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Asiatic citrus canker (ACC) foliar lesions were evaluated on progenies of 84 seed-source genotypes ("parent genotypes") from the Citrus Variety Collection (CVC) of the University of California at Riverside (UCR) of Citrus trifoliata and hybrids between C. trifoliata and other Citrus species and hybrids. Trees were planted Aug. 2013 in a completely randomized design at the Fort Pierce U.S. Department of Agriculture (USDA) grove. Plants were assessed visually Aug. 2017, Sept. 2019, and Sept. 2020 for distinctive ACC lesion incidence and severity. Progeny were compared by parent genotypes using nonparametric analysis. Incidence of ACC [percentage of leaves displaying symptoms, verified by quantitative polymerase chain reaction (qPCR) to be associated with Xanthomonas citri pv. citri] across parent genotypes ranged from 8% to 80% (mean, 49%) of leaves affected in 2017, from 4% to 58% (mean, 29%) in 2019, and 8% to 46% (mean, 25%) in 2020. In 2017, of 49 C. trifoliata parent genotypes, only four separated from the two highest ACC-incidence statistical categories [Citrus Research Center (CRC) 3345, 3484, 3888, and 4017]. whereas 29 of the 35 C. trifoliata hybrids displayed lower ACC incidence, which separated from the two highest statistical categories. In 2019, of the C. trifoliata, only six separated from the highest ACC-incidence statistical category (CRC 3330, 3484, 3547, 3549, 3876, and 3888), whereas all 35 C. trifoliata hybrids displayed lower ACC incidence and separated from the highest statistical category, and 26 hybrids separated from 18 of the C. trifoliata. In 2020, only three C. trifoliata separated from the highest ACC-incidence statistical category (CRC 2861, 3549, and 3888) and 20 hybrids separated from 18 of the C. trifoliata. By parent genotype, ACC incidence correlated substantially between each pair of the 3 years, with r2 values of 0.39, 0.57, and 0.65. Of 34 hybrids validated, similar numbers had C. trifoliata, grapefruit (C. 3aurantium var. racemosa), and sweet orange (C. 3aurantium var. sinensis) chloroplasts. Chloroplast type affected ACC incidence and severity, but not in a consistent manner. Near-isogenic groups within C. trifoliata, as determined by DNA markers, were associated with some statistically different ACC sensitivity. Overall, hybrids of C. trifoliata with other citrus types displayed markedly reduced ACC sensitivity compared with C. trifoliata, indicating that this trait is readily overcome through breeding. [ABSTRACT FROM AUTHOR]

The U.S. Department of Agriculture citrus scion breeding program is urgently working on developing huanglongbing (HLB; pathogen Candidatus Liberibacter asiaticus)-tolerant cultivars with excellent fruit quality and productivity when HLB-affected. The slow process of assessing new citrus hybrids is a major impediment to delivery of these much-needed cultivars. We generate thousands of hybrids each year, germinate the seedlings, grow them for 2 years in the greenhouse, plant them at high density in a field where the disease HLB is abundant, grow them for 5 to 10 years, and make selections based on tree performance and fruit quality of these HLB-affected trees. Based on promising reports of accelerated citrus growth when grown in a metallized reflective mulch (MRM) system, we tested the hypothesis that the MRM system may accelerate growth and selection of new hybrid seedlings compared with conventional soil culture (CSC). In the MRM system, tree rows are covered with a layer of metallized plastic film and drip irrigation is installed beneath the plastic. In 2 years of analysis, tree canopy volume was significantly greater with MRM in 2020 (27% greater than CSC) but not in 2021, and MRM tree height was greater in 2021 (7% greater than CSC). Mortality was significantly greater with MRM in both 2020 and 2021(in 2021: 32% vs. 17% under CSC), and MRM trees had more chlorotic leaves. Because of staff limitations, plant debris and soil were not routinely cleared from MRM, thus diminishing any benefit from the reflective surface. Better maintenance might have resulted in more sustained evidence of MRM growth benefits. With the current resource availability, the MRM system does not appear to accelerate the assessment of hybrid seedling trees. [ABSTRACT FROM AUTHOR]

In a Fort Pierce, FL, field planting, plant growth, and Huanglongbing (HLB) severity were assessed as indicators of HLB tolerance on progenies of 83 seed-source accessions of Citrus and Citrus relatives mainly from the Riverside, CA, genebank. The HLB-associated pathogen [Candidatus Liberibacter asiaticus (CLas)] and vector [asian citrus psyllid (ACP), Diaphorina citri] were abundant, and trees were naturally challenged for 6 years before metrics (leaf mottle, percent canopy mottle, overall health, canopy density, canopy width, canopy height, and trunk diameter) were collected in Oct. and Nov. 2015. The healthiest trees with low or no HLB symptoms were distant citrus relatives: Balsamocitrus dawei, Bergera koenigii, Casimiroa edulis, Clausena excavata, Murraya paniculata, and one accession of Severinia buxifolia. Within Citrus, most of the healthiest trees with densest canopies, little leaf loss, and greater growth were those with pedigrees that included Citrus medica (citron). These included progenies of Citrus hybrid ('Limon Real'), Citrus limetta, Citrus limettioides, Citrus limonia, C. medica, Citrus volkameriana, and some Citrus limon accessions. Trees in this category exhibited distinct leaf-mottle characteristic of HLB and substantial pathogen titers, but maintained dense canopies and exhibited good growth. Trees from seed-source accessions in the genus Citrus without citron in their background were generally among the least healthy overall with less dense canopies. The exceptions were progenies of two Citrus aurantium accessions, which were markedly healthier than progenies of other Citrus seed-source accessions not derived from citron. Linear regression analysis, between metrics collected and pedigree of seed parent, indicated that percentage of citron in the pedigree significantly correlated with measures of tolerance. Although no commercial Citrus genotypes yielded progenies with strong HLB resistance, in this field experiment several progenies maintained dense canopies and good growth, and may be useful for breeding HLB tolerant cultivars. [ABSTRACT FROM AUTHOR]

