supplemental description of paraphelenchus acontioides ...€¦ · nematology , 2011, vol. 13(8),...

Nematology 2011 Vol 13(8) 887-899

Supplemental description of Paraphelenchus acontioides(Tylenchida Aphelenchidae Paraphelenchinae) with ribosomal

DNA trees and a morphometric compendiumof female Paraphelenchus

Lynn K CARTA 1lowast Andrea M SKANTAR 1 Zafar A HANDOO 1 and Melissa A BAYNES 2

1 United States Department of Agriculture ARS-BARC Nematology Laboratory Beltsville MD 20705 USA2 Department of Forest Ecology and Biogeosciences University of Idaho Moscow ID 83844 USA

Received 30 September 2010 revised 7 February 2011Accepted for publication 7 February 2011 available online 5 April 2011

Summary ndash Nematodes were isolated from surface-sterilised stems of cheatgrass Bromus tectorum (Poaceae) in Colorado grown onFusarium (Hypocreaceae) fungus culture and identified as Paraphelenchus acontioides Morphometrics and micrographic morphologyof this species are given to supplement the original description and expand the comparative species diagnosis A tabular morphometriccompendium of the females of the 23 species of Paraphelenchus is provided as the last diagnostic compilation was in 1984 Variationsin the oviduct within the genus are reviewed to evaluate the taxonomic assignment of P deckeri a morphologically transitionalspecies between Aphelenchus and Paraphelenchus Sequences were generated for both 18S and 28S ribosomal DNA representingthe first identified species within Paraphelenchus so characterised These sequences were incorporated into phylogenetic trees withrelated species of Aphelenchidae and Tylenchidae Aphelenchus avenae isolates formed a well supported monophyletic sister group toParaphelenchus The ecology of Paraphelenchus cheat grass and Fusarium is also discussed

Keywords ndash fungivorous nematode invasive species key morphology molecular phylogeny taxonomy

Nematodes of the genus Paraphelenchus Micoletzky1922 are generally regarded as fungivorous (Hunt 1993)and are frequently found in association with plantsTwenty-three species are known including Paraphelen-chus acontioides Taylor amp Pillai 1967 P alii (Ali Fa-rooqui amp Suryawanshi 1970) Fortuner 1985 (= P afsiHunt 1993) and P micoletzkyi Ali Farooqui amp Suryawan-shi 1970 (= junior homonym) nec P micoletzkyi Steiner1941 (= junior synonym of Aphelenchus avenae) P am-blyurus Steiner 1934 P basili Das 1960 P batavicusFilipjev 1934 P crenatus Das amp Singh 1968 P deckeri(Zeidan amp Geraert 1989) Andraacutessy 2007 P fidicauda-tus Eroshenko 1966 P goodeyi Tandon amp Singh 1970P heterolineatus Haque 1967 P intermedius Thorneamp Malek 1968 P myceliophthorus JB Goodey 1958P obscurus Muchina 1988 P octolineatus Shavrov1968 P orientalis Muchina 1988 P paramonovi Haque1967 P porrectus Eroshenko 1966 P pseudoparieti-nus Micoletzky 1922 P sacchari Husain amp Khan 1967

lowast Corresponding author e-mail lynncartaarsusdagov

P tritici Baranovskaya 1958 P ussuriensis Eroshenko1966 P zeae Romaniko 1968 and P zicsii Andraacutessy1989

Many of the morphometric values of the species over-lap so qualitative morphology is particularly importantfor diagnosis The most important of these include thepresence and shape of the mucro on the female tail thenumber of lines in the lateral field (typically 4-9) andcontinuous or indented lip region Unfortunately molecu-lar data appear to be confined to an unknown species forwhich the small subunit rDNA has been derived (Holter-man et al 2009)

Many of the species have been described from Asiapart of their proposed land origin from eastern Gond-wana in the Devonian epoch (Ryss 2007) including de-scriptions of life stages and female tail papillae of Pmyceliophthorus (Ryss amp Chernetskaya 2010) Sevenspecies have been found in Europe (Andraacutessy 2007)Within the USA numerous specimens of P pseudopari-

copy Koninklijke Brill NV Leiden 2011 DOI101163138855411X560968Also available online - wwwbrillnlnemy 887

LK Carta et al

etinus and P intermedius have been deposited into theUnited States Department of Agriculture Nematode Col-lection (USDANC) database One notable species is Pacontioides which was discovered in soil around Ken-tucky bluegrass (Agrostis palustris Huds) in Urbana Illi-nois The original population of this nematode specieswas initially raised for at least 5 years on the fungusPyrenochaeta terrestris (Hansen) Gorenz JC Walker ampLarson before actually being described (Taylor amp Pil-lai 1967) Subsequently morphometrics for this nema-tode were taken for material cultured on the same fungusand seven others resulting in expanded ranges (Pillai ampTaylor 1967a) especially for the b ratio

This paper reports the discovery of a Paraphelenchuspopulation in stems of cheatgrass Bromus tectorum Lin Colorado during a survey of endophytic fungi aspossible biocontrol agents within eight mid-western andwestern United States and British Columbia CanadaBecause many other Paraphelenchus species descriptionsare incomplete and not readily comparable detailedmorphometrics of the new population were compiled andcontrasted with other isolates of P acontioides and theother species of the genus to enhance the diagnosticinformation available

Materials and methods

NEMATODE AND FUNGUS ISOLATION

Sixty-three populations of cheatgrass B tectorumwere sampled throughout North America (Idaho Wash-ington Nevada Colorado New Mexico Iowa Illinoisand British Columbia) Populations were typically aboutfive miles apart but others were 1-20 or more milesapart Twenty plants from each population were collectedA 2 cm segment was clipped around the lowest culm node(solid region on the shoot central axis that may gener-ate a leaf sheath or adventitious bud) of each plant Seg-ments were surface-sterilised in 50 ethanol for 5 minand rinsed in sterile deionised water for 1 min (Lug-inbuhl amp Muller 1980 Schulz et al 1993) To facili-tate isolation of fungi culm segments from each popu-lation were placed onto potato dextrose agar plates Im-print plates were made to ensure surface sterilisationwas successful Fungi were identified morphologicallyCultures were placed on laboratory benches at ambientroom temperature and subcultured as needed Two grasspopulations had nematodes and endophytic Fusarium spand nematodes and fungi were not found independently

of each other The cheatgrass population with P acon-tioides was isolated from near Piney River CO USA(3950prime2499primeprimeN 10638prime2685primeprimeW) south of Yampa COUSA off Highway 131

NEMATODE PRESERVATION AND IMAGING

Nematodes were rinsed from fungal plates placed in4 formalin for 24 h rinsed in 15 ml plastic tubes withwater and sent to Beltsville MD USA From here theywere placed in 4 formalin for another 24 h dehydratedin alcohol and glycerin mounted in glycerin and sealedImages and measurements were made with a Zeiss Ul-traphot II microscope (Carl Zeiss Jena Germany andBaltimore Instrument Company Baltimore MD USA)equipped with differential interference contrast (DIC) op-tics Photographs were taken of formalin-fixed specimensrinsed in water Females were measured with an ocu-lar micrometer Fixed specimens used to evaluate rela-tionships were observed from the USDANC These in-cluded type and paratype slides of P intermedius fromthe Thorne collection (USDANC Entries 31046 2974428745 31049 slides 1a 1b (type five females) 1c 1ifor different South Dakota sites of G Thorne 28744)and P pseudoparietinus Micoletzky 1922 (USDANC En-try 21084 slide G11751 from soil of Chewings fescuegrass Festuca rubra commutate WB Courtney Asto-ria Oregon 1941) identified as P intermedius here basedon six lateral incisures and morphometrics and four fe-male paratypes of P deckeri (slide T-4299p from shrubsof Abuttaraz Sudan) Type material from the old Nema-tode Slide Collection of the Department of Plant Pathol-ogy (now Crop Sciences) University of Illinois UrbanaIL USA and P amblyurus from the Steiner collection ofthe USDANC were unavailable Reproductive system ter-minology of Triantaphyllou and Fisher (1976) as used forA avenae was applied to a single gonad arm going in ananterior direction vulva vagina gonoduct (uterus sper-matheca fertilisation chamber (eight columns of two rowsof tightly packed oval cells not contained in a membrane)sphincter ndash (three rows of globular cells not contained ina membrane)) and ovary The oviduct comprises a fer-tilisation chamber and sphincter cells (Triantaphyllou ampFisher 1976) but the term has also been used to describethe sphincter cells only (Geraert 1981)

PCR AND SEQUENCING

Nematodes were rinsed from fungal plates placed in70 alcohol in 15 ml plastic tubes and sent to Beltsville

888 Nematology

Redescription of Paraphelenchus acontioides and studies on the genus

MD USA They were rinsed in water mounted on slidesfor imaging recovered and individually processed Ne-matodes were mechanically disrupted in 20 μl of extrac-tion buffer as described by Thomas et al (1997) andthen stored in PCR tubes at minus80C until needed Ex-tracts were prepared by incubating the tubes at 60Cfor 60 min followed by 95C for 15 min to deactivatethe proteinase K and centrifuged briefly prior to use inPCR Each 25 μl PCR reaction contained 1 unit Plat-inum Taq (Invitrogen Carlsbad CA USA) 1times reactionbuffer (20 mM Tris-HCl pH 84 50 mM KCl 25 mMMgCl2) 02 mM dNTP mix 03 μM of each primer and2 μl nematode extract 28S reaction contained primersD2A (5prime-ACAAGTACCGTGAGGGAAAGTTG-3prime) andD3B (5prime-TCGGAAGGAACCAGCTACTA-3prime) (Nunn1992) Cycling was performed as described by DeLey et al (2005) and Ye et al (2007) The partial 18Ssequence was amplified in two overlapping segments us-ing primers 550F (5prime-GGCAAGTCTGGTGCCAGCAGCC-3prime) with 1108R (5prime-CCACTCCTGGTGGTGCCCTTCC-3prime) and 18s12 (5prime-GGCGATCAGATACCGCCCTAGTT-3prime) with 18sr2b (5prime-TACAAAGGGCAGGGACGTAAT-3prime) Cycling conditions for the 550F1108R PCRreaction were 1 cycle of 94C for 2 min followedby 40 cycles of 94C for 20 s 65C for 30 s and72C for 30 s finishing with 1 cycle of 72C for5 min For the 18s1218sr2b primer pair cycling con-ditions were the same as above except the annealingtemperature was 59C PCR products were analysed byelectrophoresis on 1 agarose1times SB (sodium borate-EDTA) Gels were stained with ethidium bromide andvisualised using the UGenius gel documentation sys-tem (Syngene Frederick MD USA) DNA was excisedfrom the gels and purified with the QIAquick Gel Ex-traction Kit (Qiagen Valencia CA USA) PCR productswere quantified using a Nanodrop 8000 spectrophotome-ter (Thermo Fisher Scientific Pittsburgh PA USA) andsequenced directly at the University of Maryland Cen-ter for Biosystems Research DNA sequences were as-sembled using Sequencher 4101 (Genecodes Ann Ar-bor MI USA) DNA sequences were analysed using theBLASTN megablast program optimised for highly similarsequences httpwwwncbinlmnihgovblastBlastcgiSequences were submitted to GenBank under accessionnumbers HQ218322 for 28S and HQ218323 for 18S

