Bursaphelenchus willibaldi, Ne

Torrini, Giulia, Strangi, Agostino, Simoncini, Stefania, Luppino, Maria, Roversi, Pio Federico & Marianelli, Leonardo, 2020, First report of Bursaphelenchus fungivorus (Nematoda: Aphelenchida) in Italy and an overview of nematodes associated with Crocus sativus L., Journal of Nematology 52 (1), pp. 1-11 : 2-4

B. willibaldi Ne 16/05 AM180512

CharaCter Male Female

n 10 10

L 686.7 ± 31.9 735.28 ± 38.4 (645.2 – 742.0) (700.0 – 834.8)

a 28.3 ± 1.9 26.5 ± 2.0 (25.3– 30.9) (23.3– 30.3)

b 10.2 ± 0.8 10.5 ± 0.5

(9.2 – 11.8) (10.1 – 11.7)

c 20.5 ± 1.0 12.4 ± 0.5

(19.1 – 22.1) (11.8 – 13.1) c' 2.2 ± 0.2 5.4 ± 0.4

(1.9 – 2.5) (4.8 – 6.3)

T or V 64.1 ± 4.9 73.1 ± 0.4

(57.5– 72.8) (72.4– 73.7)

Max. body diam. 24.4 ± 1.8 27.9 ± 2.1 (22.4– 27.9) (24.2– 31.7)

Stylet 13.7 ± 0.8 13.4 ± 1.5 (12.6 – 14.9) (11.3 – 16.2)

Median bulb 17.4 ± 2.0 19.6 ± 1.7 length (14.3 – 20.3) (17.1 – 22.0)

Median bulb diam. 11.9 ± 1.1 14.9 ± 1.8

(10.8 – 13.3) (13.0 – 19.3) Tail length 33.7 ± 2.3 59.2 ± 2.6

(30.5– 38.4) (56.7– 65.0)

Cloacal or anal 15.5 ± 1.1 10.9 ± 0.5 body diam. (13.8 – 17.1) (10.2 – 11.8)

Spicule length* 15.3 ± 1.1 – (13.7 – 17.5)

Notes: All measurements are in μm: mean ± SD (range). *Curved along arc from bottom of capitulum depression to distal end.

Proteinase K was inactivated by heating at 96.0°C for 10 min and DNA was recovered with alcoholic precipitation adding 100.0µl of cold absolute ethanol. Pellets were air-dried and resuspended in 20.0µl of double distilled water. The amplification of Internal Transcribed Spacer locus (ITS) was performed using conditions described in Burgermeister et al. (2005). The PCR products were sequenced at the Centro di Servizi per le Biotecnologie di Interesse Agrario Chimico e Industriale, University of Florence, Italy. Species identification was obtained through two phylogenetic trees (obtained with Neighbor Joining and Maximum likelihood algorithms) based on ITS locus and focused on fUngiVOrOUS -group starting from sequences of this locus mined from GeneBank ( Table 1). Alignments were computed with a local alignment algorithm (implemented in Kalign at EBI website) and the poorly aligned regions were removed with Gblocks v. 9.1b. The choice of appropriate substitution matrix for the data set was evaluated using Jmodeltest2 v. 2.1.10 considering AICc, BI criteria and DT method. Trees were computed using MEGA 7 software, choosing GTR +G +I as nucleotide substitution matrix and 1,000 bootstrap replicates.