Plants inoculated with the huanglongbing (HLB)-associated bacterium, Candidatus Liberibacter asiaticus (CLas) typically must be monitored for 8-10 months to identify differences in susceptibility between genotypes. Continuous light is reported to accelerate development of HLB symptoms and field observations suggest that trees girdled by tags or tree ties showed greater symptoms. Therefore, an experiment was conducted assessing HLB susceptibility as influenced by light/dark periods of 12 hours: 12 hours and 24 hours: 0 hours, in combination with scoring tree trunks to disrupt phloem. Sixty trees of each of three citrus genotypes ('Kuharske', previously shown to be HLB resistant; rough lemon, previously shown to be HLB tolerant; and 'Valencia', highly HLB susceptible) were bud grafted using two CLas-infected buds (rough lemon and citron) per tree on 26 Mar. 2012, and were placed in controlled growth rooms (one 12 hour light: 12 hour dark and one constant light) on 4 June 2012. Ten trees of each genotype in each growth room were scored 10 cm above the soil (cutting through the bark but not the wood) with a knife on 18 July 2012 and the scoring was repeated at the same scoring wounds on 30 Aug. 2012. Trees were removed from growth rooms on 12 Dec. 2012 and subsequently maintained in a greenhouse. At two to three month intervals between June 2012 and May 2013, HLB symptoms and stem diameter at 5 cm above the soil were assessed, and three leaves per tree were collected for quantitative polymerase chain reaction (qPCR) determination of CLas titer. Six months after inoculation and 3 months following imposition of treatments, the 'Valencia' scored in the 12 hour light: 12 hour dark regime, the 'Valencia' non scored trees in 24 hours of light and the 'Kuharske' scored trees in 24 hours of light displayed higher CLas titers than most other trees. After an additional two months, both scored and non-scored trees of all three genotypes in 24 hours of light had significantly elevated CLas titers compared with trees in 12 hour light: 12 hour dark regime, but within most treatments all three genotypes had titers which were not statistically different from each other. Growth of 'Kuharske' and rough lemon was enhanced; whereas 'Valencia' growth was reduced when graft-inoculated plants were maintained in continuous light. Scoring enhanced early CLas development in 'Kuharske' when combined with continuous light, had no effect in rough lemon, and showed inconsistent effects in 'Valencia'. Although continuous lighting enhanced disease progression, it did not reveal differences in HLB susceptibility. [ABSTRACT FROM AUTHOR]

Assessments of the resistance of citrus germplasm to huanglongbing (HLB) can be expedited by inoculating plants under laboratory or greenhouse settings with the HLB bacterium, Candidatus Liberibacter asiaticus (CLas). Consistent rapid screening is critical to efficiently assess disease resistance among plant materials; however, a number of factors may govern the efficacy of such inoculations. Despite the rapidity at which HLB can spread in a grove, it often takes 8 to 10 months for high levels of CLas and HLB symptoms to develop even in highly susceptible sweet orange. Therefore, two experiments were conducted to assess factors that might influence efficiency in screening for HLB resistance. In one experiment, three test citrus genotypes ('Kuharske', previously shown to be HLB resistant; rough lemon, previously shown to be HLB tolerant; and 'Valencia', HLB susceptible) were bud grafted using CLas-infected buds from four different source genotypes. All bud source genotypes had similar levels of CLas titer, but citron, rough lemon, and Volkamer lemon were hypothesized to be better bud inoculum sources as they are more tolerant of HLB than 'Valencia'. Among the three test genotypes over all sources of infected buds, inoculations of 'Kuharske' resulted in lower CLas titers and fewer HLB symptoms than inoculations of rough lemon or 'Valencia'. Inoculations of rough lemon resulted in higher CLas titers and more pronounced HLB symptoms when it was inoculated using infected buds from rough lemon or 'Valencia'. Grafting 'Valencia' with infected buds from Volkamer lemon resulted in less disease than when 'Valencia' was grafted with infected citron, rough lemon, or 'Valencia' buds. Overall, these results suggest that the source of CLas-infected buds used to graftinoculate some genotypes will influence disease development. Trunk cross-sectional area increase for the year following infection was 33 higher in 'Kuharske' and rough lemon, compared with 'Valencia'. 'Kuharske' had very low levels of CLas (30 CLas/µg DNA), whereas 'Valencia' (43,000 CLas/µg DNA) and rough lemon (6700 CLas/ µg DNA) had relatively high levels. As an alternative to graft-inoculating plants with CLas-infected buds, plants can be subjected to infestations of CLas-infected Asian citrus psyllid (ACP) as occurs naturally. Of interest is if transmission rates of CLas and the development of HLB in a genotype are greater when the ACP have been feeding on the same host genotype. An experiment was therefore conducted to assess transmission of CLas by ACP reared on CLas-infected rough lemon to five different genotypes ('Carrizo', 'Flame' grapefruit, rough lemon, 'Temple', and 'Valencia'). These assessments were made using a detached leaf assay recognized as a faster method of gauging transmission rates of CLas than using whole plants. Higher percentages of ACP died when they were transferred from infected rough lemon to healthy 'Carrizo', and lower percentages died when they were transferred to rough lemon or 'Flame'. However, CLas transmission by infected ACP occurred to at least some leaves of each genotype in each of the five different assays, with 70% or more leaves of each genotype becoming infected in at least one assay. Over all assays, there was relatively little variation among genotypes in the percentage of leaves becoming CLas infected and in the titer of CLas developing in infected leaves. However, there were relatively large differences in transmission rates among individual assays unrelated to differences among test genotypes. Because of the rapidity of the detached leaf assay, efforts are merited to improve consistency of this inoculation method. [ABSTRACT FROM AUTHOR]

The citrus disease huanglongbing (HLB) has become endemic in Florida, with estimates that greater than 80% of citrus trees are currently infected. Although there are no commercial citrus varieties with strong HLB resistance, some field tolerance has been observed in trees exposed to the disease after they were mature. There is great urgency to identify citrus which may permit economic citrus production where HLB is endemic. Therefore, the objective of this study was to assess field tolerance to HLB. To expedite the trial due to urgency, nursery trees were purchased on rootstock varieties as available. The trial included the following unbalanced scion/rootstock combinations: 'Hamlin/ Kinkoji', 'Hamlin/Cleopatra', 'Temple/Cleopatra', 'Fallglo/Kinkoji', 'Sugar Belle/Sour Orange', 'Tango/Kuharske', and 'Ruby Red/Kinkoji', with most comparisons based solely on scion/rootstock combinations. A randomized complete block experiment was established at Fort Pierce, FL, in Sept. 2010. All trees exhibited symptoms of HLB and tested positive for the Candidatus Liberibacter asiaticus (CLas) bacterium by Oct. 2012, with similar titers [directly assessed as cycle threshold (Ct) using quantitative polymerase chain reaction (qPCR)] measured for all scion/rootstocks at most sample dates, but early titer development in 'Ruby Red/Kinkoji' was significantly lower than several other scion/rootstocks. Across all time-points, 'Fallglo/Kinkoji' had the lowest rating of distinctive HLB mottling and 'Ruby Red/Kinkoji' had the highest rating, but 'Sugar-Belle/Sour Orange' had the highest percentage of leaves affected. After 5 years, 'SugarBelle/Sour Orange' and 'Tango/Kuharske' had the greatest overall increase in trunk diameter, and were among the healthiest in overall appearance. In Oct. 2015, 'SugarBelle/Sour Orange' and 'Tango/Kuharske' trees had significantly greater fruit load (80-88 fruit/tree) followed by 'Temple/Cleopatra' and 'Fallglo/Kinkoji' (31-35 fruit/tree) while 'Hamlin/Kinkoji', 'Hamlin/Cleopatra', and 'Ruby Red/Kinkoji' produced less than 20 fruit per tree. Despite becoming infected by CLas in less than 2 years after planting, the trees continued to grow and all scion/rootstocks displayed increasing fruit production, although very low in 'Hamlin/Kinkoji', 'Hamlin/Cleopatra', and 'Ruby Red/Kinkoji'. Growth and fruit production in the highest performing scion/ rootstocks were likely less than would be expected for healthy trees, but these results are promising, with markedly better response of some scion/rootstocks with mandarin hybrid scions compared with trees with sweet orange or grapefruit scions. Larger fully replicated trials are underway. It is noteworthy that the most pronounced HLB symptoms and higher early pathogen titer, which are the two criteria most widely used in assessing HLB resistance, were not associated with the lowest growth and cropping, and focus on early symptomatic traits when screening for resistance may obscure important disease tolerance. [ABSTRACT FROM AUTHOR]

