scholarly journals Mitochondrial genetic variation reveals phylogeographic structure and cryptic diversity in Trioza erytreae

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Inusa Ajene ◽  
Fathiya M. Khamis ◽  
Gerhard Pietersen ◽  
Barbara van Asch

AbstractTrioza erytreae is the main vector for ‘Candidatus Liberibacter africanus’, the causative agent of African Citrus Greening disease. The insect is widespread in Africa, and has recently disseminated to Southwestern Europe. This study aimed at generating reference mitogenome sequences for T. erytreae, as a background for future genetic diversity surveys. Complete mitochondrial sequences of three specimens collected in Ethiopia, Uganda and South Africa were recovered using Ion Torrent technology. The mitogenomes of T. erytreae from Uganda and Ethiopia were highly similar, and distinct from that found in South Africa. The phylogeographic structure of T. erytreae was assessed using genetic clustering and pairwise distances, based on a dataset of public COI sequences recorded as T. erytreae. The dataset revealed ten haplotypes with strong phylogeographic structure in Africa and Europe. Three haplotypes found in Kenya on Clausena anisata belonged to pairs separated by distances as high as 11.2%, and were basal to all other sequences. These results indicate that not all sequences identified as T. erytreae belong to the same species, and that some degree of specificity with different plant hosts is likely to exist. This study provides new baseline information on the diversity of T. erytreae, with potential implications for the epidemiology of African Citrus Greening disease.

Plant Disease ◽  
2003 ◽  
Vol 87 (4) ◽  
pp. 448-448 ◽  
Author(s):  
Y. S. Ahlawat ◽  
V. K. Baranwal ◽  
Thinlay ◽  
Doe Doe ◽  
S. Majumder

During July 2002, surveys of mandarin orchards were conducted in Punakha Valley and Wangdue districts of Bhutan. Symptoms of the greening disease were observed in most of the orchard. The incidence of disease was recorded up to 30% in 24 private orchards with more than 5,000 trees total. Affected trees were generally stunted with leaves showing symptoms of mottling. Sometimes, symptoms were seen only on one part of the canopy. The greening disease is caused by a fastidious phloem restricted bacterium, “Candidatus Liberibacter asiaticus” in Asian countries and “Candidatus Liberibacter africanus” in African countries. To confirm the presence of this bacterium causing greening disease in Bhutan, 33 leaf samples were collected from seven locations in Bhutan and stored at -80°C. Petioles and midribs were used for extraction of DNA using DNeasy Plant Mini Kit (Qiagen Gmbh, Hilden, Germany). Polymerase chain reaction (PCR) was initially performed with a sample from Rimchu, Bhutan using primer pair 5′TATAAAGGTTGACCTTTCGAGTTT/5′ACAAAAGCAGAAATAGCACGAACAA previously designed for amplification of ribosomal protein genes of β-operon of two liberibacter species (1). An amplicon of approximately 700 bp was obtained. The size of the PCR product is similar to that amplified from “Candidatus Liberibacter asiaticus”. The amplicon was cloned in pGEM-T easy vector and sequenced. The clone was 703 nt long and showed 100% sequence homology with the corresponding sequence of “Candidatus Liberibacter asiaticus” confirming that “Candidatus Liberibacter asiaticus” is the cause of greening disease in Bhutan. Later, one sample from each location was analyzed and found to be positive to greening. To our knowledge, this is the first report of this bacterium and greening disease in Bhutan, and citrus greening appears to be the main cause of declining citrus in the Punakha Region of Bhutan. Reference: (1) A. Jocquellet et al. Page 363 in: Proc. Conf. Int. Organ. Citrus Virol. 14th. IOCV, Riverside, CA, 2000.


2020 ◽  
Vol 18 (3) ◽  
pp. 529-541
Author(s):  
Ho Thi Thuong ◽  
Nguyen Thi Thom ◽  
Nguyen Thi Tra ◽  
Trinh Thai Vy ◽  
Pham Bich Ngoc ◽  
...  

Citrus Greening, also known as HuangLongbing (HLB), is considered one of the most dangerous citrus diseases, and limiting the production of citrus trees all over the world. Production of antibodies against Ompa protein of Candidatus Liberibacter asiaticus (CLas) for detection of citrus greening disease is considered as promising research direction. In this study, for the purpose of producting antibodies against Ompa of CLas, we firstly used the camel VHH antibody library for screening VHH antibodies against Ompa using phage-display technique. Next, phages which had strong interaction with Ompa as shown in ELISA were selected for phagemid isolation and the DNA fragments encoding VHH antibodies were sequenced. The DNA fragment encoding the best VHH antibody was then selected and inserted into the expression vector pET-21a (+), then cloned in Ecoli DH5α strain and expressed in BL21 (DE3) strain. The expression of VHH antibodies against Ompa was optimized at different temperatures with an inductive concentration of 0.1 M IPTG. Anti-Ompa VHH antibodies were purified under denatured conditions then re-folded. The biological activity of the VHH antibody with Ompa antigen was assessed by indirect-ELISA reaction. Results indicated that the VHH antibody had a very strong interaction with the Ompa antigen. This opens up the prospect of applying VHH antibody in the detection of citrus greening disease.


2020 ◽  
Vol 21 (1) ◽  
pp. 31-35
Author(s):  
Jonathan E. Oliver ◽  
Md Emran Ali ◽  
Sumyya Waliullah ◽  
Jake Price ◽  
Jessica Warren ◽  
...  

Huanglongbing (HLB) or citrus greening disease is the most devastating disease of citrus worldwide. This disease, caused by the bacterium ‘Candidatus Liberibacter asiaticus’ (CLas), leads to low fruit quality and unproductive trees. In 2008, HLB was found in a residential citrus tree in Savannah, Georgia, and, as a result, the state has been quarantined for this disease since 2009. Nonetheless, little is known about the distribution of CLas within Georgia, even though the commercial planting of citrus in Georgia has increased exponentially in recent years. In 2019, 94 samples from commercial and residential citrus trees within 11 counties in coastal and southern Georgia were collected and tested for the presence of CLas. Molecular testing results revealed the presence of CLas in three counties where HLB had not been previously reported and in 9% of samples overall. This is the first definitive report confirming HLB in southern Georgia counties besides those along the coast.


Plant Disease ◽  
2020 ◽  
Vol 104 (5) ◽  
pp. 1535-1535 ◽  
Author(s):  
Inusa Jacob Ajene ◽  
Fathiya Khamis ◽  
Samira Mohammed ◽  
Adedapo Olutola Adediji ◽  
Gabriel Ifo Atiri ◽  
...  

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