Genetics

The objectives of this work were (i) to determine the influence of temperature on infection of citrus by ‘ Candidatus Liberibacter asiaticus’ and ‘ Candidatus Liberibacter americanus’, the two bacterial species associated with citrus huanglongbing (HLB) in Brazil, and (ii) to determine the influence of temperature on citrus colonization by ‘ Ca.  L. asiaticus’, which has taken over from ‘ Ca.  L. americanus’ as the predominant species in Brazil since 2008. Two experiments were carried out with graft‐inoculated Valencia oranges on Rangpur lime rootstocks. Immediately after inoculation the plants were maintained for 423 days in growth chambers under the following night/day temperature conditions: 17/22, 22/27 or 27/32°C, with a dark/light photoperiod of 8/16 h. Infection and colonization of plants were determined using quantitative PCR (qPCR). ‘ Candidatus Liberibacter americanus’ did not infect the plants maintained at 27/32°C; however, infection by ‘ Ca.  L. asiaticus’ occurred at all studied temperatures. Two months after inoculation, ‘ Ca.  L. asiaticus’ was distributed throughout the inoculated plants, with mean C t values in the range of 30–31 for leaves and 25–28 for roots. Over time, ‘ Ca.  L. asiaticus’ reached the highest titres in mature leaves (mean C t value = 26·7) of citrus plants maintained at 22/27°C. ‘ Candidatus Liberibacter asiaticus’ colonization of citrus plants was negatively affected by the daily temperature regime of 27/32°C (mean C t value in mature leaves = 33·6).

AbstractIn 2010, cabbages (Brassica oleracea L.) showing symptoms of proliferated axillary buds, crinkled leaves and plant stunting with shortened internodes typical to phytoplasma infection were found in a breeding facility in Beijing, China. Three symptomatic plants and one symptomless plant were collected, and total DNA was extracted from the midrib tissue and the flowers. With phytoplasma universal primers R16F2n/R16R2, a special fragment of 1247 bp (16S rDNA) was obtained from all three symptomatic cabbage plants, but not from the one symptomless cabbage plant. The 16S rDNA sequence showed 99% similarity with the homologous genes of the aster yellows group phytoplasma (16SrI group), and the phytoplasma was designed as CWBp‐BJ. Phylogenetic and computer‐simulated restriction fragment length polymorphism (RFLP) analysis of the 16S rDNA gene revealed that CWBp‐BJ belongs to subgroup 16SrI‐B. This is the first report of a phytoplasma associated with cabbage witches’‐broom in China.