PHYLOGENETIC METHODS

Nematode sequences for P acontioides were com-bined with other GenBank sequences of Tylenchidae

and Aphelenchidae namely 18S Ditylenchus dipsaci(Kuumlhn 1857) Filipjev 1936 AY593906 Tylenchus ar-cuatus Siddiqi 1963 EU306349 A avenae EU306347A avenae 1 AY284639 A avenae 2 AY284640 A ave-nae AB368918 A avenae AF036586 Aphelenchus spAY284641 Paraphelenchus sp (JH-2004 isolate)AY284642 and new P acontioides HQ218323 and 28SCoslenchus costatus (de Man 1921) Siddiqi 1978DQ328719 Ditylenchus halictus Giblin-Davis ErteldKanzaki Ye amp Center 2010 AY589364 D destructorThorne 1945 DQ328727 A avenae AB368536 A ave-nae EU325683 Aphelenchus sp DQ145664 and new Pacontioides HQ218323

Alignments were made with ClustalW2 (Larkin etal 2007) and checked by eye for consistency of con-served positions The alignment was run through PAUP4b10 (Swofford 2002) Maximum Parsimony (MP) boot-strapped searches with 1000 replicates were conductedemploying tree bisection-reconnection (TBR) branchswapping and accelerated transformation (ACCTRAN)character state optimisation PAUP was also used forgenerating sequence and tree statistics Geneious Prov 503 (Biomatters Auckland New Zealand Drummondet al 2009) was used to examine apomorphic charac-ters of Paraphelenchus Maximum likelihood (ML) treesare presented in the figures because the computationally-intensive probabilistic ML method is less affected bysampling error and infers better trees than distance orparsimony methods (Swofford et al 1996) Alignmentsin PHYLIP format were run in web-based RAxML (Sta-matakis et al 2008) with 100 bootstrap runs and ML esti-mate of 25 per site rate categories Branch support valuesabove 50 given for ML followed by those for MP andML parameters given in figure legends

Description

Paraphelenchus acontioides Taylor amp Pillai 1967(Figs 1 2A)

MEASUREMENTS

See Table 1

DESCRIPTION

Body spiral to crook-shaped after death Non-indentedlip region ca 15 stylet length high 35 μm high times9 μm wide Stylet without swellings at base although

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LK Carta et al

Fig 1 Paraphelenchus acontioides Colorado population femaleA Body B Median bulb with arrow pointing at constrictionC Tail with mucroventral tuberculus (v arrow) and obscuredorsal tuberculus (d arrow)

basal lumen sometimes slightly inflated surrounded byprominent muscles Corpus length ca 27 times isthmuslength slightly longer than isthmus pharyngeal glandsabutting intestine Median bulb with constriction ante-riorly and high collar-like sheath surrounding anteriormuscle Nerve ring just posterior to median bulb Excre-tory pore opening posterior to nerve ring Anterior gonadlength highly variable not reaching pharyngo-intestinal

Fig 2 Paraphelenchus spp gonoducts A P acontioides femalepost-vulval uterine sac (Pvs) uterus (UT) spermatheca (Spt)fertilisation chamber (Fch) sphincter (Sph) and ovary (Ovar)B P intermedius female post-vulval uterine sac uterus sper-matheca fertilisation chamber oviduct and ovary with develo-ping oocyte from USDANC slide G11751 of G Thorne AstoriaOR USA 1941

junction Post-vulval uterine sac (Pvs) length variablesometimes reaching nearly half the vulval-anal distance(VAD) VADtail = 4-45 Lateral incisures beginning atlevel of median bulb increasing to eight at mid-body end-ing at mid-tail Rectum slightly shorter than tail lengthSmall pair of ventro-lateral papillae discernible between

890 Nematology


Table 1 Morphometrics of Paraphelenchus acontioides and P intermedius females All measurements are in μm and in the formmean plusmn sd (range)

Character P acontioides P intermediusPiney River CO South Dakota G Thorne n = 6

n = 20 Astoria OR W Courtney n = 4Combined populations n = 10

L 842 plusmn 118 (540-1059) 937 plusmn 84 (835-1110)a 294 plusmn 5 (226-374) 278 plusmn 28 (217-32)b 48 plusmn 06 (33-58) 53 plusmn 04 (45-59)c 231 plusmn 22 (161-277) 253 plusmn 17 (226-279)cprime 21 plusmn 03 (17-26) 23 plusmn 04 (16-27)V 75 plusmn 2 (69-81) 76 plusmn 1 (73-78)Max body diam 29 plusmn 6 (23-43) 34 plusmn 4 (28-40)Pharynx length 176 plusmn 9 (153-199) 177 plusmn 17 (153-216)Tail length 37 plusmn 5 (28-45) 37 plusmn 3 (31-44)Anal body diam 17 plusmn 1 (15-20) 17 plusmn 3 (15-20)Stylet length 17 plusmn 2 (13-20) 17 plusmn 1 (16-19)Vulva-anal distance (VAD) 164 plusmn 294 (119-224) 185 plusmn 175 (159-214)Anterior gonad 278 plusmn 32 (185-321) 456 plusmn 72 (364-571)Post-vulval uterine sac (Pvs) 55 plusmn 10 (23-65) 58 plusmn 11 (37-74)PvsVAD 35 plusmn 7 (17-55) 35 plusmn 4 (29-39)Pvsvulval body diam 2 plusmn 04 (09-25) 25 plusmn 11 (11-43)Rectum length 27 plusmn 3 (22-33) 34 plusmn 3 (31-37)Lateral incisures 8 6

ca 60-70 of tail length Dorsal paired papillae locatedjust anterior to ventral base of asymmetrical ventrally di-rected tail mucro also known as a ventral tuberculus (d vin Fig 1C) In mature gonoducts 8-10 globular columnsof two cells squared at their edges visible on oviductsphincter and eight columns of two thinner-walled cellscomprising the fertilisation chamber Uterus and sper-matheca about equal in length to fertilisation chamber andto oviduct sphincter length Egg (n = 30) dimensions =589 plusmn 58 (51-775) times 324 plusmn 66 (24-525) μm Smallestegg 51 times 30 μm largest 775 times 525 μm in size

HOST AND LOCALITY

Isolated from the lowest node of B tectorum (cheatgrass) from Piney River CO USA (3950prime2499primeprimeN10638prime2685primeprimeW) ca 30 miles south of Yampa CO USAand ca 15 miles south of County Road 1 on Highway 131

BIONOMICS

This Colorado population of P acontioides was lo-cated ca 150 miles northwest of Colorado Springs whichwas where P pseudoparietinus and P intermedius werecollected from native prairie (USDANC entries 28758

and 31067 deposited by Gerald Thorne 1966) Paraphe-lenchus intermedius could grow and reproduce at 30C infungal culture (Thorne amp Malek 1968)

PHYLOGENY

18S sequences

For 18S sequences a ClustalW alignment of 1195alignment positions was used for a Maximum Parsimonytree (TL = 175 CI = 0914 and HI = 0086) of which56 of positions were parsimony informative It wasalso used for a Maximum Likelihood tree bootstrapped100 times using RAxML with GTR matrix Gammamodel parameters were estimated by the RAxML programand there were 115 alignment patterns Slightly differentMP bootstrap values are indicated also on the ML tree(Fig 3) This tree shows Paraphelenchus basal to theclade of fairly cohesive Aphelenchus populations thatform a monophyletic group with high bootstrap supportThe distance matrix lsquoprsquo value between Aphelenchus andParaphelenchus (0042) also visualised with relativebranch lengths was greater than the value between themost divergent Aphelenchus populations (0011)

A Geneious nucleotide alignment of 1198 total align-ment positions for all included taxa including the P acon-

Vol 13(8) 2011 891

LK Carta et al

Fig 3 18S rDNA Maximum Likelihood Tree of Aphelenchidae implemented in RAxML Proportion of gaps and completely undeterminedcharacters in alignment 0038808 Model parameter alpha 0020014 Tree Length 0188690 Final ML Optimisation Likelihoodminus2582824816 Bootstrap values are listed for 100 ML replicates and 1000 MP replicates

tioides 1061 bp sequence within a 1075 bp segment of thetotal alignment (available on request) gave the same treetopology using MrBayes as the Clustal alignment Therewere 18 Paraphelenchus-specific apomorphic nucleotidesin this alignment (position ndash bp change) most of whichoccurred between positions 98 and 176 98 ndash A vs C 105 ndashT vs G 106 ndash C vs T 124 ndash T vs C A 159 ndash A vs T C166 and 175 ndash C vs T 176 ndash A vs G 286 ndash A vs C T 490ndash T vs C 501 ndash A vs T 510 ndash A vs G 793 ndash T vs G 934 ndashG insertion differences for Paraphelenchus sp only afterthe P acontioides sequence ran out included 1109 ndash A vsT 1150 ndash A vs G 1161 ndash A vs G T 1182 ndash T vs A Therewas only one minor A vs R (= A or G) difference at po-sition 861 for P acontioides compared to Paraphelenchussp

Outgroup Tylenchus and Ditylenchus 18S sequenceshad intermediate GC sequence content values of 492 and493 respectively Sequences for Aphelenchus avenaeisolates had 496-500 GC base content whilst the moredivergent P acontioides and Paraphelenchus sp had 481-485 GC

28S sequences

For 28S sequences a ClustalW alignment (available onrequest) of 745 alignment positions was used for a Max-imum Parsimony tree (TL = 626 CI = 0821 HI =0179) of which 32 of characters were parsimony infor-mative It was also used for a ML tree bootstrapped 100times using RAxML with GTR matrix Gamma modelparameters were estimated by the RAxML program and

there were 300 alignment patterns on the ML tree (Fig 4)All MP bootstrap values were 100 and not indicated onthe tree

The GC content for Coslenchus 28S sequences was513 for D destructor 544 for Ditylenchus sp468 for two A avenae 501 for Aphelenchus sp511 and for P acontioides 482

