Yellow Decline Research Articles

Global Scenario on Phytoplasmal Diseases in Palms

Worldwide phytoplasma diseases pose great threat to the sustainability of the palm family members viz., coconut, arecanut, date palm, oil palm and ornamental palms causing setbacks in economic and livelihood security. So far, at least eight Candidatus Phytoplasma species belonging to different 16Sr groups have been reported to be associated with coconut. This includes Ca. P. asteris (lethal wilt disease in India; 16Sr IB), Ca. P. oryzae (Coconut root (wilt) disease in India, Weligama leaf wilt disease in Sri Lanka; 16Sr XIB), Ca. P. cynodontis (Coconut yellow decline in Malaysia; 16Sr XIV), Ca. P. malaysianum (Malayan yellow dwarf phytoplasma in Malaysia; 16Sr XXXIIB), Ca. P. palmicola (Cape St. Paul wilt in Ghana, Awka wilt in Nigeria; 16Sr XXIIB), Ca. P. cocostanzaniae (lethal yellowing in East Africa; 16Sr IVC), Ca. P. noviguineense (Coconut bogia syndrome in Papua new Guinea) and Ca. P. palmae (Coconut lethal yellowing in Americas and Caribbean region; 16Sr IVA,B,E). Though there are several reports on insects transmitting coconut phytoplasmal diseases, except for LY in Florida and RWD in India, the vectors remain elusive. Susceptibility of date palms to phytoplasmal diseases viz., lethal yellowing, Alwijam, White Tip Dieback (WTD) and Slow Decline (SD) or El Arkish’ are emerging concerns in Arabian countries and North Africa. Oil palm is a potential host of diverse groups (16SrI, 16SrXI, 16Sr XIV and 16SrXXXII) of phytoplasma in many parts of the world. Yellow leaf disease caused by 16SrI, 16SrXI and16SrXIV group phytoplasmas is one of the major diseases limiting the productivity of arecanut in south India. Many ornamental palm species harbour a wide array of phytoplasma groups that could serve as an intermediary mode of transmission to cultivated palms. Large scale movement of ornamental palms through international trade warrants the need for strengthening the international quarantine networks and bio-security measures to contain the spread of these phytopathogenic mollicutes. Despite rapid progress accomplished in taxonomy using molecular approaches many palm phytoplasma isolates are yet to be characterized for the assignment of specific taxonomic status. Due to its confinement to phloem, non-uniform distribution and sub-minimal titres in palms, molecular detection is many atimes intriguing. Palm-phytoplasmal interaction and transmission mechanism by auchenorrhynchan fauna are poorly understood. As a victim of climate change, phytoplasma diseases in synergy with survival superiority of insect vectors are extremely challenging to comprehend and evolve effective suppression mechanism. Management approaches involving surveillance, eradication and containment of the disease in newly emerging areas along with resistance breeding to ensure sustainable income to the farmers are discussed in this chapter.

Metabolomic study of volatile compounds emitted by lavender grown under open-field conditions: a potential approach to investigate the yellow decline disease.

Fine lavender and lavandin are perfume and medicinal plants originate from the South of France and are widely cultivated for their essential oils. Recently, cultivated plants suffered from a severe decline in France, due to the propagation of the yellow decline disease. This disease is caused by the stolbur phytoplasma, a bacterium transmitted by a sap-sucking insect, the planthopper. In order to understand the complex relationships between host plant, pest, pathogen and environment responsible for the yellow decline of lavender, we use a metabolomic approach to highlight changes in chemical emissions from asymptomatic ("healthy") and symptomatic ("infected") plants. Volatile compounds produced by fine lavender and lavandin were collected in the field using a dynamic headspace extraction approach. Afterwards, compounds trapped on Tenax adsorbent were thermodesorbed and analysed using an automated thermal desorption-gas chromatography-mass spectrometry (ATD-GC-MS). Multivariate statistical analyses was performed using principal component analysis and partial least square discriminant analyses. The untargeted screening of volatiles allowed the separation of asymptomatic and symptomatic plants according to their emissions. The approach was sufficiently accurate so as to separate the emissions according to the different stages of infection. Twelve compounds were found to be deregulated metabolites of yellow disease infection, common to fine lavender (variety 7713) and lavandin (variety abrial). The metabolomic approach allowed for the effective identification of chemical variations between infected and healthy plants in a complex field environment.

'Candidatus Phytoplasma wodyetiae', a new taxon associated with yellow decline disease of foxtail palm (Wodyetia bifurcata) in Malaysia.

