Naturalised Hakea. What species are we actually talking about in Europe?

Figures & data

Table 1. Sampling localities. All sampling from naturalized populations (species: 1. H. decurrens; 2. H. drupacea; 3. H. gibbosa; 4. H. lissosperma; 5. H. salicifolia; 6. H. sericea).

	Location	Coordinates	Species
Portugal
	Mata do Sobral	40°09’34.2“N, 8°10’29.3“W	1
	Serpins	40°09’22.2“N, 8°14’11.2“W	1
	Coimbra	40°10’38.8“N, 8°27’38.5“W	1, 5
	Vale de Canas	40°12’38“N, 8°22’40“W	5
	Entroncamento	39°27’53“N, 8°25’55“W	1
South Africa
	Outenikwa Pass	33° 52 ‘31“S, 22° 22’ 57“E	6
	Joubertina Hoek	33° 52 ‘8“S, 23° 50’ 8“E	6
	Sandrif	33° 50 ‘23“S, 23° 47’ 53“E	6
	DeVlug draai	33° 45 ‘5“S, 22° 57’ 38“E	6
	Vergelegen	34° 3 ‘23“S, 18° 55’ 0“E	6
	Pniel	33° 53 ‘60“S, 18° 56’ 52“E	6
	Caledon	34° 13 ‘5“S, 19° 24’ 12“E	6
	Karwyderskraal	34° 20 ‘56“S, 19° 11’ 36“E	3, 6
	Glenthorpe farm	33° 21 ‘36“S, 26° 30’ 9“E	6
	Grahamstown	33° 17 ‘1“S, 26° 42’ 7“E	6
	Grahamstown	33° 20 ‘1“S, 26° 35’ 42“E	6
	Fisherhaven	34° 22 ‘ 1”S, 19° 7 ‘ 22”E	2
	Sir Lowry’s pass	34° 8 ‘59“S, 18° 55’ 40“E	5
	Slanghoek	33° 32 ‘24“S, 19° 12’ 36“E	6
New Zealand
	North Island, Whangarei	35° 42 ‘37“S, 174° 20’ 27“E	6
	Gumlands, Mangaroa Stream	35° 40 ‘59“S, 174° 1’ 13“E	6
	Kawakawa, Karetu Valley	35° 22 ‘38“S, 174° 11’ 5“E	6
	Mahurangi Peninsula	36° 27 ‘0“S, 174° 43’ 60“E	6
	Orewa	36° 34 ‘0“S, 174° 40’ 0“E	6
	Sweetwater, Lake Ngatu	35° 2 ‘17“S, 173° 11’ 57“E	6
France
	Théoule-sur-Mer	43° 29 ‘47“N, 6° 56’ 39“E	1
	Saint-Raphaël, Arboretum Plan Estérel	43° 29 ‘33“N, 6° 49’ 23“E	6
	Arboretum du Caneiret	43° 29 ‘47“N, 6° 49’ 55“E	1

Table 2. Herbarium collections, yielding good quality DNA, from the native range or annotated by Barker at Naturalis Leiden (L) for verification and additional specimens from the Allan herbarium (CHR) for New Zealand.

	Subdivision	Species	Collector	Barcode
Australia
	Australian Capital Territory	H. decurrens ssp. decurrens	Taylor J 13	L.1634603
	New South Wales	H. decurrens ssp. decurrens	Coveny RG 4189	L.1634991
	New South Wales	H. lissosperma	Johnson LAS 19,281	L.1634614
	Tasmania	H. lissosperma	Eichler H 16,627	L.1634582
New Zealand
	North Auckland Land District	H. gibossa	Sykes WR 399/86	CHR 437,550
	North Auckland Land District	H. decurrens	Healy AJ s.n	CHR 70,931
	North Auckland Land District	H. salicifolia	Bell B s.n	CHR 671,880
	South Auckland Land District	H. decurrens	Inst. In Agr. s.n.	CHR 90,114
	Wellington Land District	H. decurrens	Healy AJ s.n.	CHR 33,544
	Wellington Land District	H. decurrens	Healy AJ s.n.	CHR 58,539
	Marlborough Land District	H. decurrens	Beggs JP s.n.	CHR 98,049
	Nelson Land District	H. salicifolia	Wright K s.n.	CHR 635,162
	Nelson Land District	H. decurrens	Ecroyd CE s.n.	CHR 670,269
Portugal
	Madeira	H. decurrens	Hegnauer R LEP 11,999	L.1634989

Table 3. Primers used in this study.

Locus	Primer	Primer sequence	Reference
rbcL	Forward	5′-ATG TCA CCA CAA ACA GAG ACT AAA GC-3′	Kress et al. (2007)
	Reverse	5′-GTA AAA TCA AGT CCA CCR CG-3′	Kress et al. (2009)
trnH-psbA	Forward	5′-CGC GCA TGG TGG ATT CAC AAT CC-3′	Tate (2002)
	Reverse	5′-GTT ATG CAT GAA CGT AAT GCT C-3′	Sang et al. (1997)

Figure 1. Flower and fruit of Hakea decurrens, H. sericea and H. gibbosa.

Table 4. Measurement of flower (10 per specimen) and fruit of Hakea decurrens, H. sericea and H. gibbosa.

		Length of pistil (mm)	Length Fruit (mm)	Width fruit (mm)	pale wood zone (mm)	outer layer (mm)
H. decurrens
	Marchante s.n. Coimbra WAG0452351	10	32	32	12	5
	Marchante s.n. Coimbra WAG0452210		32–34	25–26	10	4
	Marchante s.n. Serpins	10
	Marchante s.n. Mata do Sobral	10
	Beggs s.n. CHR 98,049	10.5
	Healy s.n. CHR 58,539	8.5
	Healy s.n. CHR 33,544	9.5–11
	Inst. in Agric. s.n. CHR 90,114	9
	Diadema s.n.		22–26	20–25	6.0–8.0	2.0–4.0
	Valkenburg 3860		24–29	20–25	7.0–8.0	3.0–4.0
H. sericea
	Kritzinger-Klopper 26	7
	Kritzinger-Klopper 25	4.5–6.5	24–29	25–26	8	4
	Valkenburg 4063		25–30	24–28	9	5
	Kinsler s.n.	5–7.5
H. gibbosa
	L.1634877; L.1634878	9–10.5
	Valkenburg 4056		32–35	32–36	14–16	5

Figure 2. Detail of phylogenetic tree based on concatenated matK, rbcL and trnH-psbA barcode sequences of 45 Hakea sericea specimens. A 2,425 (including gaps) alignment of 96 Hakea spp. specimens was used to construct the dendrogram with FastTree using a Jukes-Cantor substitution model and optimization of the Gamma20 likelihood. Fast global bootstrap values are shown on the internal nodes and H. drupacea (taxon level) was used to root the (taxon level) sequences. Origin of the sampled specimens is indicated with vertical bars. The full tree containing 96 Hakea spp. Is shown in S-Fig. 1.

Figure 3. Map of South Africa illustrating the distribution of the two haplotypes of Hakea sericea.

Kress WJ, Erickson DL, Shiu S-H. 2007. A two-locus global DNA barcode for land plants: the coding rbcL gene complements the non-coding trnH-psbA spacer region. PloS One. 2(6):e508. doi: 10.1371/journal.pone.0000508. PMID: 17551588; PMCID: PMC1876818.

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Data availability statement

The annotated DNA sequences will be made available on NCBI GenBank and on EPPO Q-bank (//qbank.eppo.int/plants/). A complete list of Herbarium vouchers analysed (including NCBI codes) is included in Appendix 1.