DIAGNOSIS

See Tables 1 2 Figure 5A B Morphometrics for thisColorado population of P acontioides are similar to P am-blyurus and P myceliophthorus except for the lower b ra-tio and longer stylet In a second paper by the authors ofP acontioides the b ratio was considerably lower whennematodes were grown on several different fungi (Pillai ampTaylor 1967a) The post-vulval uterine sac lengthvulvalanal distance (PvsVAD) was 39-47 vs 50 for P am-blyurus and 66 for P myceliophagus The Pvs lengthis similar for P acontioides (55 plusmn 10 23-65) and P inter-medius (58 plusmn 11 37-74 μm) The Pvsvulval body diam(Vbd) was also somewhat shorter in P acontioides com-pared to P intermedius (2 plusmn 04 09-25 vs 25 plusmn 11 11-43) (Table 1) and P pseudoparietinus (PvsVbd = 2-3 Andraacutessy 2007) The P intermedius oviduct had 8-12 cell columns and the fertilisation chamber had eightcolumns of cells (Fig 2B) whilst 8-12 cells were ob-served in the fertilisation chamber of P deckeri AlthoughP acontioides has eight lateral incisures it is otherwisesimilar to species with only six lines such as P inter-

892 Nematology


Fig 4 28S Maximum Likelihood Tree of Aphelenchidae implemented in RAxML Proportion of gaps and completely undeterminedcharacters in alignment 0288448 Model parameter alpha 0428791 Tree Length 1791170 Final ML Optimisation Likelihoodminus3800974955 Bootstrap values are listed for 100 ML replicates

medius and P amblyurus for which males are describedUnfortunately the lateral field and other features were es-pecially difficult to see in putative P pseudoparietinusfrom old specimens in the USDANC Paraphelenchus am-blyurus was described with what appeared at the time tobe a unique differentiation of the anterior median bulb(Steiner 1934) similar to the bulb in the Colorado popu-lation of P acontioides This median bulb differentiationof less striated tissue was also visible in P intermediusslides (Fig 5A) The tail of P intermedius was more at-tenuated toward the distal end (Fig 5B) than in P acon-tioides (Fig 1C)

Discussion

TAXONOMY

This population of P acontioides extended the reportedsize range with a longer body and stylet lower V andslightly lower b value The b values from the first descrip-tion were reduced substantially in a second paper describ-ing basic morphometrics on various fungal cultures (Pillaiamp Taylor 1967a) The b values of this Colorado popula-tion are more similar to those in the second paper vs theoriginal Therefore relying primarily on morphometricscould lead an identifier astray especially with species hav-ing few described populations or few replicates of a single

population As with any identification of a nematode out-side the type locality definitive assignment of this isolateto P acontioides is a hypothesis If molecular sequencesfrom this population are shown to be somewhat differentthan specimens from the type locality in the future this re-description could serve as a framework for the proposal ofa possible new cryptic or complementary species

Egg measurements extended the original range forlength and diam of P acontioides eggs ((64-74) times (28-32) μm) (Taylor amp Pillai 1967) at either end of the range((51-78) times (24-52) μm) Paraphelenchus intermedius hada similar sized egg (69times27 μm) to P acontioides whereasP myceliophthorus had larger (78times32 μm) and A avenaesomewhat smaller ((60-88) times (20-30) μm) eggs

Of the 23 Paraphelenchus species only eight lacka mucro and nine lack males Absence of males is some-times a result of the small number of specimens availablebut may also be related to temperature threshold Among39 populations of A avenae most were parthenogeneticand males were inducible in many populations only above30C (Ali et al 1999a) Both A avenae and P acon-tioides had minimum generation times of 5 and 6 days at35C (Pillai amp Taylor 1967b) Genetic diversity of A ave-nae populations did not correlate with geography fun-gal host thermal preference or presenceabsence of males(Ali et al 1999b) Paraphelenchus acontioides may bea complementary species (Osche 1954) of P pseudopari-

Vol 13(8) 2011 893

LK Carta et al

Tabl

e2

Para

phel

ench

usfe

mal

em

orph

omet

rics

Sp

ecie

sL

ab

cV

Styl

etgt

1m

ucro

cprimeM

ales

Li

Pvs

Vbd

Tail

muc

roP

acon

tioi

des

Ia71

0-88

025

-30

44-

53

20-3

073

-77

14-1

6ndash

24

ndash8

2P

acon

tioi

des

IIb

510-

960

159

-35

63

5-6

214

5-3

071

-80

ndashndash

ndash+

ndashndash

Pac

onti

oide

sC

O54

0-10

5923

-37

34-

58

16-3

37

69-7

83

13-2

3ndash

17-

24

(21

)+

81

7-2

3P

alii

790-

980

235

-29

65

2-6

514

7-2

22

73-7

613

6-1

52

ndash3

1+

61

8P

ambl

yuru

s54

0-93

023

-33

37-

59

17-2

476

17-1

9ndash

25

+6

21

Pba

sili

550-

590

234

-23

95-

53

156

-18

70-7

416

ndash3

2+

42

1P

cren

atus

680-

740

276

-30

84

8-5

119

2-2

37

746

-74

713

8-1

5ndash

34

+8

23

Pfid

icau

datu

s67

6-75

938

-39

31-

53

246

-27

475

-79

813

22

1ndash

82

8P

good

eyi

715-

766

209-

1018

-20

7412

ndash2

1ndash

63

2P

hete

roli

neat

us61

5-80

319

6-2

16

40-

48

171

-29

275

1-7

79

151

ndash2

ndash8

1P

inte

rmed

ius

700-

1000

24-2

84

3-5

623

-26

7615

-19

ndash2

4ndash

61

1-4

3P

myc

elio

phth

orus

580-

820

22-3

44

1-6

613

-24

71-7

86-

19ndash

14-

27

+6

32

Ppa

ram

onov

i58

7-93

718

9-2

64

4-5

518

1-2

52

756

-77

812

8-1

39

ndash1

5+

62

7P

porr

ectu

s70

4-77

030

8-3

12

48

207

-23

376

4-7

89

13ndash

18

+8

ndashP

sacc

hari

590-

880

30-3

95

1-6

620

-21

68-7

711

-16

ndash2

9+

43

Pps

eudo

pari

etin

us38

0-91

025

-36

(30)

41-

63

(5)

14-2

4(2

0)68

-78

(74)

13-1

4ndash

27

+9

2-3

Pzi

csii

970

365

714

7214

-15

24

3+

4+

23

2

No

tail

muc

roP

bata

vicu

s75

032

-37

7-8

2775

12-1

3ndash

22

+12

43

Pde

cker

i69

5-85

537

-39

52-

622

-25

75-7

814

5-1

75

ndash2

3-3

2ndash

10-1

22-

3P

obsc

urus

560-

630

26-4

24

2-5

220

-24

72-7

412

-13

ndash2

+5

36

Poc

toli

neat

us51

2-59

727

4-3

42

37-

46

171

-21

675

-78

913

-14

ndash2

9+

83

6P

orie

ntal

is66

6-71

225

-37

5-6

29-3

773

-77

12-1

3ndash

1ndash

83

5P

trit

ici

442-

851

268

-32

34-

59

167

-21

277

-78

12-1

3ndash

20

+ndash

2P

ussu

rien

sis

830-

841

302

-37

44

8-5

620

2-2

873

8-7

61

15ndash

24

ndash6

ndashP

zeae

658-

700

244

815

313

765

16-1

7ndash

20

ndashndash

44

Tail

muc

ropr

esen

tin

first

15sp

ecie

sndash

+ab

senc

epr

esen

ceof

mal

esL

ila

tera

linc

isur

esP

vsp

ost-

vulv

alut

erin

esa

cV

bdv

ulva

lbod

ydi

amM

easu

rem

ents

inth

efo

rmr

ange

and

or(m

ean)

a

From

Tayl

oramp

Pilla

i(19

67)

bFr

omPi

llaiamp

Tayl

or(1

967a

)

894 Nematology


Fig 5 Paraphelenchus intermedius A Median bulb B Tailfrom USDANC slide G11751 of G Thorne Astoria Oregon1941

etinus since P acontioides lacks males an indented lip re-gion and a ninth lateral incisure Otherwise these speciesgeographically overlap and have relatively unique plantstem habitats in grasslands

The molecular sequence gap between Paraphelenchusand Aphelenchus seems appropriate for these generawithin taxonomic subfamilies (Hunt 2008) rather than themore distant families of a previously taxonomic frame-work (Hunt 1993) Paraphelenchinae and its only genusParaphelenchus is distinguished from similarly mono-typic AphelenchinaeAphelenchus (Hunt 2008) by a pha-ryngeal bulb vs pharyngeal gland overlapping the intes-tine a vulval slit vs a pore male tail with 4-5 pairs ofpapillae but no bursa vs a bursa containing four papil-lae and lateral field with 4-9 lines vs lateral field withmore than six and typically 10-12 incisures (Hunt 1993)

The original description of A deckeri Zeidan amp Ger-aert 1991 mentioned transitional characters between thetwo families including a pharyngeal bulb and vulvalslit as in Paraphelenchus but 10-12 incisures in the lat-eral field and an oviduct sphincter (5-9 cell columns)slightly modified from that seen in a Paraphelenchus sp(6-10 columns although nine was most common) (Ger-aert 1981) This pattern was different from that drawnfor A avenae in another study showing eight fertilisationchamber cell columns + four globular columns of oviductcells (Triantaphyllou amp Fisher 1976) There was uncer-tainty about its generic identity in the description based onthis obscure and untested character since the oviduct wasrarely and inconsistently characterised in Paraphelenchusspp and the diagnosis was uncharacteristically madewith other Paraphelenchus species rather than with Aphe-lenchus species (Zeidan amp Geraert 1991) Hodda consid-ered this species a Paraphelenchus by generic definition(wwwentocsiroausciencenematodeschecklist 2003)a notion formalised by Andraacutessy (2007) and noted in a re-cent checklist of the group (Hunt 2008) One sugges-tion for resolution was to find or generate males (Hunt1993) Another alternative would be to update the range ofvariation of the oviduct in Paraphelenchus Considerablevariation has been reported or illustrated in the literature(Table 3) and many drawings lack clear regional limitsfor cell types The eight columns of fertilisation chambercells plus four columns of globular oviduct sphincter cellsposterior to the ovary in a population of A avenae (Tri-antaphyllou amp Fisher 1976) differed from the 5-9 oviductsphincter cell columns reported in another population ofA avenae (Geraert 1981) Unfortunately it was not pos-sible to count the number of cells in the obscure fertilisa-tion chamber of a Paraphelenchus sp in that comparativestudy (Geraert 1981) Most species of Paraphelenchushave six or more oviduct cell columns (Table 3) so the12 oviduct sphincter cells of P deckeri were consider-ably more numerous than those of either Paraphelenchusor any A avenae population Most drawings and descrip-tions are not definitively clear about the number of cells inthe fertilisation chamber especially so the values in the ta-ble should be considered estimates The relative length ofthe uterusspermatheca fertilisation chamber and oviductsphincter cell columns may be somewhat more usefulwhen characterising species In P acontioides these wereequivalent in length in three specimens but in P inter-medius the uterus and spermatheca extended to about halfthe total length of the three regions in two of three spec-imens whilst the fertilisation chamber and oviduct were