Landscape-grown foxtail palm (Wodyetia bifurcata A. K. Irvine) trees displaying symptoms of severe foliar chlorosis, stunting, general decline and mortality reminiscent of coconut yellow decline disease were observed in Bangi, Malaysia, during 2012. DNA samples from foliage tissues of 15 symptomatic palms were analysed by employing a nested PCR assay primed by phytoplasma universal ribosomal RNA operon primer pairs, P1/P7 followed by R16F2n/R2. The assay yielded amplicons of a single band of 1.25 kb from DNA samples of 11 symptomatic palms. Results from cloning and sequence analysis of the PCR-amplified 16S rRNA gene segments revealed that, in three palms, three mutually distinct phytoplasmas comprising strains related to 'Candidatus Phytoplasma asteris' and 'Candidatus Phytoplasma cynodontis', as well as a novel phytoplasma, were present as triple infections. The 16S rRNA gene sequence derived from the novel phytoplasma shared less than 96 % nucleotide sequence identity with that of each previously describedspecies of the provisional genus 'Ca. Phytoplasma', justifying its recognition as the reference strain of a new taxon, 'Candidatus Phytoplasma wodyetiae'. Virtual RFLP profiles of the R16F2n/R2 portion of the 16S rRNA gene and the pattern similarity coefficient value (0.74) supported the delineation of 'Ca. Phytoplasma wodyetiae' as the sole representative subgroup A member of a new phytoplasma ribosomal group, 16SrXXXVI.

Source of Hyalesthes obsoletus Signoret planthopper (Hemiptera: Cixiidae) in southern France and potential effects of landscape.

Cixiid planthoppers are considered of major economic importance, as they can transmit phytoplasmas responsible for many plant diseases. While thorougly studied in vineyards, the epidemiology of stolbur phytoplasma, transmitted by Hyalesthes obsoletus Signoret, was rarely investigated on minor crops as lavender, where it leads to 'yellow decline' disease and large economic losses. The objective of this paper is to understand the effect of the local landscape characteristics on the presence and density of H. obsoletus in the 'Plateau de Valensole', southern France. Potential host plants of H. obsoletus were surveyed in three contrasted zones (in terms of crops and disease intensity), by uprooting plants and capturing adults in emergence traps. The localization and potential movements of H. obsoletus from the host plants towards lavandin (infertile hybrid of lavender) were determined using yellow sticky traps. Clary sage plants were found as major hosts of H. obsoletus. Flying insects were also caught in fields of lavandin, although emergence traps and plant uprooting did not confirm this crop as a winter host, i.e., as a reservoir for the insect. Based on one zone, we showed that attractiveness may depend on crop (clary sage or lavandin) and on its age, as well as on the distance to the supposed source field. These results suggest that clary sage could be an important host of H. obsoletus, whose density largely varies between zones. Genetic studies would be required to confirm the role of clary sage in the dissemination of yellow decline of lavandin.

Transcriptomics-based analysis using RNA-Seq of the coconut (Cocos nucifera) leaf in response to yellow decline phytoplasma infection.

Invasive phytoplasmas wreak havoc on coconut palms worldwide, leading to high loss of income, food insecurity and extreme poverty of farmers in producing countries. Phytoplasmas as strictly biotrophic insect-transmitted bacterial pathogens instigate distinct changes in developmental processes and defence responses of the infected plants and manipulate plants to their own advantage; however, little is known about the cellular and molecular mechanisms underlying host-phytoplasma interactions. Further, phytoplasma-mediated transcriptional alterations in coconut palm genes have not yet been identified. This study evaluated the whole transcriptome profiles of naturally infected leaves of Cocos nucifera ecotype Malayan Red Dwarf in response to yellow decline phytoplasma from group 16SrXIV, using RNA-Seq technique. Transcriptomics-based analysis reported here identified genes involved in coconut innate immunity. The number of down-regulated genes in response to phytoplasma infection exceeded the number of genes up-regulated. Of the 39,873 differentially expressed unigenes, 21,860 unigenes were suppressed and 18,013 were induced following infection. Comparative analysis revealed that genes associated with defence signalling against biotic stimuli were significantly overexpressed in phytoplasma-infected leaves versus healthy coconut leaves. Genes involving cell rescue and defence, cellular transport, oxidative stress, hormone stimulus and metabolism, photosynthesis reduction, transcription and biosynthesis of secondary metabolites were differentially represented. Our transcriptome analysis unveiled a core set of genes associated with defence of coconut in response to phytoplasma attack, although several novel defence response candidate genes with unknown function have also been identified. This study constitutes valuable sequence resource for uncovering the resistance genes and/or susceptibility genes which can be used as genetic tools in disease resistance breeding.