A test population consisting of progenies of 92 seed-source genotypes (hereafter called "parent genotypes") of Citrus and Citrus relatives in the field in east-central Florida was assessed after natural freeze events in the winters of 2010 and 2011. Eight seedlings per parent genotype were planted in a randomized complete block design; however, as a result of mortality, the number of plants assessed in some genotype groups was reduced at some or all sampling dates. The citrus diseases huanglongbing and citrus canker were endemic in the planting and may have influenced tree response to cold temperatures. Unusually low temperatures (near -4 °C each winter) for east-central Florida were experienced during the trial period. Defoliation and dieback were significantly greater in the winter of 2011 than in the winter of 2010. The winter in 2011 was preceded by a period of extraordinarily low temperatures in mid-December with no period of cool temperatures to allow trees to acclimate. In 2010 the average defoliation was 53% ± 28% and less than 13% of the trees exhibited any noticeable dieback, whereas in 2011, the average defoliation and dieback were 93% ± 17% and 51% ± 35%, respectively. Within the genus Citrus, several progenies were identified that had 16% to 24% dieback in 2011 and these were from parent genotypes C. reticulata (CRC 2590) (23%), C. sinensis (CRC 3858) (24%), C. maxima (CRC 3945) (16%), C. hassaku (CRC 3907 and 3942) (16% and 17%), C. aurantium (CRC 628 and 2717) (18% and 7%), C. taiwanica (CRC 2588) (21%), and C. neo-aurantium (C. obovoidea + C. unshiu graft chimera) (CRC 3816) (23%). Within other genera in the Aurantiodeae, Poncirus trifoliata (CRCs 301,3957,3549, and 4007), Severinia buxifoiia (CRC 1497), Bergera koenigii (CRC 3165), and Glycosmis pentaphylla (CRC 3285) had the least amount of dieback, all at less than 23%. The two species within the Toddalioideae subfamily of the Rutaceae (Casimiroa edulis and Zanthoxylum ailanthoides) had among the least amount of dieback (1% and 8%, respectively). When considered by groups, the Citrons and Australian natives had the greatest amount of dieback in 2011, 68% and 65%, respectively. The trifoliates (Poncirus and hybrids) had the least dieback, ranging from 4% to 40%. The information from this study may be useful in germplasm enhancement and Citrus breeding targeting greater cold tolerance. [ABSTRACT FROM AUTHOR]

Xanthomonas citri ssp. citri (Xcc) is the causal agent of Asiatic citrus canker (ACC), a commercially important disease in Florida citrus as well as in many other regions. In this study we evaluated occurrence of foliar lesions from ACC on progenies of 94 seed-source genotypes (hereafter called "parent genotypes") of Citrus and Citrus relatives in the field in east-central Florida to identify the relative susceptibility to ACC. Eight seedlings per parent genotype were planted in a randomized complete block planting, but the number of plants assessed in some genotype groups was reduced by mortality at some or all sampling dates. Plants experienced ambient exposure to high Xcc inoculum pressure and plants were assessed in Sept. 2010, July 2011, Oct. 2011, May 2012, and Sept. 2012. The incidence and severity of ACC lesions were assessed and evaluated using non-parametric analyses to compare progeny from the 94 parent genotypes. Progeny of 14 parent genotypes did not exhibit ACC symptoms at any date. All were in genera other than Citrus with only Microcitrus and Eremocitrus being crosscompatible with Citrus. The kumquat hybrid C. halimii, two accessions of C. reticulata, C. nobilis, and C. sunki were the only Citrus species in the group that had a low severity (percent total leaf area showing symptoms) on each date of assessment. The aforementioned accessions had an average incidence and severity of ACC lesions of less than 4% in 2011 and 2012, but 26% to 38% in 2010 when no chemical control for ACC was applied in the adjoining citrus groves at our field site. Fourteen of the 16 progeny of C. reticulata and related parent genotypes were in the group with the lowest incidence and severity of ACC on two or more assessment dates. However, for analysis of only the most symptomatic leaves on each plant, 10 C. reticulata parent genotype progenies were in the most resistant category on the Sept. 2012 assessment despite having a low incidence and severity of ACC symptoms overall. Progeny of Poncirus and its hybrids as well as those of C. maxima, C. limon, and related species were the most severely diseased at all assessment dates. There were few instances in which progeny of different accessions of the same species had markedly different responses to ACC: progeny of C. reticulata 'Fremont' displayed more severe ACC compared with several other C. reticulata groups and C. aurantium 'Zhuluan' displayed much lower incidence and severity of ACC compared with several other accessions designated C. aurantium. Information on ACC susceptibility in diverse Citrus and Citrus relatives may prove useful for breeding programs aimed at reducing ACC susceptibility and will be of value to researchers interested in mechanisms of ACC resistance and susceptibility. [ABSTRACT FROM AUTHOR]

Transgenic solutions are being widely explored to develop huanglongbing (HLB) resistance in citrus. A critical component of a transgenic construct is the promoter, which determines tissue specificity and level of target gene expression. This study compares the characteristics of five promoters regulating the beta-glucuronidase ( GUS) reporter gene in the trifoliate hybrid rootstock US-802. Two of the selected promoters direct high levels of constitutive transgene expression in other dicotyledonous plants: 2X35S, the tandem-repeat promoter of the cauliflower mosaic virus 35S gene and bul409S, a truncation of the potato polyubiquitin promoter. Because Candidatus Liberibacter, the Gram-negative bacterium associated with HLB, infects only the phloem tissue, it may be advantageous to limit transgene expression to the vascular tissue and reduce expression in the fruit. Thus, we also tested three promoters that demonstrate phloem specificity when transformed and expressed in other plants: WDV, from wheat dwarf geminivirus; AtSUC2, the sucrose-H+ symporter gene promoter from Arabidopsis; and CsSUS, the sucrose synthase promoter from citrus. Histochemical staining for GUS activity was observed throughout leaf and stem tissues for the constitutive promoters, while the three phloem-specific promoters largely showed the expected tissue-specific staining. Expression of GUS in some individual transformants with promoters CsSUS and WDV appeared leaky, with some laminar tissue staining. Relative quantification of qRT-PCR data revealed a wide range of mRNA abundance from transgenics with each of the five promoters. Fluorometry also revealed that GUS activity differed depending on the promoter used, but mRNA levels and enzyme activity were not highly correlated. [ABSTRACT FROM AUTHOR]