Vol 13(8) 2011 895

LK Carta et al

Table 3 Paraphelenchus spp and Aphelenchus avenae female reproductive system cells interpreted from published drawings orobservations

Species Fertilisation chamber Oviduct Referencecell number sphincter cell

number

P deckeri 7-8 10-12 Zeidan amp Geraert (1991)8-12 12 This study

P batavicus 10 10 Filipjev (1934)P acontioides 8 8-10 This studyP intermedius 8 8-10 This studyP orientalis 8a 8 Muchina (1988)P crenatus ndash 8 Das amp Singh (1968)P myceliophthorus ndash 6-8 Ryss amp Chernetskaya (2010) Goodey (1958)P alii (= P micoletzkyi) 8 6-8 Ali et al (1970)

8-16b 8 Zeidan amp Geraert (1991)P tritici 8 5-6 Baranovskaya (1958)P obscurus 20 4 Muchina (1988)P zeae ndash 4 Romaniko (1968)P octolineatus 8-10 2 Shavrov (1968)A avenae 8 4 Triantaphyllou amp Fisher (1976)

5-7 5-9 Geraert (1981)

a Ambiguous cell boundaries the number represents two small rounded transitional cells and six hatched cross-sectioned cellsb Paraphelenchus micoletzkyi was described with ldquooviduct composed of fertilisation chamber only with 16-18 packed glandular cellsrdquowithout a sphincter Instead we interpreted the drawing to include a clearly defined sphincter of eight cells and a fertilisation chamberthat probably had only eight cells The discrepancy is based on an ambiguous spermathecal boundary

about equal in length Both species had the same num-ber of cells in the fertilisation chamber and oviduct (Ta-ble 3) Between specimens of the same species the lengthof the fertilisation chamber was somewhat variable al-though the relatively narrow diameter cells could curveobliquely to the body axis and bunch together Also theoviductsphincter region had atypical cells with a transi-tional appearance at either end More complete descrip-tions and variation within species may yet show the util-ity of these differences It will be especially interesting todetermine where P deckeri resides within molecular phy-logenetic trees

Ecology

Paraphelenchus pseudoparietinus was found in swell-ings on the stem base of cocklebur (Xanthium sp andX americanum Walter = X strumarium L var glabratum(DC) Cronquist) in Fayetteville AR USA and Alexan-dria VA USA While this Colorado population was notassociated with stem swelling it represents one of onlya few reports of related nematodes within plant tissueAphelenchus avenae was originally isolated by Bastian

from oat leaf sheaths (Hunt 1993) and was observed feed-ing on plant roots (Decker 1962)

Fusarium solani (Mart) Sacc and F moniliforme wereexcellent hosts of P acontioides although F monili-forme was preferred to F solani even though the popu-lation was at least four times greater on F solani af-ter two generations (Pillai amp Taylor 1967a) Recently itwas demonstrated that F mangiferae Britz Wingfield ampMarasas spores were carried by the eriophyoid mite Ace-ria mangiferae Sayed in the epidemiology of mango mal-formation disease (Gamliel-Atinsky et al 2010) Addingto speculations on evolutionary trends in aphelench-hostassociations (Ryss 2007) it is conceivable that Paraphe-lenchus could also vector pathogenic Fusarium to cropplants as well The only demonstrated nematode vectorof a fungal pathogen is that of the seed-gall nematodeAnguina tritici which was reported to vector spores ofthe fungus Dilophosphora alopecuri to cereals (Atana-soff 1925) It is more common for migratory endopar-asitic nematodes within Pratylenchidae Anguinidae andAphelenchoididae to vector bacteria (Moens amp Perry2009) such as Anguina tritici carrying Rathayibacter(= Clavibacter) toxicus (Riley amp Ophel) Sasaki Chi-

896 Nematology


jimatsu amp Suzuki to wheat (Riley 1992) and othernematode-bacteria combinations (Dorofeeva et al 2002)However most nematode-microbial associations involvewounding or host predisposition rather than a strict vectorrelationship (Powell 1963)

Cheatgrass is an invasive species that is especiallycommon in western crops such as winter wheat and alfalfa(Young 2000) and like many weeds may be capableof harbouring various plant diseases Alternatively likeA avenae (Rhoades amp Linford 1959) Paraphelenchusmay be an opportunist fungivore scavenging fungi undera variety of conditions

We look forward to future research involving moredetailed SEM images of various Paraphelenchus speciesso as to reveal clearer profiles and variability of thelateral field in various species and of the presence ofother female tail papillae homologous to those in malesas reported in P myceliophthorus (Ryss amp Chernetskaya2010) More molecular sequences and descriptions of theoviduct in more species will undoubtedly improve thesystematic knowledge of this group

Acknowledgements

We thank Maria Hult Jennifer Kramer and David Mar-tel of the Nematology Laboratory for technical assistanceWe are grateful to David Chitwood of the NematologyLaboratory for bringing to our attention the history of thegeneric status and authorities for Paraphelenchus deckeriMention of trade names or commercial products in thispublication is solely for the purpose of providing specificinformation and does not imply recommendation or en-dorsement by the US Department of Agriculture

References

ALI MR AMIN B ADACHI T amp ISHIBASHI N (1999a)Host and temperature preference male occurrence and mor-phometrics of fungivorous nematode Aphelenchus avenaeisolates from Japan Japanese Journal of Nematology 29 7-17

ALI MR YAMAGUCHI Y amp ISHIBASHI N (1999b) RAPDand PCR-RFLP analysis on genetic diversity of Aphelenchusavenae isolates collected from Kyushu and some otherdistricts of Japan Japanese Journal of Nematology 29 24-34

ALI MS FAROOQUI NM amp SURYAWANSHI MV (1970)On a new species of Paraphelenchus (Micoletzky 1922) Mi-coletzky 1925 (Nematoda Paraphelenchidae) from Marath-wada India Rivista di Parassitologia 31 139-142

ANDRAacuteSSY I (1989) Six new nematode species from SouthAmerica Acta Zoologica Hungarica 35 1-16

ANDRAacuteSSY I (2007) Free-living nematodes of HungaryIII (Nematoda errantia) Pedozoologica Hungarica No 4Budapest Hungary Hungarian Natural History Museumamp Systematic Zoology Research Group of the HungarianAcademy of Sciences 496 pp

ATANASOFF D (1925) The Dilophospora disease of cerealsPhytopathology 15 1-40

BARANOVSKAYA IA (1958) [Contribution to the knowledgeof the genus Paraphelenchus (Micoletzky 1922) Micoletzky1925 (Nematoda Aphelenchidae)] Zoologichesky Zhurnal37 13-19

BARANOVSKAYA IA (1984) [Nematodes of the genus Para-phelenchus (Micoletzky 1922) Micoletzky 1925] In Turly-gina ES (Ed) Taksonomiya i biologiya fitogelrsquomintovMoscow USSR Nauka pp 5-35

DAS VM (1960) Studies on the nematode parasites ofplants in Hyderabad (Andhra Pradesh India) Zeitschrift fuumlrParasitenkunde 19 553-605

DE LEY P TANDINGAN DE LEY I MORRIS K ABEBEE MUNDO-OCAMPO M YODER M HERAS J WAU-MANN D ROCHA-OLIVARES A BURR AHJ et al(2005) An integrated approach to fast and informativemorphological vouchering of nematodes for applications inmolecular barcoding Philosophical Transactions of the RoyalSociety B 360 1945-1958

DECKER H (1962) Zur Biologie und Oumlkologie von Aphe-lenchus avenae Bastian Nematologica 7 9 [Abstr]

DOROFEEVA LV EVTUSHENKO LI KRAUSOVA VIKARPOV AV SUBBOTIN SA amp TIEDJE JM (2002)Rathayibacter caricis sp nov and Rathayibacter festucae spnov isolated from the phyllosphere of Carex sp and the leafgall induced by the nematode Anguina graminis on Festucarubra L respectively International Journal of Systematicand Evolutionary Microbiology 52 1917-1923

DRUMMOND AJ ASHTON B CHEUNG M HELED JKEARSE M MOIR R STONES-HAVAS S THIERERT amp WILSON A (2009) Geneious v 50 Available fromhttpwwwgeneiouscom

EROSHENKO AS (1966) [Three new species of Paraphe-lenchus (Micoletzky 1922) Micoletzky 1925 (NematodaAphelenchidae)] Zoologischesky Zhurnal 45 1873-1876

FILIPJEV IN (1934) [Harmful and useful nematodes in ruraleconomy] Moscow amp Leningrad USSR Ogiz 440 pp

FORTUNER R (1985) Notes on nomenclature of plant nema-todes Revue de Neacutematologie 8 77-83

GAMLIEL-ATINSKY E FREEMAN S MAYMON MOCHOA R BAUCHAN GR SKORACKA A PENA Jamp PALEVSKY E (2010) The role of eriophyoids in fungalpathogen epidemiology mere association or true interactionExperimental and Applied Acarology 51 191-204

Vol 13(8) 2011 897

LK Carta et al

GERAERT E (1981) The female reproductive system in nema-tode systematics Annales de la Socieacuteteacute Royale Zoologique deBelgique 110 73-86

GOODEY JB (1958) Paraphelenchus myceliophthorus n spNematologica 3 1-5

HAQUE MM (1967) [Description of the new species belong-ing to the genus Paraphelenchus Micol 1922 (NematodaParaphelenchidae)] Zoologichesky Zhurnal 46 1842-1846

HOLTERMAN M KARSSEN G VAN DEN ELSEN S VAN

MEGEN H BAKKER J amp HELDER J (2009) Smallsubunit rDNA-based phylogeny of the Tylenchida sheds lighton relationships among some high-impact plant-parasiticnematodes and the evolution of plant feeding Phytopathology99 227-235

HUNT DJ (1993) Aphelenchida Longidoridae and Tricho-doridae their systematics and bionomics Wallingford UKCABI Publishing 352 pp

HUNT DJ (2008) A checklist of the Aphelenchoidea (Nema-toda Tylenchida) Journal of Nematode Morphology and Sys-tematics 10 (2007) 99-135

HUSAIN SI amp KHAN AM (1967) On the status of thegenera of the superfamily Aphelenchoidea (Fuchs 1937)Thorne 1949 with descriptions of six new species from In-dia Proceedings of the Helminthological Society of Washing-ton 34 167-174

LARKIN MA BLACKSHIELDS G BROWN NPCHENNA R MCGETTIGAN PA MCWILLIAM HVALENTIN F WALLACE IM WILM A LOPEZR et al (2007) ClustalW and ClustalX version 20Bioinformatics 23 2947-2948

LUGINBUHL M amp MULLER E (1980) [Endophytic fungusin the above-ground organs of four plants growing together atthe same locations (Buxus Hedera Ilex Ruscus)] Sydowia33 185-209

