چكيده لاتين :
Introduction: Madder plant with the scientific name of Rubia tinctorum (a member of Family Rubiaceae) is
one of the most important crops used for medical and industrial applications, is widely cultivated in Yazd
province, Iran. Madder root is widely applied in the dyeing industry. Cultivation of these plants is rather
common in Europe and Asia. In Iran madder was cultivated in ancient times in Tabriz, Urmia, Arak, Yazd, and
Fars but nowadays it is cultivated mainly in different cities in Yazd province especially in Bafq and Ardakan.
The madder plant's roots contain different polyphenolic compounds, such as 1,3-Dihydroxyanthraquinone
(purpuroxanthin), 1,4-Dihydroxyanthraquinone (quinizarin), 1,2,4-Trihydroxyanthraquinone (purpurin) and 1,2-
dihydroxyanthraquinone (alizarin).
The madder can grow up to 1.5 m in height. Their leaves produced in whorls of 4-7 star like around the
central stem with approximately 5-10 em long and 2-3 em wide. Its flowers are small with five pale yellow
petals in dense racemes followed by red to blackberries. The roots can be over a meter long, up to 12 mm thick
and the source of red dyes known as rose madder. Several factors such as fungal diseases during planting and
harvesting period infect madder plants. These factors, in addition to causing damage to madder plants reduce the
quality and quantity of color obtained from roots. Many of these fungi are associated with madder seeds and
infect plant after cultivation. Some fungi infect leaves and stems of madder. Some of these fungi such as
Fusarium, Aspergillus, and Penicillium are associated with madder seeds and in addition to a role as
phytopathogenic fungi produce dangerous mycotoxin effective on color quality of madder. So far from the aerial
parts of the madder plants such as leaves and stems, several fungi were reported. Septoriarubiae and Sporonema
punctiforme were reported as leaf spot agents. Furthermore, P/eospora ch/amydospora and Sporonea
punctiforme were isolated from Rubia spp. Fusarium species is one of the most abundant fungi that are able to
produce hazardous mycotoxin especially trichothecenes in the different growth stage of Madder. Fusarium
mycotoxins are among the main fungal mycotoxin contaminations in food and feed, and some human diseases,
such as Kashi neck diseases and esophageal cancer, have been epidemiologically associated with consumption of
trichothecenes. The most predominant trichothecenes found in crops are 8-keto trichothecenes (type B
trichothecenes). Based on the production and chemical structures of different 8-keto trichothecenes, several
trichothecene mycotoxins were identified that includes nivalenol (NIV), 4- acetylnivalenol ( 4-AcNIV),
deoxynivalenol (DON), 3-acetyldeoxynivalenol (3-AcDON) and 15-acetyldeoxynivalenol (15-AcDON). There
is a little knowledge about seed madder microtlora and given that the fungi with madder seeds can reduce the
power of viability, seedling and ultimately damage the quality and quantity of products, the present study was
aimed to the identification of fungi associated with seed madder and also fungi toxin-producing especially
Fusarium species.
Materials and Methods: During 2012, sampling was carried out form seeds of madder in important planting
areas in Yazd province, including Bafq and Ardakan. After culturing, fungal isolate was purified in potato
dextrose agar (PDA), carnation leaf agar (CLA), and CY20S (Czapek Yeast Extract Agar with 20 sucrose)
media. Fusarium, and Aspergillus species were identified by valid keys identification. The Fusarium isolates
were grown in liquid potato dextrose broth medium and total genomic DNA was extracted using the CTAB
method. Molecular identification ofF equiesti, F. semitectum and F poae was performed using specific-species
primers FEFI/2, SEM-1/2, and FpoF/R. Detection of fungi mycotoxins producing potential such as
Deoxynivalenol (DON) using Tri 13 primers was done. High-performance liquid chromatography (HPLC) was
used to confirm the produce NIV mycotoxins potential in Fusarium species.
Results and Discussion: 249 fungal strains were isolated from madder seed belonging to 6 genera of fungi
including Fusarium spp., Aspergillus spp., Penicillium spp., Alternaria spp., Rhizuctonia so/ani and Rhizopus
spp., which Fusarium isolates with 71 percent was the most frequency among fungi isolated. Among Fusarium
fungi isolated, F.solani (55 isolates) and F.oxysporum (41 isolates) were the most frequency. F. poae, F
semitectum, and F. equiseti ability to produce mycotoxins such as Deoxynivalenol (DON) that are harmful to
human health and animals as well as the effect on the yield of madder. Tri 13 gene involved in production DON
was detected in three Fusarium species (F. poae, F. semi tectum and F. equiseti). The results of HPLC showed
that all studied Fusarium fungi, have the potential to produce DON mycotoxins. The results of this study showed
that fungi associated with seeds madder are able to produce 8-keto trichothecene mycotoxins that they can be
dangerous for consumers.
Conclusions: Given that, this is the first report of fungi mycotoxins producing on seeds madder in Yazd
province, thus should be measured to control and reduce fungal agents in these products.
