TAP CHI SINH HOC 2017, 39(3): 253-263 Two new species DOI: of 10.15625/0866 plant parasitic-7160 nematodes/v39n3.10665 TWO NEW SPECIES OF PLANT PARASITIC NEMATODES Hirschmanniella bananae n.sp. (Nematoda: Pratylenchidae) AND Scutellonema tanlamense n.sp. (Nematoda: Hoplolaimidae) ASSOCIATED WITH BANANA IN VIETNAM Nguyen Ngoc Chau1,2 1Graduare University of Science and Technology, VAST 2Institute of Ecology and Biological Resources (IEBR), VAST ABSTRACT: During investigation on the plant parasitic nematodes on bananas in North and Central provinces of Vietnam carried out in 1995 two new species were found, Hirschmanniella bananae n.sp. and Scutellonema tanlamese n.sp. Hirschmanniella bananae n.sp. is characterized by having 6 annuli in lip region, areolation of entire lateral field and presence of a mid-terminal mucro at the tail end. The new species is close to H. shamini Ahmad, 1972 but differs from H. shamini by longer body length in females (1450-1750 µm vs 1190-1360 m), longer stylet longer (18-20.5 m vs 16-19 m) and longer spicule longer (28-29.5 m vs 22-25 m). Scutellonema tanlamese n.sp. is characterized by lip region with three annuli and numerous longitudinal ridges (18) in basal annulus and structure of lateral field at the tail region with inner lines extending to tail terminus. The new species is closest to S. brachyurus type B (African populations) described by Van Den Berg et al. in 2013 but can be distinguished by body length shorter (643-708 µm vs 777-820 µm), stylet length shorter (24.5-26.5 µm vs 28-29.5 m), and also by structure of lateral field with two inner lines going throught at tail region and vulval edge distinct with epiptygma appearing double and folded into vagina. Keywords: Hirschmanniella, Scutellonema, bananas, North and Central Vietnam, plant parasitic nematodes, new species, taxonomy. Citation: Nguyen Ngoc Chau, 2017. Two new species of plant parasitic nematodes Hirschmanniella bananae n.sp. (Nematoda: Pratylenchidae) and Scutellonema tanlamense n.sp. (Nematoda: Hoplolaimidae) associated with banana in Vienam. Tap chi Sinh hoc, 39(3): 253-263. DOI: 10.15625/0866-7160/v39n3.10665. *Corresponding author: [email protected] Received 19 August 2017, accepted 12 September 2017 INTRODUCTION were recorded, among them 24 species Banana (Musa paradisiaca L.) is one of the associated with banana (Eroshenko et al., 1985). important fruit crops in Vietnam and is used as Another survey to assess the occurrence and desert fruit as well as staple food. Banana is potential damage of parasitic nematodes on grown everywhere; apart from some state farms, bananas in Vietnam was carried out in 1995. every household at the countryside grows The latter investigation revealed 53 species of banana. Up to 2014 in total 50,200 ha bananas plant-parasitic nematodes associated with were planted with about 1,4 million tons of fruit bananas in North and Central Vietnam. Some produced. Most of these products is consumed species considered to be most common in domestically for dessert and food cooking, one distribution and detrimental to bananas were fourth of the production is exported to China Pratylenchus coffeae, Helicotylenchus and Japan. multicinctus, H. dihystera, Meloidogyne incognita and Rotylenchulus reniformis The preliminary surveys on plant-parasitic (Nguyen et al., 1997). Among these plant nematodes in Vietnam were carried out in North parasitic nematodes, Hirshmaniella spec. n. and Vietnam during 1977-1978, by joined Scutellonema spec. n were revealed as new collaboration of Russian-Vietnamese scientists. species but were not putting scientific named Ninety species of plant-parasitic nematodes for official publication so far. This paper deals 253 Nguyen Ngoc Chau with the description of these two new species length; m = length of conus as percentage of named as Hirschmanniella bananae n.sp. and total stylet length; O = distance between stylet Scutellonema tanlamese n.sp. base and orifice of dorsal esophageal gland in Materials and Methods µm. All measurements are presented in micrometers and expressed as the mean ± Survey and sampling. A total of 47 standard deviation followed by the range. combination samples that included soils and roots associated with planed banana varieties SEM photographs. The nematodes used for AAA and AAB were collected from five SEM observation were fixed in a 4% localities from Northern and Central Vietnam formaldehyde solution and mounted on slide in including: Yen Nghia and Ba Vi districts dehydrated glycerin following Seinhorst’s rapid (surround Hanoi city), Tam Thien Mau farming method (1959). When glycerin