First Report of Phytoplasma (16SrI) Associated with Yellow Decline Disease of Royal Palms [Roystonea regia (Kunth) O. F. Cook] in Malaysia

AbstractRoyal Palms (Roystonea regia) with symptoms such as severe chlorosis, stunting, collapse of older fronds and general decline were observed in the state of Selangor, Malaysia. Using polymerase chain reaction (PCR) amplification with phytoplasma universal primer pair P1/P7 followed by R16F2N/R16R2 and fU5/rU3 as nested PCR primer pairs, all symptomatic plants tested positively for phytoplasma. Results of phylogenetic and virtual RFLP analysis of the 16S rRNA gene sequences revealed that the phytoplasma associated with Royal Palm yellow decline (RYD) was an isolate of ‘Candidatus Phytoplasma asteris’ belonging to a new 16SrI‐subgroup. These results show that Roystonea regia is a new host for the aster yellows phytoplasma (16SrI). This is the first report on the presence of 16SrI phytoplasma on Royal Palm trees in Malaysia.

First Report of Two Distinct Phytoplasma Species, 'Candidatus Phytoplasma cynodontis' and 'Candidatus Phytoplasma asteris,' Simultaneously Associated with Yellow Decline of Wodyetia bifurcata (Foxtail Palm) in Malaysia.

The foxtail palm (Wodyetia bifurcata), an Australian native species, is an adaptable and fast-growing landscape tree. The foxtail palm is most commonly used in landscaping in Malaysia. Coconut yellow decline (CYD) is the major disease of coconut associated with 16SrXIV phytoplasma group in Malaysia (1). Symptoms consistent with CYD, such as severe chlorosis, stunting, general decline, and death were observed in foxtail palms from the state of Selangor in Malaysia, indicating putative phytoplasma infection. Symptomatic trees loses their green and vivid appearance as a decorative and landscape ornament. To determine the presence of phytoplasma, samples were collected from the fronds of 12 symptomatic and four asymptomatic palms in September 2012, and total DNA was extracted using the CTAB method (3). Phytoplasma DNA was detected in eight symptomatic palms using nested PCR with universal phytoplasma 16S rDNA primer pairs, P1/P7 followed by R16F2n/R16R2 (2). Amplicons (1.2 kb in length) were generated from symptomatic foxtail palms but not from symptomless plants. Phytoplasma 16S rDNAs were cloned using a TOPO TA cloning kit (Invitrogen). Several white colonies from rDNA PCR products amplified from one sample with R16F2n/R16R2 were sequenced. Phytoplasma 16S rDNA gene sequences from single symptomatic foxtail palms showed 99% homology with a phytoplasma that causes Bermuda grass white leaf (AF248961) and coconut yellow decline (EU636906), which are both members of the 16SrXIV 'Candidatus Phytoplasma cynodontis' group. The sequences also showed 99% sequence identity with the onion yellows phytoplasma, OY-M strain, (NR074811), from the 'Candidatus Phytoplasma asteris' 16SrI-B subgroup. Sequences were deposited in the NCBI GenBank database (Accession Nos. KC751560 and KC751561). Restriction fragment length polymorphism (RFLP) analysis was done on nested PCR products produced with the primer pair R16F2n/R16R2. Amplified products were digested separately with AluI, HhaI, RsaI, and EcoRI restriction enzymes based on manufacturer's specifications. RFLP analysis of 16S rRNA gene sequences from symptomatic plants revealed two distinct profiles belonging to groups 16SrXIV and 16SrI with majority of the 16SrXIV group. RFLP results independently corroborated the findings from DNA sequencing. Additional virtual patterns were obtained by iPhyclassifier software (4). Actual and virtual patterns yielded identical profiles, similar to the reference patterns for the 16SrXIV-A and 16SrI-B subgroups. Both the sequence and RFLP results indicated that symptoms in infected foxtail palms were associated with two distinct phytoplasma species in Malaysia. These phytoplasmas, which are members of two different taxonomic groups, were found in symptomatic palms. Our results revealed that popular evergreen foxtail palms are susceptible to and severely affected by phytoplasma. To our knowledge, this is the first report of a mixed infection of a single host, Wodyetia bifurcata, by two different phytoplasma species, Candidatus Phytoplasma cynodontis and Candidatus Phytoplasma asteris, in Malaysia.

