Wyss, U.

Zunke, U.

Horizon / Pleins textes

Notes brèves 
N-pinky 
N-tropic 
Sioux 
Tr,ope GRO-7 
Table 1 (cont.) 
~~ 
1 O00 4.60  (29.23)  0.691  0.365  7.3 
L.S.D. (1 '/O) 1.276 0.009 0.005 
L.S.D.  (5 "/O) 0.842 0.006 0.003 
O 6.20 - 0.450 0.215 
10 5.50  (11.29) 0.520 0.242 104.1 
100 4.60  (25.80) 0.565 0.270 36.0 
1 O00 4.10  (33.87) 0.625 0.325 9.4 
- 
L.S.D. (1 '/O) 0.584 0.010 0.005 
L.S.D.  (5 '/O) 0.386 0.006 0.003 
O 5.70 - 0.365 0.178 
10 4.70  (17.54) 0.410 0.198 140.3 
100 3.60  (36.84) 0.455 0.220 49.9 
1 O00 3.00  (47.36) 0.500 0.268 10.9 
- 
L.S.D. (1 O/') 0.201 0.007 0.003 
L.S.D. (5 %) O. 133 0.004 0.002 
O 6.30 - 0.435 0.212 
10 
- 
5.55 (11.90) 0.503  0.237  108.8 
100  4.65 (26.19) 0.548  0.265  36.2 
1 O00 4.15 (34.12) 0.596  0.322  9.4 
L.S.D. (1 O/') 0.259  0.009 0.003 
L.S.D.  (5 ' /O) O. 170  0.006  0.002 
O 5.80 - 0.400  0.193
10  4.90  (15.51)  0.455  0.216 
- 
125.1 
100 3.80  (34.48) 0.500 0.240  43.6 
1 O00 3.30  (43. IO) 0.550 0.293  10.3 
L.S.D.  (1 '/O) 0.279  0.007  0.006 
L.S.D.  (5 '/O) 0.184  0.004  0.004 
Each value is a mean of five  replicates. 
In  parenthesis are  given per  cent reduction over  control. 
R = Reproduction  factor of the nematode, Pf = Final population, Pi = Initial Population, O.D. = O-dihydroxy  phenols. 
REFERENCES GIEBEL, J. (1974). Biochemical  mechanism  of  plant  resistance. 
A review. J. Nematol., 6 : 175-184. 
BIEHN, W. L., KUC, J. &WILLIAM, E. B. (1968). Accumulation JOHNSON, G. SCHAAL, L* (1954). RelationshiP of 
of  phenols  in  resistant  plant-fungi  nteractions. Phgo- genic  acid to the  scab  resistance  in  potatoes. Science, 115 : 
pathology, 58 : 1255-1260. 627-629. 
BRAY, H. G. & THORPE, W. V. (1954). Analysis of phenofic REBOIS, R. S., ELDRIDGE, B. P. &WEBB, R. E. (1977). Rotylen- 
compounds of interest  in  metabolism. Meth. Biochem.,Anal., chulus renijomzis parasitism of potato and its effect on 
1 : 27-52. yields. Pl. Dis. Repts; 62 : 520-523. 
Accepté pour  publication le 14 mai 1985. 
THE  POTENTIAL OF HIGH  RESOLUTION  VIDEO-ENHANCED  CONTRAST 
MICROSCOPY  IN  NEMATOLOGICAL  RESEARCH 
Urs Wyss and Ulrich ZUNKE* 
High power interference  contrast ciné-microscopy is tional  morphology (e.g. Robertson & Wyss, 1979; Sey- 
an  important tool for  studying  nematode  behaviour  and mour, 1983a, b). Video recordings  are very useful  for  an 
is, perhaps,  indispensable  for  detailed analysis of func- immediate analysis, provided the resolution is of suf- 
* Institut f i r  Phytopathologie der Christian-Albrechts-Universitat Kiel, Olshausenstr. 40, 0-2300 Kiel 1, W. Gennany. 
Revue Nématol. 9 (1) : 91-94 (1986) 91 
U. Wyss & U. Zunke 
ficient standard. Seymour, Minter and Doncaster (1978) cells with their video-enhanced contrast, differential 
used a low-light remote  recording system to study interference  contrast (AVEC-DIC) microscopy.  For Our 
nematode feeding behaviour at very low light levels. observations  on nematodes, we applied a less sophisti- 
Great advances in video recording  techniques were cated and therefore less  expensive  system  which still gives 
made  by Allen,  Allen and  Travis (1981) Who optimised excellent results and may be considered a full substitute 
visual and  photographic  contrast  in  human  and  animal for cine-micrography when  used  for  analytical  purposes. 
Fig. 1 A. Components of the hi& resolution video-enhanced mimscopy system (see t a ) .  B : An infective J 2 juvede of Heterodem 
