Professional Documents
Culture Documents
Can. J. Microbiol. Downloaded from www.nrcresearchpress.com by "Institute of Vertebrate Paleontology and Paleoanthropology,CAS" on 06/05/13
For personal use only.
Acknowledgements
A . P. SINGH
Forest Research Institute, Private Bag, Rotoura, New Zealand
Received April 13, 1987
Accepted June 24, 1987
DANIEL,
G. F., NILSSON,
T., and SINGH,A. P. 1987. Degradation of lignocellulosics by unique tunnel-forming bacteria. Can.
J. Microbiol. 33: 943-948.
Transmission electron microscopic observations on a wide range of decaying wood samples obtained from both field situations
and laboratory exposure tests have confirmed bacteria to have a capacity to degrade intact highly lignified substrates including
preservative-treated and naturally durable woody tissues. Studies have shown a number of bacterial forms to be involved and
have provided morphological evidence for in situ lignin degradation confirming I4c-labelled experiments with synthetic and
natural lignins. A unique type of bacterial attack (tunnelling) characterized by the development of tunnels containing peculiar
cross-tunnel wall secretions has been recognized. Cytochemical studies have shown the extracellular tunnel secretions to contain
negatively charged constituents, while transmission electron microscopic energy-dispersive X-ray spectroscopy has shown these
tunnels to bind heavy metals during decay of timbers treated with metal-containing preservatives. The tunnelling bacteria are
motile nonflagellated Gram-negative rods. These bacteria have highly plastic cell envelopes which can produce vesicles. Our
studies provide evidence that tunnelling bacteria can remove total cell wall material, including lignin, and show a relationship
between their motility and the unique tunnels they produce.
' ~ u t h o rto whom reprint requests should be addressed.
943
Can. J. Microbiol. Downloaded from www.nrcresearchpress.com by "Institute of Vertebrate Paleontology and Paleoanthropology,CAS" on 06/05/13
For personal use only.
944
DANIEL,
G. F., NILSSON,T., et SINGH,A. P. 1987. Degradation of lignocellulosics by unique tunnel-forming bacteria. Can.
J. Microbiol. 33 : 943-948.
Des observations ont CtC faites en rnicroscopie Clectronique a transmission sur une grande variCtC d'Cchantillons de ligneux
pourrissants qui provenaient de situations de champs ainsi que de tests d'exposition en laboratoire. Ces Cchantillons ont permis
de confirmer que es bactCries ont la capacitC de ddgrader des substrats intacts hautement lignifiCs, incluant des ligneux traitCs de
fason prkventive et d'autres durables naturellement. Des Ctudes ont dCmontrC qu'un certain nombre de formes bactdriennes sont
impliquCes dans ce processus et ont fourni I'Cvidence morphologique de la dkgradation de la lignine in situ, confirmant les
espkriences du marquage de lignines synthktiques et naturelles auI4c. Un type unique d'attaque bactkrienne caractCrisC par le
dkveloppement de tunnels ("tunnelling") a CtC reconnu; des sCcrCtions pariCtales particulieres occupent ces tunnels. Des Ctudes
cytochimiques ont dCmontrC que les sCcrCtions extracellulaires des tunnels contenaient des substances chargdes nkgativement,
alors qu'en rnicroscopie Clectronique a transmission 1'Cnergie de dispersion analysCe par spectromCtrie de rayons X a montrC que
ces substances servaient a lier les mktaux lourds au cours du pourrissement de poutres traitCes avec des agents de conservation
contenant des mktaux. Les bactkries qui forment des tunnels sont des bitonnets non-flagellCs, mobiles et Gram nCgatifs. Ces
bactCries ont des enveloppes cellulaires hautement plastiques qui peuvent produire des vCsicules. Les prksents travaux
fournissent l'kvidence que les bactkries qui forment des tunnels peuvent dCgrader tout le matkriel pariCtal cellulaire, incluant la
lignine, et qu'il existe une relation entre leur mobilitC et les tunnels uniques qu'elles produisent.
[Traduit par la revue]
Can. J. Microbiol. Downloaded from www.nrcresearchpress.com by "Institute of Vertebrate Paleontology and Paleoanthropology,CAS" on 06/05/13
For personal use only.
