Abstract

Endophytes are the plant mutualists that live asymptomatically inside plant tissue and are found in nearly whole plant kingdom. Endophytic fungi receive shelter and nutrition from host plants and in return provide great advantages to the host. Grasses are a useful forage species and are of great agricultural and socio-economic value. The presence of endophytes in these grasses provide protection, persistence and improved yield against herbivores, insects, pathogens, drought and several other biotic and abiotic stresses. This review summarizes traditional and modern molecular techniques to identify endophytes from turf and forage grasses. Traditional approaches include direct observation, staining, laser micro dissection and pressure catapulting and cultivation-dependent methods that provide a morphological identification of endophytic mycobiota in grass tissues. Earlier studies on endophytes using these methods resulted in several technical implications which molecular approaches are able to solve now-a-days. Molecular approaches include DNA extraction, PCR based DNA Fingerprinting techniques, Denaturing Gradient Gel Electrophoresis, Sanger sequencing, Pyrosequencing, Immunoblot assay, Biosensors, DNA Barcoding and Molecular Phylogenetics etc. A comparison of these detection techniques will facilitate other researchers as well to develop new ways for the detection of endophytes that will contribute to the improvement of grassland in future.

Highlights

  • Fungal endophytes are the organisms living in a plant for at least a part of its life, without causing any apparent disease

  • This review summarizes traditional and modern molecular techniques to identify endophytes from turf and forage grasses

  • We can say that mutualistic root inhibited or mycorrhizal fungi that associates themselves with plants and they belong to Ericaceae and Orchidaceae are called as endophytes (Bayman, Lebron et al, 1997)

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Summary

A Brief History of Endophyte Detection Techniques in Grasses

Rana Zaheer Ahmad, Fuad Ameen , Rida Khalid, Mousa A. Alghuthaymi, Reem Alsalmi6 & Chunjie Li1 1College of Pastoral Agriculture Science and Technology, Lanzhou University, Lanzhou, China 2Department of Botany and Microbiology, College of Science, King Saud University, Riyadh 11451, Saudi Arabia 3Department of Marine Biology, College of environmental & Marine Science, Al-Hodeidah University, Al-Hodeidah, Yemen 4University of the Punjab, Lahore, Pakistan 5Biology Department, Science and Humanities College, Shaqra University, Saudi Arabia 6Biology Department, College of science, Al-Baha University, Saudi Arabia. Received: June 3, 2019 Accepted: June 28, 2019 Online Published: July 27, 2019 doi:10.5539/sar.v8n3p66

Introduction
Morphological Identification of Endophytic Fungi from Grasses
Direct Observation Method
Staining Methods
Cultivation Dependent Method
Laser Micro Dissection and Pressure Catapulting Method
DNA Extraction
PCR based DNA Fingerprinting Techniques
RAPD Analysis
RFLP and AFLP Analysis
SSR Analysis
Denaturing Gradient Gel Electrophoresis
DNA Sequencing Methods
Sanger Sequencing
Pyrosequencing
DNA Barcoding
Tissue Print Immunoblot Assay
Biosensors
Molecular Phylogenetics
Conclusions and Future Studies
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