SYSTEMATICS OF SOME GILLED

MUSHROOMS FROM SHANGLA DISTRICT,
KHYBER PAKHTUNKHWA, PAKISTAN

Anum Saeed
MSF1800122

Supervisor:
Dr. Sana Jabeen

Department of Botany
Division of Science and Technology
University of Education, Township campus, Lahore.
INTRODUCTION
 GILLED VS NON-GILLED
MUSHROOMS

http://www.asergeev.com/pictures/archives/compress/ https://en.wikipedia.org/wiki/Boletus_pallidoroseus#/media/
File:Boletus_pallidoroseus_245628.jpg
2017/2154/14x.htm
 IMPORTANCE

• Gilled mushrooms are ecologically and economically important.

• Decomposers, symbiotic partners and pathogens.

• Forestry and agriculture.

 SPECIES DIVERSITY

Gilled
Mushrooms

Agaricales
Families Species
+
32 15,000 approx.
Russulales

Genera 500
approx.
(Mueller & Schmit, 2007; Thiers & Halling, 2017).
 METHODS OF
IDENTIFICATION

• In past, the identification and naming of fungi was based on morphological description

• In addition to morphological features certain microscopic features were also used for
identification

• Morphological data is insufficient for the characterization and identification of gilled

mushrooms.

• Now-a-days molecular techniques are used for the investigation of problems related to
fungal taxonomy

• This research work deals with the identification of gilled mushrooms collected from
different areas of Shangla district, Khyber Pakhtunkhwa, Pakistan
(Schoch et al., 2012 ; Hibbett & Taylor, 2013; Ullah et al., 2017; Hussain et al., 2018; Jabeen et al., 2019)
RESEARCH METHODOLOGY

Sampling Site
 MORPHOLOGICAL STUDY

• Morphological characteristics were based on the pictures of samples.

• Significant characters include:
• Size
• Shape
• Color of pileus and stipe
MUNSELL COLOR CHART
• Compound electric microscope
• KOH (5%) for rehydrating the tissues
• Congo red for staining tissues
• Pileus anatomical study includes:
• Cystidia
• Basidia
• Basidiospores from gills
• Stipe anatomical study includes:
• Hyphae and clamp connection
• Drawing were made using the pictures that were taken directly from microscope.
• Measurements were recorded using calibrated Motic Images Plus 2.0 software.
 ANATOMICAL STUDY

MOUNTING MEDIA
KOH (5%)
Congo red (1%)
 ANATOMICAL FEATURES

Basidia Stipitipellis

Cystidia
Basidiospores
MEASUREMENTS WERE RECORDED USING CALIBRATED MOTIC IMAGES
PLUS 2.0
 MOLECULAR STUDY

DNA Extraction
Extract-N-Amp kit used for DNA extraction.

PCR and Sequencing

Fungal specific primer was used for
amplification and sequencing of ITS region of
nrDNA by BGI China.
BIOEDIT SEQUENCE ALIGNMENT EDITOR VERSION 7.2.5
 BLAST AT NCBI

BLAST at NCBI (http://www.ncbi.nlm.nih.gov/).

(http://www.ncbi.nlm.nih.gov/).
MULTIPLE ALIGNMENTS
PHYLOGENETIC ANALYSIS
MEGA 6 SOFTWARE
 RESULTS

Total samples
18

Total genera New identified New records

species
13 4 12
Conocybe Fayod
Conocybe juniana (Velen.) Hauskn. & Svrček, Österr. Z. Pilzk. 8, 46 (1999)
Cortinarius (Pers.) Gray
Cortinarius uliginosus Berk., Outlook British Fungi (London) 190 (1860)
Cortinarius (Pers.) Gray
Cortinarius ahmadii nom. prov.

