Bulletin OEPPIEPPO Bulletin 23, 5 17-520 (1993)

Spread of verticillium wilt by nursery plants in olive groves in

the Halkidiki area (Greece)’

by C. C. THANASSOULOPOULOS
Faculty of Agriculture, Aristotelian University of Thessaloniki, 540 06 Thessaloniki (Greece)

Olive groves have been established during the last decade in the coastal area of Halkidiki and
in the northern part of the Kassandra peninsula (GR). Visual symptoms of verticillium wilt
were observed recently all over this area, particularly in young trees of cv. Hondrolia
Halkidikis but not in the occasional old trees already in the orchards. The symptoms appeared
regardless of the previous crop (cotton or not), so the nurseries supplying the farmers with
young trees were assumed to be the primary source of infection. A survey of olive nurseries in
the area was conducted during 1991. Eleven out of the total 18 nurseries in the area were
located in the Verticilhm duhliue-infested zone and 9 of these were sampled. Three-year-old
olive seedlings were selected and isolations were made from the xylem onto PDA acidified with
0.25% lactic acid. V . duhliae-infected seedlings were detected in 4 out of the 9 nurseries
examined. The presence of infected young trees in nurseries covering 15.6% of the total
nursery area provided strong evidence that nurseries may be responsible for the recent
expansion of verticillium wilt in the area.

introduction
Olive has been cultivated on hilly and sloping sites in the Halkidiki area since early times. The
main cultivar in the area is Hondrolia Halkidikis, which bears large fruits used for both table
consumption and oil production. Olive cultivation has expanded during the last decade,
especially since 1985,to the coastal area between the Kassandra and Sithonia peninsulas, in the
region of the villages Nea Moudania, Agios Mamas, Olynthos and Gerakini, and to the southern
part of Kassandra peninsula. New olive groves were established on fields previously cultivated
with cereals, cotton or other field crops. Recently, symptoms of verticillium wilt have been
observed all over the coastal area, in which irrigation of olive trees is also a new practice.
A survey of olive orchards during I991 and subsequent isolation from random trees showing
wilt symptoms revealed that verticillium wilt disease (caused by Verticillium dahlia) was present
in all the coastal area. In the same area, V . dahliae-infected pistachio and apricot trees were also
found. Examinations of suspected old olive trees pre-existing in the highland area showed the
absence of verticillium wilt; similar symptoms were the result of other disease attacks, mainly
root o r hard wood rots.
In the I/. dahliae-infested area, the problem was more severe in relatively newly planted trees.
Old trees already in the area rarely showed symptoms. V. dahliue was found in the groves
irrespective of the cultivation history of the field on which they had been established, whether
cotton or other, and it was therefore difficult to confirm the primary source of infection. On the
other hand, it was obvious that irrigation water was responsible for carrying the disease through
the orchards.
The most probable source of primary infection seemed to be the olive nurseries supptying the
farmers with young trees. These nurseries had been established and were operating all over the
Halkidiki area by the time olive cultivation started to expand. T o investigate whether this
assumption was true, a disease survey of olive nurseries in the area was conducted in 1991.
Paper presented at the Joint MPU/EPPO Conference on Olive Diseases, Sounion (GR), 1992-05-05/08,

517
518 C. C. Thanassoulopoulos

Methods and results

The nurseries usually held I-3-year-old olive seedlings. The 3-year-old seedlings are those
that have not been sold the previous year, most young plants being delivered to the growers
when 2 years old. In these young plants, symptoms of verticillium wilt were seen similar to
those observed in olive orchards. Samples were taken from these trees, o r in most cases the
whole tree was taken, and isolations from the xylem were made in the laboratory onto PDA
acidified with 0.25% lactic acid, as reported elsewhere (Thanassoulopoulos & Kitsos, 1972;
Thanassoulopoulos et ul., 1979).
A total of 18 nurseries, covering an area of 70.5 ha, operate in the area, with an approximate
production of more than 500,000 young trees per year, part of which is also directed to other
regions of Greece where olive trees are grown.
Seven of the 18 nurseries were located in areas where, up to now, no V . dahliae had been
isolated from olive trees. In six of them, located in the highland area, the seedlings are produced
by grafting the desired cultivar on young plants, 1-year-old, derived from root cuttings of wild
indigenous olive trees found in nearby forested land. In the seventh nursery, leafy cuttings were
taken from selected trees of the desired cultivar. In five out of these seven, no V. duhliue could be
detected (Table I).
The remaining 11 nurseries were located in the western plain of Halkidiki (Nea Moudania,
Agios Mamas and Olynthos), where verticillium wilt was present. Nine of them, covering almost
half of the total nursery area, were surveyed. In all these nurseries, seedlings are produced by mist
propagation. In four of them, covering 33% of the above nursery area and 15.6% of the total
area, V . duhliue was isolated from several 3-year-old seedlings (Table 1).

