Insight into resistance to 'Candidatus Liberibacter asiaticus,' associated with Huanglongbing, in Oceanian citrus genotypes.

Huanglongbing (HLB), the most destructive citrus disease, is associated with unculturable, phloem-limited Candidatus Liberibacter species, mainly Ca. L. asiaticus (Las). Las is transmitted naturally by the insect Diaphorina citri. In a previous study, we determined that the Oceanian citrus relatives Eremocitrus glauca, Microcitrus warburgiana, Microcitrus papuana, and Microcitrus australis and three hybrids among them and Citrus were full-resistant to Las. After 2 years of evaluations, leaves of those seven genotypes remained Las-free even with their susceptible rootstock being infected. However, Las was detected in their stem bark above the scion-rootstock graft union. Aiming to gain an understanding of the full-resistance phenotype, new experiments were carried out with the challenge-inoculated Oceanian citrus genotypes through which we evaluated: (1) Las acquisition by D. citri fed onto them; (2) Las infection in sweet orange plants grafted with bark or budwood from them; (3) Las infection in sweet orange plants top-grafted onto them; (4) Las infection in new shoots from rooted plants of them; and (5) Las infection in new shoots of them after drastic back-pruning. Overall, results showed that insects that fed on plants from the Oceanian citrus genotypes, their canopies, new flushes, and leaves from rooted cuttings evaluated remained quantitative real-time polymerase chain reaction (qPCR)-negative. Moreover, their budwood pieces were unable to infect sweet orange through grafting. Furthermore, sweet orange control leaves resulted infected when insects fed onto them and graft-receptor susceptible plants. Genomic and morphological analysis of the Oceanian genotypes corroborated that E. glauca and M. warburgiana are pure species while our M. australis accession is an M. australis × M. inodora hybrid and M. papuana is probably a M. papuana × M. warburgiana hybrid. E. glauca × C. sinensis hybrid was found coming from a cross between E. glauca and mandarin or tangor. Eremocitrus × Microcitrus hybrid is a complex admixture of M. australasica, M. australis, and E. glauca while the last hybrid is an M. australasica × M. australis admixture. Confirmation of consistent full resistance in these genotypes with proper validation of their genomic parentages is essential to map properly genomic regions for breeding programs aimed to generate new Citrus-like cultivars yielding immunity to HLB.

Evidence That 'Candidatus Liberibacter asiaticus' Moves Predominantly Toward New Tissue Growth in Citrus Plants.

'Candidatus Liberibacter asiaticus' (Las) is an unculturable, phloem-limited, insect-transmitted bacterium associated with the Asiatic form of huanglongbing (HLB), the most destructive citrus disease. In Asia and the Americas, it is transmitted by the Asian citrus psyllid (Diaphorina citri Kuwavama). Despite considerable research, little is known about the processes involved in plant infection and colonization by Las. This study was conducted to determine whether the basal portion (below girdling) of the plant is an important route for Las to move laterally from a point of inoculation on a branch to pathogen-free branches elsewhere in the canopy, and to quantify the influence of actively growing tissues on vertical upward (acropetally) or downward (basipetally) movement of Las. Nongirdled and fully or partially girdled stems of potted plants of 'Pera' sweet orange, graft-inoculated above or below girdling, were sampled in distinct regions and assessed by qPCR, 6 months postinoculation. Las invaded all regions of partially and nongirdled plants but remained restricted to the inoculated regions of fully girdled plants, evidence that in planta bacterium movement is limited to the phloem. In fully girdled plants, starch accumulated above the girdling site, probably because of changes in flow of phloem sap. To study the influence of actively growing tissues, inoculated 'Valencia' sweet orange plants were kept intact or were top- or root-pruned to force production of new tissues, and sampled at 15-day intervals. Las migrated rapidly and most predominantly toward newly developing root and leaf tissues. The rapid and predominant movement of Las to newly developed shoots and roots would explain failures of canopy heat treatments and pruning to cure HLB-affected trees, and reinforces the need to protect rapidly growing new shoots from feeding by D. citri in order to minimize transmission and spread of the pathogen by the vector within and between orchards.

Resistance to 'Candidatus Liberibacter asiaticus,' the Huanglongbing Associated Bacterium, in Sexually and/or Graft-Compatible Citrus Relatives.

