13: 127-142, 2004

Xiphinema insigne

1 2 3 2 4,5

1 2 3 4 5 [email protected] : +886-4-22876712 93 5 13

. 2004. insigne . 13: 127-142.

1998 2002 10 (populations) Xiphinema insigne 4

10 (group) rDNA ITS-1 ITS-2 (Xins1, 2 Xins7, 8) 10 10 rDNA 0-3.02 % X. insigne 5-8 X. insigne

DNA

mm 160 m X. insigne 137 - 161 m 150 m Xiphinema insigne Loss, 1949 (Type Tarjan locality) (Sri - Lanka, , Luc X. insigne Ceylon) Kurunegala (Type habitat) (lectotype) Loos X. insigne (soursop, Anona muricala L.) (31) (coconut, Cocos nucifera L.) (grasses, ) (holotype) (29) X. indicum Siddiqi, 1959 X. insigne 161 m X. indicum Aligarh X. indicum (tail) Grewia asiatica L. (41) Siddiqi X. insigne ( 80 97 m) C' ( (total stylet) 3.9 4.4 ) Loos 7 (158 - 167 146 - 159 mm) (caudal pores) (3 7 ) Tarjan Luc (43) Siddiqi 4 X. indicum (paratypes) Loss 4 X. insigne (syntypes) Tarjan Luc (43) X. indicum X. indicum 155 - 164 X. insigne (synonym) 128 13 2 2004

Cohn Sher (11) Loof Maas (28) (35) (14,23) (7) (22,48) (22) (1,25,26,45,46) rDNA Cohn (10) (Israel) X. (gene marker) insigne Saigusa Yamamoto (40) X. insigne rDNA Southey (4) Tarjan Luc rDNA X. insigne "indium type" "long-tail type" Siddiqi X. indicum 16 3 82 130 m Bajaj Xiphinema insigne Jairajpuri (4) 23 X. insigne (functional odontostyle) DNA rDNA 2 (group) 18 rDNA (consensus "indicum-form" sequence) ITS-1 ITS-2 (2) 5 "insigne-form" 2 4 SAS GLM (1999, V8.2) (lip region) 2 Bajaj Jairajpuri X. insigne (4) Luc Southey (31) X. insigne (type Xiphinema insigne specimens) (population complex) X. insigne Loss sensu stricto 5 10 X. insigne X. insigne (type locality) (morphometrics) X. insigne (L) 2.13 mm (Xins3) DNA (ribosomal DNA, rDNA) 2.41 mm (Xins6) (18,19,23,32,33,34) Powers (36) Vrain (45) a b c 10 18S 28S 3' 5' c (Xins4, 7, 8, 9) c' (universal primers) Xins8 (4.75) (genus) ITS-1 ITS-2 5.8S Xins4, 7, 8, 9 4 c' 18S 28S rDNA (V %) Xins1 - 7 7 30.0 - 32.0 % 26.4 - (Pratylenchus spp.) 34.6 % Xins8 - 10 3 (Tylenchorhynchus spp.) 34.2 - 34.7 % (Heterodera spp., Globodera spp.) 32.0 - 40.1 % Xins1, 5, 9 3 rDNA (ITS-1, ITS-2) 7 149.7 - 151.5 m PCR-RFLP 158.3 m (isolates) (6,15,16,42,44) 157.4 - 163.0 m Xins1, 5, 9 3 (intra-individual) rDNA 7 88.6-90.3 m (heterogenicity) (6) 95.3-102.0 m 10 (50) (21) 60.1-64.0 m Xiphinema insigne 129

Xiphinema insigne Table 1. The list of geographical populations of Xiphinema insigne investigated in this study Population Host Locality (Code) Xins1 Litchi (Litchi chinensis Sonn.) Tsautuen Jen, Nantou County Xins2 Litchi (Litchi chinensis Sonn.) Taichung City (1) Xins3 Litchi (Litchi chinensis Sonn.) Taichung City (2) Xins4 Bamboo (Bambusa sp.) Taichung City Xins5 Bamboo (Bambusa sp.) Shinhua Jen, Tainan County Xins6 Loquat (Eriobotrya japonica Lindl.) Shinshe Shiang, Taichung County Xins7 Loquat (Eriobotrya japonica Lindl.) Guoshing Shiang, Nantou County Xins8 Loquat (Eriobotrya japonica Lindl.) Luye Shiang, Taitung County Xins9 Bermuda-grass (Cynodon dactylon Pers.) Daya Shiang, Taichung County (On green of golf course) Xins10 Pear (Pyrus pyrifolia var. yokoyama) Heping Shiang, Taichung County

Xins8 (109.0 m) Xins4, 7, 8, 9 4 Xins10 71, 79, 63 87 m (anal body width) X. insigne ( G, H, I, J) Xins8, 9 2 A 4 8 (lip region) (hemispherical) (slight constriction) B 10 (elongate-conical) C, D, E, F, Xiphinema insigne Xiphinema insigne Xins2, 4, 7, 8, 9 5 10 X. insigne 15 2 4 4 Xins8

6 (Xins2, 5, 6, 9, 10) 4 Xins8 1.69 mm 10 a b 6 c c' Xins2, 4, 7, 8 a, b, c c' a b 4 11 Xins9 c Xins9 2 Xins8 c' Xins5, 145.8 m 155.7 6 2 8 m Xins2, 4, 7, Xins8, 9 2 8 4 Xins9 c' 4 4 Xins8 Xins9 Xins1, 5, 9 3 Xins8 7 40-79 m 4 Xins4, 5, 6, 9 4 Xins8 Xins9 6 Xins1, 5, 9 3 (supplements) 130 13 2 2004