Incidence and severity of Huanglongbing (HLB) disease were assessed in Apr. 2010 among eight citrus cultivars representing diverse scion types growing in commercial groves in Florida's Indian River region, an area with a high incidence of HLB. In each grove, 20 trees of each cultivar were rated for visual HLB symptoms and leaves were collected for quantitative polymerase chain reaction quantification of Candidatus Liberibacter asiaticus (CLas), the presumptive causal agent of HLB. There was a strong correlation between HLB rating and CLas titer (titer represented by Ct, r² = 0.37 and 0.40, for whole tree and leaf sample, respectively, both with P < 0.0001) across all cultivars and groves. Although incidence and severity of HLB varied considerably among the groves, scion-specific differences were apparent, even when analyses excluded potentially confounding grove effects. 'Temple' tangor showed the most consistently low incidence of HLB symptoms and CLas titer; in contrast, 'Murcott' tangor and 'Minneola' tangelo had the highest incidence of HLB symptoms and highest CLas titer. These results suggest useful resistance to HLB with reduced symptoms and reduced CLas titer may be found in conventional scion cultivars and further work is needed to assess this potential and its commercial value. [ABSTRACT FROM AUTHOR]

The article offers information on the fruit pomegranate (Punica granatum). It mentions that the fruit has a deep association with the cultures of the Mediterranean region and Near East. It notes that the pomegranate plant inherently develops numerous trunks. It details the health benefits of pomegranate.

The article offers information on the fig fruit. It mentions the association of the fruit with horticulture in the Mediterranean region. It provides an overview of the structure of the fig fruit. The types of figs that are described based on cropping/pollination characteristics include common figs and caprifigs.

In spring 1999, a commercial NAA (1-naphthaleneacetic acid) preparation for trunk sprout inhibition was compared with a corrugated plastic trunk wrap, aluminum foil wrap, bimonthly hand removal of sprouts, use of NAA preparation plus bimonthly hand removal when sprouts appeared, and a nontreated control. Three recently planted groves on three different rootstocks [Midsweet' orange (Citrus sinensis)] un Swingle citrumelo (Citrus paradisi x Poncirus trifoliata), 'Valencia' orange on Volkamer lemon (Volk, Citrus limon), and 'Minneola' tangelo (Citrus paradisi x C. reticulata) on Smooth Flat Seville (SFS, Citrus hybrid) received each of the treatments in a randomized complete block experimental design with trees blocked by initial height and circumference. Every 2 months, sprouts were counted on each tree and removed from the hand removal treatments. After 1 year, all sprouts were removed and counted and height and circumference of trees was determined. Across all experiments, 82% to 100% of nontreated trees produced trunk sprouts and all sprout control methods significantly reduced sprouts per tree. NAA treatments were never significantly less effective at sprout suppression than the wraps at the P = 0.05 level, although in two experiments, wraps were more effective than NAA at P = 0.10. Time of sprout appearance varied between the three experimental blocks. Plastic and foil trunk wraps enhanced development of trunk circumference compared with nontreated controls in 'Midsweet'/Swingle and 'Valencia'/ Volk. Greater trunk circumference resulted from use of wraps versus NAA in all three experiments, which appeared unrelated to differential sprout suppression. In these experiments, it appears that either wraps enhanced tree development beyond the suppression of sprouts or NAA influence on tree metabolism somewhat reduced trunk growth. The economics of the sprout suppression methods are also discussed. [ABSTRACT FROM AUTHOR]

Six trials were conducted to determine whether lower spray volumes or inclusion of different surfactants would permit adequate thinning of mandarin hybrids (Citrus reticulata hybrids) at a much lower cost per hectare. Sprays were applied using a commercial airblast orchard sprayer during physiological drop when fruitlets averaged 8 to 16 mm in diameter. Surfactant was always included at 0.05% v/v. NAA always reduced fruit per tree, increased fruit size, and decreased production of smallest size fruit. However, in only three experiments, contrast of all NAA treatments vs. controls indicated increased production of the largest (80-100 fruit per carton) and most valuable fruit. In four of five experiments, comparison of spray volumes of 600 (only examined in three of four experiments), 1200, or 2300 L·ha-1 demonstrated significant fruit size enhancement from all NAA applications. Most individual NAA treatments resulted in fewer fruit per tree, but there were no statistically significant differences between NAA treatments at different spray volumes. In only one of the four experiments, there was a marked linear relationship between spray volume and fruit per tree, yield, mean fruit size, and production of largest fruit sizes. The effects of surfactants (Activator, a nonionic, Silwet L-77, and LI-700) on NAA thinning were tested in both 'Murcott' and 'Sunburst'. In comparisons between Silwet L-77 and Activator surfactant, one experiment with 'Murcott' showed greater fruit per tree and yield reduction from using Silwet, but with a smaller increase in production of largest fruit sizes, whereas in another 'Murcott' experiment, Silwet L-77 reduced numbers of smaller fruit size with no increase in production of larger fruit. Based on these findings, current recommendations for NAA thinning of Fla. mandarins are use of spray volume of ≈1100-1400 L·ha-1 on mature trees with proportionally lower volume on smaller trees. These data appear to support use of a nonionic surfactant rather than other tested surfactants in NAA thinning of Florida mandarins. Because experience with NAA thinning of Florida citrus is limited, it is only recommended where the disadvantages of overcropping are perceived to substantially outweigh the potential losses from overthinning. [ABSTRACT FROM AUTHOR]

Wind-induced blemishing known as windscar and lesions from the disease melanose (caused by Diaporthe cirri) are two of the most important causes of fresh grapefruit (Citrus paradisi) cullage in Florida. Copper hydroxide fungicides are the primary means of controlling melanose, but high air velocities from passing sprayers have been suspected of increasing windscar. In 1998 and 1999, airblast applications of Cu(OH)2 (1.7 kg⋅ha-1 Cu) were made at a range of early fruit development stages to a fresh grapefruit orchard in the Indian River region of Florida. These applications supplemented aerial sprays of Cu(OH)2 that were made uniformly across the entire experimental site at biweekly intervals beginning near full bloom. During the commercial harvest period fruit were sampled from three regions (interior, upper exterior, and lower exterior) of each treatment tree and were evaluated for percentage of fruit surface covered by windscar and severity of melanose. Airblast applications did not affect windscar in either year, but windscar was significantly greater from the upper exterior of the canopy, which is likely to experience the highest natural wind velocities. From these data, it appears unlikely that airblast applications significantly contribute to windscar of Indian River grapefruit. In 1998, no trees receiving airblast applications had significantly lower melanose incidence than the trees sprayed only via aircraft; however, trees receiving four airblast applications were scored as having higher apparent melanose on exterior samples than trees receiving most other treatments. This is consistent with high levels of Cu injury on these fruit which can superficially resemble melanose. Following treatment in 1999, trees receiving four airblast applications of Cu(OH)2 had significantly lower melanose scores than trees receiving either no or only early airblast applications, but were not significantly... [ABSTRACT FROM AUTHOR]

