces members, K. aestuarii and K. nonfermentans, have been isolated associated with marine environments (Araujo et al., 1995; Nagahama et al., 1999). K. dobzhanskii is found in diverse substrates globally (Sukhotina et al., 2006), with no reported animal or plant pathogenicity. K. wickerhamii has been suggested as a bio-preservative in the wine industry due to its ability to produce compounds that counteract undesired spoilage yeasts during the wine aging process (Comitini et al., 2004). Our experimentation will focus primarily on K. marxianus and K. lactis, with the objective of assessing their use in large scale heterologous production of a selected bioactive compound. Sarocladium zeae Domain: Fungi Phylum: Ascomycota Class: Sordariomycetes Order: Hypocreales Family: N/A Genus: Sarocladium Species: Sarocladium zeae (W. Gams & D.R. Sumner) Summerbell, Studies in Mycology 68: 158 (2011) Category 2 (spore forming) Description: Sarocladium zeae, previously known as Acremonium zeae (Summerbell, 2005) has been characterized as a protective endophyte of maize (Zea mays). There are no reports of S. zeae causing any mycotoxicosis in cultivated cereals. It is capable of producing the antibiotics pyrrocidines A and B, which inhibit the growth of Aspergillus flavus and Fusarium verticillioides that may produce the mycotoxins aflatoxin and fumonisin respectively, thus gaining recognition as a potential biocontrol agent in maize (Wicklow et al., 2005). The proposed research seeks to enhance and exploit the positive attributes of S. zeae by genetic modification. Serendipita indica Domain: Fungi Phylum: Basidiomycota Class: Agaricomycetes / Hymenomycetes Order: Sebacinales Family: Serendipitaceae Genus: Serendipita Species: Serendipita indica (Sav. Verma, Aj. Varma, Rexer, G. Kost & P. Franken) M. Weiß, Waller, A. Zuccaro & Selosse (2016) Category 2 (spore forming) Description: Serendipita indica (synonym Piriformospora indica), although discovered only a decade ago (Verma et al., 1998; Weiß et al., 2016), is already considered a model to study symbiotic root interactions of fungi due to being an axenically-culturable root endophyte (Varma et al., 1999; Ye et al., 2014). It has a growth-promoting effect May 2012 EPA0168 6 Proposal to amend a new organism approval on a number of plants, including the commercially valuable crops maize (Zea mays L.) and tobacco (Nicotiana tabaccum L.) (Varma et al., 1999). In barley (Hordeum vulgare), S. indica has been shown to improve yield, induce systemic resistance to pathogens, and abolish the detrimental effect of moderate salt stress (Waller et al., 2005). The proposed research seeks to genetically manipulate this fungus, which has been recommended as a good candidate for growth and yield enhancement of commercial plants. Umbelopsis isabellina Domain: Fungi Phylum: Mucoromycota Class: Incertae sedis Order: Mucorales Family: Mucoraceae Genus: Umbelopsis Species: Umbelopsis isabellina (Oudem.) W. Gams, Mycological Research 107 (3): 349 (2003) Category 2 (spore forming) Description: Umbelopsis isabellina (synonym Mortierella isabellina) belongs to a clade of fungi that are rhizosphere inhabitants commonly associated with woody roots of Douglas-fir (Pseudotsuga menziesii) and ponderosa pine (Pinus ponderosa) (Hoff et al., 2004; Spatafora et al., 2016). In addition to being a root endophyte (Hoff et al., 2004; Terhonen et al., 2014), U. isabellina is an oleaginous fungus, holding industrial importance as a producer of ɣ-linolenic acid (Fakas et al., 2009; Takeda et al., 2014; Zhang et al., 2007). We propose to genetically manipulate standard strains for heterologous production of bioactive compounds. Ustilago maydis Domain: Fungi Phylum: Basidiomycota Class: Ustilaginomycetes Order: Ustilaginales Family: Ustilaginaceae Genus: Ustilago Species: Ustilago maydis (DC.) Corda, Icones fungorum hucusque cognitorum 5: 3 (1842) Category 2 (spore forming) Description: Ustilago maydis is the causative agent of corn smut (Kämper et al., 2006), a disease that is arguably economically unimportant to maize (Zea mays) (Brefort et al., 2009; Dean Ralph et al., 2012). However, the Ustilago- maize system has
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