E Xins2 2 2 10 bp 5.8S 6 C, D Xins9 2 4 160 bp 8 7 C, D Xins8 ITS-1 3.02 ITS-2 2.23 5.8S 29 5-8 Xins7 Xins8 2 0 2.5 ITS-1 ITS-2 Xiphinema insigne rDNA 4 Xins1, 2 Xins7, 8 2 Xins1 Xins2 Xins7 Xins8 4 18S 2 rDNA Xins2 2087 (base pair, bp) 28S 4 3 2088 bp 4 ITS-1 5.8S ITS-2 X. insigne 2 rDNA ITS-1 5.8S ITS-2 ITS-1 Xins1 Xins2 2 GenBank Xins7 Xins8 10 11 bp ITS-2 (Accession number) AY553980 (Xins2) AY563427 (

A B

C D E F

Xiphinema insigne A, ( Xins2 ) B, (Xins2) C D (Xins2), E (Xins8) F (Xins9), G, H, I J (Xins2) 90 Fig. 1. Xiphinema insigne (female and juvenile) from Taiwan: A, Female habitus (Xins2); B, Anterior region of female (Xins2); C and D (Xins2), E (Xins8) and F (Xins9), female tails; G, H, I and J (Xins2), Tails of the first, second, third and fourth stage of juveniles, respectively. Scale bar = 90 m.

GHIJ Xiphinema insigne A, ( Xins2 ) B, (Xins2) C D (Xins2), E (Xins8) F (Xins9), G, H, I J (Xins2) 90 Fig. 1. Xiphinema insigne (female and juvenile) from Taiwan: A, Female habitus (Xins2); B, Anterior region of female (Xins2); C and D (Xins2), E (Xins8) and F (Xins9), female tails; G, H, I and J (Xins2), Tails of the first, second, third and fourth stage of juveniles, respectively. Scale bar = 90 m. Xiphinema insigne 131

AB

CDEF Xiphinema insigne A (Xins2, ), B (Xins7), C D (Xins2), E (Xins1), F (Xins7), 90 Fig. 2. Xiphinema insigne male from Taiwan: A, (Xins2, flatted), B (Xins7), Anterior region; C & D (Xins2), E (Xins1), F (Xins7), Shape of tail and spicule; Scale bar = 90 m.

AB

CD

Xiphinema insigne (Xins9) A, B, C D, 90 Fig. 3. Xiphinema insigne male from Taiwan (Xins9): A, Male habitus; B, Anterior region; C and D, Shape of tail and spicule; Scale bar = 90 m. 132 13 2 2004

Xiphinema insigne (Paralectotypes) Table 2. Morphometrics and comparisons of Xiphinema insigne females collected in Taiwan and Sri-Lanka Locality Host (Code) Character1 Tsautuen Taichung Taichung Taichung Shinhua Litchi Litchi Litchi Bamboo Bamboo (Xins1) (Xins2) (Xins3) (Xins4) (Xins5) n2530153435 L (mm) 2.212 efgh3 2.26 defg 2.13 gh 2.39 abcd 2.33 bcdef (2.00-2.41) (2.05-2.51) (1.88-2.26) (2.13-2.67) (2.09-2.54) a 61.7 bcdef 58.0 efg 59.9 cdefg 62.7 abcdef 65.3 abc (49.4-73.6) (51.8-64.9) (54.0-64.6) (51.6-75.5) (59.2-71.5) b 6.48 cdef 6.58 bcdef 6.41 cdef 6.78 abcd 6.87 abc (5.85-7.03) (5.91-7.35) (5.95-6.97) (5.92-7.84) (6.19-8.47) c 16.4 fg 16.3 fg 17.2 defg 18.5 cdef 19.7 bcde (13.7-18.7) (13.6-17.6) (14.5-19.1) (15.9-31.3) (17.54-22.0) c' 6.25 ab 6.33 ab 5.89 ab 6.07 ab 5.45 bcd (5.36-7.14) (5.59-7.52) (5.09-6.57) (3.38-7.33) (4.50-7.26) V % 31.2 bcd 30.3 cde 30.5 cde 30.1 de 30.0 de (28.8-33.2) (26.4-32.9) (28.5-33.4) (27.5-33.1) (26.9-32.9) Total stylet 150.3 e 158.3 cd 158.8 bcd 157.4 cd 151.5 e ( m) (143.3-157.5) (146.7-166.7) (153.3-165.8) (146.7-166.7) (143.3-158.3) Functional 90.2 f 97.2 cde 98.5 bcd 95.3 de 90.3 f odontostyle (85.8-95.8) (85.0-105.0) (94.2-102.5) (87.5-101.7) (80.8-94.1) ( m) Odontophore 60.1 de 61.0 cde 60.3 de 62.1 cd 61.2 cde ( m) (57.5-63.3) (59.2-64.2) (55.0-64.2) (58.3-65.0) (57.5-64.2) Tail 136.0 abcd 139.0 abc 125.0 defgh 131.0 bcdef 119.0 efgh ( m) (118.0-150.0) (119.0-173.0) (108.0-143.0) (71.0-148.0) (105.0-138.0) Anal body 22.0 cde 22.0 cde 21.0 de 22.0 de 22.0 cde width ( m) (20.00-23.0) (20.0-24.0) (19.0-23.0) (18.0-25.0) (19.0-24.0)

Xins8) 10

4 Xiphinema insigne 10 (31) (2)

10 X. insigne (38) (V %)

(13,24,30) 10 2 2 Xins1 - 7 (2) Xins8 - 10 (30.0-32.0 % 34.2-34.7 %) Xins2, 3, 4, 6, 7, 8, 10 Xins1, 5, 9 (157.4 - 163.0 m 149.7 - 151.5 m) Xins1, 2, 3, 4, 6, 7, 10 Xins5, 8, 9 (125 - 139 m (3,8,12) 109 - 119 m) Xiphinema insigne 133

