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Incidence and Severity of Huanglongbing and Candidatus

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Incidence and Severity of Huanglongbing and Candidatus HORTSCIENCE 46(10):1344–1348. 2011. P. trifoliata and its hybrid, ‘Carrizo’ citrange, developed less severe HLB symptoms and among the lowest titer of CLas among the Incidence and Severity of genotypes evaluated in a recent greenhouse study (Folimonova et al., 2009). This is strong Huanglongbing and Candidatus evidence that variation for HLB susceptibility exists within the citrus gene pool, and it is Liberibacter asiaticus Titer among possible that scion cultivars may display use- ful resistance. The majority of citrus production in Florida Field-infected Citrus Cultivars is represented by sweet orange (C. sinensis, Ed Stover1 and Greg McCollum 86% of the 200,000 ha of Florida citrus) USDA, ARS, US Horticultural Research Laboratory, 2001 S. Rock Road, followed by grapefruit (C. paradisi, 10% of total Florida citrus; NASS, 2008). In the Ft. Pierce, FL 34945 Folimonova et al. (2009) study on citrus sus- Additional index words. greening, resistance, tolerance, PCR ceptibility to HLB, sweet orange was in the most susceptible category and grapefruit Abstract. Incidence and severity of Huanglongbing (HLB) disease were assessed in Apr. showed slightly lower susceptibility. ‘‘Other 2010 among eight citrus cultivars representing diverse scion types growing in commer- citrus’’ grown in Florida is mostly mandarin cial groves in Florida’s Indian River region, an area with a high incidence of HLB. In hybrids (tangors, tangerines, and tangelos), each grove, 20 trees of each cultivar were rated for visual HLB symptoms and leaves which represent 9300 ha or 4% of all were collected for quantitative polymerase chain reaction quantification of Candidatus Florida citrus (NASS, 2008). Tangors are nat- Liberibacter asiaticus (CLas), the presumptive causal agent of HLB. There was a strong ural hybrids of C. reticulata and C. sinensis; correlation between HLB rating and CLas titer (titer represented by Ct, r2 = 0.37 and tangelos are hybrids between C. reticulata and 0.40, for whole tree and leaf sample, respectively, both with P < 0.0001) across all C. paradisi; whereas tangerines range from cultivars and groves. Although incidence and severity of HLB varied considerably among pure C. reticulata to complex hybrids that have the groves, scion-specific differences were apparent, even when analyses excluded been created by plant breeders and have a phe- potentially confounding grove effects. ‘Temple’ tangor showed the most consistently low notype similar to C. reticulata.Thereismuch incidence of HLB symptoms and CLas titer; in contrast, ‘Murcott’ tangor and greater genetic diversity among the mandarin ‘Minneola’ tangelo had the highest incidence of HLB symptoms and highest CLas titer. group than sweet oranges and grapefruit, These results suggest useful resistance to HLB with reduced symptoms and reduced CLas which are each represented essentially by bud titer may be found in conventional scion cultivars and further work is needed to assess sports from single genotypes. this potential and its commercial value. Although greenhouse studies using graft inoculation have been conducted to compare susceptibility to HLB among cultivars, it is Huanglongbing is a very destructive citrus tree removed. The ability to grow replanted unlikely that greenhouse results can be ex- disease (Bove, 2006) and is currently consid- trees near HLB-affected groves is uncertain trapolated to ultimate tree performance in the ered to be the greatest threat facing the Florida as a result of the presence of inoculum and field. Replicated field studies of HLB sus- and U.S. citrus industry (Gottwald, 2010). frequent initiation of new leaf growth by ceptibility require years to complete. This HLB is associated with a phloem-limited bac- young trees, which attracts ACP (Keiji et al., study was conducted to detect differences in terium, CLas, and its insect vector, Diaphorina 2005). A more sustainable approach for the HLB severity and incidence as well as CLas citri, the Asian citrus psyllid (ACP). As HLB management of HLB would be the cultivation titer among diverse citrus cultivars in extant progresses, trees become increasingly debil- of citrus cultivars that are unaffected, or less field plantings known to be affected by HLB itated, leading to loss of yield and poor fruit affected, by the disease rather than more sus- to provide guidance in selection of cultivars quality followed eventually by tree death ceptible types (Gottwald, 2010). for long-term field tests. (Bove, 2006; Gottwald, 2010). Since HLB Several studies have been conducted to was first confirmed in Florida in 2005, it has determine if there are differences in HLB Materials and Methods now been found in all 32 citrus-producing susceptibility among citrus cultivars and/or counties in the state as well as in dooryard cit- species. Compared with other tested cultivars Plant material. Our objective was to quan- rus in non-citrus-producing counties (Halbert within individual experiments, lower suscep- tify the incidence and