MICOLETZKY H (1922) Die freilebenden Erd-Nematodenmit besonderer Berucksichtigung der Steiermark und derBukowina zugleich mit einer Revision samtlicher nichtmariner freilebender Nematoden in Form von Genusndash Beschreibunger un Bestimmungsschlusseln Archiv fuumlrNaturgeschlichte 87 1-650

MICOLETZKY H (1925) Die Freilebenden Susswasser- undMoornematoden Danemarks Nebst Anhang eber Amo-bosporidien und andere Parasiten bei freilebenden Nemato-den Kongelige Danske Videnskabernes Selskab Skrifter (8)10 57-310

MOENS M amp PERRY RN (2009) Migratory plant endopar-asitic nematodes a group rich in contrasts and divergenceAnnual Review of Phytopathology 47 313-332

MUCHINA TI (1988) [Two new species of nematodes (Para-phelenchidae) with a description of anomalies in their repro-ductive system] Zoologischesky Zhurnal 67 1240-1245

NUNN G (1992) Nematode molecular evolution An investiga-tion of evolutionary patterns among nematodes based upon

DNA sequences PhD Dissertation University of Notting-ham Nottingham UK

OSCHE G (1954) Uumlber dei gegenwaumlrtig ablaufende Entste-hung von Zwillings- und Komplementaumlrarten bei Rhabditi-den (Nematodes) (Foumltalisation und Artbildung) ZoologischeJahrbuumlcher Abteilung fuumlr Systematik Oumlkologie und Geogra-phie der Tiere 82 618-654

PILLAI JK amp TAYLOR DP (1967a) Influence of fungi onhost preference host suitability and morphometrics of fivemycophagous nematodes Nematologica 13 529-540

PILLAI JK amp TAYLOR DP (1967b) Effect of temperatureon the time required for hatching and duration of life cycle offive mycophagous nematodes Nematologica 13 512-516

POWELL NT (1963) The role of plant-parasitic nematodes infungus diseases Phytopathology 53 28-35

RHOADES HL amp LINFORD MB (1959) Control of Pythiumroot rot by the nematode Aphelenchus avenae Plant DiseaseReporter 43 323-328

RILEY IT (1992) Anguina tritici is a potential vector ofClavibacter toxicus Australasian Plant Pathology 21 147-149

ROMANIKO VI (1968) [A new nematode species of the genusParaphelenchus (Micoletzky 1922) Micoletzky 1925] TrudyBiologicheskii Chelyabinskaia Gosudarstvennogo Pedagog-ickej Institut 23 40-42

RYSS AYU (2007) [Main evolution lines of plant parasiticnematodes of the order Aphelenchida Siddiqi 1980] Parazi-tologiya 41 484-511

RYSS AYU amp CHERNETSKAYA AYU (2010) [Life cycleof Paraphelenchus myceliophthorus Goodey 1958 (Nema-toda Aphelenchida)] Parazitologiya 44 105-127

SCHULZ B WANKE U DRAEGER S amp AUST HJ (1993)Endophytes from herbaceous plants and shrubs effectivenessof surface sterilization methods Mycological Research 971447-1450

SHAVROV GN (1968) [New species of Paraphelenchus(Micoletzky 1922) Micoletzky 1925 (Nematoda Aphe-lenchinae)] Soobshcheniya dalrsquonevost Filial VL KomarovaSibirskoe Otdelenie Akademiya 26 135-136

STAMATAKIS A HOOVER P amp ROUGEMONT J (2008)A rapid bootstrap algorithm for the RAxML web-serversSystematic Biology 75 758-771

STEINER G (1934) Observations on nematodes parasitic in tu-bers of the cinnamon-vine Proceedings of the Helmintholog-ical Society of Washington 1 15-17

STEINER G (1941) Nematodes parasitic on and associatedwith roots of marigolds Tagetes hybrids Proceedings of theBiological Society of Washington 54 31-34

SWOFFORD DL (2002) PAUP phylogenetic analysis usingparsimony ( and other methods) Version 4 SunderlandMA USA Sinauer Associates

SWOFFORD DL OLSEN GJ WADDELL PJ ampHILLIS DM (1996) Phylogenetic inference In HillisDM Moritz C amp Mable BK (Eds) Molecular systemat-

898 Nematology


ics 2nd edition Sunderland MA USA Sinauer Associatespp 407-513

TANDON RS amp SINGH SP (1970) On two new nematodes(Aphelenchoidea) from tobacco roots in India Journal ofHelminthology 44 323-328

TAYLOR DP amp PILLAI JK (1967) Paraphelenchus acon-tioides n sp (Nematoda Paraphelenchidae) a mycophagousnematode from Illinois with observations on its feedinghabits and a key to the species of Paraphelenchus Proceed-ings of the Helminthological Society of Washington 34 51-54

THOMAS WK VIDA JT FRISSE LM MUNDO Mamp BALDWIN JG (1997) DNA sequences from formalinfixed nematodes integrating molecular and morphologicalapproaches to taxonomy Journal of Nematology 29 250-254

THORNE G amp MALEK RB (1968) Nematodes of the North-ern Great Plains Part I Tylenchida (Nemata Secernen-tea) Technical Bulletin 31 Agricultural Experiment StationSouth Dakota State University 111 pp

TRIANTAPHYLLOU AC amp FISHER JM (1976) Gemeto-genesis in amphimictic and parthenogenetic populations ofAphelenchus avenae Journal of Nematology 8 168-177

YE W GIBLIN-DAVIS RM DAVIES KA PURCELLMF SCHEFFER SJ TAYLOR GS CENTER TDMORRIS K amp THOMAS WK (2007) Molecular phy-logenetics and the evolution of host plant associations inthe nematode genus Fergusobia (Tylenchida Fergusobiinae)Molecular Phylogenetics and Evolution 45 123-141

YOUNG J (2000) Bromus tectorum L In Bossard CCRandall JM amp Hoshovsky MC (Eds) Invasive plants ofCaliforniarsquos wildlands Berkeley CA USA University ofCalifornia Press pp 76-80

ZEIDAN AB amp GERAERT E (1991) Aphelenchoides Aphe-lenchus and Paraphelenchus from Sudan with the descriptionof two new species Nematologica 37 420-438

Vol 13(8) 2011 899

LK Carta et al

etinus and P intermedius have been deposited into theUnited States Department of Agriculture Nematode Col-lection (USDANC) database One notable species is Pacontioides which was discovered in soil around Ken-tucky bluegrass (Agrostis palustris Huds) in Urbana Illi-nois The original population of this nematode specieswas initially raised for at least 5 years on the fungusPyrenochaeta terrestris (Hansen) Gorenz JC Walker ampLarson before actually being described (Taylor amp Pil-lai 1967) Subsequently morphometrics for this nema-tode were taken for material cultured on the same fungusand seven others resulting in expanded ranges (Pillai ampTaylor 1967a) especially for the b ratio

This paper reports the discovery of a Paraphelenchuspopulation in stems of cheatgrass Bromus tectorum Lin Colorado during a survey of endophytic fungi aspossible biocontrol agents within eight mid-western andwestern United States and British Columbia CanadaBecause many other Paraphelenchus species descriptionsare incomplete and not readily comparable detailedmorphometrics of the new population were compiled andcontrasted with other isolates of P acontioides and theother species of the genus to enhance the diagnosticinformation available

Materials and methods

NEMATODE AND FUNGUS ISOLATION

Sixty-three populations of cheatgrass B tectorumwere sampled throughout North America (Idaho Wash-ington Nevada Colorado New Mexico Iowa Illinoisand British Columbia) Populations were typically aboutfive miles apart but others were 1-20 or more milesapart Twenty plants from each population were collectedA 2 cm segment was clipped around the lowest culm node(solid region on the shoot central axis that may gener-ate a leaf sheath or adventitious bud) of each plant Seg-ments were surface-sterilised in 50 ethanol for 5 minand rinsed in sterile deionised water for 1 min (Lug-inbuhl amp Muller 1980 Schulz et al 1993) To facili-tate isolation of fungi culm segments from each popu-lation were placed onto potato dextrose agar plates Im-print plates were made to ensure surface sterilisationwas successful Fungi were identified morphologicallyCultures were placed on laboratory benches at ambientroom temperature and subcultured as needed Two grasspopulations had nematodes and endophytic Fusarium spand nematodes and fungi were not found independently

of each other The cheatgrass population with P acon-tioides was isolated from near Piney River CO USA(3950prime2499primeprimeN 10638prime2685primeprimeW) south of Yampa COUSA off Highway 131

NEMATODE PRESERVATION AND IMAGING

Nematodes were rinsed from fungal plates placed in4 formalin for 24 h rinsed in 15 ml plastic tubes withwater and sent to Beltsville MD USA From here theywere placed in 4 formalin for another 24 h dehydratedin alcohol and glycerin mounted in glycerin and sealedImages and measurements were made with a Zeiss Ul-traphot II microscope (Carl Zeiss Jena Germany andBaltimore Instrument Company Baltimore MD USA)equipped with differential interference contrast (DIC) op-tics Photographs were taken of formalin-fixed specimensrinsed in water Females were measured with an ocu-lar micrometer Fixed specimens used to evaluate rela-tionships were observed from the USDANC These in-cluded type and paratype slides of P intermedius fromthe Thorne collection (USDANC Entries 31046 2974428745 31049 slides 1a 1b (type five females) 1c 1ifor different South Dakota sites of G Thorne 28744)and P pseudoparietinus Micoletzky 1922 (USDANC En-try 21084 slide G11751 from soil of Chewings fescuegrass Festuca rubra commutate WB Courtney Asto-ria Oregon 1941) identified as P intermedius here basedon six lateral incisures and morphometrics and four fe-male paratypes of P deckeri (slide T-4299p from shrubsof Abuttaraz Sudan) Type material from the old Nema-tode Slide Collection of the Department of Plant Pathol-ogy (now Crop Sciences) University of Illinois UrbanaIL USA and P amblyurus from the Steiner collection ofthe USDANC were unavailable Reproductive system ter-minology of Triantaphyllou and Fisher (1976) as used forA avenae was applied to a single gonad arm going in ananterior direction vulva vagina gonoduct (uterus sper-matheca fertilisation chamber (eight columns of two rowsof tightly packed oval cells not contained in a membrane)sphincter ndash (three rows of globular cells not contained ina membrane)) and ovary The oviduct comprises a fer-tilisation chamber and sphincter cells (Triantaphyllou ampFisher 1976) but the term has also been used to describethe sphincter cells only (Geraert 1981)

PCR AND SEQUENCING

Nematodes were rinsed from fungal plates placed in70 alcohol in 15 ml plastic tubes and sent to Beltsville

