Molecular identification of a new phytoplasma associated with alfalfa witches'-broom in Oman

A. J. Khan, S. Botti, A. M. Al-Subhi, D. E. Gundersen-Rindal, A. F. Bertaccini

Research output: Contribution to journalArticle

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Abstract

Alfalfa (Medicago sativa) plants showing witches'-broom symptoms typical of phytoplasmas were observed from Al-Batinah, Al-Sharqiya, Al-Bureimi, and interior regions of the Sultanate of Oman. Phytoplasmas were detected from all symptomatic samples by the specific amplification of their 16S-23S rRNA gene. Polymerase chain reaction (PCR), utilizing phytoplasma-specific universal primer pairs, consistently amplified a product of expected lengths when DNA extract from symptomatic samples was used as template. Asymptomatic plant samples and the negative control yielded no amplification. Restriction fragment length polymorphism profiles of PCR-amplified 16S-23S rDNA of alfalfa using the P1/P7 primer pair identified phytoplasmas belonging to peanut witches'-broom group (16SrII or faba bean phyllody). Restriction enzyme profiles showed that the phytoplasmas detected in all 300 samples belonged to the same ribosomal group. Extensive comparative analyses on P1/P7 amplimers of 20 phytoplasmas with Tru9I, Tsp509I, HpaII, TaqI, and RsaI clearly indicated that this phytoplasma is different from all the other phytoplasmas employed belonging to subgroup 16SrII, except tomato big bud phytoplasma from Australia, and could be therefore classified in subgroup 16SrII-D. The alfalfa witches'-broom (AlfWB) phytoplasma P1/P7 PCR product was sequenced directly after cloning and yielded a 1,690-bp product. The homology search showed 99% similarity (1,667 of 1,690 base identity) with papaya yellow crinkle (PapayaYC) phytoplasma from New Zealand. A phylogenetic tree based on 16S plus spacer regions sequences of 35 phytoplasmas, mainly from the Southern Hemisphere, showed that AlfWB is a new phytoplasma species, with closest relationships to PapayaYC phytoplasmas from New Zealand and Chinese pigeon pea witches'-broom phytoplasmas from Taiwan but distinguishable from them considering the different associated plant hosts and the extreme geographical isolation.

Original languageEnglish
Pages (from-to)1038-1047
Number of pages10
JournalPhytopathology
Volume92
Issue number10
Publication statusPublished - Oct 1 2002

Fingerprint

witches' broom
Oman
Phytoplasma
alfalfa
16SrII (Peanut WB group)
polymerase chain reaction
papayas
Chinese pigeon pea witches'-broom phytoplasma
Tomato big bud phytoplasma
sampling
faba beans
Medicago sativa
signs and symptoms (plants)
peanuts
Taiwan
molecular cloning

ASJC Scopus subject areas

  • Plant Science

Cite this

Khan, A. J., Botti, S., Al-Subhi, A. M., Gundersen-Rindal, D. E., & Bertaccini, A. F. (2002). Molecular identification of a new phytoplasma associated with alfalfa witches'-broom in Oman. Phytopathology, 92(10), 1038-1047.

Molecular identification of a new phytoplasma associated with alfalfa witches'-broom in Oman. / Khan, A. J.; Botti, S.; Al-Subhi, A. M.; Gundersen-Rindal, D. E.; Bertaccini, A. F.

In: Phytopathology, Vol. 92, No. 10, 01.10.2002, p. 1038-1047.

Research output: Contribution to journalArticle

Khan, AJ, Botti, S, Al-Subhi, AM, Gundersen-Rindal, DE & Bertaccini, AF 2002, 'Molecular identification of a new phytoplasma associated with alfalfa witches'-broom in Oman', Phytopathology, vol. 92, no. 10, pp. 1038-1047.
Khan AJ, Botti S, Al-Subhi AM, Gundersen-Rindal DE, Bertaccini AF. Molecular identification of a new phytoplasma associated with alfalfa witches'-broom in Oman. Phytopathology. 2002 Oct 1;92(10):1038-1047.
Khan, A. J. ; Botti, S. ; Al-Subhi, A. M. ; Gundersen-Rindal, D. E. ; Bertaccini, A. F. / Molecular identification of a new phytoplasma associated with alfalfa witches'-broom in Oman. In: Phytopathology. 2002 ; Vol. 92, No. 10. pp. 1038-1047.
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