Xiphinema insigne (Paralectotypes) ( ) Table 2. Morphometrics and comparisons of Xiphinema insigne females collected in Taiwan and Sri-Lanka (Cont’d) Locality Host (Code) Character Shinshe Guoshing Luye Daya Heping Sri Lanka4 Loquat Loquat Loquat Bermuda-grass Pear (paralectotypes) (Xins6) (Xins7) (Xins8) (Xins9) (Xins10) n29302047154 L (mm) 2.41 abc 2.31 bcdef 2.28 cdefg 2.33 bcdef 2.36 abcde 2.24, 2.02 (2.07-2.84) (1.95-2.51) (2.02-2.49) (2.14-2.51) (2.15-2.51) 2.20, 2.32 a60.3 cdefg 63.4 abcde 57.5 efg 59.3 efg 60.8 bcdefg 55, - (52.8-66.2) (56.3-76.1) (51.9-64.3) (49.4-67.8) (53.8-66.0) 50, 66 b 6.74 abcde 6.73 abcde 6.39 def 6.57 bcdef 6.38 def - (5.89-7.82) (5.74-7.61) (5.80-7.32) (5.83-7.77) (5.83-7.17) c 19.3 bcde 17.4 defg 21.4 a 20.0 bcd 18.8 bcdef 20, 20 (14.3-22.4) (14.4-28.6) (18.0-32.1) (15.7-26.2) (17.5-20.4) 26, 21 c' 5.39 cd 6.25 ab 4.75 e 5.25 cd 5.33 cd 4.8, - (4.26-7.24) (4.16-7.47) (3.00-5.74) (3.95-6.73) (4.48-5.78) 3.8, 5.1 V % 32.0 bc 30.8 cde 34.5 a 34.7 a 34.2 a 30, 29.0 (28.4-34.2) (28.1-34.6) (32.9-36.9) (32.2-37.1) (32.0-40.1) 29.6, 32.4 Total stylet 162.9 ab 161.1 abc 163.0 ab 149.7 e 162.3 ab 161, 160 ( m) (150.0-168.3) (154.2-165.0) (158.3-167.5) (144.2-158.3) (158.3-169.2) 154, 156 Functional 99.4 bc 98.0 bcde 102.0 a 88.6 f 98.3 bcd 102, 98 odontostyle (90.0-105.0) (92.5-102.5) (95.9-106.6) (83.3-95.0) (94.1-104.2) 95, 94 ( m) Odontophore 63.5 ab 63.1 abc 61.0 cde 61.1 cde 64.0 ab 59, 62 ( m) (60.0-68.3) (60.0-69.2) (51.7-63.3) (56.7-65.0) (61.7-67.5) 59, 62 Tail 126.0 defg 134.0 abcde 109.0 i 117.0 efgh 126.0 cdefg 110, 101 ( m) (104.0-152.0) (79.0-155.0) (63.0-132.0) (87.0-148.0) (112.0-139.0) 84, 109 Anal body 23.0 ab 22.0 de 23.0 abc 22.0 bcd 24.0 ab - width ( m) (19.0-28.0) (19.0-23.0) (21.0-25.0) (20.0-25.0) (22.0-26.0) 1 n = no. of ; L = body length; a = body length / the maximum body width; b = body length / the distance from lips to the base of esophageal gland;c = body length / the length of tail; c' = tail length / anal body width; V (%) = the distance of vulva from anterior end / body length and then multiplied by 100; total stylet = length of functional odontostyle plus odontophore; tail = length of anus position to the body posterior end; anal body width = body width at the anus position. 2 Measurements in the form: mean (min. - max.), "-" = No data. 3 Means in each row with the same letter or letters were not significantly different at 1% level by LSMEANS/TDIFF. 4 Luc, M., and Southey, J. F. 1980 (31)