Huanglongbing (HLB) is the most destructive, yet incurable disease of citrus. Finding sources of genetic resistance to HLB-associated 'Candidatus Liberibacter asiaticus' (Las) becomes strategic to warrant crop sustainability, but no resistant Citrus genotypes exist. Some Citrus relatives of the family Rutaceae, subfamily Aurantioideae, were described as full-resistant to Las, but they are phylogenetically far, thus incompatible with Citrus. Partial resistance was indicated for certain cross-compatible types. Moreover, other genotypes from subtribe Citrinae, sexually incompatible but graft-compatible with Citrus, may provide new rootstocks able to restrict bacterial titer in the canopy. Use of seedlings from monoembryonic species and inconsistencies in previous reports likely due to Las recalcitrance encouraged us to evaluate more accurately these Citrus relatives. We tested for Las resistance a diverse collection of graft-compatible Citrinae species using an aggressive and consistent challenge-inoculation and evaluation procedure. Most Citrinae species examined were either susceptible or partially resistant to Las. However, Eremocitrus glauca and Papua/New Guinea Microcitrus species as well as their hybrids and those with Citrus arose here for the first time as full-resistant, opening the way for using these underutilized genotypes as Las resistance sources in breeding programs or attempting using them directly as possible new Las-resistant Citrus rootstocks or interstocks.

Unsuitability of indigenous South American Rutaceae as potential hosts of Diaphorina citri.

BACKGROUND: 'Candidatus Liberibacter asiaticus' is transmitted by Diaphorina citri, an insect with a wide range of hosts in Rutaceae. Species related to Citrus occur in Brazilian forests where they may serve as hosts for psyllids and infested citrus orchards. RESULTS: The suitability of plants as hosts of D. citri was classified into four groups. Group I (high suitability): Citrus × aurantium 'Valencia', 'Citrus limonia', Murraya paniculata (syn. Murraya exotica L.) (Aurantioideae: Aurantieae) and Bergera koenigii (Aurantioideae: Clauseneae). Group II (intermediate to low suitability): Citrus (Poncirus) trifoliata 'Pomeroy', Citrus wintersii, Swinglea glutinosa (Aurantieae) and Clausena lansium (Clauseneae). Group III (not suitable): Aegle marmelos, Atalantia buxifolia, Citrus ('Microcitrus') sp. (Aurantieae) and Helietta apiculata (Amyridoideae). Group IV (non-hosts): Glycosmis pentaphylla (Clauseneae), Balfourodendron riedelianum, Casimiroa edulis, Esenbeckia febrifuga, Esenbeckia leiocarpa, Metrodorea stipularis, Zanthoxylum rhoifolium (Amyridoideae) and Dictyoloma vandellianum (Cneoroideae). Insects survived longer on newly differentiated leaves compared with fully expanded soft leaves. Psyllids either did not develop or did not survive for long on most Group IV species, all of which, with the exception of G. pentaphylla, occur naturally in Brazilian forests. CONCLUSION: Citrus relatives occurring in forests near citrus orchards are not suitable hosts of D. citri and, therefore, do not contribute to huanglongbing spread. © 2018 Society of Chemical Industry.

Grafting height does not affect Fusarium wilt control or horticultural performance of Passiflora gibertii N.E.Br. rootstock.

The influence of the grafting height (5, 10, 20 and 30 cm above the root collar) of P. edulis on P. gibertii was evaluated on the incidence of Fusarium wilt and horticultural performance. Plants of P. gibertii grafted on P. edulis and non-grafted plants of both species were also studied. In addition, histopathological studies were also performed on the roots of non-grafted P. edulis collected at three severity stages of Fusarium wilt. In greenhouse, the graft take was inversely related to the grafting height in general. In the field conditions, the plant growth of P. gibertii grafted on P. edulis was superior to its reciprocal grafting, even though the former combination was susceptible to Fop. Plants of P. edulis grafted on P. gibertii at all grafting heights did not present symptoms of Fop, and the number of fruit yield and quality were equivalent, but plant growth was decreased in relation to the non-grafted plants. Starch depletion in the root system of P. edulis was directly related to the severity of the Fusarium wilt. P. gibertii was confirmed as a Fusarium wilt resistant rootstock of P. edulis, with minimal influence of the grafting height for the control of the disease.