NDLI: Sensitive Detection of a Phytoplasma Associated with Little Leaf Symptoms in Withania somnifera

Content Provider	Springer Nature Link
Author	Khan, J. A. Srivastava, P. Singh, S. K.
Copyright Year	2006
Abstract	Withania somnifera is an important medicinal plant native to the Indian-sub continent. Owing to the presence of a number of precious alkaloids, flavonoids and withanolides, it is widely used in the Indian and African systems of medicines. It is severely affected by phytoplasma present in the sieve tubes of phloem. With a view to micropropagate phytoplasma-free W. somnifera plants, an efficient and effective nested PCR-based system was developed for detection of associated phytoplasmas. Universal primers, designed from the 16S rDNA sequences of phytoplasmas, were applied in direct/nested-PCR. Total DNA extracts from leaf tissues of 33 suspected symptomatic and 11 non-symptomatic plants were subjected to direct PCR. The direct PCR products were subsequently employed as templates in nested PCR. The nested PCR could reamplify direct PCR products yielding a DNA fragment of 1.4 kb. A phytoplasma was detected in all the diseased plants and not from the healthy looking plants. Further, it was sensitive enough to amplify phytoplasma DNA obtained from crude DNA diluted up to 2500 times from naturally infected plants and also from various stages of in vitro-propagated diseased plants. Identical restriction fragment polymorphism enzyme profiles were obtained following restriction enzyme digestion of nested PCR products, obtained from five different plants, by EcoRI, AluI and RsaI restriction endonucleases. The developed nested PCR based system should facilitate indexing of the phytoplasma in different stages of in vitro-generated plants and probably identification of, as yet unknown, hosts and vectors of phytoplasma associated with phytoplasma disease of W. somnifera.
Starting Page	401
Ending Page	408
Page Count	8
File Format	PDF
ISSN	09291873
Journal	European Journal of Plant Pathology
Volume Number	115
Issue Number	4
e-ISSN	15738469
Language	English
Publisher	Kluwer Academic Publishers
Publisher Date	2006-06-20
Publisher Place	Dordrecht
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	detection elimination phytoplasma Withania somnifera Plant Pathology Plant Sciences Ecology
Content Type	Text
Resource Type	Article
Subject	Plant Science Horticulture Agronomy and Crop Science

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