'Mclntosh' apples (Malus x domestica Borkh.) display a rapid increase in ethylene production as they ripen, resulting in more preharvest drop and accelerated softening compared with other major cultivars. Economic considerations often dictate a choice between delaying harvest to achieve color development or harvesting earlier to avoid excessive fruit softening and drop. We have evaluated the effects of plant growth regulators (PGRs) and summer pruning on this balance. Treatments were applied to trees in the Mid-Hudson region in New York state in 1995 and 1996, and a subset of treatments was applied in the Champlain Valley region in 1996. NAA, applied at 10 mg⋅L[sup-1] in 1995 and 20 mg⋅L[sup-1] in 1996, reduced drop on only one sample date in only one of the three trials. Ethephon at 150 mg⋅L[sup-1] plus 10 mg⋅L[sup-1] NAA, accelerated ripening and permitted harvest before substantial drop occurred. However, earlier harvest resulted in smaller fruit size, and if ethephon-treated fruit were not picked within a narrow window, rapid drop ensued, and fruit developed a high senescent breakdown incidence during storage. ReTain, containing AVG, at 124 g⋅ha[sup-1] a.i. delayed drop in all three trials, but its use resulted in firmer fruit after storage in only two of seven comparisons. Use of ethephon on AVG-treated trees enhanced red color but accelerated drop, although it was reduced less than when ethephon was used alone. Severe late summer pruning accelerated red color development, drop and ripening in both years of the study. AVG was more effective for management of 'McIntosh' harvest in the cooler Champlain Valley region than in the Mid-Hudson Valley region. Chemical names used: naphthalene acetic acid (NAA); 2-chloroethylphosphonic acid (ethephon); aminoethoxyvinylglycine (AVG). [ABSTRACT FROM AUTHOR]

Tropical fruits have long excited the imaginations and palates of people living in temperate countries. Bananas became commonplace in the few decades ending the 19th century since they were so amenable to shipping and marketing. Pineapples made the transition from canned-only to routine fresh availability and avocados, mangoes, and papayas rapidly joined them as routine grocery products in the last two decades of the 20th century. Affluence and affordable jet freight have made it possible for many U.S. consumers to periodically sample delights such as Annona species (cherimoyas and atemoyas), carambolas, lychees, longans, pitayas, and passionfruit. In addition, distinctive cultivars of the staple tropicals are finding their way to market. The future seems likely to offer an even greater variety of tropical fruits which will be explored in this session. The workshop will be followed by a tasting of some of these fruits and the American Pomological Society business meeting. [ABSTRACT FROM AUTHOR]

Sweet orange (Citrus sinensis) exhibits limited genetic diversity and high susceptibility to Huanglongbing (HLB). Breeding HLB-tolerant orange-like hybrids is in dire need. However, our understanding of the key compounds responsible for orange flavor and their genetic regulation remains elusive. Evaluating 179 juice samples, including oranges, mandarins, Poncirus trifoliata, and hybrids, distinct volatile compositions were found. A random forest model predicted untrained samples with 78% accuracy and identified 26 compounds crucial for orange flavor. Notably, seven esters differentiated orange from mandarin flavor. Cluster analysis showed six esters with shared genetic control. Differential gene expression analysis identified C. sinensis alcohol acyltransferase 1 (CsAAT1) responsible for ester production in orange. Its activity was validated through overexpression assays. Phylogeny revealed the functional allele was inherited from pummelo. A SNP-based DNA marker in the coding region accurately predicted phenotypes. This study enhances our understanding of orange flavor compounds and their biosynthetic pathways and expands breeding options for orange-like cultivars.k [ABSTRACT FROM AUTHOR]

Huanglongbing (HLB) is the most devastating disease for citrus worldwide. Candidatus Liberibacter asiaticus (CLas), vectored by Asian citrus psyllid (ACP, Diaphorina citri Kuwayama), is the most common pathogen causing the disease. Commercial citrus varieties are highly susceptible to HLB, whereas trifoliate orange (Poncirus trifoliata) is considered highly tolerant to HLB. An F1 segregating population and their parent trifoliate orange and sweet orange, which had been exposed to intense HLB pressure for three years, was evaluated for disease symptoms, ACP colonization, CLas titer and tree vigor repeatedly for two to three years. Trifoliate orange and sweet orange showed significant differences for most of the phenotypic traits, and the F1 population exhibited a large variation. A high-density SNP-based genetic map with 1 402 markers was constructed for trifoliate orange, which exhibited high synteny and high coverage of its reference genome. A total of 26 quantitative trait locus (QTLs) were identified in four linkage groups LG-t6, LG-t7, LG-t8 and LG-t9, of which four QTL clusters exhibit a clear co-localization of QTLs associated with different traits. Through genome-wide analysis of gene expression in response to CLas infection in 'Flying Dragon' and 'Larger-Flower DPI-50-7' trifoliate orange, 85 differentially expressed genes were found located within the QTL clusters. Among them, seven genes were classified as defense or immunity protein which exhibited the highest transcriptional change after CLas infection. Our results indicate a quantitative genetic nature of HLB tolerance and identified candidate genes that should be valuable for searching for genetic solutions to HLB through breeding or genetic engineering. [ABSTRACT FROM AUTHOR]

Citrus Huanglongbing (HLB) associated with ‘Candidatus Liberibacter asiaticus’ (Las) and citrus canker disease incited by Xanthomonas citri are the most devastating citrus diseases worldwide. To control citrus HLB and canker disease, we previously screened over forty antimicrobial peptides (AMPs) in vitro for their potential application in genetic engineering. D2A21 was one of the most active AMPs against X. citri, Agrobacterium tumefaciens and Sinorhizobium meliloti with low hemolysis activity. Therefore, we conducted this work to assess transgenic expression of D2A21 peptide to achieve citrus resistant to canker and HLB. We generated a construct expressing D2A21 and initially transformed tobacco as a model plant. Transgenic tobacco expressing D2A21 was obtained by Agrobacterium-mediated transformation. Successful transformation and D2A21 expression was confirmed by molecular analysis. We evaluated disease development incited by Pseudomonas syringae pv. tabaci in transgenic tobacco. Transgenic tobacco plants expressing D2A21 showed remarkable disease resistance compared to control plants. Therefore, we performed citrus transformations with the same construct and obtained transgenic Carrizo citrange. Gene integration and gene expression in transgenic plants were determined by PCR and RT-qPCR. Transgenic Carrizo expressing D2A21 showed significant canker resistance while the control plants showed clear canker symptoms following both leaf infiltration and spray inoculation with X. citri 3213. Transgenic Carrizo plants were challenged for HLB evaluation by grafting with Las infected rough lemon buds. Las titer was determined by qPCR in the leaves and roots of transgenic and control plants. However, our results showed that transgenic plants expressing D2A21 did not significantly reduce Las titer compared to control plants. We demonstrated that transgenic expression of D2A21 conferred resistance to diseases incited by P. syringae pv. tabaci and X. citri but not Las. Our results underscore the difficulty in controlling HLB compared to other bacterial diseases. [ABSTRACT FROM AUTHOR]

