Presented by

Lt. Deore P.B.

Reg. No. 2018A/15P

DEPARTMENT OF PLANT PATHOLOGY

COLLEGE OF AGRICULTURE,
VNMKV, PARBHANI 431 402 (M.S.)
 Martinus Willem Beijerinck
 Born: March 16, 1851in Amsterdam, Holland
 Chemical Engineer
 1873: Lecturer, Agricultural School,
Wageningen, Holland
 1877: Doctor of Science at Leyden
 1887: Microbiologist,
Netherland Yeast & Alcohol Factory,
Delft
 1893: Professor of Bacteriology,
Technical School, Delft
 1897: Founded Microbiological Laboratory
Martinus Willem Beijerinck

 His interests in TM was fostered by Adolf Mayer

 Famous as a soil microbiologist
 Professionally a botanist
 Studies on plant galls, fermentation, sulphur
bacteria, azotobacter, & denitrification
 Nutrition studies on algae, amoebae, yeasts,
fungi, & bacteria
 Isolated: Bacillus radicicola
Concerning a contagium viwm fluidum as
cause of spot disease of tobacco leaves
M. W. Beijerinck
 Beijerinck was at that time Mr. Mayer’s colleague at
Agricultural School, Wageningen
 Mayer showed him his experiments
 Carried out cultural experiments pertaining to aerobes
 Possibility: anaerobes in plant tissue affecting
surrounding plant tissue with poisons, like tetanus
bacteria, which is soluble, non-living, i. e., unable to
reproduce itself.
 Taken great pains to find anaerobes connected with
disease: negative results
 Conclusion: spot disease is an infectious one that is
not caused by microbes
 1897: Erected bacteriological laboratory, Delft
Carried out a series of experiments
Concerning a contagium viwm fluidum as
cause of spot disease of tobacco leaves
1. Infection is not caused by microbes,
but by a contagium vivum fluidum
 Carried out experiments to determine presence
of anaerobes in filtered juice: negative results
Conclusion:
 The quantity of candle filtrate necessary for
infection is extremely small
 A small drop put in plant with a Pravaz syringe
could infect numerous leaves & branches
 Contagium, although fluid, reproduces itself in
living plant
Concerning a contagium viwm fluidum as
cause of spot disease of tobacco leaves
1. Infection is not caused by microbes,
but by a contagium vivum fluidum
 Experiments using candle filtrate still open to
criticism: possibility of presence of anaerobes
because of corpuscular nature of contagium
Diffusion experiments
 Drops of extracted juice of diseased leaves put on
surface of thick agar plates & left to diffuse with water
for several days
 To separate virus from raw leaf substance & bacteria
Concerning a contagium viwm fluidum as
cause of spot disease of tobacco leaves
1. Infection is not caused by microbes,
but by a contagium vivum fluidum
Results:
 Substance causing infection may penetrate into agar
plate to no small depth
a) A layer of agar about 1/2mm thick was removed from
outer surface of spot
b) Mass lying immediately below was then removed in
two successive layers & both parts used for infection
Results:
 In both cases: Characteristic symptoms of infection
Very intensively by upper
More weakly by lower layer of agar.
 After 10 days distance covered by virus ≥2mm
 Proved: virus must really be regarded as liquid or
soluble & not as corpuscular
Concerning a contagium viwm fluidum as
cause of spot disease of tobacco leaves
1. Infection is not caused by microbes,
but by a contagium vivum fluidum

