Partial purification of Thai isolate of citrus huanglongbing (greening) bacterium and antiserum production for serological diagnosis

• Published in: Journal of general plant pathology : JGPP 2002-12, Vol.68 (4), p.372-377
• Main Authors: Ohtsu, Y. (National Inst. of Fruit Tree Science, Tsukuba, Ibaraki (Japan)), Prommintara, M, Okuda, S, Goto, T, Kano, T, Nakashima, K, Koizumi, M, Imada, J, Kawashima, K
• Format: Article
• Language: English
• Subjects: BACTERIA; CITRUS; Citrus fruits; DIAGNOSIS; Fruits; GREENING; IMMUNE SERUM; IMMUNOLOGICAL TECHNIQUES; Leaves; PATHOGENS; Plant pathology; Plant populations; PURIFICATION; THAILAND
• ISSN: 1345-2630; 1610-739X
• DOI: 10.1007/PL00013105

We aimed to improve the purification of citrus Huanglongbing (greening) bacterium (HB), Candidatus Liberobacter asiaticum and to produce an antiserum against HB. Periwinkle plants Catharantus roseum L. graft-inoculated with HB were used to produce an antiserum. All young leaves of new shoots incubated at 20-25 degrees C and 25-30 degrees C, a few mature leaves incubated at 20-25 degrees C, and all mature leaves incubated first at 25-30 degrees C and later transferred to 20-25 degrees C developed yellowing symptoms and were then used to prepare immunogen. The HB was partially purified from these leaves by an improved method that included a macerating enzyme treatment of the midribs of infected leaves and homogenization of infected phloem sieve tissues. An antiserum raised against partially purified HB reacted clearly at a dilution of 1/16 with HB-infected citrus extract prepared at a concentration of 40 times, but did not react with healthy or tristeza virus-infected citrus extract in microprecipitin tests. [PUBLICATION ABSTRACT]