World Applied Sciences Journal 32 (2): 193-200, 2014
ISSN 1818-4952
© IDOSI Publications, 2014
DOI: 10.5829/idosi.wasj.2014.32.02.14520
Corresponding Author: M.Y. Awad, Department of Botany, Faulty of Science, Benha University, Benha, Qalubia, Egypt.
193
Finger Printing of Certain Seed-Borne Fungi on Imported Tomato Seeds in Egypt
H.M. Emara, E.E. Hafez, M.A. El-shafey, A.A. Khattab and M.Y. Awad
1 2 1 1 1
Department of Botany, Faulty of Science, Benha University, Benha, Qalubia, Egypt
1
City of Scientific Research and Technology Applications,
2
Arid Lands Cultivation Research Institute (ALCRI), Plant Protection and
Biomolecular Diagnosis Department, New Borg El Arab City, Alexandria, Egypt
Abstract: Egypt imported different cultivars of tomato seeds from different companies. Different microorganisms
associated these seeds, among them the seed borne fungi. Seeds of five different companies were collected and
subjected to fungal isolation using Standard blotter Technique and agar plate technique. Results revealed that
five species were observed. The observed fungi were morphologically identified under microscope and the
results were confirmed using specific PCR with primers (ITS-1 & ITS-4). Amplicones with molecular size 600 bp
was observed. The bands were purified and subjected to DNA sequence. The sequence analysis revealed that
the five fungi are Trichoderma longibrachiatum, Aspergillus fumigatus, A. fumigatus strain TFR-8, A. flavus
and A. oryzae. The sequences were submitted into the Gene Bank and were deposited under accession no.
KJ424352, KJ424353, KJ734707, KJ734708 and KJ742820. T. longibrachiatum was used as biocontrol agent
against several plant pathogenic fungi and the results showed significant growth inhibition for the treated fungi
with percentage ranging from (49.7 % to 73.4%). Using T. longibrachiatum strain as abiological control agent
is effective, applicable, safe and commercial.
Key words: Biological control Egypt Seed borne fungi Specific PCR Tomato Trichoderma
longibrachiatum
INTRODUCTION alternative to the use of synthetic fungicides and has
Tomato (Lycopersioon esoulentum Mill.) is an decades [9]. Various fungi (e.g. Trichoderma have shown
important vegetable crop cultivated over an (216,400,000) good biocontrol potential [10, 11]. Biological control is
hectare area in Egypt. Tomato seeds such as all the seeds considered a sustainable and safe [12]. It is relative free
play an important role in crop production, however the from problems of pest resistance and is extremely safe to
seeds are attacked by different fungi either human health [12]. Biopesticides often are effective in
saprophytically or within the tissue of embryo [1]. The very small quantities and often decompose quickely,
disease caused by fungi may lead to seed abortion or thereby resulting in lower exposure and largely
elimination of germination capacity [2]. At least 10 avoiding the pollution problems caused by conventional
different fungi have been reported as seed-borne fungi pesticides [13]. In general, cultural practices are less
on tomato from different parts of the world, which likely to be in conflict with natural enemies than pesticide
include Alternaria alternate, A. solani, Aspergillus spp., [13]. In contrast to chemical control, biological control
Botrytis cinerea, Cheatomium globosum, Cladosporium lessens long-term damage to the environment by
spp., Colletotrichum dematium, Curvularia lunata, persistent chemicals [13]. It may be a more economical
F. equiseti, F. moniliforme, F. oxysporum, F. solani, alternative to some insecticide and fungicides
Macrophomina phaseolina, Penicillium spp., Phoma applications [13, 14]. Unlike most fungicides biological
destouctiva, Pythium aphanidermatum, Rhizoctonia control is often very specific for a particular pathogen,
solani, Verticillium spp. [3-8]. Biological control offers an thus other beneficial microorganisms can go completely
become a well established principle over the last few