Daniel Gargani

Kinetoplast DNA permits characterization of pathogenic plant trypanosomes of economic importance

Summary— Twelve Phytomonas isolates were obtained from different plants originating from several countries and cultured in vitro in complex media. The kinetoplast DNA (kDNA) was purified and observed by electron microscopy. The structure of kDNA from all isolates appeared as a large network of interlocked minicircles with some maxicircles extruding from the network, as has often been shown for Trypanosomatidae. Topoisomerase II resolved the kDNA network into free minicircles which were then analyzed by electron microscopy and by electrophoresis in agarose gel. The minicircle sizes varied from 1.3 to 2.8 kilobase pairs according to the Phytomonas isolate. The analysis by restriction endonucleases revealed a base sequence heterogeneity in the minicircles of 10 of these Phytomonas isolates. By contrast, in 2 Phytomonas isolates, more than 90% of their minicircle content was found to be homogeneous. Most interestingly, the minicircle cleavage patterns were found to be different between Phytomonas isolates and thus could be used to distinguish them.

Variability in the phloem restricted plant trypanosomes (Phytomonas spp.) associated with wilts of cultivated crops. Isoenzyme comparison with the lower trypanosomatids.

Plant trypanosomatids (Phytomonas spp) have been isolated from the crude sap of coconut tress and oil palm trees affected with Hartrot or Marchitex diseases in South America andAlpinia purpurata affected with wilt in the Caribbean. They are also commonly isolated from the fruits and latex of Euphorbiaceae or Asclepiadaceae. Thirty-fourPhytomonas stocks were studied by isoenzyme electrophoresis (11 loci) in order to investigate genetic variability in the intraphloemic group. Our results showed that variability in phloem restrictedPhytomonas is very high, and that Marchitez or Hartrot associated trypanosomatids are not readily separated into two distinct groups. Moreover, the two isolates fromAlpinia are very close to the other intraphloemic isolates. The results confirm the existence of several distinct groups, comprising of at least 2 groups of phloem restricted trypanosomatids, not related to the host species, 3 groups of latex trypanosomatids, and one group of fruit trypanosomatids.

Classification of plant trypanosomatids (Phytomonas spp.): parity between random-primer DNA typing and multilocus enzyme electrophoresis

The genetic polymorphism of 30 isolates of plant trypanosomatids colloquially referred to as plant trypanosomes was assayed by means of RAPD. The principle objectives of this study were to assess the discriminative power of RAPD analysis for studying plant trypanosomes and to determine whether the results obtained were comparable with those from a previous isoenzyme (MLEE) study. The principle groups of plant trypanosomes identified previously by isoenzyme analysis--intraphloemic trypanosomes, intralaticiferous trypanosomes and trypanosomes isolated from fruits--were also clearly separated by the RAPD technique. Moreover, the results showed a fair parity between MLEE and RAPD data (coefficient of correlation = 0.84) and the two techniques have comparable discriminative ability. Most of the separation revealed by the two techniques between the clusters was associated with major biological properties. However, the RAPD technique gave a more coherent separation than MLEE because the intraphloemic isolates, which were biologically similar in terms of their specific localization in the sieve tubes of the plant, were found to be in closer groups by the RAPD. For both techniques, the existence of the main clusters was correlated with the existence of synapomorphic characters, which could be used as powerful tools in taxonomy and epidemiology.

Virus of Plant Trypanosomes (Phytomonas Spp.)

Trypanosomes have been known to exist in the laticifers of latex-bearing plants since 1909 (15). The pathogenicity of these organisms in plants has been the subject of much discussion, but it can now be said that no plant diseases exist with which intralaticiferous trypanosomes are specifically associated (5). On the other hand, intraphloemic trypanosomes are specifically associated with decay in cultivated plants in South America: coffee phloem necrosis (25), coconut Hartrot (21), oil palm Marchitez (8), and in the Caribbean: decay in Alpiniapurpurata (P. Hunt, M. Dollet, 1991, not published).