Detection and Identification of Aster Yellows Phytoplasma Associated with Lipstick Yellow Frond Disease in Malaysia

AbstractYellowing symptoms similar to coconut yellow decline phytoplasma disease were observed on lipstick palms (Cyrtostachys renda) in Selangor state, Malaysia. Typical symptoms were yellowing, light green fronds, gradual collapse of older fronds and decline in growth. Polymerase chain reaction assay was employed to detect phytoplasma in symptomatic lipstick palms. Extracted DNA was amplified from symptomatic lipstick palms by PCR using phytoplasma‐universal primer pair P1/P7 followed by R16F2n/R16R2. Phytoplasma presence was confirmed, and the 1250 bp products were cloned and sequenced. Sequence analysis indicated that the phytoplasmas associated with lipstick yellow frond disease were isolates of ‘Candidatus Phytoplasma asteris’ belonging to the 16SrI group. Virtual RFLP analysis of the resulting profiles revealed that these palm‐infecting phytoplasmas belong to subgroup 16SrI‐B and a possibly new 16SrI‐subgroup. This is the first report of lipstick palm as a new host of aster yellows phytoplasma (16SrI) in Malaysia and worldwide.

Phytoplasmas associated with disease of coconut in Malaysia: phylogenetic groups and host plant species

Coconut palm (Cocos nucifera), oil palm (Elaeis guineensis), Bermudagrass (Cynodon dactylon) and Madagascar periwinkle (Catharanthus roseus) with symptoms indicative of phytoplasma disease were collected from different locations in Malaysia. PCR assays employing phytoplasma universal rRNA gene primers P1/P7 alone or P1/P7 followed by R16F2n/R16R2 detected phytoplasmas in eight out of 20 Malayan Red Dwarf (MRD), nine out of 12 Malayan Yellow Dwarf (MYD) and 12 out of 12 Malayan Tall (MT) coconut palms displaying coconut yellow decline symptoms. Positive detections were also obtained from six out of six oil palm seedlings showing symptoms of yellowing and necrosis, from 10 out of 10 Bermudagrass samples with white leaf symptoms, and from eight out of eight periwinkle plants showing phyllody, virescence, little leaf, proliferation and foliar yellowing. Phytoplasmas were not detected in any of the symptomless plants tested. Sequencing and phylogenetic analysis of PCR products determined that phytoplasmas infecting both MRD and MT coconuts and Bermudagrass in Serdang, Selangor State, were all members of the 16SrXIV ‘Candidatus Phytoplasma cynodontis’ group, whereas isolates in periwinkle in Serdang were all members of the 16SrI ‘Ca. Phytoplasma asteris’ group. However, the phytoplasmas detected in MYD coconuts and oil palms from Banting, Selangor State, and in periwinkle from Putrajaya were collectively very similar (99%), but shared <97·5% similarity with 16S rDNA sequences of all other known phytoplasmas, indicating that they represent a novel taxonomic group. Thus, at least two phylogenetically distinct phytoplasmas are associated with the coconut yellow decline syndrome in Malaysia, both of which were also detected in other plant species.

Molecular Characterization of a Phytoplasma Associated with Coconut Yellow Decline (CYD) in Malaysia

Problem statement: Molecular methods have been used to detect phytoplasma in Malayan Red Dwarf (MRD) ecotype of coconut for the first time in Malaysia. Detect phytoplasma from coconut showing yellowing symptoms by nested PCR and Classify phytoplasma associated with disease of coconut palms, based on analysis of 16S rRNA gene operon sequences and virtual RFLP. Approach: Twenty MRD symptomatic palms were tested. Leaf spear, inflorescence and wood shavings from trunks of coconut palms showing yellowing symptoms were harvested from Serdang located in Selangor state. PCR assays and sequence analysis were carried out. Results: Nested PCR with primer pairs R16F2n/R16R2 and fU5/rU3 resulted in amplification of products of approximately 1.2 kb and 890 bp respectively, from 8 out of 20 MRD symptomatic palms tested. Sequence analysis of the 16S rDNA PCR products determined that the phytoplasma strain associated with Coconut Yellow Decline (CYD) in MRD ecotype belongs to the 'Candidatus Phytoplasma cynodontis' (16SrXIV) group of phytoplasmas. In addition, nested R16F2n/R16R2 PCR products from 6 spear leaves and 2 inflorescences from MRD palms showed high sequence similarity to the 16S rRNA gene from coconut chloroplasts, with a similar size (approximately 1.3 kb) and a further 5 R16F2n/R16R2 PCR products from MRD inflorescences showed high sequence similarities to Bacillus spp. 16S rRNA gene sequences. Conclusion: These results indicate that sequencing is a reliable method for the detection. Furthermore, trunk borings are the most reliable source of DNA for phytoplasma detection in coconuts using 16S rRNA gene primers, since there is less co-amplification of PCR products from other organisms when compared to spear leaves and inflorescences, nor from the spear leaves and inflorescences of MRD palms when primers fU5/rU3 were used in nested PCR.