2 Xins2 10 10 2 X. insigne Xins8 2 X. diffusum 2 3 X. insigne Xins4, 7, 9 3 X. elongatum

10 1 5 Xins9 2 X. 2 elongatum 7 8 X. 134 13 2 2004 0.96 2.63 7.0 0.10 4.64 0.46 0.45 2.20 1.35 3.05 1.0 0.67 13.61 2.03 138.1 4.0 128.0 0.08 1.73 4.61 58.5 0.47 5.42 0.53 6.97 1.69 82.7 0.88 55.4 2.64 99.8 1.0 18.0 0.58 11.75 0.05 1.36 2.80 52.9 0.29 4.88 0.68 7.03 2.58 118.0 2.16 69.3 1.15 48.7 1.87 83.9 4.0 116.0 1.0 17.0 0.04 1.02 3.04 47.8 0.29 4.25 0.47 10.25 0.64 7.07 1.18 97.5 1.11 55.5 1.09 42.1 1.21 70.1 4.0 99.0 1.0 14.0 Locality Host (Code) 0.80 9.32 3.84 74.4 9.0 80.0 0.11 0.74 5.21 41.0 0.55 3.84 0.42 6.95 3.29 40.4 2.22 34.0 2.74 54.6 1.0 12.0 isolated from Taiwan isolated from Taiwan 2.87 130.5 0.55 13.24 6.0 128.0 0.07 1.69 5.46 55.9 0.33 5.22 0.39 6.77 2.10 77.7 2.04 52.8 1.80 93.6 1.0 19.0 Xiphinema insigne 0.55 11.36 0.05 1.26 3.19 51.1 0.27 4.54 0.49 7.32 1.81 109.8 1.84 64.4 1.58 45.4 2.73 76.6 4.0 111.0 1.0 15.0 0.97 2 0.05 2.68 44.7 0.27 4.14 0.58 10.18 0.49 7.14 2.05 91.7 2.12 52.2 0.91 39.5 1.43 65.8 4.0 95.0 1.0 13.0 sautuen Guoshing 22 20 23 23 26 26 18 18 (69.2-78.3) (89.2-95.0) (105.8-116.7)(31.7-35.0) (123.3-136.7) (71.7-76.7) (36.7-42.5) (91.7-101.7) (40.8-48.3)(75.0-88.0) (115.0-121.70) (48.3-56.7) (134.2-145.0) (90.0-100.0)(10.0-14.0) (32.5-35.8) (98.0-123.0) (12.0-15.0) (39.2-43.3) (112.0-143.0) (14.0-18.0) (73.00-90.0) (47.5-50.0) (18.0-20.0) (91.0-107.0) (51.7-57.5) (108.0-125.0) (10.0-13.0) (108.0-138.0) (12.0-16.0) (14.0-18.0) (18.0-20.0) (0.69-0.87)40.2 (0.88-1.07)(35.0-45.8)3.88 (1.17-1.45) (37.5-50.0)(3.48-4.39)9.22 (1.48-1.86) (42.3-60.5) (3.80-4.65) (0.68-0.85) (48.0-69.2) (3.93-5.15)(6.25-7.80) (0.93-1.09) (35.5-46.5) (4.43-6.79) (6.27-7.92) (1.22-1.50) (41.6-53.0) (3.48-4.72) (6.53-7.86) (1.54-1.92) (43.3-60.0) (3.70-4.95) (6.00-7.53) (50.3-69.1) (4.06-5.64) (6.00-8.30) (4.28-6.17) (6.07-8.67) (6.22-8.00) (6.00-7.67) T Litchi (Xins1) Loquat (Xins7) (8.37-10.61) (9.00-11.06) (10.64-12.76) (11.56-14.75) (8.21-10.12) (9.49-11.98) (10.40-13.04) (12.42-15.56) Xiphinema insigne 1 m) ( m) m) m) m) m) ail 82.0 otal stylet 73.3 ( ( ( odontostyle (36.7-46.6) (50.0-57.5)odontostyle( (60.8-68.4) (50.0-55.0) (73.3-83.4) (60.8-71.7)width (38.3-42.5) (73.3-80.0) (51.7-59.2) (88.3-98.3) (66.6-73.4) (52.5-56.7) (79.2-89.2) (65.5-74.2) (77.5-90.0) (95.8-105.0) Character Stagesn J1 J2 J3( J4Odontophore 33.5 J1T J2 J3 J4 L (mm) 0.75 Functional 39.9 Replacement 53.0 Anal body 12.0 a b c T c' 6.85 able 3. Morphometrics of the 4 juvenile stages T Xiphinema insigne 135 2.82 5.0 0.13 3.97 0.30 1.11 0.30 3.20 1.16 1.80 1.0 2.20 130.0 0.11 1.87 3.31 54.3 0.42 5.70 2.02 16.9 0.53 5.52 2.05 75.4 0.97 54.6 1.80 91.2 7.0 111.0 1.0 20.0 17 15 0.06 1.40 2.95 49.5 0.25 5.05 0.70 14.6 0.33 5.57 2.58 109.1 2.05 62.3 1.68 46.8 1.90 76.0 3.0 97.0 1.0 17.0 0.05 1.00 2.06 41.9 0.33 4.15 0.55 5.90 1.48 90.2 0.91 50.5 0.72 39.7 0.87 62.7 6.0 84.0 1.0 14.0 0.74 12.0 Locality 1.95 70.4 4.49 51.4 7.0 69.0 0.07 0.76 2.71 37.5 0.36 3.95 1.10 11.1 0.46 5.32 2.00 37.9 1.82 32.5 1.0 13.0 46 16 isolated from Taiwan (Cont'd ) isolated from Taiwan 4.0 111.0 1.91 141.1 0.07 1.71 3.43 52.8 0.31 5.21 0.61 15.4 0.32 5.55 1.83 86.8 1.49 54.3 1.97 104.2 1.0 20.0 Xiphinema insigne 0.07 1.34 2.96 47.9 0.38 4.87 0.44 6.00 2.03 116.7 1.43 70.7 1.03 45.9 2.93 86.1 7.0 103.0 1.0 17.0 0.47 13.1 SD (min.- max.) 0.05 0.98 3.77 43.4 0.30 4.25 0.62 11.5 0.41 5.84 1.33 93.8 1.44 54.7 0.96 39.1 2.50 70.8 5.0 86.0 1.0 15.0 yue Daya (0.67-0.82)39.4 (0.88-1.12)(33.5-50.0)3.91 (1.19-1.54) (39.1-48.7)(3.45-4.39)10.2 (1.57-1.83) (41.6-57.8) (3.72-4.91)(8.9-11.3) (0.68-0.83) (48.0-57.0) (4.25-5.48)(5.54-7.09) (10.9-12.8) (0.86-1.09) (34.0-39.5) (4.58-5.87) (5.27-6.77) (11.9-14.6) (1.18-1.74) (37.4-46.7) (3.71-4.61) (5.42-6.47) (13.77-17.81) (10.1-12.3) (1.59-2.07) (45.67-55.2) (3.58-4.54) (4.57-6.47) (46.8-61.3) (10.4-13.2) (4.07-6.00) (4.43-5.85) (12.2-20.0) (5.24-6.41) (5.25-6.54) (15.0-18.8) (4.22-6.47) (4.86-6.00) L Litchi (Xins8)(73.3-77.0) Loquat (Xins9) (90.8-99.2) (112.5-120.0)(30.0-33.3) (138.3-143.3) (68.3-72.5) (37.5-40.8) (85.8-93.3) (44.2-50.0)(67.0-83.0) (105.8-113.3) (50.8-57.5) (125.0-134.2) (75.0-98.0)(11.0-14.0) (31.7-33.3) (94.0-110.0) (13.0-18.0) (36.7-43.3) (96.0-123.0) (15.0-19.0) (45.0-48.3) (62.0-76.0) (18.0-22.0) (51.7-56.7) (77.0-92.0) (12.0-14.0) (76.0-103.0) (13.0-16.0) (103.0-122.0) (15.0-18.0) (18.0-22.0) Xiphinema insigne m) ( m) m) m) m) m) ail 73.0 otal stylet 74.4 ( ( ( ( ( odontostyle (40.0-45.4) (52.5-58.4)odontostyle (67.5-74.2) (45.3-56.7) (83.3-90.9) (66.7-75.0)width (36.6-39.2) (82.5-90.0) (46.6-54.1) (94.2-111.7) (59.1-66.6) (50.0-52.5) (68.3-80.8) (58.3-66.7) (74.2-80.0) (89.20-95.0) Stagesn1718241 J1 J2 J3 J4Odontophore 31.5 J1T J2 J3 J4 CharacterL (mm) 0.74 Host (Code) a b c c'T 5.99 Functional 42.9 Replacement 53.9 Anal body 12.0 stages = the first (J1), second (J2), third(J3) and fourth(J4) developing of juveniles, others see table 2. able 3. Morphometrics of the 4 juvenile stages Measurements in the form: mean T 1 2 136 13 2 2004 0.15 5 4.1 0.5 0.4 5.3 4.8 1.3 10 1.1 1.6 4 China 3 rinidad T 2.27 2.58 5 4.01 1612.19 165 61 63 1.00 78 69 0.20 4.47 63.10.36 7.09.05 71 29.10.43 6.7 3.0 38 3.63 2.4 1001 102 1.0 -2.34 50 29 49 2 Locality Host (Code) males isolated from Taiwan, Trinidad and Mainland China Trinidad males isolated from Taiwan, Xiphinema insigne (2.45-2.52) (2.28-2.38) (2.34-2.42) (1.69-2.58) (2.32-2.80) (161.7-166.7) (160.0-166.7) (160.0-163.3)(60.8-66.7) (62.5-65.0)(51.0-55.0) (145.8-166.7) (60.0-63.3) (55.0-60.0)(29.0-32.0) (63.0-80.0) (28.0-30.0)(42.5-50.0) (27.0-30.0) (45.0-50.8) (156-171) (47.5-52.5) (56.7-63.3) (40.0-79.0) (24.0-31.0) (45.0-53.3) (61-65) (57-83) (27-31) (47-52) (58.4-68.1) (61.6-68.0)(6.45-7.00) (59.0-63.2) (5.95-6.63)(44.6-49.2) (6.05-6.37) (39.7-41.5)(1.59-1.96) (29.3-37.5) (1.83-2.14) (2.25-2.96) (53.3-72.7) (5.28-6.73) (24.9-56.5) (1.48-3.16) (63-81) (6.1-7.7) (30-49) (1.9-3.0) SD (min. - max.), "-" = No data . (20) aichung Taichung Guoshing Luye Daya Luye T Litchi(Xins2) Bamboo (Xins4) Loquat (Xins7) Loquat (Xins8) Bermuda-grass (Xins9) Loquat Citrus Magnolia 55.1, 53.6 62.6 65.1 60.9 54.9, 63.0 64.2 6.24, 6.6333.4, 42.0 6.74 46.8 6.31 40.7 6.19 33.8 6.29, 6.47 35.1, 43.2 6.16 38.2 (39) Xiphinema insigne ., 2000 m) 1 ( et al m) m) m) m) m) ail 71.0, 60.0 53.0 57.0 71.00 61.0, 54.0 59.0 otal stylet 160.0, 168.3 163.9 162.7 161.7 146.7, 148.3 155.7 width ( ( ( ( ( odontostyle (98.3-100.9) (97.5-102.5) (96.7-101.7) (85.0-103.4) (93-107) Character n235322919 L (mm) 2.37, 2.52 2.49 2.33 2.37 2.14, 2.33 2.15 a Functional 95.8, 100.0 99.7T Anal body 98.8Spicule 30.0, 29.0 30.0 99.7 45.0, 49.2 47.2 29.0 85.9, 86.6 95.0 48.2 28.0 50.3 25.0, 27.0 27.0 45.8, 45.0 49.1 Odontophore 64.2, 68.3 64.2 63.8 61.9 60.8, 61.7 60.73 b c c'T 2.37, 2.07 1.81 1.99 2.52 2.44, 2.00 2.19 See table 2. able 4. Morphometrics and comparisons of Xins8 subcultured on two potted citrus in green house of TARI from 2002/ 8/15 to 2003/9/15. Xins8 subcultured on two potted citrus in green house of TARI Hunt, D. J., and Singh, N. 1984 Robbins Measurements in the form: mean 5 T 1 2 3 4 Xiphinema insigne 137