Table 1. Olive nurseries infested by Verticillium duhliue in the Halkidiki area

P6piniBre.s d’olivier contaminees par V . dnhliue dans la region de Halkidiki

Number of nurseries Percent

Locations total examined a’ b

A Nurseries in area without V. dahliue

I . Seedlings derived from grafted wild
indigenous olive trees 6(1.85)* 4(0.85) 45.9 0
2. Seedlings derived by mist propagation
of leafy cuttings l(0.80) l(0.80) 100 0
Occurrence of V . dahliae 0 0
Total 7 (2.65) 5 (1.65) 62.3 0

B Nurseries in areas with V. dahliae

(seedlings derived by mist propagation)
1. No verticillium wilt 5 (2.20) 66.7
2. Seedlings infected with V. dahliae 4 (1.10) 33.3
Total 11 (4.40) 9 (3.30) 75.0 25.0

’Percentages were calculated on area basis. Column a is the %, of the column ‘examined’ over column ‘total’
Column b is the % infected of those examined.
Number in parentheses indicates the cultivated area in ha.
 Spread of verticillium wilt by nursery plants 519

Discussion
The results of this survey seem to indicate that the source of primary infection was the nurseries,
since in 4 of the 9 nurseries examined the presence of V. dahliue-infected seedlings was verified.
This explained satisfactorily the observed fact that the disease was mainly present in young olive
groves and not in the occasional old trees pre-existing in the area.
There are two reasons which may account for the incidence of the disease in the nurseries of
the area. Firstly, the nursery growers do not take the necessary precautions in nursery
preparation, and use infected or improperly sterilized soil. Secondly, they select leafy cuttings
from old trees that are already infected but symptomless. It is also very interesting to note that
the disease is prevalent in irrigated areas, where over-irrigation is in most cases a common
practice. This facilitates disease spread (Thanassoulopoulos & Kitsos, 1972), which is more
pronounced in groves where the previous crop was susceptible to V. dahliae, e.g. cotton
(Thanassoulopoulos et al., 1980).
Under these conditions, it seems necessary to take drastic measures to eliminate fungus
infection from the nurseries. The presence of infected seedlings in the nurseries, even at low
percentages, constitutes a potential hazard, particularly in cases where the young seedlings are
transplanted into areas where verticillium wilt probably does not exist.

Dissemination de Verticillium dahliae par les plants dans les oliveraies de

Halkidiki
La culture de l’olivier a etk etablie durant les 10 dernieres annees dans la region littorale de
Halkidiki et dans le nord de la peninsule de Kassandra (GR). Des sympt6mes visuels de
verticilliose (due a Verticillium dahliae) ont ete observes recemment dans toute cette region,
notamment sur de jeunes oliviers du cv. Hondrolia Halkidikis mais non sur les arbres bgks qui
existent sporadiquement dans les oliveraies. Ces sympt6mes sont apparus independamment de
la culture precedente (coton ou non). Par consequent, il est supposC. que les pepinikres qui
fournissent les plants aux oleiculteurs ont constitue la source primaire de l’infection. Nous avons
effectue en 1991 une prospection dans les pepinieres d’olivier de la region. Sur 18 dentre elles, 11
se situaient dans la zone contaminee par V. dahliae et 9 ont &ti.examinees. A partir d’echantillons
de plants de 3 ans, des prelevements ont ete effectues pour isolement sur milieu nutritif P D A
acidifik a l’acide lactique a 0,25”/0.Des plants contaminks par V. dahliae ont etk reperks dans 4 des
9 pepinieres etudiees. La presence de plants infectes dans des etablissements representant 15,6%
de la surface totale de pepinieres confirme notre hypothese que ces dernieres sont probablement
responsables de la dissemination recente de V . dahliae dans la region.

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520 C . C. Thanassoulopoulos

References

THANASSOULOPOULOS, C.C. & KITSOS,G.T. (1972) Verticillium wilt in Greece. Plant Disease Reporter 56,
264-267.
THANASSOULOPOULOS, C.C., BIRIS,D.A. & TjAMOS, E.C. (1979) Survey of verticillium wilt of olive trees in
Greece. Plant Disease Reporter 63, 936-940.
THANASSOULOPOULOS, C.C., BIRIS,D.A. & TJAMOS, E.C. (1980) Dissemination of Verticilliunz propagules in
olive orchards by irrigation water. In Proceedings of the 5th Congress of the Mediterraneun
Phytopathological Union, pp. 52-53. Patras (GR).