First report of a 16SrXIV, ‘Candidatus Phytoplasma cynodontis’ group phytoplasma associated with coconut yellow decline in Malaysia

The coconut palm (Cocos nucifera) is an important perennial oil crop cultivated throughout Malaysia and is used for its aesthetic appeal in landscapes. Symptoms of yellow decline have been observed in a plantation in Selangor, similar to those caused by phytoplasma groups of the lethal yellowing (16SrIV) type recorded from the Americas, Caribbean and Africa, and similar also to Stolbur (16SrXII) associated with Kalimantan wilt in Indonesia. Although not widespread, this is the first report on such symptoms in coconut in Malaysia. The lower canopy foliage initially turns light yellow and eventually light-brown, spreading rapidly to the younger leaves. Also severe chlorosis of the emerging spear leaf, inflorescence necrosis, premature nut fall, and gradual collapse of fronds occur, and immature palms generally die within 5 months of initial symptom appearance. Samples were collected from 20 affected and five symptomless Malayan red dwarf coconut palms. Malayan yellow dwarf and some Malayan tall coconuts exhibited similar symptoms, but were not tested in this study. Total DNA was extracted and assayed in a nested PCR with universal phytoplasma primer pairs P1/P7 followed by either R16F2/R16R2 or fU5/ rU3. Products of the expected size (approximately 1200 bp and 880 bp, respectively) were amplified from 8/20 plants with symptoms but not from any symptomless coconut palms. Amplification also failed when the LY (16SrIV) specific primers LY16Sf/LY16Sr were used for nested PCR on P1/P7 first round PCR samples (Harrison et al., 2002), as well as when LY non-ribosomal specific primers LYC24F/LYC24R (Harrison et al ., 1994) were used, indicating that the phytoplasma was not from the 16SrIV group. The phytoplasma 16SrDNA was cloned and sequenced (GenBank Acc. No. EU328159) and showed the highest homology (99%) with those of Bermuda grass white leaf phytoplasma from Thailand (AF248961) and Malaysia (EU294011), members of the 16SrXIV ‘ Candidatus Phytoplasma cynodontis’ group. This group has previously been found associated with a decline disease in date palm in Sudan (Cronje et al ., 2000). However, this is the first report of a 16SrXIV phytoplasma associated with yellow decline in Malaysia.

First Report of a Novel Fusarium Species Causing Yellowing Decline of Sugar Beet in Minnesota.

In the United States, yellows disease of sugar beet (Beta vulgaris), which causes wilt, early death, and yield reduction, is caused primarily by Fusarium oxysporum f. sp. betae (3,4), but F. graminearum (2) has also been implicated. During the past 3 years, a similar disease causing yellowing and severe decline appeared in some sugar beet fields of central and southwest Minnesota planted with cultivars resistant to yellows. The disease has become a concern to the local sugar beet industry, which produces 56% of sugar beets in the United States. From 2005 to 2007, isolations were made from sugar beets collected in commercial fields and from a Fusarium screening nursery showing symptoms of yellowing, interveinal chlorosis, scorching, stunting, vascular discoloration of the taproot, and early death of plants. Of 96 Fusarium isolates recovered and used in root-dip inoculation trials in the greenhouse, 58 were pathogenic to sugar beets. On the basis of morphology, 12 were identified as F. oxysporum, 6 as F. graminearum, and 40 as a novel Fusarium species. The remaining 38 isolates were nonpathogenic. All three pathogenic Fusarium species were isolated from taproots, but only the novel Fusarium was isolated from petioles. In culture, the novel Fusarium exhibited a bright orange color on the underside of potato dextrose agar medium and produced micro- and macroconidia sparsely. Hyphal tip isolates of all novel Fusarium isolates were pathogenic, causing typical yellowing symptoms and plant death to the Fusarium yellows susceptible sugar beet cv. VDH46177 in replicated greenhouse trials. Isolates were successfully reisolated from the symptomatic plants, fulfilling Koch's postulates. Restriction fragment length polymorphism (RFLP) endonuclease digestion patterns (Alu1, Fnu4HI, HaeIII, and HhaI) of the internal transcribed spacer (ITS) region of 40 pathogenic novel isolates showed a distinct pattern compared with known Fusarium species. Thin layer chromatography analysis of 13 novel isolates detected the type A trichothecenes neosolaniol and 4,15-diacetoxyscirpenol. Partial sequences of the translation elongation factor 1-α (TEF) from 12 single-spored novel Fusarium isolates were generated. BLAST analysis of the TEF sequence against the FUSARIUM-ID (1) and GenBank databases did not match any known Fusarium species. On the basis of pathogenicity, morphology, RFLP patterns, mycotoxin production, and TEF sequence analysis it appears that this is a new species of Fusarium, but additional multilocus phylogenetic analyses are warranted. The natural occurrence of this novel Fusarium pathogen in sugar beet may have implications in breeding for resistance to Fusarium yellows, since yellow decline has been observed in purportedly Fusarium-tolerant cultivars in the Minnesota and North Dakota production regions.