888 Nematology







Description


MEASUREMENTS

See Table 1

DESCRIPTION


Vol 13(8) 2011 889

LK Carta et al





890 Nematology







HOST AND LOCALITY


BIONOMICS



PHYLOGENY

18S sequences



Vol 13(8) 2011 891

LK Carta et al




28S sequences




DIAGNOSIS


892 Nematology




Discussion

TAXONOMY





Vol 13(8) 2011 893

LK Carta et al

Tabl

e2

Para

phel

ench

usfe

mal

em

orph

omet

rics

Sp

ecie

sL

ab

cV

Styl

etgt

1m

ucro

cprimeM

ales

Li

Pvs

Vbd

Tail

muc

roP

acon

tioi

des

Ia71

0-88

025

-30

44-

53

20-3

073

-77

14-1

6ndash

24

ndash8

2P

acon

tioi

des

IIb

510-

960

159

-35

63

5-6

214

5-3

071

-80

ndashndash

ndash+

ndashndash

Pac

onti

oide

sC

O54

0-10

5923

-37

34-

58

16-3

37

69-7

83

13-2

3ndash

17-

24

(21

)+

81

7-2

3P

alii

790-

980

235

-29

65

2-6

514

7-2

22

73-7

613

6-1

52

ndash3

1+

61

8P

ambl

yuru

s54

0-93

023

-33

37-

59

17-2

476

17-1

9ndash

25

+6

21

Pba

sili

550-

590

234

-23

95-

53

156

-18

70-7

416

ndash3

2+

42

1P

cren

atus

680-

740

276

-30

84

8-5

119

2-2

37

746

-74

713

8-1

5ndash

34

+8

23

Pfid

icau

datu

s67

6-75

938

-39

31-

53

246

-27

475

-79

813

22

1ndash

82

8P

good

eyi

715-

766

209-

1018

-20

7412

ndash2

1ndash

63

2P

hete

roli

neat

us61

5-80

319

6-2

16

40-

48

171

-29

275

1-7

79

151

ndash2

ndash8

1P

inte

rmed

ius

700-

1000

24-2

84

3-5

623

-26

7615

-19

ndash2

4ndash

61

1-4

3P

myc

elio

phth

orus

580-

820

22-3

44

1-6

613

-24

71-7

86-

19ndash

14-

27

+6

32

Ppa

ram

onov

i58

7-93

718

9-2

64

4-5

518

1-2

52

756

-77

812

8-1

39

ndash1

5+

62

7P

porr

ectu

s70

4-77

030

8-3

12

48

207

-23

376

4-7

89

13ndash

18

+8

ndashP

sacc

hari

590-

880

30-3

95

1-6

620

-21

68-7

711

-16

ndash2

9+

43

Pps

eudo

pari

etin

us38

0-91

025

-36

(30)

41-

63

(5)

14-2

4(2

0)68

-78

(74)

13-1

4ndash

27

+9

2-3

Pzi

csii

970

365

714

7214

-15

24

3+

4+

23

2

No

tail

muc

roP

bata

vicu

s75

032

-37

7-8

2775

12-1

3ndash

22

+12

43

Pde

cker

i69

5-85

537

-39

52-

622

-25

75-7

814

5-1

75

ndash2

3-3

2ndash

10-1

22-

3P

obsc

urus

560-

630

26-4

24

2-5

220

-24

72-7

412

-13

ndash2

+5

36

Poc

toli

neat

us51

2-59

727

4-3

42

37-

46

171

-21

675

-78

913

-14

ndash2

9+

83

6P

orie

ntal

is66

6-71

225

-37

5-6

29-3

773

-77

12-1

3ndash

1ndash

83

5P

trit

ici

442-

851

268

-32

34-

59

167

-21

277

-78

12-1

3ndash

20

+ndash

2P

ussu

rien

sis

830-

841

302

-37

44

8-5

620

2-2

873

8-7

61

15ndash

24

ndash6

ndashP

zeae

658-

700

244

815

313

765

16-1

7ndash

20

ndashndash

44

Tail

muc

ropr

esen

tin

first

15sp

ecie

sndash

+ab

senc

epr

esen

ceof

mal

esL

ila

tera

linc

isur

esP

vsp

ost-

vulv

alut

erin

esa

cV

bdv

ulva

lbod

ydi

amM

easu

rem

ents

inth

efo

rmr

ange

and

or(m

ean)

a

From

Tayl

oramp

Pilla

i(19

67)

bFr

omPi

llaiamp

Tayl

or(1

967a

)

894 Nematology






Vol 13(8) 2011 895

LK Carta et al



number




5-7 5-9 Geraert (1981)



Ecology




896 Nematology





Acknowledgements


References


















Vol 13(8) 2011 897

LK Carta et al

































898 Nematology











Vol 13(8) 2011 899







Description


MEASUREMENTS

See Table 1

DESCRIPTION


Vol 13(8) 2011 889

LK Carta et al





890 Nematology







HOST AND LOCALITY


BIONOMICS



PHYLOGENY

18S sequences



Vol 13(8) 2011 891

LK Carta et al




28S sequences




DIAGNOSIS


892 Nematology




Discussion

TAXONOMY





Vol 13(8) 2011 893

LK Carta et al

Tabl

e2

Para

phel

ench

usfe

mal

em

orph

omet

rics

Sp

ecie

sL

ab

cV

Styl

etgt

1m

ucro

cprimeM

ales

Li

Pvs

Vbd

Tail

muc

roP

acon

tioi

des

Ia71

0-88

025

-30

44-

53

20-3

073

-77

14-1

6ndash

24

ndash8

2P

acon

tioi

des

IIb

510-

960

159

-35

63

5-6

214

5-3

071

-80

ndashndash

ndash+

ndashndash

Pac

onti

oide

sC

O54

0-10

5923

-37

34-

58

16-3

37

69-7

83

13-2

3ndash

17-

24

(21

)+

81

7-2

3P

alii

790-

980

235

-29

65

2-6

514

7-2

22

73-7

613

6-1

52

ndash3

1+

61

8P

ambl

yuru

s54

0-93

023

-33

37-

59

17-2

476

17-1

9ndash

25

+6

21

Pba

sili

550-

590

234

-23

95-

53

156

-18

70-7

416

ndash3

2+

42

1P

cren

atus

680-

740

276

-30

84

8-5

119

2-2

37

746

-74

713

8-1

5ndash

34

+8

23

Pfid

icau

datu

s67

6-75

938

-39

31-

53

246

-27

475

-79

813

22

1ndash

82

8P

good

eyi

715-

766

209-

1018

-20

7412

ndash2

1ndash

63

2P

hete

roli

neat

us61

5-80

319

6-2

16

40-

48

171

-29

275

1-7

79

151

ndash2

ndash8

1P

inte

rmed

ius

700-

1000

24-2

84

3-5

623

-26

7615

-19

ndash2

4ndash

61

1-4

3P

myc

elio

phth

orus

580-

820

22-3

44

1-6

613

-24

71-7

86-

19ndash

14-

27

+6

32

Ppa

ram

onov

i58

7-93

718

9-2

64

4-5

518

1-2

52

756

-77

812

8-1

39

ndash1

5+

62

7P

porr

ectu

s70

4-77

030

8-3

12

48

207

-23

376

4-7

89

13ndash

18

+8

ndashP

sacc

hari

590-

880

30-3

95

1-6

620

-21

68-7

711

-16

ndash2

9+

43

Pps

eudo

pari

etin

us38

0-91

025

-36

(30)

41-

63

(5)

14-2

4(2

0)68

-78

(74)

13-1

4ndash

27

+9

2-3

Pzi

csii

970

365

714

7214

-15

24

3+

4+

23

2

No

tail

muc

roP

bata

vicu

s75

032

-37

7-8

2775

12-1

3ndash

22

+12

43

Pde

cker

i69

5-85

537

-39

52-

622

-25

75-7

814

5-1

75

ndash2

3-3

2ndash

10-1

22-

3P

obsc

urus

560-

630

26-4

24

2-5

220

-24

72-7

412

-13

ndash2

+5

36

Poc

toli

neat

us51

2-59

727

4-3

42

37-

46

171

-21

675

-78

913

-14

ndash2

9+

83

6P

orie

ntal

is66

6-71

225

-37

5-6

29-3

773

-77

12-1

3ndash

1ndash

83

5P

trit

ici

442-

851

268

-32

34-

59

167

-21

277

-78

12-1

3ndash

20

+ndash

2P

ussu

rien

sis

830-

841

302

-37

44

8-5

620

2-2

873

8-7

61

15ndash

24

ndash6

ndashP

zeae

658-

700

244

815

313

765

16-1

7ndash

20

ndashndash

44

Tail

muc

ropr

esen

tin

first

15sp

ecie

sndash

+ab

senc

epr

esen

ceof

mal

esL

ila

tera

linc

isur

esP

vsp

ost-

vulv

alut

erin

esa

cV

bdv

ulva

lbod

ydi

amM

easu

rem

ents

inth

efo

rmr

ange

and

or(m

ean)

a

From

Tayl

oramp

Pilla

i(19

67)

bFr

omPi

llaiamp

Tayl

or(1

967a

)

894 Nematology






Vol 13(8) 2011 895

LK Carta et al



number




5-7 5-9 Geraert (1981)



Ecology




896 Nematology





Acknowledgements


References


















Vol 13(8) 2011 897

LK Carta et al

































898 Nematology











Vol 13(8) 2011 899

LK Carta et al





890 Nematology







HOST AND LOCALITY


BIONOMICS



PHYLOGENY

18S sequences



Vol 13(8) 2011 891

LK Carta et al




28S sequences




DIAGNOSIS


892 Nematology




Discussion

TAXONOMY





Vol 13(8) 2011 893

LK Carta et al

Tabl

e2

Para

phel

ench

usfe

mal

em

orph

omet

rics

Sp

ecie

sL

ab

cV

Styl

etgt

1m

ucro

cprimeM

ales

Li

Pvs

Vbd

Tail

muc

roP

acon

tioi

des

Ia71

0-88

025

-30

44-

53

20-3

073

-77

14-1

6ndash

24

ndash8

2P

acon

tioi

des

IIb

510-

960

159

-35

63

5-6

214

5-3

071

-80

ndashndash

ndash+

ndashndash

Pac

onti

oide

sC

O54

0-10

5923

-37

34-

58

16-3

37

69-7

83

13-2

3ndash

17-

24

(21

)+

81

7-2

3P

alii

790-

980

235

-29

65

2-6

514

7-2

22

73-7

613

6-1

52

ndash3

1+

61

8P

ambl

yuru

s54

0-93

023

-33

37-

59

17-2

476

17-1

9ndash

25

+6

21

Pba

sili

550-

590

234

-23

95-

53

156

-18

70-7

416

ndash3

2+

42

1P

cren

atus

680-

740

276

-30

84

8-5

119

2-2

37

746

-74

713

8-1

5ndash

34

+8

23

Pfid

icau

datu

s67

6-75

938

-39

31-

53

246

-27

475

-79

813

22

1ndash

82

8P

good

eyi

715-

766

209-

1018

-20

7412

ndash2

1ndash

63

2P

hete

roli

neat

us61

5-80

319

6-2

16

40-

48

171

-29

275

1-7

79

151

ndash2

ndash8

1P

inte

rmed

ius

700-

1000

24-2

84

3-5

623

-26

7615

-19

ndash2

4ndash

61

1-4

3P

myc

elio

phth

orus

580-

820

22-3

44

1-6

613

-24

71-7

86-

19ndash

14-

27

+6

32

Ppa

ram

onov

i58

7-93

718

9-2

64

4-5

518

1-2

52

756

-77

812

8-1

39

ndash1

5+

62

7P

porr

ectu

s70

4-77

030

8-3

12

48

207

-23

376

4-7

89

13ndash

18

+8

ndashP

sacc

hari

590-

880

30-3

95

1-6

620

-21

68-7

711

-16

ndash2

9+

43

Pps

eudo

pari

etin

us38

0-91

025

-36

(30)

