First Record of Lepista sordida (Schumach) Singer in Eastern North Africa

Abstract

A NEW record of Lepista sordida was collected from a lemon fruit farm in El-Sinania at Damietta District of North delta of Egypt in December, 2014. It was identified using morphological (macro and microscopic) and molecular techniques. Complete description was preformed for the collected fresh fruiting bodies and isolated pure culture. Radial growth rate of culture was estimated on Potato dextrose and Malt extract agar media (8.5 & 7.8 mm/ day, respectively). Lepista sordida and L. nuda are very close in morphological characters; hence, the identification was confirmed by DNA sequence analysis of the ribosomal 5.8S rRNA gene including the flanking internal transcribed spacers (ITS). Then, its taxonomic position among some genera of Tricholomataceae and its relation with some other Lepista species was discussed. The surrounding habitat was also observed and environmental conditions were recorded as Temperature degree (29.7oC), relative humidity (RH: 44.6) and soil moisture was (5.56). Lepista sordida was reported from South Africa and Nigeria (in the South), Algeria and Tunisia (in the North-west) and this is first record in North-East Africa.

Full text

A
NEW record of Lepista sordida was collected from a lemon fruit farm in El-Sinania at
             
morphological (macro and microscopic) and molecular techniques. Complete description
was preformed for the collected fresh fruiting bodies and isolated pure culture. Radial growth
rate of culture was estimated on Potato dextrose and Malt extract agar media (8.5 & 7.8 mm/
day, respectively). Lepista sordida and L. nuda are very close in morphological characters;


Lepista species was
          
oC), relative humidity (RH: 44.6) and soil moisture was
(5.56). Lepista sordida

Keywords: , Lepista sp.    

9
First Record of Lepista sordida (Schumach) Singer in Eastern North
Africa
A.A. El-Fallal, A.K.A. El-Sayed and H.M. El-Gharabawy#
Botany & Microbiology Department, Faculty of Science, Damietta University,
Damietta El-Gededa, PO 34517, Egypt.
Introduction
Lepista 
with many edible species such as L. sordida, L.
nuda and L. saeva 

     
       

     
Lepista sordida (Schumach) Singer was reported
     
in Q. ilex     
    
      Eucalyptus
L. sordida in

Some of Lepista species have commercial
potentials. L. nuda presents antioxidant properties
(Murcia et al., 2002) and antibiotic activities
against many bacteria (Dulger et al., 2002).
 L. sordida produces two diterpenes
     
     
extracted from L. sordida possessed potent ant
proliferative effect on mice and human laryngeal
carcinoma Hep-2 cells (Miao et al., 2013), also
had antioxidant activity and retard aging effects

polysaccharides could be used as a potential
natural antitumor drug and attenuate age-related
diseases in humans.

phylogenetic relationships in the largest order;
(Moncalvo et al., 2002; Garnica et al.,

assessment of evolutionary relationships within
this order is necessary. Lepista

     
descriptions. So, molecular techniques such
      

Stott et al., 2005).


of an Egyptian Lepista mushroom that grows in
El-Sinania Farms at Damietta – Egypt.
The 7th Inter. Conf.”Plant & Microbial Biotech. & their Role in the Development of the Society”pp.111 -118 (2017)
#Corresponding author email: 

©2017 National Information and Documentaion Center (NIDOC)

112
Egypt. J. Bot. (2017)
A.A. EL-FALLAL, A.K.A. EL-SAYED AND H.M. EL-GHARABAWY
Materials and Methods
Morphology and identication
Descriptions of basidiomes were made according
to their external and internal morphology. For
external morphology the material was observed for
colour, texture, gills morphology, margin, and pileal
surface of basidiocarp. For internal morphology,
       
through gills, and mounted in also in Melzer’s


compound microscope having a combination of
10x eyepiece and 10x, 45x and oil immersion
    
using Canon digital camera. Measurements
     
micrometer or calibrated ocular.
Isolation into pure culture was carried out
      
according to the method of El-Gharabawy et al.
(2016). Small pieces of either inner layers of the
fruiting body tissue were cultured on to plates of

     
were incubated at 25°C and pure cultures were

       

growing cultures at 25°C.
DNA extraction
    
        
     
washed with sterile water and frozen with liquid
nitrogen followed by grinding with sterilized
   µl extraction buffer (Equal
        
       
Sulphate) was added to the micro tube sample and
       
the same volume of (25 Phenol: 24 Chloroform:
1 Isoamyl-alcohol) mixture was added, mixed by
inverting the tubes and centrifuged at 4°C for 10

was precipitated by isopropanol, washed with

and then stored at -20°C until used for PCR

PCR amplication and sequencing of ITS-5.8S
rRNA region
5´-
3´   5’-
3’) used for ampli-
      
        -
ONEER, South Korea.
PCR reaction was carried out using a thermal
     
mixture consisted of 4 µl of each primer (20 pmole
ml-1

  µl. PCR was performed with
initial denaturation at for 5 min, followed



      

Alignment and phylogenetic analyses
    
    
database (http://www.ncbi.nlm.nih.gov/) to match
       
       
      

      
    
and the sequences from the same species and
    

      
joining was performed using the maximum
     

only considered and represented next to the

estimated by 1000 bootstrap replicates.
Results
Morphological characteristics
    


     
gregarious and form a fairy ring, on soil and in
mixed lawn and usually in areas where leaf litter
collects  Lepista isolate had the

      
 
at Damietta district in 16th of December 2014, at
oC,
relative humidity (RH) was 44.6 and the soil
moisture was 5.56.

