CSIRO PUBLISHING www.publish.csiro.au/journals/apdn Australasian Disease Notes, 2009, 4,9–11

First record of Turnip mosaic virus in Cook’s scurvy grass ( oleraceum agg.) ” an endangered native plant in

J. D. Fletcher A,C, S. Bulman A, P. J. Fletcher A and G. J. Houliston B

ANew Zealand Institute for Plant and Food Research Limited, Private Bag 4704, Christchurch, New Zealand. BManaaki Whenua-Landcare Research, PO Box 40, Lincoln 7640, New Zealand. CCorresponding author. Email: fl[email protected]

Abstract. Cook’s scurvy grass (Lepidium oleraceum agg.) is an endangered species of native that is considered threatened with extinction. Virus-like disease symptoms were observed in a newly introduced plant of L. oleraceum at Stony Bay, Banks Peninsula, Canterbury, New Zealand. This is the first record of a virus in L. oleraceum and the first report of a Turnip mosaic virus infection in a New Zealand native host.

Of the indigenous Lepidium species in the Brassicaceae family, (DAS-ELISA) (AS0132, DSMZ, Germany). TuMV infection six are coastal, of which five are endemic to New Zealand was further confirmed by reverse transcriptase polymerase (Lepidium banksii, Lepidium oleraceum agg., Lepidium chain reaction (RT–PCR). RNA was extracted from ~1 cm2 obtusatum, Lepidium tenuicaule, Lepidium naufragorum), leaf discs of five TuMV-infected (SB, LD, LF, LG, and whereas Lepidium flexicaule also occurs in Tasmania (Hewson LJ) using the RNeasy Plant Mini Kit (Qiagen, Germantown, MD, 1981). Cook’s scurvy grass (L. oleraceum) is an endangered USA). RNA was reverse transcribed using the SuperScript III species considered to be threatened with extinction (Norton and First-Strand Synthesis System for RT–PCR (Invitrogen, de Lange 1999). Auckland, New Zealand), according to the manufacturer’s There are few early (1840–42 and 1921) records of instructions. RT–PCRs were carried out using the TuMVF1 L. oleraceum from Banks Peninsula in Canterbury, (50-GCACAAGAAGAGGGAAAACG-30)andTuMVR1 New Zealand (Hugh Wilson, pers. comm. 2007). However, in (50-TGCACCAGACCATTAAACCA-30) primers, designed December 2000, a population of L. oleraceum was discovered from the sequence of a New Zealand TuMV isolate (GenBank on Island Rock (2499350E, 5701240N: 4353043.840S accession AY995214). Cycling conditions were a denaturation 1725202.900E) off Island Bay on the south side of Banks step of 94C for 2 min, followed by 35 cycles of 94C (20 s), Peninsula. Plants were collected from this site and then 55C (20 s), 72C (30 s). The expected ~290 bp DNA fragment propagated at Landcare Research in the glasshouse for was amplified from all five samples, while RT-negative and restoration plantings by the Department of Conservation H2O-negative PCR reactions did not give DNA amplification. Mahaanui Area Office (DOC), Christchurch. A plant from this In subsequent work, PCR products from the five samples were population was re-introduced to Stony Bay (4351041.980S sequenced and were confirmed to be from more than one TuMV 1732039.050E) on the north side of the peninsula. During isolate; phylogenetic analysis of these isolates will be presented September 2007, this plant was examined and found to have elsewhere. mosaic mottling symptoms typical of a plant virus infection. Material from the five selected symptomatic plants was used to Tissue was removed for further examination. In addition, mechanically inoculate appropriate herbaceous indicator plants, 10 plants of L. oleraceum recently collected from Island Rock including Brassica pekinensis cvv. ‘Chi hi li’ and ‘Pak Choi’, as part of the DOC re-establishment program were also examined B. oleracea var. capitata cv. ‘Flower of Spring’, B. oleracea var. for virus symptoms. botrytis cv. ‘White Acre’, C. amaranticolor, Chenopodium The Lepidium at Stony Bay (SB) exhibited inter-veinal quinoa, Nicotiana glutinosa, Raphinus sativus cv. ‘Gentle yellow mottling, mosaic, distortion and twisting of the leaves Giant’, R. sativus cv. ‘French Breakfast’ and Sinapsis alba. (Fig. 1). Of the 10 further plants (LALJ) seven showed similar Symptoms on all hosts appeared characteristic of those for symptoms (Fig. 2). In general, symptoms were more pronounced TuMV infection as summarised in Provvidenti (1996). Some in the glasshouse-grown plants collected for conservation. differences in host response between isolates were observed. In Crushed leaf sap from leaf specimens from seven of the particular, B. pekinensis cv. ‘Pak Choi’ exhibited a clear mottled 10 plants tested positive for Turnip mosaic virus (TuMV) mosaic in LF and LG compared with a milder vein banding in LD, (Potyviridae) (mean of controls + 3 sd) using double LJ and SB infected plants, indicating that two virus strains may antibody sandwich enzyme-linked immunosorbent assay be present.

