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Description and SEM Observations of Meloidogyne Chitwoodi N. Sp

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Description and SEM Observations of Meloidogyne Chitwoodi N. Sp Description and SEM Observations of Meloidogyne chitwoodi n. sp. (Meloidogynidae), a Root-knot Nematode on Potato in the Pacific Northwest A. Morgan Golden, J. H. O'Bannon, G. S. Santo, and A. M. Finley 1 Abstract: Meloidogyne chitwoodi n. sp. is described and illustrated from potato (Solanum tuberosum) originally collected from Quincy, Washington, USA. This new species resembles M. hapla, but its perineal pattern is basically round to oval with distinctive and broken, curled, or twisted striae around and above the anal area. The vulva is in a sunken area devoid of striae. Vesicles or vesicle-like structures are present in the median bulb of females. The larva tail, being short and blunt with a hyaline tail terminal having little or no taper to its rounded terminus, is distinctively different from M. hapla. SEM observations revealed the nature of the perineal pattern and details of the head of larvae and males, and showed the spicules to have dentate tips ventrally. Hosts for M. chitwoodi n. sp. include potato, tomato, corn, and wheat but not straw- berry, pepper, or peanut. The latter three crops are excellent hosts for M. hapla. The known distribntion of this new root-knot species presently involves certain areas of Idaho, Washington, and Oregon. The common name "Columbia root-knot nematode" is proposed for M. chitwoodi n. sp. Key Words: taxonomy, morphology, Meloidogyne, root-knot, new species, SEM ultra- structure, potato, Solarium tuberosum, hosts. In December 1974 an infected 'Russet Larvae for morphological examination Burbank' potato and dissected root-knot were generally recovered from fresh infected specimens which had been tentatively iden- roots, or egg sacs, that were kept in petri tified as Meloidogyne hapla Chitwood, 1949 dishes with a small amount of water. Fe- were received from Aberdeen, Idaho, for males were later dissected from the roots deposit in the USDA Nematode Collection, after fixation overnight in 3% formalde- Beltsville, Maryland. The specimens were hyde solution. Males were obtained in this filed for later study as part of the "M. hapla same manner or by soil sieving followed by complex" (2). When examined critically in Baermann funnel extraction. The pro- 1977, they were found to represent an unde- cedures used for measuring, drawing, and scribed species o[ Meloidogyne. Concur- preparing specimens were essentially the rently in 1977, and subsequently, this new same as those used by Golden and Birch- Meloidogyne species has been found and field (3), except that some fixed females identified in several areas of the Pacific were cut and mounted in lactophenol solu- Northwest, specifically in Idaho, Washing- tion. Photomicrographs of perineal patterns ton, and Oregon (4). The present taxonomic and eggs were made with an automatic 35- report on the description of this new root- ram camera attached to a compound micro- knot species represents only a part of the scope having an interference contrast sys- extensive research being conducted in the tem, and those of infected roots and whole Pacific Northwest on this destructive pest females were made with a 8.3 × 10.8-cm of potato (4). sheet-film camera attached to a