41-

63

(5)

14-2

4(2

0)68

-78

(74)

13-1

4ndash

27

+9

2-3

Pzi

csii

970

365

714

7214

-15

24

3+

4+

23

2

No

tail

muc

roP

bata

vicu

s75

032

-37

7-8

2775

12-1

3ndash

22

+12

43

Pde

cker

i69

5-85

537

-39

52-

622

-25

75-7

814

5-1

75

ndash2

3-3

2ndash

10-1

22-

3P

obsc

urus

560-

630

26-4

24

2-5

220

-24

72-7

412

-13

ndash2

+5

36

Poc

toli

neat

us51

2-59

727

4-3

42

37-

46

171

-21

675

-78

913

-14

ndash2

9+

83

6P

orie

ntal

is66

6-71

225

-37

5-6

29-3

773

-77

12-1

3ndash

1ndash

83

5P

trit

ici

442-

851

268

-32

34-

59

167

-21

277

-78

12-1

3ndash

20

+ndash

2P

ussu

rien

sis

830-

841

302

-37

44

8-5

620

2-2

873

8-7

61

15ndash

24

ndash6

ndashP

zeae

658-

700

244

815

313

765

16-1

7ndash

20

ndashndash

44

Tail

muc

ropr

esen

tin

first

15sp

ecie

sndash

+ab

senc

epr

esen

ceof

mal

esL

ila

tera

linc

isur

esP

vsp

ost-

vulv

alut

erin

esa

cV

bdv

ulva

lbod

ydi

amM

easu

rem

ents

inth

efo

rmr

ange

and

or(m

ean)

a

From

Tayl

oramp

Pilla

i(19

67)

bFr

omPi

llaiamp

Tayl

or(1

967a

)

894 Nematology






Vol 13(8) 2011 895

LK Carta et al



number




5-7 5-9 Geraert (1981)



Ecology




896 Nematology





Acknowledgements


References


















Vol 13(8) 2011 897

LK Carta et al

































898 Nematology











Vol 13(8) 2011 899







HOST AND LOCALITY


BIONOMICS



PHYLOGENY

18S sequences



Vol 13(8) 2011 891

LK Carta et al




28S sequences




DIAGNOSIS


892 Nematology




Discussion

TAXONOMY





Vol 13(8) 2011 893

LK Carta et al

Tabl

e2

Para

phel

ench

usfe

mal

em

orph

omet

rics

Sp

ecie

sL

ab

cV

Styl

etgt

1m

ucro

cprimeM

ales

Li

Pvs

Vbd

Tail

muc

roP

acon

tioi

des

Ia71

0-88

025

-30

44-

53

20-3

073

-77

14-1

6ndash

24

ndash8

2P

acon

tioi

des

IIb

510-

960

159

-35

63

5-6

214

5-3

071

-80

ndashndash

ndash+

ndashndash

Pac

onti

oide

sC

O54

0-10

5923

-37

34-

58

16-3

37

69-7

83

13-2

3ndash

17-

24

(21

)+

81

7-2

3P

alii

790-

980

235

-29

65

2-6

514

7-2

22

73-7

613

6-1

52

ndash3

1+

61

8P

ambl

yuru

s54

0-93

023

-33

37-

59

17-2

476

17-1

9ndash

25

+6

21

Pba

sili

550-

590

234

-23

95-

53

156

-18

70-7

416

ndash3

2+

42

1P

cren

atus

680-

740

276

-30

84

8-5

119

2-2

37

746

-74

713

8-1

5ndash

34

+8

23

Pfid

icau

datu

s67

6-75

938

-39

31-

53

246

-27

475

-79

813

22

1ndash

82

8P

good

eyi

715-

766

209-

1018

-20

7412

ndash2

1ndash

63

2P

hete

roli

neat

us61

5-80

319

6-2

16

40-

48

171

-29

275

1-7

79

151

ndash2

ndash8

1P

inte

rmed

ius

700-

1000

24-2

84

3-5

623

-26

7615

-19

ndash2

4ndash

61

1-4

3P

myc

elio

phth

orus

580-

820

22-3

44

1-6

613

-24

71-7

86-

19ndash

14-

27

+6

32

Ppa

ram

onov

i58

7-93

718

9-2

64

4-5

518

1-2

52

756

-77

812

8-1

39

ndash1

5+

62

7P

porr

ectu

s70

4-77

030

8-3

12

48

207

-23

376

4-7

89

13ndash

18

+8

ndashP

sacc

hari

590-

880

30-3

95

1-6

620

-21

68-7

711

-16

ndash2

9+

43

Pps

eudo

pari

etin

us38

0-91

025

-36

(30)

41-

63

(5)

14-2

4(2

0)68

-78

(74)

13-1

4ndash

27

+9

2-3

Pzi

csii

970

365

714

7214

-15

24

3+

4+

23

2

No

tail

muc

roP

bata

vicu

s75

032

-37

7-8

2775

12-1

3ndash

22

+12

43

Pde

cker

i69

5-85

537

-39

52-

622

-25

75-7

814

5-1

75

ndash2

3-3

2ndash

10-1

22-

3P

obsc

urus

560-

630

26-4

24

2-5

220

-24

72-7

412

-13

ndash2

+5

36

Poc

toli

neat

us51

2-59

727

4-3

42

37-

46

171

-21

675

-78

913

-14

ndash2

9+

83

6P

orie

ntal

is66

6-71

225

-37

5-6

29-3

773

-77

12-1

3ndash

1ndash

83

5P

trit

ici

442-

851

268

-32

34-

59

167

-21

277

-78

12-1

3ndash

20

+ndash

2P

ussu

rien

sis

830-

841

302

-37

44

8-5

620

2-2

873

8-7

61

15ndash

24

ndash6

ndashP

zeae

658-

700

244

815

313

765

16-1

7ndash

20

ndashndash

44

Tail

muc

ropr

esen

tin

first

15sp

ecie

sndash

+ab

senc

epr

esen

ceof

mal

esL

ila

tera

linc

isur

esP

vsp

ost-

vulv

alut

erin

esa

cV

bdv

ulva

lbod

ydi

amM

easu

rem

ents

inth

efo

rmr

ange

and

or(m

ean)

a

From

Tayl

oramp

Pilla

i(19

67)

bFr

omPi

llaiamp

Tayl

or(1

967a

)

894 Nematology






Vol 13(8) 2011 895

LK Carta et al



number




5-7 5-9 Geraert (1981)



Ecology




896 Nematology





Acknowledgements


References


















Vol 13(8) 2011 897

LK Carta et al

































898 Nematology











Vol 13(8) 2011 899

LK Carta et al




28S sequences




DIAGNOSIS


892 Nematology




Discussion

TAXONOMY





Vol 13(8) 2011 893

LK Carta et al

Tabl

e2

Para

phel

ench

usfe

mal

em

orph

omet

rics

Sp

ecie

sL

ab

cV

Styl

etgt

1m

ucro

cprimeM

ales

Li

Pvs

Vbd

Tail

muc

roP

acon

tioi

des

Ia71

0-88

025

-30

44-

53

20-3

073

-77

14-1

6ndash

24

ndash8

2P

acon

tioi

des

IIb

510-

960

159

-35

63

5-6

214

5-3

071

-80

ndashndash

ndash+

ndashndash

Pac

onti

oide

sC

O54

0-10

5923

-37

34-

58

16-3

37

69-7

83

13-2

3ndash

17-

24

(21

)+

81

7-2

3P

alii

790-

980

235

-29

65

2-6

514

7-2

22

73-7

613

6-1

52

ndash3

1+

61

8P

ambl

yuru

s54

0-93

023

-33

37-

59

17-2

476

17-1

9ndash

25

+6

21

Pba

sili

550-

590

234

-23

95-

53

156

-18

70-7

416

ndash3

2+

42

1P

cren

atus

680-

740

276

-30

84

8-5

119

2-2

37

746

-74

713

8-1

5ndash

34

+8

23

Pfid

icau

datu

s67

6-75

938

-39

31-

53

246

-27

475

-79

813

22

1ndash

82

8P

good

eyi

715-

766

209-

1018

-20

7412

ndash2

1ndash

63

2P

hete

roli

neat

us61

5-80

319

6-2

16

40-

48

171

-29

275

1-7

79

151

ndash2

ndash8

1P

inte

rmed

ius

700-

1000

24-2

84

3-5

623

-26

7615

-19

ndash2

4ndash

61

1-4

3P

myc

elio

phth

orus

580-

820

22-3

44

1-6

613

-24

71-7

86-

19ndash

14-

27

+6

32

Ppa

ram

onov

i58

7-93

718

9-2

64

4-5

518

1-2

52

756

-77

812

8-1

39

ndash1

5+

62

7P

porr

ectu

s70

4-77

030

8-3

12

48

207

-23

376

4-7

89

13ndash

18

+8

ndashP

sacc

hari

590-

880

30-3

95

1-6

620

-21

68-7

711

-16

ndash2

9+

43

Pps

eudo

pari

etin

us38

0-91

025

-36

(30)

41-

63

(5)

14-2

4(2

0)68

-78

(74)