Hybrids of Poncirus trifoliata L. Raf. with Citrus have shown degrees of tolerance to the deadly citrus greening disease, hence prompting interest as potential commercial varieties. Although P. trifoliata is known to produce fruit that is inedible, fruit from many advanced hybrid trees have not been evaluated for their quality potential. The sensory quality of selected Citrus hybrids with varying degrees of P. trifoliata in their pedigrees is reported herein. Four Citrus × P. trifoliata hybrids developed through the USDA Citrus scion breeding program—1‐76‐100, 1‐77‐105, 5‐18‐24, and 5‐18‐31—had acceptable eating quality and sweet and sour taste, with mandarin, orange, fruity–noncitrus, and floral flavors. On the other hand, hybrids with higher proportion of P. trifoliata in their pedigrees, US 119 and 6‐23‐20, produced a juice characterized by green, cooked, bitter, and Poncirus‐like flavor and aftertaste. Partial least square regressions revealed that the Poncirus‐like off‐flavor is likely due to a combination of higher than typical amounts of sesquiterpene hydrocarbons (woody/green odor), monoterpenes (citrus/pine), and terpene esters (floral) and a lack of aldehydes with typical citrus odor (octanal, nonanal, and decanal). Sweetness and sourness were mostly explained by high sugars and acids, respectively. Further, carvones and linalool contributed to sweetness in the samples from early and late seasons, respectively. In addition to highlighting chemical contributors to sensory descriptors in Citrus × P. trifoliata hybrids, this study provides useful information on sensory quality for future citrus breeding efforts. Practical Application: The relationships between the sensory quality and secondary metabolites of Citrus × P. trifoliata hybrids described in this study help identify disease‐resistant Citrus scion hybrids with acceptable flavor and help mobilize this resistance in future breeding efforts. It also shows potential of such hybrids to be commercialized. [ABSTRACT FROM AUTHOR]

Avocado (Persea americana Mill.) is a high-value fruit that continues to increase in consumer demand. A population of 'Hass'-'Bacon' hybrids was planted at USDA-ARS, Fort Pierce, as part of a study to find selections with good horticultural and postharvest quality traits for Florida. Extensive phenotypic data on quality were collected over 3 years. Ten selections were identified in 2014 and 2015 with promising fruit quality and postharvest shelf life characteristics and were tested in sensory panels using store-bought 'Hass' as the standard. In general, the selections had fruit quality similar to commercial 'Hass'. Avocados that were most liked were described as creamy in texture with buttery and nutty flavor. Only one selection (R7T54 in 2014) and one store-bought control ('Hass' in 2015) were disliked, which was associated with greater firmness at the time of evaluation, likely relating to insufficient postharvest conditioning. Furthermore, CA 'Hass' commercial requirements for minimum dry matter (20.8%) were generally achieved by these selections under Florida conditions, ranging from 18.4% to 25.7%. This study identified 10 selections with composition and sensory quality similar to 'Hass' that are suitable for further testing and development in Florida. [ABSTRACT FROM AUTHOR]

The level of anthocyanins in plants vary widely among cultivars, developmental stages and environmental stimuli. Previous studies have reported that the expression of various MYBs regulate anthocyanin pigmentation during growth and development. Here we examine the activity of 3 novel R2R3-MYB transcription factor (TF) genes,PamMybA.1, PamMybA.3andPamMybA.5fromPrunus americana. The anthocyanin accumulation patterns mediated byCaMV double35Spromoter (db35Sp) controlled expression of the TFs in transgenic tobacco were compared with citrus-MoroMybA, Arabidopsis-AtMybA1and grapevine-VvMybA1transgenics during their entire growth cycles. Thedb35Sp-PamMybA.1anddb35Sp-PamMybA.5 constructs induced high levels of anthocyanin accumulation in both transformed tobacco calli and the regenerated plants. The red/purple color pigmentation induced in thePamMybA.1andPamMybA.5 lines was not uniformly distributed, but appeared as patches in the leaves, whereas the flowers showed intense uniform pigmentation similar to theVvMybA1expressing lines.MoroMybAandAtMybA1showed more uniform pink coloration in both vegetative and reproductive tissues. Plant morphology, anthocyanin content, seed viability, and transgene inheritance were examined for thePamMybA.5transgenic plants and compared with the controls. We conclude that these TFs alone are sufficient for activating anthocyanin production in plants and may be used as visible reporter genes for plant transformation. Evaluating these TFs in a heterologous crop species such as citrus further validated that these genes can be useful for the metabolic engineering of anthocyanin production and cultivar enhancement. [ABSTRACT FROM AUTHOR]

The article provides a historical overview placing Mediterranean fruits into cultural context discussed at a workshop presented by the American Pomological Society (APS) held in the U.S. The event also provided information on the current cultivars and their place in modern commerce. It notes that the APS is an organization affiliated with the American Society for Horticultural Science (ASHS).

Asiatic huanglongbing (HLB) is a devastating disease of citrus associated in North America with the bacterium 'Candidatus Liberibacter asiaticus' (LAS) vectored by the Asian citrus psyllid (ACP), Diaphorina citri Kuwayama. ACP management is considered a vital component of a program aimed at reducing the incidence and spread of HLB. Considerable research has been published comparing the efficacy of different insecticides for ACP control as well as on seasonal strategies for applying ACP insecticides. However, published information was largely lacking for even the most intense insecticide programs on their effectiveness for keeping HLB out of a new citrus planting in Florida citrus. We therefore conducted two replicated (individual plots 0.2 to 0.4 ha in size) experiments on protecting young citrus from HLB using different ACP management programs. An intensive insecticide program was evaluated in each experiment: eight annual calendar applications of traditional insecticides (hereafter referred to as the "complete" program). In one experiment, citrus was either planted alone and subjected to the complete program or citrus was interplanted with orange jasmine, Murraya exotica L. (a favored ACP host plant) and subjected to a reduced insecticide program (four calendar sprays of traditional insecticides). There was one set of plots in which both jasmine and citrus were treated with insecticides and one set in which jasmine was not treated at all. In the second experiment, citrus was either subjected to the complete program or to one of two other programs: a reduced insecticide program consisting of five calendar applications of traditional insecticides or a mineral oil program (oil applications every 3 weeks plus one dormant insecticide spray). The results of the two experiments were similar. Relatively good ACP control was achieved under each ACP management program during the first year but, as the experiments progressed and trees increased in size, ACP outbreaks occurred regardless of the psyllid management program. Little HLB developed under any ACP management program during the first year, but thereafter HLB increased and large percentages of the trees in each experiment became LAS-infected in less than two to three years. The combined results of the experiments indicated that up to eight monthly pesticide treatments per year applied on a calendar schedule were ineffective for preventing young citrus from becoming diseased. Of important significance is that the orchard within which the two experiments were conducted was subjected to a minimal psyllid management program and contained many older trees known to be infected by the HLB pathogen, a challenging situation for getting young trees into production without contracting the disease. Thus, the ACP management programs we evaluated might have been more effective if ACP in the surrounding areas had been more aggressively controlled and diseased trees in the surrounding areas removed to reduce inoculum loads. Also, the programs might have been more effective for slowing the spread of HLB if our research plots had been larger. [ABSTRACT FROM AUTHOR]