dissecting Specimens used in this description were microscope. In making the scanning elec- obtained from cultures originating at tron micrographs, living specimens were Quincy, Washington, and grown on potato fixed in 3% glutaraldehyde solution with in a growth chamber and in an isolated area 0.05 M phosphate buffer (pH 6.8), dehy- o[ a greenhouse. drated in a graded series of ethanol, critical- Received for publication 27 May 1980. point dried from liquid CO2, sputter-coated ~Respectiveiy: Nematologist, Nematology Laboratory, with 20-30 nm layer of gold-palladium, and Plant Protection Institute, USDA, SEA, AR, Beltsville, MD 20705; Nematologist, USDA, SEA, AR, Prosser, WA 99350; examined with a scanning electron micro- Associate Nematologist, Washington State University, Irri- scope (SEM). gated Agriculture Research and Extension Center, Prosser, WA 99350; and Plant Pathologist, University of Idaho, Meloidogyne chitwoodi n. sp. Moscow, ID 83843. University of Idaho Publication No. 8076. Appreciation for technical assistance is extended to Females (60): Length 430-740/~m (mean Donna M. S. Ellington, Support Scientist, and Paula Crow- ley, Laboratory Technician (in laboratory of senior author), 591 tzm, standard deviation [SD] 60 /zm); and to Bruce Ingber, Laboratory Technician, Electron width 344-518 /zm (422 /~m, SD 42); a = Microscopy Laboratory, USDA, SEA, AR, Beltsville, MD 20705, for SEM technical assistance, 1.1-1.8 (1.4, SD 0.2); b = 3.8-5.0 (4.4, SD 319 320 Journal o[ Nematology, Volume 12, No. 4, October 1980 ! ;i; ,~,il ~,i h' i ,, i II ] /,# I':7 4-1 i. t!. i4 I I /' - Iv/ / I • ..~: 1 • T T 5 L'7 I • / H. / (' ,/..f,. "I" gz:~, ~1'" • u. #" X •,i"/ I I ^ ~ ," /~t" ,...]" • i~ I. I I I "T- ~ r//'. i. i.t T tit /. I. /~ t%" • , " " . ~\I • iL I / ' .:..; ",,' .. ~$,'. ,/ ,.;~ , • ~', N\ l " ' ~ ~ / " tzf/F,i-Ts.' q 'l • • . , , . '& ,, , , .-¢,' • ,"'-..,.: :,, ",.'~l;'...'.' .,,.I, ,," ', " ,,,.,,X"~. ~,' oIlI -) ' ' "' ..... --I ...... ~,:'/ [ ." ~,". / "- ,t" .......... .~Wt/.#~.z.. ' " I ' ",:I ~ • -r ,-" ' • ' v" - ./. ,~ \ \. ., , , • I ~,,,\. /,-', / ,. >/ ~t" v ,.,,~ . / ,.::.., . • ,,,% .I ]. • -,, ' '*'. / '.. 11. .\' .".~," • ", ~'~ "[ - 'X....." ', / ---' ~' ', / . #/ . ', . ," . \, ~/ It .1". • :~ ~>" \ l':',/ . " " ' ~-:-" ~7' "-'--'J ' • ~ '\")., ('-TI I,.. ,',,~. ' ' \' I"l' I <:, . :::i:, " tk~. / • ' . -- .=..._.T-:.~.,...J.,t,---: - • e L.' \,,3-, br I.":...:.~." \V .i. s # >- ,..~...:c...f~.... .~,.,,.. <-': ..,7-:-.. • ". ... ~ -:,,.~. :" ----:,,...-..: -- 'm .---, ~. .~-~ ' . • \ ,~~ • ,L r I'i~,. 'A " " \ ' " '.."-":"' .... • ....... - "'.': ~- • ........ :.: ...... ~.,..~1.1:!,.:' • :1 ..' ." Ii. 1 :1'~ ~: .: '' (:" " " @::.'"'~ <:: '.- :., .... <:>"-, i:....:~ ~ir... ~:::I : ,':: - ....., ..'...... (,:-". ' ~ " . " ....,:. i . '......... ......"''" ." \ " t-r'i i.. t!~.: ;i . ". i, • ~,. '~' ~:~ '" ' . "" * * , . ",."::, I .:, +~ '...... ':: :::, "";" "+""~..... " "" "~ ~'/-i l*" ' :,''": ":1 . .,...:....... :,,. I ...... • • ~ .............. , . .- <, ,,.,'::. .:i;i ' 81-'1 Figs. 1-8. Drawings of Meloidogyne chltwoodi n. sp. 1) Anterior region of female (note vesicles or vesicle- like structures in the median bulb, clustered around the lumen anterior to the center valve plates). 2-6) Larvae:: Consecutively, anterior region, cuticular annulation and lateral field at midbody, tails showing shapes with blunt hyaline tail terminals. 