schachtii on  its  migration  through a cortical  cell, three ce11 layers  beneath the root surface. S : stylet; MB : median  oesophageal 
bulb;  arrow  points to the  border of the  picture  monitor  screen  (Nr. 6 in  Fig. 1 A) from  which  the  picture  was  taken. 
92 Revue Nématol. 9 (1) : 91-94 (1986) 
High resolution video-enhanced contrast nticroscopy 
Fig. 2 A. In vivo observation of cytoplasm  dynamics  around the feeding tube (FT) of a  female J 3 Hetemdera schachtii within the 
initial  syncytial ce11 (SY). Note  the  modified zone of  cytoplasm  free  of  organelles  around the feeding  tube. CW : stub of  a partially 
dissolved ce11 wall; N : head of the  nematode;  arrow  points to flow of cytoplasm past the ce11 wall stub. B-C : The posterior 
oesophageal bulb of an Acrobeles species during  food  ingestion. C : Bulb  flaps (F> start to invert, 6 frames (0.12 sec.)  after B. 
Revue Néwzatol. 9 (1) : 91-94  (1956) 93 
U. Wyss & U. Zunke 
In addition, it enables a careful study of nematode 
behaviour  and ce11 responses  deep  inside roots. Events 
that otherwise remain  obscure  when viewed through  the 
differential interference contrast microscope, become 
well defined by video  contrast  enhancement. The system 
we use  consists of the following components (Fig. 1A) : 
Reichert  differential  interference  contrast  micro- 
scope '' Polyvar " equipped with 100 x /1.32 N.A. 
and 40 x Il.00 N.A. planapochromatic oil immer- 
sion objectives and  optovar 0.8 x 11 x 11.25 x 
12 X? 
Monochrome  camera system Grundig FA 76 1, con- 
verted  to  875 lines/50 Hz vertical frequency.  Equip- 
ped  with  a 1" Newvicon  pickup tube (light sensitivity 
O. 1 lx) and  switch  to select withlwithout line interlace 
to  permit  magnetic single frame recording. 
Remote control Grundig FB 400 E to control all 
functions. With  pulse  generator  fitted  to  set  recor- 
ding  and  playback  sequence (1 ... 300 sec. in 11  sta- 
ges). Single  frame release is either  manually or from 
an external  source. 
Picture  monitor  Grundig  BG 330 (screen diagonal 
31 cm), converted to 875 lines/50 Hz vertical fre- 
quency. 
Professional 2.5 cm video recorder Grunding BK 
411 HE, 10 MHz resolution,  converted  to  875 
lines/50 Hz. 110 min. playing time (2.5 cm  chromium 
dioxide BASF  tapes CV 26 R). With electronic edi- 
ting to assemble and insert various scenes; slow 
motion  fonvard  and reverse and additional  module 
to  record  individual  frames  with  stationary  magnetic 
tape. 
6) Picture monitor Grundig BG 12 (screen diagonal 
12.8  cm), converted  to 875 lines/50 Hz vertical fre- 
quency. 
Relevant  scenes  from  tape  recordings  are analysed by 
single frame release (50 pictures/sec.). For documen- 
tation,  photographs (1 sec.  exposure)  are  taken  with a 
camera (7) from  the small picture  monitor (6). 
Figures  1 B and  2 A-C illustrate the potential of this 
high  resolution  video-enhanced  contrast system for 
nematological research. Figure 1 B shows an infective 
Heterodera  schachtii juvenile during its  intracellular 
migration  through  cortical cells towards  its  final  feeding 
site  within  the  vascular cylinder. The photograph was 
taken  after the juvenile had  reached  the third layer of 
cortical cells in roots of rape-seed (Brussica  napus var. 
oleifera) seedlings  grown in sterile nutrient agar culture. 
Figure 2 A shows, at about 3 800 x magnification, 
the  feeding  tube (FI') of a female fourth stage H. schachtii 
juvenile in  the initial syncytial cell within the  vascular 
