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Section 4.2
Virus Eradication: Tissue
Culture of Meristems,
Thermotherapy, and
Chemotherapy
Introduction
Due to the fact that it takes many months for meristems to become
plantlets, some researchers have tested the application of chemical
products (chemotherapy) that reduce or inhibit viral multiplication. In the
case of potato, three of the most important viruses, PVX, PVS, and PVY,
have been eradicated by adding Ribavirin to the culture media and
isolating the axillary buds (Griffiths et al., 1990).
The disease does not depend only on the presence of the host, but on
environmental conditions as well, especially humidity and temperature,
which play an important role. The disease can be defined as the product
of the interaction among the host, pathogen, and environment.
Not all the cells in a diseased plant become infected with pathogens. The
meristematic tissue of the root and the terminal sprouts of an infected
plant are sometimes pathogen-free. Sometimes, such as in potato with
PVX and TRV (tobacco rattle virus), only the apical dome and the first
young primordial leaves are free of virus. The reason for this is unknown.
Nevertheless, it is believed that one or more of the following factors are
responsible:
Thermotherapy
Experiments carried out with viruses and their host plants have shown
that when plants are treated at high temperatures (thermotherapy) the
virus concentration is reduced (Kassanis, 1957; Quak, 1977). There are
different explanations for this phenomenon. One explanation is that
competition among the rapidly dividing host cells and the virus particles
for the places where nucleic acids and proteins synthesize results in a
change in the balance between the synthesis and degradation of virus
particles. Another explanation is that under high temperatures, the union
of the protein subunits that protect the nucleic acid of the virus becomes
weaker and temporal fissures appear, allowing the attack of nucleases,
which inactivate the virus and decreases its concentration.
Culture of meristems
The meristem is the active growing point of the plant shoot. It is a small
zone composed of cells (meristematic), which divide very fast.
The dome of the apical bud contains the real meristematic cells and is
surrounded by leaf primordia and primary leaves. Due to the fact that the
most differentiated vascular tissues are far away from the meristems
The process used at CIP for the eradication of potato viruses includes the
following steps (Fig. 1):
The culture media used for this purpose is based in Murashige and
Skoog salts (1962), supplemented with 2 mg/l glycine, 0.5 mg/l
nicotinic acid, 0.5 mg/l piridoxine, 0.4 mg/l thiamine, 0.1 mg/l
gibberellic acid, 0.04 mg/l kinetine, and 2.5% sucrose. The media
is turned to a gel with agar (0.6%) and then sterilized in an
autoclave at 15 pounds pressure and 121°C for 15 minutes.
Chemotherapy
Recommended Literature
CIAT (Centro Internacional de Agricultura Tropical). 1991. Cultivo de
tejidos en la agricultura: Fundamentos y aplicaciones. W.M. Roca
and L.A. Mroginski (eds.). Cali, Colombia. P. xii, 970.
Espinoza, N., R. Lizárraga, C. Sigueñas, F. Buitrón and J.H. Dodds.
1991. Cultivo de tejidos: Micropropagación, conservación y
exportación de germoplasma de papa. Guía de Investigación CIP 1.
Centro Internacional de la Papa. Lima, Perú. 19 p.
Griffiths, H.M., S.A. Slack and J.H. Dodds. 1990. Effect of chemical and
heat therapy on virus concentration in in vitro plantlets. Can. J. Bot.
68:1515–1521.
Kassanis, B. 1957. The use of tissue culture to produce virus-free clones
from infected potato varieties. Annals of Applied Biology 45:422–
427.
International Potato Center (CIP). 1993. Basic techniques in plant
virology. U. Jayasinghe y L.F. Salazar (eds.). CIP, Lima (Technical
Training Unit 1).
Lizárraga, R., A. Panta, U. Jayasinghe and J.H. Dodds. 1991. Tissue
culture for elimination of pathogens. CIP Research Guide 3.
International Potato Center (CIP). Lima, Peru.
Lizárraga, R.E., L.F. Salazar, W.H. Roca and L. Schilde-Rentscheler.
1980. Elimination of potato spindle tuber viroid by low temperature
and meristem culture. Phytopathology 70:754–755.
Lizárraga, R.E. and L.F. Salazar. 1982. Effect of meristem size on
eradication of potato spindle tuber viroid. In: Hooker, W.J. (ed.).
Research for the potato in the year 2000. International Potato
Center. Lima, Peru. P. 118–119.
Mellor, F.C. and R. Stace-Smith. 1977. Virus-free potatoes by tissue
culture. In: Plant cell, tissue, and organ culture. J. Reinert and Y.P.S.
Bajaj (eds.). Springer-Verlag, Berlin. P. 616–635.