Etymology: The specific epithet “ahmadii” refers to the Sultan Ahmad, pioneer pakistani
mycologist.
 97 AY669665 Cortinarius hinnuleus
64 AY083184 Cortinarius hinnuleus
AY083183 Cortinarius hinnuleus
1
MF696141 Cortinarius conicoumbonatus
93 MF696139 Cortinarius conicoumbonatus
MG786242 Cortinarius cf. hinnuleus
9 74 MG739379 Cortinarius sp
SJ169 Cortinarius ahmadii
AJ534713 Cortinarius sp.
21 MT112168 Cortinarius sp
NR 166355 Cortinarius brunneocarpus
27
71 MN738695 Cortinarius sp
MT112145 Cortinarius sp
15
DQ499462 Cortinarius hinnuleoarmillatus
DQ499463 Cortinarius hinnuleoarmillatus
89 DQ499460 Cortinarius hinnuleoarmillatus
DQ499464 Cortinarius hinnuleoarmillatus
NR 131790 Cortinarius hinnuleoarmillatus
DQ499461 Cortinarius hinnuleoarmillatus
MG136827 Cortinarius hinnuleocervinus
65 KU041730 Cortinarius badioflavidus
KU041731 Cortinarius badioflavidus
82 KU041733 Cortinarius badioflavidus
17
MH266412 Cortinarius sp
KU041732 Cortinarius badioflavidus
57 KU041724 Cortinarius badioflavidus
NR 153055 Cortinarius badioflavidus
KU041727 Cortinarius badioflavidus
7 KU041726 Cortinarius badioflavidus
53 KU041728 Cortinarius badioflavidus
MF696139 Cortinarius conicoumbonatus
AY669658 Cortinarius umbrinolens
HQ604701 Cortinarius cf. umbrinolens
97 KX449461 Cortinarius umbrinolens
14 JQ711811 Cortinarius sp
KY287710 Cortinarius sp.
29 KY287725 Cortinarius sp.
81 NR 154868 Cortinarius fulvopaludsus
MG136823 Cortinarius fulvopaludosus
KX897435 Cortinarius sp.
DQ822800 Cortinarius sp.
2 44 FJ039538 Cortinarius sp
59 HQ604721 Cortinarius leucopus
95 FJ039539 Cortinarius vernus
JQ711860 Cortinarius sp.
5
EU821655 Cortinarius sp.
70 DQ102683 Cortinarius cf. saniosus
84
84 AJ534712 Cortinarius sp
NR 131829 Cortinarius politus
KP165545 Cortinarius uraceisporus
5 41 KP165544 Cortinarius uraceisporus
MT112163 Cortinarius sp99
98 DQ497190 Cortinarius rubrovioleipes
MG773850 Cortinarius rubrovioleipes
FJ717563 Cortinarius boulderensis
FJ039559 Cortinarius
EU652364 Cortinarius boulderensis
75 NR 121207 Cortinarius boulderensis
44
DQ499466 Cortinarius boulderensis
EU652362 Cortinarius boulderensis
99 EU652363 Cortinarius boulderensis
EU852803 Cortinarius boulderensis
EU837218 Cortinarius boulderensis
99
FJ039558 Cortinarius boulderensis
EU837216 Cortinarius boulderensis
NR 131791 Cortinarius pseudobovinus
99 DQ499465 Cortinarius pseudobovinus
DQ367900 Cortinarius traganus
FJ039568 Cortinarius traganus
FJ157105 Cortinarius traganus
FJ717573 Cortinarius traganus
99 FJ039567 Cortinarius traganus
FJ039570 Cortinarius traganus
FJ717574 Cortinarius traganus
FJ717571 Cortinarius traganus
95 JX045678 Cortinarius neosanguine
FJ157089 Cortinarius neosanguine Outgroup
KP087973 Cortinarius birkebakii
77 FJ039593 Cortinarius birkebakii

0.01
Cortinarius (Pers.) Gray
Cortinarius orellanus Fr., Epicrisis Systematis Mycologici (Upsaliae) 288 (1838)
Cortinarius (Pers.) Gray
Cortinarius rubellus Cooke. Grevillea, 16, 44 (1887)
Hygrophorus Fr.
Hygrophorus sordidus Velen. České Houby 5, 912 (1922)
Hygrophorus Fr.
Hygrophorus gliocyclus Fr., Öfversigt Kungliga Svenska vetenskaps Akademiens
Förhandlingar 18, 27 (1861)  
Hypholoma Fr.
Hypholoma capnoides (Fr.) Pers. Kreisel Handbuch für Pilzfreunde 72 (1871)
Inocybe (Fr.) Fr.
Inocybe acrystallis nom. prov.
Etymology: The specific epithet refers to the absence of crystals on apex of cystidia.
Inosperma (Kühner) Matheny & Esteve-Rav.
Inosperma maculatum (Boud.) Matheny & Esteve-Rav., Hobbs & Esteve
Raventós, Mycologia 21 (2019)
Lepiota (Pers.) Gray
Lepiota magnispora Murrill, Mycologia 4, 237 (1912)
Lepista (Fr.) W.G. Sm.
Lepista tarda (Peck) Murrill, North American Flora (New York) 10, 110 (1917)
Lepista VS Infundibulicybe

Peck, C. H. (1897). New species of fungi. Bulletin of the torrey Botanical Club, 24(3),

137-147.
Leucoagaricus Locq. ex Singer 
Leucoagaricus albiceratus nom. prov.
Etymology: The specific epithet refers to the pale yellow colour of the pileus.
Leucoagaricus Locq. ex Singer
Leucoagaricus brunneogriseonus nom. prov.
Etymology: The specific epithet refers to the brownish grey colour of the pileus.
Mycena (Pers.) Roussel
Mycena capillaripes Peck, Trustees State Museum of Natural History, New York 41,
63 (1888)
Psathyrella (Fr.) Quél.
Psathyrella candolleana (Fr.) Maire, Mémoirs of the Society of Natural Sciences of
Morocco 45, 112 (1937)
Tricholomopsis Singer
Tricholomopsis flammula Métrod ex Holec, Journal of the National Museum
(Prague), Natural History Series 178, 8 (2009)
Volvariella Speg.
Volvariella bombycina (Schaeff.) Singer, Lilloa 22, 401 (1951)
 CONCLUSION

• During this investigation 18 species were identified among these 4 represented new species
and 12 new records from Pakistan.

• From this data, it is concluded that Shangla district is very rich in diversity of gilled
mushrooms with many unidentified taxa. More exploration is needed to identify and report
these taxa up to saturation point.
 ACKNOWLEDGEMENTS

• Dr. Sana Jabeen (Research Supervisor)

• Prof. Mohammad Alam Saeed (Director of Division of Science and Technology)

• Dr. Zahra Noreen (Ex Chairperson Department of Botany)

• Prof. Dr. Abrar Hussain (Chairperson Department of Botany)

• Dr. Sadiq Ullah

• Lab fellows, class fellows and friends

• Family
Thank
You