13-1

4ndash

27

+9

2-3

Pzi

csii

970

365

714

7214

-15

24

3+

4+

23

2

No

tail

muc

roP

bata

vicu

s75

032

-37

7-8

2775

12-1

3ndash

22

+12

43

Pde

cker

i69

5-85

537

-39

52-

622

-25

75-7

814

5-1

75

ndash2

3-3

2ndash

10-1

22-

3P

obsc

urus

560-

630

26-4

24

2-5

220

-24

72-7

412

-13

ndash2

+5

36

Poc

toli

neat

us51

2-59

727

4-3

42

37-

46

171

-21

675

-78

913

-14

ndash2

9+

83

6P

orie

ntal

is66

6-71

225

-37

5-6

29-3

773

-77

12-1

3ndash

1ndash

83

5P

trit

ici

442-

851

268

-32

34-

59

167

-21

277

-78

12-1

3ndash

20

+ndash

2P

ussu

rien

sis

830-

841

302

-37

44

8-5

620

2-2

873

8-7

61

15ndash

24

ndash6

ndashP

zeae

658-

700

244

815

313

765

16-1

7ndash

20

ndashndash

44

Tail

muc

ropr

esen

tin

first

15sp

ecie

sndash

+ab

senc

epr

esen

ceof

mal

esL

ila

tera

linc

isur

esP

vsp

ost-

vulv

alut

erin

esa

cV

bdv

ulva

lbod

ydi

amM

easu

rem

ents

inth

efo

rmr

ange

and

or(m

ean)

a

From

Tayl

oramp

Pilla

i(19

67)

bFr

omPi

llaiamp

Tayl

or(1

967a

)

894 Nematology






Vol 13(8) 2011 895

LK Carta et al



number




5-7 5-9 Geraert (1981)



Ecology




896 Nematology





Acknowledgements


References


















Vol 13(8) 2011 897

LK Carta et al

































898 Nematology











Vol 13(8) 2011 899




Discussion

TAXONOMY





Vol 13(8) 2011 893

LK Carta et al

Tabl

e2

Para

phel

ench

usfe

mal

em

orph

omet

rics

Sp

ecie

sL

ab

cV

Styl

etgt

1m

ucro

cprimeM

ales

Li

Pvs

Vbd

Tail

muc

roP

acon

tioi

des

Ia71

0-88

025

-30

44-

53

20-3

073

-77

14-1

6ndash

24

ndash8

2P

acon

tioi

des

IIb

510-

960

159

-35

63

5-6

214

5-3

071

-80

ndashndash

ndash+

ndashndash

Pac

onti

oide

sC

O54

0-10

5923

-37

34-

58

16-3

37

69-7

83

13-2

3ndash

17-

24

(21

)+

81

7-2

3P

alii

790-

980

235

-29

65

2-6

514

7-2

22

73-7

613

6-1

52

ndash3

1+

61

8P

ambl

yuru

s54

0-93

023

-33

37-

59

17-2

476

17-1

9ndash

25

+6

21

Pba

sili

550-

590

234

-23

95-

53

156

-18

70-7

416

ndash3

2+

42

1P

cren

atus

680-

740

276

-30

84

8-5

119

2-2

37

746

-74

713

8-1

5ndash

34

+8

23

Pfid

icau

datu

s67

6-75

938

-39

31-

53

246

-27

475

-79

813

22

1ndash

82

8P

good

eyi

715-

766

209-

1018

-20

7412

ndash2

1ndash

63

2P

hete

roli

neat

us61

5-80

319

6-2

16

40-

48

171

-29

275

1-7

79

151

ndash2

ndash8

1P

inte

rmed

ius

700-

1000

24-2

84

3-5

623

-26

7615

-19

ndash2

4ndash

61

1-4

3P

myc

elio

phth

orus

580-

820

22-3

44

1-6

613

-24

71-7

86-

19ndash

14-

27

+6

32

Ppa

ram

onov

i58

7-93

718

9-2

64

4-5

518

1-2

52

756

-77

812

8-1

39

ndash1

5+

62

7P

porr

ectu

s70

4-77

030

8-3

12

48

207

-23

376

4-7

89

13ndash

18

+8

ndashP

sacc

hari

590-

880

30-3

95

1-6

620

-21

68-7

711

-16

ndash2

9+

43

Pps

eudo

pari

etin

us38

0-91

025

-36

(30)

41-

63

(5)

14-2

4(2

0)68

-78

(74)

13-1

4ndash

27

+9

2-3

Pzi

csii

970

365

714

7214

-15

24

3+

4+

23

2

No

tail

muc

roP

bata

vicu

s75

032

-37

7-8

2775

12-1

3ndash

22

+12

43

Pde

cker

i69

5-85

537

-39

52-

622

-25

75-7

814

5-1

75

ndash2

3-3

2ndash

10-1

22-

3P

obsc

urus

560-

630

26-4

24

2-5

220

-24

72-7

412

-13

ndash2

+5

36

Poc

toli

neat

us51

2-59

727

4-3

42

37-

46

171

-21

675

-78

913

-14

ndash2

9+

83

6P

orie

ntal

is66

6-71

225

-37

5-6

29-3

773

-77

12-1

3ndash

1ndash

83

5P

trit

ici

442-

851

268

-32

34-

59

167

-21

277

-78

12-1

3ndash

20

+ndash

2P

ussu

rien

sis

830-

841

302

-37

44

8-5

620

2-2

873

8-7

61

15ndash

24

ndash6

ndashP

zeae

658-

700

244

815

313

765

16-1

7ndash

20

ndashndash

44

Tail

muc

ropr

esen

tin

first

15sp

ecie

sndash

+ab

senc

epr

esen

ceof

mal

esL

ila

tera

linc

isur

esP

vsp

ost-

vulv

alut

erin

esa

cV

bdv

ulva

lbod

ydi

amM

easu

rem

ents

inth

efo

rmr

ange

and

or(m

ean)

a

From

Tayl

oramp

Pilla

i(19

67)

bFr

omPi

llaiamp

Tayl

or(1

967a

)

894 Nematology






Vol 13(8) 2011 895

LK Carta et al



number




5-7 5-9 Geraert (1981)



Ecology




896 Nematology





Acknowledgements


References


















Vol 13(8) 2011 897

LK Carta et al

































898 Nematology











Vol 13(8) 2011 899

LK Carta et al

Tabl

e2

Para

phel

ench

usfe

mal

em

orph

omet

rics

Sp

ecie

sL

ab

cV

Styl

etgt

1m

ucro

cprimeM

ales

Li

Pvs

Vbd

Tail

muc

roP

acon

tioi

des

Ia71

0-88

025

-30

44-

53

20-3

073

-77

14-1

6ndash

24

ndash8

2P

acon

tioi

des

IIb

510-

960

159

-35

63

5-6

214

5-3

071

-80

ndashndash

ndash+

ndashndash

Pac

onti

oide

sC

O54

0-10

5923

-37

34-

58

16-3

37

69-7

83

13-2

3ndash

17-

24

(21

)+

81

7-2

3P

alii

790-

980

235

-29

65

2-6

514

7-2

22

73-7

613

6-1

52

ndash3

1+

61

8P

ambl

yuru

s54

0-93

023

-33

37-

59

17-2

476

17-1

9ndash

25

+6

21

Pba

sili

550-

590

234

-23

95-

53

156

-18

70-7

416

ndash3

2+

42

1P

cren

atus

680-

740

276

-30

84

8-5

119

2-2

37

746

-74

713

8-1

5ndash

34

+8

23

Pfid

icau

datu

s67

6-75

938

-39

31-

53

246

-27

475

-79

813

22

1ndash

82

8P

good

eyi

715-

766

209-

1018

-20

7412

ndash2

1ndash

63

2P

hete

roli

neat

us61

5-80

319

6-2

16

40-

48

171

-29

275

1-7

79

151

ndash2

ndash8

1P

inte

rmed

ius

700-

1000

24-2

84

3-5

623

-26

7615

-19

ndash2

4ndash

61

1-4

3P

myc

elio

phth

orus

580-

820

22-3

44

1-6

613

-24

71-7

86-

19ndash

14-

27

+6

32

Ppa

ram

onov

i58

7-93

718

9-2

64

4-5

518

1-2

52

756

-77

812

8-1

39

ndash1

5+

62

7P

porr

ectu

s70

4-77

030

8-3

12

48

207

-23

376

4-7

89

13ndash

18

+8

ndashP

sacc

hari

590-

880

30-3

95

1-6

620

-21

68-7

711

-16

ndash2

9+

43

Pps

eudo

pari

etin

us38

0-91

025

-36

(30)

41-

63

(5)

14-2

4(2

0)68

-78

(74)

13-1

4ndash

27

+9

2-3

Pzi

csii

970

365

714

7214

-15

24

3+

4+

23

2

No

tail

muc

roP

bata

vicu

s75

032

-37

7-8

2775

12-1

3ndash

22

+12

43

Pde

cker

i69

5-85

537

-39

52-

622

-25

75-7

814

5-1

75

ndash2

3-3

2ndash

10-1

22-

3P

obsc

urus

560-

630

26-4

24

2-5

220

-24

72-7

412

-13

ndash2

+5

36

Poc

toli

neat

us51

2-59

727

4-3

42

37-

46

171

-21

675

-78

913

-14

ndash2

9+

83

6P

orie

ntal

is66

6-71

225

-37

5-6

29-3

773

-77

12-1

3ndash

1ndash

83

5P

trit

ici

442-

851

268

-32

34-

59

167

-21

277

-78

12-1

3ndash

20

+ndash

2P

ussu

rien

sis

830-

841

302

-37

44

8-5

620

2-2

873

8-7

61

15ndash

24

ndash6

ndashP

zeae

658-

700

244

815

313

765

16-1

7ndash

20

ndashndash

44

Tail

muc

ropr

esen

tin

first

15sp

ecie

sndash

+ab

senc

epr

esen

ceof

mal

esL

ila

tera

linc

isur

esP

vsp

ost-

vulv

alut

erin

esa

cV

bdv

ulva

lbod

ydi

amM

easu

rem

ents

inth

efo

rmr

ange

and

or(m

ean)

a

From

Tayl

oramp

Pilla

i(19

67)

bFr

omPi

llaiamp

Tayl

or(1

967a

)

894 Nematology






Vol 13(8) 2011 895

LK Carta et al



number




5-7 5-9 Geraert (1981)



Ecology




896 Nematology





Acknowledgements


References


















Vol 13(8) 2011 897

LK Carta et al

































898 Nematology











Vol 13(8) 2011 899






Vol 13(8) 2011 895

LK Carta et al



number




5-7 5-9 Geraert (1981)



Ecology




896 Nematology





Acknowledgements


References


















Vol 13(8) 2011 897

LK Carta et al

































898 Nematology











Vol 13(8) 2011 899

LK Carta et al



number




5-7 5-9 Geraert (1981)



Ecology




896 Nematology





Acknowledgements


References


















Vol 13(8) 2011 897

LK Carta et al

































898 Nematology











Vol 13(8) 2011 899





Acknowledgements


References


















Vol 13(8) 2011 897

LK Carta et al

































898 Nematology











Vol 13(8) 2011 899

LK Carta et al

































898 Nematology











Vol 13(8) 2011 899











Vol 13(8) 2011 899

supplemental description of paraphelenchus acontioides ...€¦ · nematology , 2011, vol. 13(8),...

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