Huanglongbing (HLB) is a serious and devastating disease of citrus caused by Candidatus Liberibacter spp. and vectored by the Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae). The disease has the potential to greatly limit the production of citrus in Florida and other citrus-growing regions worldwide. Current control of D. citri and HLB is inadequate, but the identification and incorporation of D. citri resistance traits from uncultivated Citrus spp. and Citrus relatives is seen as a potential disease management strategy. In this study, seedlings of 87 Rutaceae seed-source genotypes, primarily in the orange subfamily Aurantioideae, were assessed in the field for their propensity in a free-choice situation for infestations of natural south Florida populations of D. citri. The majority of test populations surveyed hosted all three life stages of D. citri; however, there were significant differences among the test populations in the mean ranks for 19. citri eggs (F = 3.13, df = 86, P < 0.0001), nymphs (F = 9.01, df 86, P < 0.0001), and adults (F = 4.21, df 86, P < 0.0001). The only sample test population that was completely avoided by all life stages of D. citri was seedlings of Casimiroa edulis Llave et Lex, commonly known as white sapote, which was one of the few plants included in the study belonging to the Rutaceae subfamily Toddalioideae. Although not completely avoided, very low levels of D. citri were found on two surveyed test populations of Poncirus trifoliata (L.) Raf, seedlings of 'Simmon's trifoliate' and 'Little-Leaf'. Poncirus trifoliata, the trifoliate orange, readily forms hybrids with Citrus spp., is commonly incorporated into rootstock varieties, and has been used in breeding-advanced scion material. The identification of partial resistance in P. trifoliata to D. citri could prove useful in future citrus breeding efforts aimed at reducing the incidence and spread of HLB. Determining if there is true resistance to D. citri in this and other germplasm sources with a low propensity for infestations in free-choice situations will require no-choice experiments. [ABSTRACT FROM AUTHOR]

The year 2005 marked the 25th anniversary of the establishment of the U.S. Department of Agriculture (USDA) National Plant Germplasm System (NPGS), repositories devoted to clonally propagated, horticultural fruit and nut crops. During this quarter century, facilities in Hilo, Hawaii; Mayaguez, PR,; Miami, Fla.; and Riverside, Calif. were developed to preserve collections of tropical and subtropical fruit and nut crops; facilities in Brownwood, Texas; Corvallis, Ore.; Davis, Calif. and Geneva, N.Y. preserve the temperate crops. Each of these facilities now has internationally recognized, globally diverse collections of genetic resources for their assigned genera. Germplasm of unique genotypes are maintained as growing plants, evaluated for phenotypic and genotypic traits, documented in a national public germplasm database, and freely distributed as clonal propaggules to researchers and other germplasm users around the world. Seed collections represent wild populations for some crop relatives. These 8 genebanks maintain 30,000 accessions representing 1600 species of fruit and nut crops and their wild relatives. The genebanks distribute more than 15,000 accessions annually to international researchers. Although originally conceived as working collections for crop improvement, NPGS genebanks have also become invaluable in providing the raw materials for basic plant genetic research, reservoirs tot rare or endangered species or vulnerable landraces, archives of historic cultivars, and field classrooms for educating the public. These collections preserve botanical treasures as well as the American horticultural heritage for now and for future generations. [ABSTRACT FROM AUTHOR]

The Asian citrus psyllid (Hemiptera: Liviidae) transmits a bacterium responsible for huanglongbing, a serious citrus disease. Insecticidal control of the psyllid is a key tactic used to manage the disease, but host plant resistance may hold some promise and clues in the search for alternative tactics. Results of a field survey revealed that relatively large infestation densities of the psyllid developed on conventional Citrus (Rutaceae) and citrange (Rutaceae) cultivars but not on any of 6 Poncirus trifoliata (L.) Raf. (Rutaceae) cultivars. Poncirus trifoliata is a species closely related to Citrus, and citranges are hybrids of sweet orange and P. trifoliata. Reduced colonization by the psyllid on P. trifoliata was largely a result of reduced rates of oviposition. Poncirus trifoliata resistance to oviposition was not observed in 4 citrange cultivars. [ABSTRACT FROM AUTHOR]

Genetic traits in Poncirus trifoliata (Rutaceae) are known to confer resistance to colonization by the Asian citrus psyllid, but field surveys indicated these traits were not expressed in any of 24 simple hybrids between P. trifoliata and sweet orange. No resistance to colonization by the psyllid was observed in 14 other types of P. trifoliata hybrids. Three citrumelo hybrids appeared to be resistant in 1 survey, but in another survey, none of 6 citrumelos exhibited resistance. Interestingly, colonization was somewhat reduced on a hybrid with P. trifoliata in the pedigree of both parents, suggesting the possibility that traits suppressing oviposition may be recovered through similar crosses. [ABSTRACT FROM AUTHOR]

Genetic transformation is a powerful means for the improvement of crop plants, but requires labor- and resource-intensive methods. An efficient method for identifying single-copy transgene insertion events from a population of independent transgenic lines is desirable. Currently, transgene copy number is estimated by either Southern blot hybridization analyses or quantitative polymerase chain reaction ( qPCR) experiments. Southern hybridization is a convincing and reliable method, but it also is expensive, time-consuming and often requires a large amount of genomic DNA and radioactively labeled probes. Alternatively, qPCR requires less DNA and is potentially simpler to perform, but its results can lack the accuracy and precision needed to confidently distinguish between one- and two-copy events in transgenic plants with large genomes. To address this need, we developed a droplet digital PCR-based method for transgene copy number measurement in an array of crops: rice, citrus, potato, maize, tomato and wheat. The method utilizes specific primers to amplify target transgenes, and endogenous reference genes in a single duplexed reaction containing thousands of droplets. Endpoint amplicon production in the droplets is detected and quantified using sequence-specific fluorescently labeled probes. The results demonstrate that this approach can generate confident copy number measurements in independent transgenic lines in these crop species. This method and the compendium of probes and primers will be a useful resource for the plant research community, enabling the simple and accurate determination of transgene copy number in these six important crop species. [ABSTRACT FROM AUTHOR]

Citrus huanglongbing (HLB) is a destructive disease with no known cure. To identify sources of HLB resistance in the subfamily Aurantioideae to which citrus belongs, we conducted a six-year field trial under natural disease challenge conditions in an HLB endemic region. The study included 65 Citrus accessions and 33 accessions belonging to 20 other closely related genera. For each accession, eight seedling trees were evaluated. Based on quantitative polymerase chain reaction analysis of the pathogen titers and disease symptoms, eight disease-response categories were identified. We report two immune, six resistant, and 14 tolerant accessions. Resistance and tolerance observed in different accessions may be attributed to a multitude of factors, including psyllid colonization ability, absence of pathogen multiplication, transient replication of the bacterium. lack of pathogen establishment in the plant, delayed infection, or recovery from infection. Most citrus cultivars were considered susceptible: 15 citrons, lemons, and limes retained leaves in spite of the disease status. Resistance and high levels of field tolerance were observed in many noncitrus genera. Disease resistance/tolerance was observed in Australian citrus relative genera Eremocitrus and Microcitrus, which are sexually compatible with citms and may be useful in future breeding trials to impart HLB resistance to cultivated citrus. [ABSTRACT FROM AUTHOR]

