Molecular characterization of Candidatus Phytoplasma aurantifolia isolate infecting chickpea (Cicer arietinum) in Dharwad, Karnataka

Chickpea plants showing phytoplasma symptoms were observed in the research plots at University of Agricultural Sciences, Dharwad, Karnataka, India. The symptoms included phyllody, pale green leaves, bushy appearance and excessive axillary proliferation. The causal agent of the phyllody disease was identified based on symptoms, amplification of 16S rDNA of the phytoplasma by nested PCR with primers P1/P7 and R16F2n/R16R2 and 1,800 bp and 1,200 bp size products were amplified in first round PCR and nested-PCR respectively. The PCR product was sequenced and compared with the reference phytoplasma sequences collected from the database (NCBI). 16S rDNA sequences of Dharwad chickpea phytoplasma shared the highest nucleotide identity of (>98%) with Periwinkle phyllody16SrII-E (EU096500). This study indicated the association of ‘Candidatus Phytoplasma aurantifolia’ the 16SrII-E group infecting chickpea from Northern Karnataka.

Catharanthus roseus (Apocynaceae) naturally infected with diverse phytoplasmas in Costa Rica

Phytoplasmas (class Mollicutes) are causal agents of plant diseases with an economic impact on crops or threatening local biodiversity. A survey was conducted from 2012 to 2016 on infected Catharanthus roseus plants that exhibited symptoms reminiscent of phytoplasma infection throughout Costa Rica. A total of 73 plants were collected exhibiting symptoms such as virescence, phyllody, axillary proliferation, little leaf, leaf malformation, chlorosis, or yellowing. All samples were tested by nested PCR using phytoplasma universal and specific primer pairs. Phytoplasma infection was detected in 52 (71.2 %) of the plants collected. Phytoplasmas of six subgroups belonging to 16Sr groups I, III, IX, XIII and XV were identified based on sequencing and in silico RFLP analyses. ´Candidatus Phytoplasma asteris´ (16SrI) was the predominant group among the positive samples (n = 30) showing variety of symptoms and wide distribution from sea level to ca. 1 400 masl in six of the seven Costa Rican provinces. Group 16SrIII was the second most abundant (14 samples); and the remaining three groups were seldom found in C. roseus (8 samples). Moreover, group 16SrXIII phytoplasma was detected for the first time in the country. To the best of our knowledge, this is the first report of natural infection of C. roseus with phytoplasma subgroups 16SrI-B, 16SrI-P, 16SrIII-F, 16SrIX-F, 16SrXIII-A, and 16SrXV-B in Costa Rica and Central America.

Assessment of clonal stability of in vitro regenerated shoots of Macadamia tetraphylla by RAPD analysis

Macadamia nuts constitute an important part of the world nut industry and are highly valued for their health-promoting properties. Macadamia is an open-pollinated crop that takes 8–12 years to bear fruit when multiplied via seeds. The yield and nut quality in seedling plantations are often highly variable, and grafting is currently the most common method for producing nursery trees with reduced variability. We have previously reported on the tissue-culture propagation of macadamia, and in the present study we assessed the clonal integrity of the regenerated shoots. The RAPD profiles of 3 macadamia stock plants and 10 in vitro regenerated lines from each stock plant were analysed to assess the clonal integrity of the shoots regenerated in vitro for micropropagation purposes. The extent of genetic variation between the stock plants and 9 randomly selected seedlings was also assessed. There was no difference in clonal identity between the stock plants and their micropropagated progeny, indicating that clonal micropropagation was possible using enhanced axillary proliferation in macadamia. In contrast, there was a large genetic variation among the seedlings and between the seedlings and stock plants, with genetic distance estimates ranging from 0.121 to 0.637 among seedlings, indicating rampant out-crossing of the macadamia plant.