 The candle filtrate had a somewhat weaker effect on

plant than extracted juice that not filtered
 Conclude:
 Virus is held back in filter pores, at least at beginning
of filtration process
 Virus is non-corpuscular nature
Concerning a contagium viwm fluidum as
cause of spot disease of tobacco leaves
2. Only those organs of plant that are growing & whose
cells are dividing are capable of being infected; here
only does virus reproduce itself
 Virus attacks only actively growing tissues & organs in which
cells division is still in full progress
 All tissue reached its full growth is immune but transport
virus
 If growing Leaves beyond expansion stage inoculated: not
infect, but still suitable for transmission of virus to stem
 If stem is inoculated: only young leaf-buds & leaves newly
developed from these growing points are infected
If young leaves are inoculated: virus returns to stem from leaf
& infects either axillary buds or rise to infect terminal bud
Concerning a contagium viwm fluidum as
cause of spot disease of tobacco leaves
2. Only those organs of plant that are growing & whose
cells are dividing are capable of being infected; here
only does virus reproduce itself
 If fully matured organs used for infection:
o Virus remains in matured cells without having any effect
o Virus moves out of matured parts into surrounding new
tissues & affect them
 In any case: virus in plant is capable of
o reproduction & infection only in dividing cell/tissues
o Matured & expanded tissues unsuitable
o Unable to grow independently
o No ability to reproduce outside of plant
 On storage
o Not losing its virulence or decrease it.
But an increase of infectivity is not observed
Concerning a contagium viwm fluidum as
cause of spot disease of tobacco leaves
3. Flow of virus inside plant. Various ways of infecting:
Local & general infection
 Virus move with flow of water through xylem during
strong transpiration
 Through xylem able to infect meristematic leaf-buds
 Large quantity of virus at tip through transpiration
stream along xylem bundles.
 Ascending & descending flow through phloem.
 Buds take up virus less easily,
healthy organs may be produced from it
 Seedlings injured in root:
Show symptoms sooner after infection.
 Plants infected from soil :
Diseased leaves were all around stem
 Plants infected through wounds:
local infection,
which later developed into a general infection
Concerning a contagium viwm fluidum as
cause of spot disease of tobacco leaves
4. Virus may be dried without loss in strength of infection.
 Dried diseased leaves
o Capable of inducing infection even after 2 years
o Virulence is always less than that of fresh materials
o Partial destruction of virus in drying
o Not modified, weaker form
 Same is true of pieces of filter paper
 Alcohol precipitate of virulent sap retains its virulence
after drying at 40oC
5. Virus may winter in its dry state outside of plant in soil
 Flower pot experiment:
 Virus can retain its full virulence after wintering in
air-dried soil
Concerning a contagium viwm fluidum as
cause of spot disease of tobacco leaves
6. Other attempts at infection through roots.
 Normal roots are capable of taking up virus from soil
through their closed outer epidermis
 Admitted that this conclusion may not be reliable bcz
animals living underground facilitated entrance of virus
by means of root injuries
7. Virus becomes ineffective in boiling temperatures. Effect
of Formalin
 Carried out Several experiments with freshly extracted
heated juices & juices after filtration through candles
 Boiling completely destroy virus (90OC)
 Shortest period of heating is sufficient to destroy virus.
Weak solutions of Formalin mixed with virus do not
destroy it, but lengthen period of incubation
Concerning a contagium viwm fluidum as
cause of spot disease of tobacco leaves
8. Different symptoms of disease. Development of
malformed leaf tissues through large quantities of virus
 Spot disease (mosaic):
Milder form
Disease of chlorophyll particles
 More intensive forms:
General disease of living protoplasm
 Contagium increases through growth
Concerning a contagium viwm fluidum as
cause of spot disease of tobacco leaves
9. Albinism or “bunt” as an incidental result of
artificial infection
 Albinism: in a mixture infection of a bacterium with virus
 Isolated two types of bacteria
 Suspension was prepared in tap water & inoculated
a) Non or weakly-liquefying & non-fermenting bacterium,
B. anglomerans
 Noticed a beginning of disease
 Not develop in regular manner but produced albino
b) The second variety of bacteria
 Without effect in infection
 Albinism produced through infection
 With virus combined with formalin
 From soil
b) There is some connection between virus of spot disease
& bunt (mosaic)
Concerning a contagium viwm fluidum as
cause of spot disease of tobacco leaves
10. Other infectious plant diseases caused by a contagium
fluidum & not by parasites
 Infectious diseases of chlorophyll bodies
 Spot disease
 Albinism or bunt (mosaic)
 Difference in mode of transmission of contagium
consider them as separate kinds of diseases
 Albinism (variegation):
 Transmit only by grafting or budding
 Closer relationship to protoplasm of
plant than contagium of spot disease
Concerning a contagium viwm fluidum as
cause of spot disease of tobacco leaves
10. Other infectious plant diseases caused by a contagium
fluidum & not by parasites
 Peach yellows
 Transmit through grafting or budding
 Peach rosette
 Transmit through budding & root grafting
 Beijerinck: virus moves laterally with difficulty but
upwards with ease
 So that a tree may become diseased on side on which
rosette bud is grafted, while opposite side remains
healthy for years
 Yellows & rosette much more closely spot disease
Concerning a contagium viwm fluidum as
cause of spot disease of tobacco leaves
10. Other infectious plant diseases caused by a contagium
fluidum & not by parasites
 Distinguish a contagium fluidum
 Independent contagium:
 Capable of existence outside of plant
 Eg. Leaf-spot disease of tobacco
 Contagium that exists only in living tissues:
 Eg. Variegation
Transmissible through graftage only
Martinus Willem Beijerinck
 Spot disease:
 Infectious
 Not caused by microbes
 Cause: Cantagium vivum fludum
 Fluid?
 Viruses: liquid or soluble
 Virus reproduce in living plant
 Virus attacks only on living cells/tissues
 On storage not losing virulence
 Virus moves with flow of water
 Partial destruction of virus on drying
 Boiling destroy virus
 Albinism transmitted by grafting