Almond witchesÕ broom (AlmWB) is a destructive disease in several provinces in Iran. Association of phytoplasma with the disease has been established previously. In the present work two phytoplasmas from Khafr (KAlmWB) and Neyriz (NAlmWB) in the Fars Province were compared by biological and molecular analysis. Both infected bitter almond, wild almond, peach and nectarine but not apple and pear, by grafting. In bitter almond the symptoms induced by KAlmWB consisted of severe proliferation, internode shortening and leaf size reduction. In contrast, NAl-mWB caused leaf necrosis, dieback and death. KAlmWB was transmitted to periwinkle and eggplant and from experimentally infected periwinkle to almond by dodder. It was also transmitted from eggplant to eggplant, ornamental eggplant and tomato by grafting. Under similar test conditions, NAlmWB was not transmitted to herbaceous plants by dodder. Phylogenetic analysis of 16S-23S rDNA spacer region (SR) sequences placed both strains in the pigeon pea wit-chesÕ broom (PPWB) group. However, based on phylogenetic and putative restriction site analyses and sequence homology, NAlmWB was identical with the Lebanese AlmWB phytoplasma, while KAlmWB was closer to the Knautia arvensis phyllody (KAP) agent. Clustering of KAlmWB with KAP was confirmed by analysis of full length 16S rDNA sequence. On the basis of host range, dodder transmission, host range, symptomatology and molecular analyses of 16S rDNA and SR, two different phytoplasmas related to PPWB group were associated with AlmWB disease in Iran. KAlmWB phytoplasma is being reported as a new phytoplasma of AlmWB disease. Graft inoculationOne-year-old seedlings of fruit trees and 8-week-old herbaceous plants were used as test indicators and grafted with small auxillary shoots from infected or www.blackwell-synergy.com
During 2010-14 surveys in the major sesame growing areas of Fars, Yazd and Isfahan provinces (Iran), genetic diversity and vector transmission of phytoplasmas associated with sesame phyllody were studied. Virtual RFLP, phylogenetic, and DNA homology analyses of partial 16S ribosomal sequences of phytoplasma strains associated with symptomatic plants revealed the presence of phytoplasmas referable to three ribosomal subgroups, 16SrII-D, 16SrVI-A, and 16SrIX-C. The same analyses using 16S rDNA sequences from sesame phyllody-associated phytoplasmas retrieved from GenBank database showed the presence of phytoplasmas clustering with strains in the same subgroups in other Iranian provinces including Bushehr and Khorasan Razavi. Circulifer haematoceps and Orosius albicinctus, known vectors of the disease in Iran, were tested for transmission of the strains identified in this study. C. haematoceps transmitted 16SrII-D, 16SrVI-A, and 16SrIX-C phytoplasmas, while O. albicinctus only transmitted 16SrII-D strains. Based on the results of the present study and considering the reported presence of phytoplasmas belonging to the same ribosomal subgroups in other crops, sesame fields probably play an important role in the epidemiology of other diseases associated with these phytoplasmas in Iran.
Phyllody is a destructive disease of sesame (Sesamum indicum L.) in Iran. The major symptoms of the disease are floral virescence, phyllody and proliferation. Other symptoms which sometimes accompany the disease are yellowing, cracking of seed capsules, germination of seeds in the capsules and formation of dark exudates on the foliage. Light microscopy of hand‐cut sections of sesame and colza (Brassica napus L. cv. Oro) stems treated with Dienes' stain showed blue areas in the phloem region of phyllody infected plants. Mycoplasma‐like bodies were found in the sieve cells of infected sesame stems when thin sections were examined m an electron microscope. Sesame phyllody was successfully transmitted from sesame to sesame by grafting. Among various leafhoppers collected in sesame fields only Neoaliturus haematoceps transmitted the disease. This is the first report on the identification of a Mycoplasma‐like organism (MLO) as the cause of sesame phyllody and N. haematoceps as an MLO vector in Iran. In host range studies using the leafhopper vector, only B. napus cv. Oro, Lepidium sativum, Catharanthus roseus, Lactuca sp. and Portulaca oleracea, but not 17 other species, developed symptoms. The species of vector and host range of MLO indicate that sesame phyllody in Iran is different from that reported in India and Upper Volta.
In 2001, a disease tentatively named Iranian cabbage yellows (ICY) was observed in cabbage fields of Zarghan (Fars Province, Iran). The major symptoms of the disease were yellowing, little leaves, plant stunting, opening of the head, and proliferation of the buds at the base of the stem into a witches'-broom. Among leafhoppers collected in cabbage fields, only Circulifer haematoceps transmitted the ICY agent. The disease agent was transmitted by the leafhopper from cabbage to cabbage, cauliflower, rape, and periwinkle, causing phytoplasma-type symptoms in these plants. Polymerase chain reaction (PCR) using phytoplasma-specific primer pair P1/P7 and nested PCR using P1/P7 and R16F2n/R16R2 primer pairs amplified products of expected size (1.8 and 1.2 kb, respectively) from symptomatic cabbage plants. Both restriction fragment length polymorphism (RFLP) of nested PCR products (1.2 kb) and phylogenetic analyses of 16S–23S rDNA spacer region sequence indicated that the ICY phytoplasma had the closest relationship to subgroup A members of the clover proliferation group, including beet leafhopper-transmitted virescence agent, ‘Candidatus Phytoplasma trifolii’, Columbia Basin potato purple top phytoplasma, and vinca virescence phytoplasma. Cabbage is reported as a new natural host to the 16SrVI group of phytoplasmas.
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