NOTES
945
TABLE1. Relative count ratesa (K,) (background corrected) and ratios between the major elements
detected in the bacterial secretions and in the adjacent wood cell wall
Can. J. Microbiol. Downloaded from www.nrcresearchpress.com by "Institute of Vertebrate Paleontology and Paleoanthropology,CAS" on 06/05/13
For personal use only.
Ca
Cu
Cr
As
4866(2.82)
1406(1.06)
467 l(2.87)
269(0.36)
1223(0.63)
7847(3.15)
1089(0.57)
396(0.16)
756(0.70)
4554(1.64)
72(0.16)
2475(1.OO)
1899(1.00)
2332(1.00)
1067(1.OO)
2768(1 .OO)
441(1.00)
14090(5.29)
4416(2.16)
103(0.04)
3 164(2.75)
927 l(3.11)
7265(3.42)
3 159(1.94)
62(0.03)
2278(2.49)
4520(1.90)
'Count rates represent mean values obtained from energy-dispersive X-ray spectroscopy point analyses (100 s) of 10 different
regions for each category. Spot size for analysis was kept constant at 50nm.
bRatios are given in parentheses.
'ppts., precipitates.
FIGS.1-5. Electron micrographs showing features of tunnel formation and reaction with preservatives. Figs. 1-3, and 5. TEM. Fig. 1.
Stages in bacterial penetration of the fibre cell wall of Laurelia novae-zelandiae;a tropical hardwood. (a) Initial settlement and adhesion of an
encapsulated bacterial cell (B) to the fibre lumen S3layer. (6) Penetration of the fibre wall by a characteristic pear-shaped bacterium (B) and the
development of characteristic cross-tunnel secretions (CT) into the lumen. ( c )Change in direction of decay tunnel on entry to the secondary S2
fibre wall layer. Continuity between extracellular material sealing the entrance and that forming the cross-tunnel walls (CT) is apparent (arrow).
Fig. 2. Tunnel formation in a vessel wall from the hardwood Homalium foetidum (lignin content, ca 32.0%). (a) Compact concentrically aligned
cross-tunnel wall (CT) secretions are developed periodically behind the bacteria during degradation. (6) High magnification to show the fibrils
which form the wall secretions (arrowhead). Fig. 3. Lateral tunnel formation in Pinus radiata (lignincontent, ca. 28%) and release of vesicles (V)
and fibrillar materials to form the cross wall (CT) secretions. Fig. 4. SEM showing the pointed ends of both tunnels (arrowheads)and bacteria (B)
and the characteristic 3-D nature of cross-tunnel secretions (CT) produced in Pinus sylvestris (lignin content, ca. 27%). Fig. 5. Tunnels formed
within the S2wall layer of Tanalith NCA treated Pinus radiata showing aggregation of electron-dense preservative (Cu, Cr, As) particles with the
cross-tunnel (CT) and tunnel wall (TW) extracellular secretions. Unosmicated unstained preparation as used for energy-dispersive X-ray analysis
TEM. (Bars = 0.5 Fm, except in Fig. 26,O. 1 Fm).
Can. J. Microbiol. Downloaded from www.nrcresearchpress.com by "Institute of Vertebrate Paleontology and Paleoanthropology,CAS" on 06/05/13
For personal use only.
NOTES
947
948
Can. J. Microbiol. Downloaded from www.nrcresearchpress.com by "Institute of Vertebrate Paleontology and Paleoanthropology,CAS" on 06/05/13
For personal use only.
TABLE2. 14c02
produced by cultures containing tunnelling or erosion
bacteria from synthetic lignins labelled in the side chain, aromatic ring,
or methoxyl groups
Days
Culture
no.
Type of
bacteria
Label
Tunnelling
Tunnelling
Tunnelling
Tunnelling
Tunnelling
Tunnelling
Erosion
Erosion
Ring
Ring
Side chain
Side chain
Methoxyl groups
Methoxyl groups
Ring
Ring
11
16
Acknowledgements
This research was supported partly by the Swedish Board for
Technical Development (STU) and by a fellowship (to T .
Nilsson) from the Organization for Economic Co-operation and