The Genetic Relationship Between Mycoplasma‐like Organisms Causing Diseases in the Sudan and Different Continents Revealed by Polymerase Chain Reaction (PCR) Amplification of the 16S rRNA Gene Followed by Restriction Fragment Length Polymorphism Analysis

AbstractThe genetic relationship between faba bean (Vicia faba L.) phyllody and other mycoplasma‐like organism (MLO) diseases has been studied by amplification of the conserved region of the 16S rRNA gene followed by restriction fragment length polymorphism (RFLP) analysis using Alu I restriction endonuclease. The restriction patterns produced by faba bean phyllody MLO were smilar to that of Crotalaria saltiana phyllody MLO which persists throughout the year in the Sudan. These, and serological results clearly confirmed that C. saltiana is a reservoir of faba bean phyllody MLO in the Sudan. Moreover, restriction patterns have also shown that MLOs of other diseases have the same RFLP fragment pattern as faba bean phyllody MLO, including C. juncea witches'broom (Thailand) and tomato big‐bud (Australia), which differs from the other selected MLO diseases (Gladiolus aster yellow, clover phyllody and yellow decline of lavender, aqll from France).Fragment patterns also revealed the existence of genetically diverse MLO strains in the Sudan. Faba bean phyllody may be placed in group III including WX, apricot chlorotic leaf roll, golden flaveswcence dorée of grapevine, plum leptonecrosis of Prunus salciana, peachy yellow leaf roll, sunnhemp phyllody from Thailand, and blueberry witches' broom.

Molecular Cloning, Detection of Chromosomal DNA of the Mycoplasmalike Organism (MLO) Associated with Faba Bean (Vicia faba L.) Phyllody by Southern Blot Hybridization and the Polymerase Chain Reaction (PCR)

AbstractDNA from faba bean (Vicia faba L.) phyllody‐diseased periwinle plants was separated from host plant DNA by bisbenzimid‐CsCl buoyant‐density gradient centrifugation. The mycoplasmalike organism (MLO) DNA was used for the construction of DNA probes. Two probes, 1.45 and 1.35 kbp, were selected and used for the detection of MLO DNA associated with faba pean (FBP) and for assessing the genetic relatedness of FBP‐MLO with other mollicutes. The 1.45 kbp DNA probe hybridized with all MLO strains and, with Spiroplasma citri. The 1.35 kbp DNA probe specifically detected the MLO associated with FBP.Moreover, a specific primer pair (E1 and E2) selected from the partially sequenced 1.35 kbp probe allowed amplification of the 1.35 kbp fragment. DNA amplification was obtained also with Crotaltiana saltiana phyllody (Sudan), C. juncea, witches' broom (Thailand), and tomato big‐bud (Australia), but no amplification was obtained in the cases of the healthy control, C. roseus phyllody (isolate n0) from Sudan, clover phyllody, Gladiolus aster yellow and yellow decline of lavender from France. The very strong signal observed in the case of FBP and C. saltiana phyllody agrees with previous results indicating that FBP and C. saltiana phyllody are caused by an identical MLO, and hence, C. saltiana acts as a reservoir of FBP‐MLO in the Sudan. The weak signal obtained in the case of C. juncea witches' broom and tomáto big‐bud indicates partial nucleotide homology. The major interest of this primer pair is the low quantity (as little as 100 pg) of the total DNA of diseased plant required for the detection of the FBP disease and the possibility of detecting genetic relatedness with other MLOs.