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Witches'-Broom Disease of Lime Affects Seed Germination and Seedling Growth But Is Not Seed Transmissible

Witches'-broom disease of lime (WBDL), caused by ‘Candidatus Phytoplasma aurantifolia’, has devastated many Mexican lime orchards and is currently a threat to lime production in neighboring provinces of southern Iran. Several reports have suggested transmission and spread of WBDL phytoplasma through the seed of infected plants. In the present study, claims of seed transmission of this phytoplasma were examined. Fruit were collected from infected trees in the infested areas of Minab (Hormozgan Province) and from symptomless trees in noninfested areas. Lime seed from symptomless and witches'-broom–affected trees were sown in separate beds in an insect-proof screenhouse and the resulting seedlings were examined for phytoplasmal infection. Leaf, stem, and root samples were collected from both groups of seedlings every 3 months for 2 years and tested for WBDL phytoplasma using direct and nested polymerase chain reaction (PCR). Repeated PCR tests on the seedlings did not reveal the presence of phytoplasmal DNA. Likewise, symptoms of the disease were not observed on these seedlings after 2 years. PCR assays detected the phytoplasma in coats of some seed from infected trees; however, no excised embryos were positive for the phytoplasma. All positive PCR results were confirmed by restriction fragment length polymorphism assay. One-year-old seedlings derived from seed of noninfected plants appeared more vigorous in terms of height, number of leaves, and fresh weight of shoot compared with those from infected trees. The germination percentage, mean daily germination, peak value, and germination value were significantly higher for seed of fruit from noninfected trees and seed from fruit on asymptomatic branches of infected trees than those from fruit on symptomatic branches of infected trees.