severity of HLB among et al., 2008). tibility to HLB associated with CLas has been citrus types growing in commercial groves Currently, the strategies for management reported for limes (de Lange et al., 1985; with disease pressure typical of the region. of HLB include insecticide applications for Schwarz et al., 1973; Shokrollah et al., 2009), Because Florida citrus production is dominat- vector control, roguing trees affected by the pummelos (Koizumi et al., 1997; Schwarz ed by sweet orange and grapefruit, there are disease to reduce inoculum, and production et al., 1973), lemons (Cheema et al., 1982; few groves with diverse cultivars. As a conse- and use of nursery stock free of CLas/ACP Nariani, 1982; Schwarz et al., 1973), some quence, it was impossible to develop a bal- (Gottwald, 2010). Insecticide applications for mandarin types (e.g., ‘Ladu’ and ‘Som Pan’ anced sample set with all scions represented in ACP control have escalated in Florida in- in Thailand; Koizumi et al., 1997), and var- all groves. The eight commercial citrus groves creasing the likelihood of ACPs developing ious non-cultivated citrus or related species. selected for survey each contained four or insecticide resistance (e.g., Srinivasan et al., However, reports are inconsistent which may more scions (Table 1) planted within 1 km of 2008). Scouting to identify trees expressing reflect genotypes selected, interactions be- each other. Groves were located in the Florida HLB symptoms and subsequent tree removal tween the host genotype and strains of HLB counties of Indian River, St. Lucie, Martin, and are costly and reduce fruit yield with each pathogens studied (e.g., Kiritani and Su, Palm Beach (lat. 26.7 to 27.6° N and long. 80.2 1999), or the methods used to evaluate dis- to 80.4° W). Trees in each grove were mature ease. Reports before routine use of molecular bearing trees on standard industry rootstocks diagnostic tools may also reflect confusion of and had been maintained using local commer- Received for publication 14 July 2011. Accepted for publication 27 July 2011. apparent HLB with other diseases. Poncirus cial practices regarding fertilization, irrigation, The technical assistance of Gregory Brock, Angela trifoliata (proposed as Citrus trifoliata in a and pest control. Citrus scions evaluated in this Ledger, and Luc Overholt is gratefully acknowledged. recent revision of citrus taxonomy; Bayer study included sweet orange (cultivars Hamlin, 1To whom reprint requests should be addressed; et al., 2009) appears to be less susceptible to Valencia, and navel), grapefruit (‘Marsh’, e-mail [email protected]. HLB than are cultivated citrus scion varieties. ‘Ruby Red’, and ‘Flame’), tangors (‘Temple’, 1344 HORTSCIENCE VOL. 46(10) OCTOBER 2011 | BREEDING,CULTIVARS,ROOTSTOCKS, AND GERMPLASM RESOURCES Table 1. Percentage of trees (20 samples per cultivar per grove) with one or more CLas genomes per 100 ng titer (Ct values less than the overall mean). nucleic acid sample from diagnostic leaf extraction as assessed in quantitative polymerase chain Sweet orange is intermediate among cultivars reaction.z tested. This analysis does not consider the Grove influence of individual grove differences in Type Cultivar 12345678HLB incidence and cultivar composition, Sweet orange C. sinensis 45 20 NA 56 20 30 0 0 which are potentially confounding in analyz- Grapefruit C. paradisi 20 45 5 55 NA 5 0 0 ing overall means. Tangor Murcott NA 70 NA 65 5 NA NA NA Differences between cultivars in Candidatus Tangor Temple 10 NA 70 10 5 0 0 5 Liberibacter asiaticus titer after adjusting for Tangor Ortanique NA NA NA NA NA NA 30 NA grove differences. Because individual groves Tangelo Minneola 55 NA 90 50 25 30 5 0 varied greatly in CLas titer (Fig. 1A), it is Mandarin hybrid Fallglo 25 NA 20 NA NA NA NA NA Mandarin hybrid Sunburst 0 35 NA NA NA NA 0 NA possible that distribution of cultivars across groves may influence apparent susceptibility zData are presented for each Citrus scion type/cultivar in each grove assessed in this study. CLas = Candidatus Liberibacter asiaticus; NA = not available. to CLas. When Ct for each tree was normal- ized by individual grove means (‘‘grove- adjusted Ct all’’ in Fig. 2A; ‘Ortanique’ is ‘Murcott’, and ‘Ortanique’), and mandarin-type Data analysis. Data were analyzed using omitted from this analysis because presence hybrids (‘Fallglo’, ‘Minneola’, and ‘Sunburst’) SAS (Cary, NC) with all sweet orange culti- in a single grove resulted in a large SE), (Table 1). In each grove, 20 trees per cultivar vars combined in one group and all grapefruit ‘Temple’, ‘Fallglo’, ‘Sunburst’, and grape- were evaluated; however, in no case were all cultivars combined in another group. Analyses fruit show markedly lower CLas titers (high- cultivars available in all groves (Table 1). A included Ct values from qPCR (with a Ct value er relative Ct) compared with other cultivars, total of 720 trees were included in the study. of 40 assigned when there was no detectable whereas ‘Minneola’ and ‘Murcott’ showed Because there tends to be a higher than expected amplicon replication), grove-normalized Ct substantially higher CLas titers.
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