Ó Australasian Plant Pathology Society 2009 10.1071/DN09004 1833-928X/09/010009 10 Australasian Plant Disease Notes J. D. Fletcher et al.

Fig. 1. Virus symptoms on Lepidium oleraceum from Stony Bay, Banks Peninsula, New Zealand, September 2007.

Fig. 2. Leaf from Lepidium LG (left) with TuMV leaf mosaic, and leaf from LA (right) with no virus infection.

TuMV is found in New Zealand in a wide range of hosts been recorded on other Lepidium spp. (Norton and de (Pearson et al. 2006). In Canterbury, large areas of Brassica rapa Lange 1999). Cavariella aegopodii, Myzus cerasi and (turnip) and Brassica napus var. napobrassica (swede) have Myzus ornatus are also reported to colonise L. oleraceum been grown regularly for sheep and cattle fodder. This is the (Teulon et al. 2004; www.crop.cri.nz/home/plant-synz/ first record of a virus in L. oleraceum and the first report of a database/databasehome.php). TuMV infection in a New Zealand native host. TuMV is It is of concern that virus-infected clones of this rare and transmitted by species that may be found on these hosts. endangered plant may be accidentally introduced into protected Myzus persicae and Brevecoryne brassicae are the two most areas. We have been working with DOC to screen and select common vectors of TuMV (Karl 1971), of which the latter has suitable TuMV-free clones for re-introduction. Turnip mosaic virus in Cook’s scurvy grass – first record Australasian Plant Disease Notes 11

Acknowledgements Pearson MN, Clover GRG, Guy PL, Fletcher JD, Beever REB (2006) A review of the plant virus, viroid and mollicute records for We thank Sonia and Mark Armstrong for allowing access to their property at New Zealand. Australasian Plant Pathology 35, 217–252. Stony Bay. We also thank Hugh Wilson for his notes detailing L. oleraceum doi: 10.1071/AP06016 and its history on Banks Peninsula. Provvidenti R (1996) Turnip mosaic potyvirus. In ‘Viruses of plants. Descriptions and lists from the VIDE database’. (Eds AA Brunt, References K Crabtree, MJ Dallwitz, AJ Gibbs, L Watson). pp. 1340–1343. Hewson HJ (1981) The genus Lepidium L. (Brassicaceae) in Australia. (CAB International: Wallingford, UK) Brunonia 4, 217–308. Teulon DAJ, James D, Fletcher JD (2004) Documenting invasive on Karl E (1971) New vectors for some non-persistent viruses. Archiv fur indigenous flora. New Zealand Crop & Food confidential report no. 936. Pflanzenschutz 7, 337–342. Norton DA, de Lange PJ (1999) Threatened species recovery plan no. 26. Biodiversity Recovery Group, Department of Conservation, Wellington. Manuscript received 9 January 2009, accepted 27 January 2009

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