7-8) Male. 0.4); stylet 11.2-12.5 ffm (11.9 t~m, SD 0.3); /~m (63 /~m, SD 7) from anterior end; ex- width of stylet knobs 3.4-4.3 /~m (3.8 /~m, cretory pore 10-27 ffm (18 /~m, SD 5) from SD 0.3); dorsal esophageal gland orifice anterior end; vulva slit length 19-32 /~m (DGO) 3.4-5.5 #m (4.2 /~m, SD 0.6) from (27 /~m, SD 3); distance from vulva slit to base of stylet; center of median bulb 52-80 anus 13-22/~m (18/~m, SD 2). Meloidogyne Taxonomy, Morphology,SEM: Golden et al. 321 Figs. 9-17. Photomicrographs of nine females of Meloidogyne chitu,oodi n. sp. showing representative perineal patterns. 322 Journal of Nematology, Volume 12, No. 4, October 1980 Figs. 18-25. Scanning electron micrographs of perineal patterns and vulva areas of females of Meloidogyne chitwoodi n. sp. Holotype (female): Length 598 t~m; /~m; distance from vulva slit to anus 18.1 width 474 t~m; a = 1.3; b = 4.3; styler 12 ptm. #m; DGO 4.3/zm from base of stylet; width Description. Females: Body pearly white, of stylet knobs 3.9/~m; vulva slit length 29.2 globular to pear-shaped, with slight pos- Meloidogyne Taxonomy, Morphology, SEM: Golden et al. 323 terior protuberance visible occasionally, and monly appear as illustrated (Fig. 8). Testis with distinct neck situated anteriorly on a one or two. Spicules arcuate, and under median plane with terminal vulva (Fig. 36). SEM can be seen to have dentate tips ven- Esophageal and anterior region often ap- trally (Fig. 33). Phasmids located at or pearing as illustrated (Fig. 1). Several small anterior to cloaca. Tail short, rounded (Fig. vesicles or vesicle-like structures usually 7). present within the median bulb and are Second-stage larvae (60); Length 336--417 clustered around the lumen anterior to the ~tm (390 ~m, SD 16); a -~ 24.5-29.8 (27.5, valve plates of the median bulb (Fig. 1). SD 1.2); b = 3.3-3.8 (3.6, SD 0.2); c = 7.9-- Head with distinct but weak cephalic frame- 9.6 (8.9, SD 0.4); stylet 9.0-10.3 t~m (9.9 ttm, work, offset from neck but variable in exact SD 0.3); DGO 2.6-3.9 ~tm (3.2 ~m, SD 0.2) shape, and bearing a labial cap and usually from base of stylet; center of median bulb one cephalic annule. Excretory pore clearly 43-56 ttm (51 ,m, SD 3) from anterior end; visible and commonly located at a distance head width 4.7-5.2/~m (5/zm, SD 0.2); head equal to about 1 1/2 stylet lengths from height 1.7-2.6 /ml (2.3 /zm, SD 0.2); hw/hh anterior end. Stylet small but strong, having ratio 1.8-2.7 (2.3, SD 0.2); tail length 39-47 a dorsal curvature and rounded knobs that ~m (43 t~m, SD 1.8); hyaline tail terminal slope posteriorly. Perineal pattern (Figs. 8.6-13.8 tzm (11 tzm, SD 1); caudal ratio A 9-17, 18-25) quite distinctive with striae 1.9-3.9 (2.5, SD 0.3); caudal ratio B 3.0-5.7 around and above anal area being broken, (4.2, SD 0.7). curved, twisted, or curled as illustrated; Description. Larvae: Body small, vermi- overall shape of pattern appearing round form, 12.5-15.5 tzm (14.2 #m, SD 0.6) in to oval, sometimes with inner striae form- width at midbody, tapering at both ex- ing an arch of varying degrees. Punctation tremities but more so posteriorly. Head not not observed. Vulva sunken in an area offset, with weak cephalic framework, and variable in shape and devoid of striae. bearing a labial disc and a large post-labial Males (30): Length 887-1268 /~m (1068 annule without striations (Figs.
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