Publications

4368

Citrus huanglongbing (HLB) is a devastating disease for the citrus industry. The previous studies demonstrated that oxytetracycline and penicillin are effective antibiotics against Candidatus Liberibacter asiaticus (CLas). However, since CLas is uncultured, the mechanisms of action of antibiotics against CLas are still unclear. It was recently reported that the endophytic microbial communities are associated with the progression of citrus HLB after oxytetracycline and penicillin treatment. Therefore, we hypothesize that penicillin has greater antibacterial activity against CLas than oxytetracycline, which may be associated with the alteration of the structure and function of endophytic microbial communities in HLB-affected citrus in response to these antibiotics. To test this hypothesis, the microbiome of HLB-affected citrus leaves treated with these two antibiotics was analyzed using a metagenomic method. Our results indicate that the microbial structure and function in HLB-affected citrus were altered by these two antibiotics. The relative abundance of beneficial bacterial species, including Streptomyces avermitilis and Bradyrhizobium, was higher in penicillin-treated plants compared to those treated with oxytetracycline, and the relative abundance of the bacterial species (such as Propionibacterium acnes and Synechocystis sp PCC 6803) associated with CLas survival was lower for penicillin-treated plants compared to oxytetracycline-treated plants. These results indicate that penicillin has greater antibacterial activity against CLas. Based on the metagenomic analysis, this study elucidated the mechanism for the observed increase in antibacterial activity of penicillin against CLas. The data presented here are not only invaluable for developing eco-friendly and effective biocontrol strategies to combat citrus HLB, but also provide a method for revealing mechanism of antimicrobial against uncultured bacteria in host.

Health, growth, fruit production, and fruit quality of citrus crops are severely affected by tree infection with Candidatus Liberibacter asiaticus (CLas) and subsequent development of the disease huanglongbing (HLB). The use of HLB-tolerant rootstocks is one strategy that is used to ameliorate the effects of HLB in commercial production. Although there is a clear long-term rootstock effect to improve tree performance, such field trials take many years for establishment and evaluation, and this long time-period is very limiting to expeditious evaluation of new rootstocks. In this study, we have conducted a 50-week greenhouse experiment to evaluate rootstock influences on Valencia sweet orange tree response to CLas infection. The infection of trees with CLas reduced scion and rootstock growth, increased leaf yellowing, and reduced the number of leaves per tree and leaf area, regardless of rootstock. There were clear rootstock influences on some traits during the 50-week study. In general, infected trees on US-942 rootstock had lower CLas root titers, less reduction of the number of leaves, less reduction of leaf area, and less leaf yellowing, as compared with some of the other rootstocks. The 50-week greenhouse evaluation method provided results that corresponded well with results from long-term field testing, indicating this may be a useful tool to accelerate evaluation and selection of new rootstocks, as well as in testing other HLB management strategies.

Abstract Rickettsia are obligate intracellular pathogens transmitted by arthropod vectors. The re‐emergence of several rickettsioses imposes severe global health burden. In addition to the well‐established rickettsial pathogens, newer rickettsial species and their pathogenic potentials are being uncovered. There are many reports of spotted and typhus fever caused by rickettsiae in India. Hence, in this study we screened the ectoparasites of pet and domestic animals for the presence of rickettsia using polymerase chain reaction. Nine cat flea samples ( Ctenocephalides felis felis ), that tested positive for the presence of rickettsia were subjected to Multi Locus Sequence Typing. Nucleotide sequencing and Phylogenetic analysis of gltA, ompB and 16rrs genes revealed that the rickettsiae detected in cat fleas was Rickettsia asembonensis . Further studies are required to assess Rickettsia asembonensis pathogenic potential to human and its enzootic maintenance of in various hosts and vectors.

Phytoplasmas have been associated with a disease that affects trees of at least 11 species from different botanic families in Bogotá, Colombia. ‘Candidatus Phytoplasma asteris’ and ‘Candidatus Phytoplasma fraxini’ are the major groups of phytoplasma in the area of Bogotá. In this study, the genetic diversity within ‘Ca. P. asteris’ and ‘Ca. P. fraxini’ was studied in five urban tree species: Croton species (Euphorbiaceae), Fraxinus uhdei (Oleaceae), Magnolia grandiflora (Magnoliaceae), Populus nigra (Salicaceae) and Quercus humboldtii (Fagaceae). Analyses of the 16S rRNA gene using nested PCR, RFLP and sequencing showed that phytoplasmas of ‘Ca. P. asteris’ could be assigned to: subgroup 16SrI-B; a new subgroup named 16SrI-AF, with a restriction pattern similar to that of 16SrI-B; and a new subgroup named 16SrI-AG, with a restriction pattern similar to that of 16SrI-K and 16SrI-AH with a restriction pattern similar to that of 16SrI-AC. ‘Ca. P. fraxini’ isolates belonged to a new subgroup named 16SrVII-G, with a restriction pattern similar to that of 16SrVII-A. To complement the identification of the phytoplasma strains, we amplified nonribosomal genes such as leuS and secA. Unexpectedly, it was observed that in 16 trees in which 16S rRNA gene analysis showed the presence of ‘Ca. P. fraxini’ only, the leuS or secA primers amplified sequences exclusively affiliated to ‘Ca. P. asteris. In those plants, sequences belonging to ‘Ca. P. fraxini’ leuS or secA genes were not amplified. The present work contributes to the identification of novel strains of both species in Colombia, and supports previous suggestions that phytoplasmas in South America are highly variable.

AbstractCandidatus Liberibacter solanacearum haplotype B (CLsoB) is an economically important pathogen of tomato (Solanum lycopersicum L.) crops in New Zealand and Central and North America. Currently, resistant cultivars of tomato are not available as a management tactic because breeding programmes lack sources of resistance. Therefore, the objective of this study was to identify sources of resistance in tomato to CLsoB. Forty‐six landraces of tomato were collected from several states in Mexico and were inoculated with CLsoB using 20 infested adults of Bactericera cockerelli per plant. Two assays were done over two years under greenhouse conditions. In the first trial, landraces FC22 and FC44 showed a significantly higher proportion of resistant plants, less symptom severity and longer incubation time compared with the other forty‐four landraces and two susceptible cultivars. In the second assay, resistance to CLsoB of the landraces FC22 and FC44 was confirmed because they had again significantly greater numbers of resistant plants, less symptom severity, relative lower CLsoB titers and longer incubation time relative to the other genotypes. All plants considered resistant from both assays had DNA of CLsoB. Results indicate that all resistant plants from these landraces are promising sources for the development of tomato cultivars with resistance to CLsoB.