Xiphinema insigne rDNA (ITS-1, 5.8S, ITS-2) Table 5. The size of rDNA ITS-1, 5.8S, ITS-2 and the pairwise percentage dissimilarities among the four geographical populations of Xiphinema insigne isolated form Taiwan ITS-1 (%) 5.8S (%) ITS-2 (%) Population (Code) No. of Xins1 Xins2 Xins7 No. of Xins1 Xins2 Xins7 No. of Xins1 Xins2 Xins7 bases bases bases Xins1 1112 - - - 160 - - - 508 - - - Xins2 1111 0.63 - - 160 1.88 - - 508 0.79 - - Xins7 1122 2.66 2.93 - 160 2.5 1.63 - 498 2.23 1.42 - Xins8 1122 2.75 3.02 0.09 160 2.5 1.63 0 498 2.23 1.42 0 elongatum 2 X. insigne Xins4, 7, 8, 9 4 Xins8 29 (63, 71, 79, 87, 87, 97 m ) 5 8 indicum-form 4 10 X. insigne (intra-population) X. elongatum (2) X. insigne 'indicum-form' 4 rDNA Bajaj Jairajpuri indicum-form ITS-1 ITS-2 18 138 4 2 (Xins1, 2 Xins7, 8) X. indicum (104, 94 - 110 mm 102, 98 - 105 mm) 3 2 X. indicum X. insigne 4 ITS-1 ITS-2 0-3.02 % X. (topotypes) X. elongatum (0 - 1.34 %)(2) Xins8 X. insigne rDNA elongatum (2083 bp, GenBank Accession No: AY524971) Razak Loof (37) 32.73 42.66 % (Royal palm) X. insigne 5.8S 0.63 % rDNA short-tailed long-tailed ( ITS-2 3.02 % 92 vs 113 m) Xins9 X. insigne (96 m) 10 2 10 X. insigne 4 10 X. (paralectotypes) insigne Wang (47) c' (Sichuan) Jiangjin 16 X. insigne 4 c' L=2.30 (1.79 - 2.59) V% = 33 (30 - 37) c'= 5.7 (4.7 - 6.3); functional odontostyle = 96.1 Xins8 X. indicum 9 (91.4 - 101.9) mm total stylet = 157.4 (150.2 - 163.8) m X. insigne tail = 127.8 (99.1 - 145.8) m [77 (74 - 83) m] Bajaj Jairajpuri 10 Zheng Brown (49) 23 X. insigne indicum-form (Zhejiang) (Hangzhou) 5 (populations 1 - 18) insigne-form (population 19 - 23) 2 X. insigne (Yulan magnolia) 10 (4) (2.57 m) insigne-form 3 4 (unidentified host) V% Xins1, 5, 9 3 [37.9 (34.2 - 42.1)] [170 10 10 (166.4 - 172.8) m] 138 13 2 2004