embedded (Ha Bac province), Van Giang district (Hai nematodes were used for SEM. Specimens were Hung province) and Tanlam Faming (Quang Tri first transferred into a small dish filled with a province). Each combination sample was taken drop of glycerin. Distilled water is added drop 250 mL soil and roots within a rhizosphere with by drop until the nematodes are in pure distilled facial profile 20 20 20 cm at base of banana water. Particles which might adhere to the tree (Speijer & De Waele, 1997). nematodes are removed by ultrasonic treatment for about 5 min. The nematodes are dehydrated Nematode extraction. Nematodes from 250 using a gradual series of ethanol concentration mL soil samples were extracted by a modified of 30%, 50%, 75% and 95%. After this initial Cobb’s sieving-decanting technique (ref?). dehydration which lasts for about 8 hours in Firstly, the 250 ml soil sample was transferred total. The specimens are left overnight in 100% into a 5L capacity container in 3 liters of water, ethanol. The standard critical point drying gently stirred to homogenize the sample in the procedure is applied with CO2 this temperature water, and then poured over a standard sieve (22 of 31.1oC and the pressure at 72.9 bar. During cm diameter) with a mesh size of 0.5 mm into a the transportation the specimens are kept into second container to remove coarse soil and absolute ethanol. The dried nematodes are plant particles. This decantation was repeated individually mounted on standard specimens about 5-7 times, and then the soil solution is stubs and sputter-coated with gold. The filtered over a standard sieve with mesh size 75 nematodes are picked by hand one by one and µm, so as to collect sediment with nematodes. put on the specimen table, so that the head of The latter is rinsed over a small sieve described nematodes lies on the glass rod and can be by Nguyen & Nguyen (1993). Nematodes were o studied en face. The surface of the specimen killed in water at 65-70 C, fixed in TAF and table consists of an auto-adhesive conducting mounted in anhydrous glycerin using the slow aluminum tape stick to the nickel-print (glue). method of Hooper & Evans (1993). The scanning of nematode specimens were All morphometrics were performed with a observed and taken photographs with the JSM- camera lucida drawing tube. In the 840 (Eisenback, 1991). measurement and description of the species, the All examined nematode specimens was De Man's formulae (I884) are used. Of these deposited at the Nematode Collection of the some abbreviated letter used as: L = total body Department of Nematology, Institute Ecology length in µm; a = body length/maximum body and Biological Resources (IEBR) Vietnam width; b = body length/esophageal length; b' = Academy of Science and Technology, 18 Hoang body length/distance from anterior end to Quoc Viet Road, Cau Giay District, 10000 posterior end of esophageal glands; c = body Hanoi, Vietnam. length/anal body width; c’ = body length/body width at anus; V = distance from anterior end to Descriptions of new species vulva x 100/body length; T = distance from Hirschmanniella bananae n.sp. (Figs 1 & 2) cloacal apeature to anterior of testis 100/body Measurements 254 Two new species of plant parasitic nematodes Holotype (female). L = 1600 m; a = 67.2; 0.1 (2.2-2.6) m. b = 12.8; c = 17; c’ = 6.8; V (%) = 52.6; stylet Male paratypes (n = 12). L= 1250 ± 87.6 length = 19.1 m; m = 50. (1150-1400) m; body width 20 ± 1.4 (18-22.5) Female paratypes (n = 17). L = 1582 ± 98.7 m; a = 62.6 ± 3 (57.5-65.5); oesophageal (1450-1750) m; body width 23.6 ± 1 (22.0- length = 114 ± 13 (93-133) m; b = 10.9 ± 0.7 24.5) m; a = 66.8 ± 2.4 (64.0-71.0); (10.3-12.3); tail length = 84 ± 4.5 (77-90) m; c esophageal length = 133.1 ± 9.5 (122.5-144.0) = 14.4 ± 1.0 (13.0-15.5); c’ = 6.6 ± 1.0 (5.5- m; b = 11.9 ± 0.8 (10.5-13.0); tail length = 8.5); testis length = 34.7 ± 2.7 (30-37.5) m; 88.9 ± 6.0 (77.5-93.5) m; c = 17.9 ± 2.4 (16.0- stylet length = 18 ± 0.4 (18-19) m; m = 49 ± 1 22.5); c’ = 6.7 ± 0.9 (5.5-8.1); V = 53.7 ± 1.6 (47-50); O = 2 ± 0.5 (1.5-3) m; spicule length (52.5-57.0); stylet length = 19.2 ± 0.8 (18.0- = 28 ± 0.6 (28-29.5) m; gubernaculum length 20.5) m; m = 50.0 ± 0.6 (49.0-51.0); O = 2.5 ± = 10 ± 0.5 (10-11) m. Figure 1. Hirschmanniella bananae n. sp. A. Body habitus of heat-relaxed specimens; B. anterior region; C. lateral view at the vulva region with showing infecting of parasitic bacteria (Pasteuria penetrans); D. female tail; E. male tail; F, G, H. variation of the tail terminus. 255 Nguyen Ngoc Chau Figure 2. SEM photographs of Hirschmanniella bananae n.sp.