Morpho-molecular identification of Alternaria alternata causing leaf spot on loquat in Pakistan
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Morpho-molecular identification of Alternaria alternata causing leaf spot on loquat in Pakistan Muhammad Fahim Abbas 1
&
Farah Naz 1 & Sana Batool 2 & Misbah Iqbal Qamar 3 & Anam Moosa 4
Received: 19 April 2019 / Accepted: 21 January 2020 # Società Italiana di Patologia Vegetale (S.I.Pa.V.) 2020
Keywords Loquat . Pakistan . Alternaria . First evidence
Loquat (Eriobotrya japonica) is an important sub-tropical fruit of Pakistan. During the year 2014-15, circular reddish brown leaf spots with concentric rings surrounded by yellow halo were observed in Taxila (33.74° N, 72.78° E), Wah Cantt (33.77° N, 72.75° E), Khanpur (33.80° N, 72.93° E), Tret (33.83° N, 73.30° E), Chatar (33.77° N, 73.24° E), Murree (33.90° N, 73.39° E), Kalar Kahar (32.78° N, 72.70° E) and Choa Saidan Shah (32.71° N, 72.98° E). The leaves (3 cm2) were sterilized by 75% ethanol, rinsed twice in sterile distilled water, blotted dry and transferred in PDA. The samples were incubated at 25 ± 2°C for 7 days and golden brown colonies which later turned black were observed. The conidia were single or in short chains, obpyriform, obclavate measuring ranged from 11 to 41 x 6.4 to 18 μm with 0 to 3 longitudinal and 2 to 6 transverse septa. Spore suspensions (106 conidia ml–1) of PAK38 and PAK39 isolates were sprayed on three healthy asymptomatic detached leaves and sterile distilled water was used as negative control. The experiment was repeated twice, leaves were incubated at 25 ± 2°C in sterile glass chambers. Symptoms were recorded 5 days after inoculation, while no symptoms were recorded on negative control. Fungal isolates were re-isolated from the diseased portions and isolated fungi were identified as Alternaria alternata. For the purpose of molecular identification, the sequences from internal * Muhammad Fahim Abbas [email protected] 1
Department of Plant Pathology, Pir Mehr Ali Shah Arid Agriculture University Rawalpindi, Rawalpindi, Pakistan
2
Department of Plant Protection, Northwest Agriculture and Forest University, Xianyang, China
3
Department of Plant Pathology, College of Agriculture, University of Sargodha, Sargodha, Pakistan
4
Department of Plant Pathology, University of Agriculture, Faisalabad, Pakistan
transcribed spacer regions (ITS) and Beta tubulin (BT) gene of PAK38 (GenBank accession Nos. KT280011 and KY126829) and PAK39 (KT312818 and KY113113) were submitted to National Center for Biotechnology Information (NCBI). Both isolates were exhibiting 99% genetic homology with previously reported ITS region (KT280009, KF039678 and KT280010) and BT (KU512287 and KT345697) gene sequences of A. alternata in the public database of NCBI. A. alternata has been reported to cause leaf spots and fruit rot of loquat in Taiwan, Greece and Iran (Tziros 2013; Mirhosseini et al. 2015) but to our knowledge this is the first report of A. alternata causing leaf spots and fruit rot on loquat in Pakistan.
References Tziros GT (2013) Alternaria alternata causes leaf spot and fruit rot on loquat (Eriobotrya japonica) in Greece. A
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