3 Xins8 Xins10 Xins2, 4, 7, 8, 9 5 15 2 (33.5-35.0 %) Xins8 ' ' Robbins (39) Yulan magnolia 25 [2.61 (2.24 - 2.87) m] [165 (158 - 171) m] (145.80 - 166.70 m) 9 Luc Southey (31) (143.30 - 169.20 m) (population 9 (host adaptation) Xins10 (Population 6) (population genetic structure, PGS)(5) [2.68 (2.44 - 3.09) m] (evolution) ('bonsai' tree, population 3) 6 Robbins (39) [2.74 (2.58 - 2.94) m] Yulan Magnolia 9 [(171 (167 - 175) m) Choi (9) 1: 13.1 10 118 (genital tracts) 'bonsai' Luc Southey (North Carolina, N.C.) Raleigh (31) (population 4) (California, Ca.) Luc (30) X. (Fresno) (population 5) neodimorphicaudatum Khan, 1982 X. tugewai Darekar and Khan, 1983 2 X. insigne (Mauritius)(population 7) 1 2 (Trinidad)(20) 50 m [2.52 (2.30 - 2.65) 12 (24) 40 - mm] [149 (136 - 171) m] (17) 55 m 2 2.78 2.72 mm V% 39.9 113 116 m 4 40.4 85 - 92 m(13) Zheng (49) X. insigne X. savaryi Lamberti, Troccoli & Agostinelli, 1997 X. insigne 15 2.6 (2.4 - 2.7) mm 66.0 (64.0 - 68.7) m(27) X. insigne 10 X. insigne

