DNA barcoding, Phylogenetic Analysis and disease transmission study in Medicago sativa (16Sr-II-D) infected by Phytoplasma bacteria | [Abstract Book on International Conference on Food and Applied Sciences (ICFAS-23) 3-5 August 23 • 2023]

Author(s):

1. Samina Tanwir: Dr. Jam Laboratory, Dept. of Entomology, University of Agriculture Faisalabad; Plant Stress Physiology and Molecular Biology Lab, Dept. of Botany, University of Agriculture Faisalabad, Pakistan

2. Jam N Ahmad: Dr. Jam Laboratory, Dept. of Entomology, University of Agriculture Faisalabad; Plant Stress Physiology and Molecular Biology Lab, Dept. of Botany, University of Agriculture Faisalabad, Pakistan

3. Muhammad Z Sharif: Dr. Jam Laboratory, Dept. of Entomology, University of Agriculture Faisalabad, Pakistan

4. Piotr Trebicki: Agriculture Victoria Research, Victoria, Australia

Abstract:

Medicago sativa known as lucerne or alfalfa is a good fodder crop in Pakistan. Its continuous cultivation and sustainable production is important for the better growth and productivity of cattle. Phytoplasma is an emerging disease worldwide affecting various crops including lucerne. The study was conducted to identify phytoplasma and the potential insect vectors responsible for its transmission. The sampling was done from Multan, RY Khan and Faisalabad, Punjab during 20172020. The detection of phytoplasma in infected samples through staining and electron microscope indicated the presence of phytoplasma bodies in sieve tube cells. It was further confirmed by Nested Polymerase Chain Reaction (Nested-PCR) by the amplification of 1.8 kb and 1.2 kb DNA fragment of 16SrDNA using the primer pairs P1/P7 and RI6F2n/R2 respectively. Restriction fragment length polymorphism (RFLP) analysis also showed a similar pattern of bands formation associating with 16S ribosomal-DNA of 16SrII-D subgroup linked with sesame phyllody (16SrIID) group. DNA sequencing and phylogenetic analysis exhibited >99% homology of Pakistani isolates (MT614018.1 and MT614019.1) with submitted phytoplasma strain “Ca. P. australasia” of 16SrII-D subgroup. The potential insect vectors, Orosius orientalis, Orosius argentatus, and Laudelphax striatellus captured from infected plants were detected positive. The transmission study confirmed their vector status for alfalfa phyllody diseases transmission. To our information, this is first detection of phytoplasma infestation and the insect vectors associated with Medicago sativa in Pakistan. The 16Sr-II D group of phytoplasma is spreading widely in many crops so, control is essential to stop it from entering into other economically important crops.

Page(s): 155-155

DOI: DOI not available

Published: Journal: Abstract Book on International Conference on Food and Applied Sciences (ICFAS-23) 3-5 August 23, Volume: 0, Issue: 0, Year: 2023

Keywords:

sequencing Nested PCR Phylogenetic analysis , Alfalfa phytoplasmaMedicago sativaInsect vectors 16SrIID phytoplasma DNA

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