cylinder of an oil radish (Raphanus  sativus var. oleifor- 
mis) root. The in vivo formation  and  detection of these 
tubes  can only be  seen  through  the microscope by video 
contrast  enhancement. In  an electron microscopic study, 
Accepté pour publication le 14 octobre 1985. 
94 
a zone of modified cytoplasm free of organelles was 
found to surround  the  feeding  tubes of H. schachrii in 
initial syncytial cells of the  same  host (Wyss, Stender & 
Lehmann, 1984). The in  vivo observations  confirm  this 
finding  and  support  the  assumption  that  feeding  tube 
material may help to " predigest " cytoplasm for an 
efficient withdrawal of nutrients  from  the  dense cyto- 
plasm. For  Figures  1 B and 2 A, observation  chambers 
were used as described by Wyss (1973). 
Figures  2 B and 2 C show the potential of the system 
to  evaluate  functional morphology, here  within  the 
posterior bulb of an Acrobeles species during  food in- 
gestion. In  Figure  2 C the  bulb flaps start  to invert, six 
pictures (0.12 sec.) after  2 B. Single  frames (438 lines) 
from  the video tape  are close to  the quality of 16 mm 
ciné  film recordings. 
Apart  from  the  immediate analysis of motion  studies, 
important scenes of tape  recordings  can  be fïlmed 
directly  from the small picture monitor  with  a 35 mm 
ciné  film camera, equipped  with  a  phase  shift  adaptor. 
In this way a  16 mm research film*  on  the  behaviour of 
H. schachtii inside  roots of host  seedlings was produced 
(Wyss, Zunke & Inst. wiss. Film,  1985) which shows hm- 
cellular  migration of J 2 juvenile, induction  and  main- 
tenance of syncytia within  the  vascular cylinder as  well 
as characteristic  events during salivation and 'ingestion. 
REFERENCES 
ALLEN, R. D., ALLEN, N. S. & TRAWS, J. L. (1981). Video- 
enhanced  contrast,  differential  nterference  contrast 
(AVEC-DIC)  microscopy : A  new  method  capable  of  ana- 
lyzing microtubule-related motility in the reticulopodial 
network  of Allogromia laticollaris. Ce11 hlotility, 1 : 291-302. 
ROBERTSON, W. M. & WYSS, U. (1979).  Observations  on  the 
ultrastructure  and  function of the dorsal  oesophageal  gland 
ce11 in Xiphinema index.  Nematologica, 25 : 391-396. 
SEYMOUR, M. K. (1983~). Action of the feeding pump in 
Longidorus caespiticola Hooper. Nematologica, 29 : 65-77. 
SEYMOUR, M. K. (1983b). The feeding pump of Ditylenchus 
dipsnci (Nematoda : Tylenchida). Nematologica, 29 : 
SEYMOUR, M. K., MINTER, B. A. & DONCASTER, C.  C. (1978). 
A low-light  remote  recording  system  used to study  nema- 
tode behavior. Nematologica, 24 : 167-174. 
WYSS, U. (1973).  Feeding  of Tylenchorhynchus dubius. Nemc. 
tologicu, 19 : 125-136. 
WYSS, U., STENDER, C. & LEHMANN, H.  (1984). Ultrastructure 
of feeding  sites of the cyst  nematode Heteroderu schachtii 
Schmidt in roots of susceptible and resistant Raphanus 
sativus L. var. oleifonnis Pers.  cultivars. Physiol. Pl. Pathol., 
-WYSS, u., ZUNKE Ü. & ïNST. WISS. FILM (1985). Héterodera 
schachtii (Nematoda). Behaviour inside roots (rape). Film 
E 2904 der Enc.  Cin., Gottinnen. 
* The film  (see  reference)  has  an  English  commentary  and 
can  be  ordered  from : Institut  fiir den wissenschaftlichen  Film, 
Nonnenstieg  72,  D-3400  Gottingen, W. Germany. 
171-189. 
25 : 21-37. 
Revue Nématol. 9  (1) : 91-94 (1986) 


The potential of high resolution video-enhanced contrast microscopy in nematological research

Abstract

Similar works

Full text

Available Versions

Horizon / Pleins textes