4 rDNA [2.68 (2.44 - 3.09) mm] X. insigne Xins1, 5, 9 3 (49) 3

[2.57 (2.35 - 2.80) mm] 3 7 3

(parthenogenesis)(31) X. insigne 1. . 2003. rDNA-RFLP . Xiphinema insigne 139

12: 235-241 15. Ferris, V. R., Ferris, J. M., and Faghihi, J. 1993. Variation 2. . 2004. in spacer ribosomal DNA in some cyst-forming species of Xiphinema elongatum . plant parasitic nematodes. Fundam. Appl. Nematol. 16: 13: 45-60 177-184. 3. Alkemada, J. R. M., and Loof, P. A. A. 1989. 16. Ferris, V. R., Ferris, J. M., Faghihi, J., and Ireholm, A. Observations on the ontogeny of some Xiphinema species 1994. Comparisons of isolates of Heterodera avenae (Nematoda: ). Med. Fac. Landbouww. using 2-D PAGE protein patterns and ribosomal DNA. J. Rijksuniv. Gent 54: 1177-1186. Nematol. 26: 144-151. 4. Bajaj, H. K., and Jairajpuri, M. S. 1977. Variability within 17. Heyns, J. 1991. The genus Xiphinema in southern Africa. Xiphinema insigne populations from India. Nematologica XXII. X. miekeae spec. nov., with notes on X. clavatum 23: 33-46. Heyns, 1965, X. setariae Luc, 1958 and X. insigne Loos, 5. Blouin, M. S., Yowell, C. A., Courtney, C. H., and Dame, 1949 (Nematoda: ). Phytophylactica 23:207- J. B. 1995. Host movement and the genetic structure of 211. populations of parasitic nematodes. Genetics 141: 1007- 18. Hoste, H., Chilton, N. B., Beveridge, I., and Gasser, R. B. 1014. 1998. Differences in the first internal transcribed spacer 6. Bulman, S. R., and Marshall, J. W. 1997. Differentiation of ribosomal DNA among five species of of Australasian potato cyst (PCN) populations Trichostrongylus. Int. J. Parasitol. 28: 1251-1260. using the polymerase chain reaction (PCR). New Zealand 19. Hoste, H., Chilton, N. B., Gasser, R. B., and Beveridge, I. J. Crop Horti. Sci. 25, 123-129. 1995. Differences in the second internal transcribed 7. Cherry, T., Szalanski, A. L., Todd, T. C., and Powers, T. spacer (ribosomal DNA) between five species of O. 1997. The internal transcribed spacer region of Trichostrongylus (Nematoda: Trichostrongylidae). Int. J. Belonolaimus (Nemata: Belonolaimidae). J. Nematol. Parasitol. 25: 75-80. 29:23-29. 20. Hunt, D. J., and Singh, N. D. 1984. Xiphinema 8. Cho, M. R. and Robbins, R. T. 1990. Observations of all costaricense Lamberti & Tarjan, 1974 and X. insigne postembryonic stages of Xiphinema coxi coxi (Nematoda: Loos, 1949, from Trinidad. Rev. Nematol. 7:19-22. Longidoridae). J. Nematol. 22:69-78. 21. Ibrahim, S. K., Baldwin, J. G., Roberts, P. A., and Hyman, 9. Choi, Y. E., Choi, Y. S., and Cho, M. R. 1992. Nematodes B. C. 1997. Genetic variation in Nacobbus aberrans: an associated with forest trees in Korea III. A new species of approach toward taxonomic resolution. J. Nematol. 29: Xiphinemella Loos, 1950, and four unrecorded species of 241-249. Xiphinema Cobb, 1913. Korean J. Appl. Entomol. 31:416- 22. Ibrahim, S. K., Perry, R. N., Burrows, P. R. and Hooper, 426. D. J. 1994. Differentiation of species and populations of 10. Cohn, E. 1969. The occurrence and distribution of species Aphelenchoides and of Ditylenchus angustus using a of Xiphinema and Longidorus in Israel. Nematologica fragment of ribosomal DNA. J. Nematol. 26:412-421. 15:179-192. 23. Kaplan, D. T., Thomas, W. K., Frisse, L. M., Sarah, J. L., 11. Cohn, E., and Sher, S. A. 1972. A contribution to the Stanton, J. M., Speijer, P. R., Marin, D. H., and taxonomy of the genus Xiphinema Cobb, 1913. J. Opperman, C. H. 2000. Phylogenetic analysis of Nematol. 4:36-65. geographically diverse Radopholus similis via rDNA 12. Coomans, A., and De Coninck, L. 1963. Observations on sequence reveals a monomorphic motif. J. Nematol. spear-formation in Xiphinema. Nematologica 9:85-96. 32:134-142. 13. Darekar, K. S., and Khan, E. 1983. Xiphinema tugewai sp. 24. Khan, E. 1982. Inagreius gloriosus gen. n. sp. n. and n. and Longidorus paramirus sp. n. (Longidoroidea: descriptions of three new species of Xiphinema Cobb, Nematoda) from Maharashtra, India. Ind. J. Nematol. 1913 along with report on X. radicicola T. Goodey, 1936 12:153-157. and X. elongatum Schuurmans Stekhoven & Teunissen, 14. Fallas, G. A., Hahn, M. L., Fargette, M., Burrows, P. R., 1938 (Nematoda: Longidoroidea) from India. Ind. J. and Sarah, J. L. 1996. Molecular and biochemical Nematol. 11: 189-204. diversity among isolates of Radopholus spp. from 25. Knoetze, R., Burger, J. T. and Meyer, A. J. 2000. different areas of the world. J. Nematol. 28:422-430. Discrimination of some Xiphinema species from South 140 13 2 2004

Africa by rDNA-RFLP analysis. African Plant Prot. 6:25- Xiphinema Cobb, 1913 (Nematoda: Longidoridae) in 30. western Malaysia. Fundam. Appl. Nematol. 21:413-428. 26. Lamberti, F., Sabova, M., DE Luca, F., Molinari, S., 38. Robbins, R. T., Brown, D. J. F., Halbrendt, M., and Vrain, Agostinelli, A., Coiro, M. I., and Valocka, B. 1999. T. C. 1996. Compendium of juvenile stages of Xiphinema Phenotypic variations and genetic characterization of species (Nematoda: Longidoridae). Russ. J. Nematol. Xiphinema populations from Slovakia (Nematoda: 4:163-171. Dorylaimida). Nematol. Medit. 27: 261-275. 39. Robbins, R. T., Zheng, J. W., and Brown, D. J. F. 2000. 27. Lamberti, E., Troccoli, A., and Agostinelli, A. 1997. Two Observations on a Xiphinema insigne population with new species of Xiphinema (Nematoda, Dorylaimida) from several males from Hangzhou, China. J. Nematol. 32:253- Thailand. Nematol. Medit. 25:239-247. 257. 28. Loof, P. A. A., and Maas, P. W. Th. 1972. The genus 40. Saigusa, T., and Yamamoto, Y. 1971. Distribution and Xiphinema (Dorylaimida) in Surinam. Nematologica hosts of a dagger nematode, Xiphinema insigne Loos, 18:92-119. detected on export Lily bulb. Res. Bull. Pl. Prot. Serv. 29. Loos, C. A. 1949. Notes on free living and plant-parasitic Japan 9:27-38. nematodes of Ceylon. No: 5. J. Zool. Soc. India 1: 23-29. 41. Siddiqi, M. R. 1959. Studies on Xiphinema spp. 30. Luc, M., Loof, P. A. A. and Brown, D. J. F. 1984. On the (Nematoda: Dorylaimoidea) from Aligarh (North India) systematics of eleven Xiphinema species (Nematoda: with comments on the genus Longidorus Micoletzky, Longidoridae) described from India. Rev. Nematol. 1922. Proc. Helminth. Soc. Wash. 26:151-163. 7:399-405. 42. Szalanski, A. L., Sui, D. D., Harris, T. S., and Powers, T. 31. Luc, M., and Southey, J. F. 1980. Study of biometrical O. 1997. Identification of cyst nematodes of agronomic variability in Xiphinema insigne Loos, 1949, and X. and regulatory concern with PCR-RFLP of ITS-1. J. elongatum Schuurmans Stekhoven & Teunissen, 1938; Nematol. 29: 255-267. description of X. savanicola n. s.p (Nematoda: 43. Tarjan, A. C., and Luc, M. 1963. Observations on Longidoridae) and comments on thelytokous species. Rev. Xiphinema insigne Loos, 1949 and Xiphinema elongatum Nematol. 3:243-269. Schuurmans Stekhoven & Teunissen, 1938 (Nematoda: 32. Nasmith, C. G., Speranzini, D., Jeng, R. and Hubbes, M. Dorylaimidae). Nematologica 9:163-172. 1996. RFLP analysis of PCR amplified ITS and 26S 44. Thiery, M., and Mugniery, D. 1996. Interspecific rDNA ribosomal RNA genes of selected entomopathogenic restriction fragment length polymorphism in Globodera nematodes (Steinernematidae, Heterorhabditidae). J. species, parasites of Solanaceous plants. Fundam. Appl. Nematol. 28:15-25. Nematol. 19: 471-479. 33. Newton, L. A., Chilton, N. B., Beveridge, I., and Gasser, 45. Vrain, T. C., Wakarchuk, D. A., Levesque, C. A., and R. B. 1998. Genetic evidence indicating that Cooperia Hamilton, R. I. 1992. Intraspecific rDNA restriction surnabada and Cooperia oncophora are one species. Int. fragment length polymorphism in the Xiphinema J. Parasitol. 28: 331-336. americanum group. Fundam. Appl. Nematol. 15:563-573. 34. Nguyen, K. B., Maruniak, J., and Adams, B. J. 2001. 46. Wang, X., Bosselut, N., Castagnone, C., Voisin, R., Abad, Diagnostic and phylogenetic utility of the rDNA internal P., and Esmenjaud, D. 2003. Multiplex polymerase chain transcribed spacer sequences of Steinernema. J. Nematol. reaction identification of single individuals of the 33:73-82. Longidorid nematodes Xiphinema index, X. 35. Orui, Y. 1996. Discrimination of the main Pratylenchus diversicaudatum, X. vuittenezi, and X. italiae using species (Nematoda: Pratylenchidae) in Japan by PCR- specific primers from ribosomal genes. Phytopathology RFLP analysis. Appl. Entomol. Zool. 31:505-514. 93:160-166. 36. Powers, T. O., Todd, T. C., Burnell, A. M., Murray, P. C. 47. Wang, S., Chiu, W. F., Yu, C., Li, C., and Robbins, R. T. B., Fleming, C. C., Szalanski, A. L., Adams, B. A., and 1996. The occurrence and geographical distribution of Harris, T. S. 1997. The rDNA internal transcribed spacer longidorid and trichodorid nematodes associated with region as a taxonomic marker for nematodes. J. Nematol. vineyards and orchards in China. Russ. J. Nematol. 4:145- 29:441-450. 153. 37. Razak, A. R., and Loof, P. A. A. 1998. The genus 48. Wendt, K. R., Vrain, T. C., and Webster, J. M. 1993. Xiphinema insigne 141