113
Egypt. J. Bot. (2017)
FIRST RECORD OF LEPISTA SORDIDA (SCHUMACH) SINGER IN EASTERN NORTH AFRICA
     
        
not distinguishable from Lepista nuda. Cap is
2 to 6 cm across; initially convex sometimes
with a slight umbo (Fig. 1C),   
developing a central depression at maturity (Fig.
1 D, E), usually with a slightly a wavy margin
and in-rolled; deep lilac, turning brown from the
centre in dry weather. Cap surface is smooth not
       
emarginated and crowded (Fig. 1E & F), the gills
are initially greyish lilac fading to buff with age.
Stem is 2 to 4 cm long and 4 to 10 mm in diameter;

     
parallel trama, 2 or 4 spored. Spores are ellipsoidal,
µm (Fig. 1H) nonamyloid, colourless,
hyaline, thin walled, roughened and ornamented
with tiny spines. Spore print is creamy-white.
Fig. 1. Fruit body external morphology of L. sordida



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A.A. EL-FALLAL, A.K.A. EL-SAYED AND H.M. EL-GHARABAWY

  
        
colonized all the Petridish within 8 days of
incubation. Culture showed more aggressive
Fig.2. Culture of Lepista sordida 
      
     
cottony surface. It also pins readily and easily on
agar with pale violet reverse.
ITS based identication

L. sordida 


L. sordida

L. sordida L. sordida
(KJ137272). Some other different L. sordida and
L. tarda strains showed less similarities reached
    
with L. nuda, L. saevaand L. personata strains.
Tricholoma mongolicum isoletes aligned with our
 
Clitocybe, Collybia and Lyophyllum
     
    


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FIRST RECORD OF LEPISTA SORDIDA (SCHUMACH) SINGER IN EASTERN NORTH AFRICA
      
sequence (Fig.3) clustered L. sordida 
in one clade with some other isolates of the
same L. sordida species possessing approximate
dissimilarity distance reached 0.015 with
Tricholoma mongolicum clade.
Fig. 3. Phylogenetic tree analysis based on the ITS1-5.8S-ITS2rDNA sequence alignment for L. sordida EGDA2
(AC: LN827702) with some other related genera and lepista species which possessed the best similarity.
The ITS sequence of Agaricussp (AC: AM930985) was used as outgroup to root the tree. The bootstrap
values 50 or above were represented next to the phylogenetic tree branches.
Discussion
Lepista sordida (Schumach) Singer is a
basidiomycete fungus that produces an excellent
tasting light purple mushroom. L. sordida usually
occurs so late in the season than other mushroom,
it was observed growing in December and late
     
only represented by Lyophyllum buxeum in the
North East Nile delta (El-Fallal, 2003, 2013),
until we recorded L. sordida   
2014. However, Lyophyllum is now belonging to
Lyophyllaceae (Species Fungorum  
Lepista
Tricholoma or a Clitocybe.
          


Tricholoma species typically have gills
notched at the stem, as do several Lepista species,
the spores of Tricholoma are white, never colored,
whereas L. sordida produces roughened pale buff
   Tricholoma caps
never change color on drying; in several Lepista
Tricholoma
cap margins typically incurve, young Lepista
cap margins typically inroll. Tricholoma species
      

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A.A. EL-FALLAL, A.K.A. EL-SAYED AND H.M. EL-GHARABAWY
are grayish while whitish Lepistas are typically
brownish or buff tinged.
L. sordida


fade from the centre toward the margin with age.

      L. sordida is
fairly easy to identify. It is probably most similar
in morphological characters to the closely related
L. nuda, however, L. sordida is more slender
than L. nuda and has more of a tendency to grow
in clusters, as is seen in Fig. 1. Hence, more
       
differentiation.
Interestingly, the phylogenetic tree based on the
L. sordida 
Tricholoma mongolicum, which is morphologically
different, in the same Lepista   
       
who renamed T. mongolicum to Leucocalocybe
mongolicum which clustered with Lepista irina.
      
fungi showed also that L. sordida and L. nuda
were in the same clade with L. mongolicum and
closely clustered with some species of Clitocybe
and Collybia (Cooper, 2014). However, Clitocybe
species typically have gills running down the
stem and in-rolled cap margins. Lepista species
with such characteristics may not be readily
distinguished, except that if gills run down stem,
they are nearly always short decurrent. White,
        
genera. Furthermore, Clitocybe species have no
purple colors. Most Lepista species have a growth
habit that is clustered, sub-clustered or at least
gregarious. Only C. dilatata in genus Clitocybe
appears to have a clustered growth habit. In the
   Clitocybe has white
spores while Lepista     

     L. tarda and L. sordida

morphologically similar except that L. tarda stem

Lepista. sordida as a valuable edible and
medicinal mushroom is widespread in northern

& Fujiie, 2005). L. sordida was successfully
        

     
molecular technique and a temperature of 25 - 30
°C was the best for mycelial growth. L. sordida,
as a fairy-ring-forming fungus, was examined
for its effect on the growth of turfgrass in Japan
      

       
the growth of plant roots (Choi et al., 2010).
Furthermore, L. sordida proved active role in lignin
degradation, dye removal and other industrial
applications. Laccases produced by L. sordida was
characterized by Cavallazzi et al. (2004).
      
sequence analysis and morphological characters
   Lepista sp. isolated from
Damietta District at North East Nile delta
is belonging to L. sordida species. It is also
more related to L. tarda and T. mongolicum (L.
mongolicum) than Clitocybe and Collybia spesies.
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A.A. EL-FALLAL, A.K.A. EL-SAYED AND H.M. EL-GHARABAWY
  Lepista sordida (Schumach) Singer  



Lepista sordida
2014

7.8 8.5


Lepista 
Lepista sordida5.5644.6


(Received 21/4/2017;
accepted 2/10/2017 )
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