Separation of three species of Ditylenchus and some host Lamberti, Troccoli & Agostinelli, 1997 with X. insigne, races of D. dipsaci by restriction fragment length and X. siamense Lamberti, Troccoli & Agostinelli, 1997 polymorphism. J. Nematol. 25:555-563. with X. radicicola Goodey, 1936 (Nematoda: 49. Zheng, J. and Brown, D. J. F. 1999. Xiphinema insigne Longidoridae). Russ. J. Nematol. 7:127-137. Loos, 1949 and X. hunaniense Wang & Wu, 1992 from 50. Zijlstra, C., Lever, A. E. M., Uenk, B. J. and Van Silfhout, Hangzhou, China, and synonymization of X. savaryi C. H. 1995. Differences between ITS regions of isolates 142 13 2 2004

ABSTRACT

Chen, D. Y.1, Ni, H. F.2, Yen, J. H.3, Cheng, Y. H.2, and Tsay, T. T.4, 5. 2004. Variability within Xiphinema insigne populations in Taiwan. Plant Pathol. Bull. 13:127-142. (1 Plant Pathology Division, Agricultural Research Institute, Wufeng, Taichung, Taiwan, R.O.C.; 2 Department of Plant Protection, Chia Yi Agricultural Experiment Station, ARI, Taiwan; R.O.C.; 3Agricultural Extension Center, National Chung Hsing University, Taichung, Taiwan, R.O.C.; 4 Department of Plant Pathology, National Chung Hsing University, Taichung, Taiwan, R.O.C.; 5 Corresponding author, E-mail: [email protected], Fax: +886-4- 22876712)

Ten geographical populations of Xiphinema insigne were isolated from the rhizosphere of litchi, loquat, pear, Bermuda-grass and bamboo using modified Baermann funnel method during 1998 to 2002 in Taiwan. According to the matchable length of replacement odontostyle and functional odontostyle, the evident gap of replacement odontostyle and total stylet length between the four successive stages of juvenile and its female, it was assured that examined specimens of each population were not mixed with other morphological similar Xiphinema species. Although, based on each of the three items of the position of vulva (V%), the length of total stylet and tail of the morphometrics of females, the 10 populations could be divided into two groups, respectively, but which were different to each other. Comparing the size and pairwise percentage dissimilarity of rDNA (ITS-1 and ITS-2), the four tested populations also were separated into two groups (Xins1, 2 vs Xins7, 8), but however it didn't coincide with the results conducted by each of the above three items. Also the morphometrics for the first and second stages of juvenile among the 10 populations were very similar to each other. After all, judging from the above three viewpoints, we recognized that these 10 populations with somehow morphological variance were one species X. insigne, and which with intraspecific variation of ITS-1 and ITS-2 ranged from 0 to 3.02 %. The shape of lip region and the morphometrics of X. insigne males were almost identical with the females, but its tail shape with more variance, and the numbers of supplements were 5 to 8. Compared to some published reports, the morphometrics of X. insigne specimens identified in Taiwan had showed two main differences, the length of female body and total stylet. However, collecting more morphometric data of the juveniles, males and the sequences of rDNA of X. insigne around the world should be the only feasible method to decide that whether these differences were intraspecific or interspecific variation.

Key words: internal transcribed spacer (ITS), ribosomal DNA (rDNA), Variability, Xiphinema insigne

of root-knot nematodes Meloidogyne hapla and M.

chitwoodi. Phytopathology 85:1231-1237.