Background: Citrus represents a crop of global importance both in economic impact and significance to nutrition. Citrus production worldwide is threatened by the disease Huanglongbing (HLB), caused by the phloem-limited pathogen Liberibacter spp.. As a source of stable HLB-resistance has yet to be identified, there is Candidatus considerable interest in characterization of novel disease-associated citrus genes. Results: A gene family of Small Cyclic Amphipathic Peptides (SCAmpPs) in citrus is described. The citrus genomes contain 100-150 SCAmpPs genes, approximately 50 of which are represented in the citrus EST database. These genes encode small ~50 residue precursor proteins that are post-translationally processed, releasing 5-10 residue cyclic peptides. The structures of the SCAmpPs genes are highly conserved, with the small coding domains interrupted by a single intron and relatively extended untranslated regions. Some family members are very highly transcribed in specific citrus tissues, as determined by representation in tissue-specific cDNA libraries. Comparison of the ESTs of related SCAmpPs revealed an unexpected evolutionary profile, consistent with targeted mutagenesis of the predicted cyclic peptide domain. The SCAmpPs genes are displayed in clusters on the citrus chromosomes, with apparent association with receptor leucine-rich repeat protein arrays. This study focused on three SCAmpPs family members with high constitutive expression in citrus phloem. Unexpectedly high sequence conservation was observed in the promoter region of two phloem-expressed SCAmpPs that encode very distinct predicted cyclic products. The processed cyclic product of one of these phloem SCAmpPs was characterized by LC-MS-MS analysis of phloem tissue, revealing properties consistent with a K+ ionophore. Conclusions: The SCAmpPs amino acid composition, protein structure, expression patterns, evolutionary profile and chromosomal distribution are consistent with designation as ribosomally synthesized defense-related peptides. [ABSTRACT FROM AUTHOR]

Six 'Ambersweet'-derived hybrids, similar to sweet orange fruit size, color, and taste and potential as new sweet orange cultivars, were selected to determine their fruit categorization by comparison of their volatile profiles with the parent and 'Hamlin', a typical sweet orange. All hybrids are at least ½ sweet orange and varying amounts of mandarin, grapefruit, Poncirus trifoliata, and sour orange in each pedigree. In total, 135 volatiles were detected in the eight hybrid lines/commercial cultivars over two harvests, and 20 compounds were detected in all samples, including terpenes (limonene, β-myrcene, α-pinene, α-terpinene, α-terpineol, and linalool), esters (ethyl butanote, ethyl pentanoate, and ethyl acetate), aldehydes (acetaldehyde, hexanal, and nonanal), and alcohols (ethanol and hexanol). Total abundance of volatiles in January-harvested fruits averaged 30% higher than for fruits of the same trees harvested in November. 'Ambersweet' contained the highest total amount of volatiles (mainly as a result of very high levels of monoterpenes), and of them, nootkatone and six other compounds were not detected in any of the hybrids, and some of them were also not detected in 'Hamlin'. On the other hand, 12 compounds, including pentanal, ethyl 2-butenoate, and ethyl nonanoate, were not detected in 'Ambersweet' but were found in 'Hamlin' and some of the hybrids. Cluster analysis separated the cultivar/hybrid and harvest time combinations into three clusters. FF-1-76-50, FF-1-76-52 and January FF-1-75-55, all with the same parents ('Ambersweet' x FF-1-30-52), were close to FF-1-65-55, but they were separated from 'Hamlin' and further separated from 'Ambersweet'. The cluster containing 'Hamlin' has three subclusters: January 'Hamlin' and November FF-1-74-14, a hybrid with one-eighth P. trifoliata, which includes a slight off-flavor frequently found in P. trifoliata hybrids, independent of each other, and both were separated from a group of November 'Hamlin', FF-1-64-97, and FF-1-75-55. The cluster containing 'Ambersweet' included January FF-1-64-97. A principle component analysis (PCA) separated 'Ambersweet' from all hybrids and 'Hamlin' along the PC1 axis and separated November harvests from January harvests along PC2. This volatile analysis supports the classification of the hybrids as sweet orange. [ABSTRACT FROM AUTHOR]

Background Candidatus Liberibacter solanacearum (Lso) is a phloem-limited alphaproteobacterium associated with the devastating zebra chip disease of potato (Solanum tuberosum). Like other members of Liberibacter, Lso-ZC1 encodes a flagellin domain-containing protein (FlaLso,) with a conserved 22 amino-acid peptide (flg22Lso). To understand the innate immune responses triggered by this unculturable intracellular bacterium, we studied the pathogenassociated molecular patterns (PAMPs) that triggered immunity in Nicotiana benthamiana, using the flg22Lso peptide and the full length flaLso gene. Results Our results showed that the expression of flaLso via Agrobacterium-mediated transient expression induced a slow necrotic cell death in the inoculated leaves of N. benthamiana, which was coupled with a burst of reactive oxygen species (ROS) production. Moreover, the expression of several representative genes involved in innate immunity was transiently upregulated by the flg22Lso in N. benthamiana. The FlaLso, however, induced stronger upregulation of these representative genes compared to the flg22Lso, especially that of flagellin receptor FLAGELLIN SENSING2 (FLS2) and respiratory burst oxidase (RbohB) in N. benthamiana. Although neither cell death nor ROS were induced by the synthetic flg22Lso, a weak callose deposition was observed in infiltrated leaves of tobacco, tomato, and potato plants. Conclusion The flagellin of Lso and its functional domain, flg22Lso share characteristics of pathogenassociated molecular patterns, and trigger unique innate immune responses in N. benthamiana. Slow and weak activation of the innate immune response in host plants by the flagellin of Lso may reflect the nature of its intracellular life cycle. Our findings provide new insights into the role of the Lso flagellin in the development of potato zebra chip disease and potential application in breeding for resistance. [ABSTRACT FROM AUTHOR]

Huanglongbing (HLB) is one of the most destructive diseases of citrus worldwide. The three known causal agents of HLB are species of α-proteobacteria: 'Candidatus Liberibacter asiaticus', 'Ca. L. africanus', and 'Ca. L. americanus'. Previous studies have found distinct variations in temperature sensitivity and tolerance among these species. Here, we describe the use of controlled heat treatments to cure HLB caused by 'Ca. L. asiaticus', the most prevalent and heat-tolerant species. Using temperature-controlled growth chambers, we evaluated the time duration and temperature required to suppress or eliminate the 'Ca. L. asiaticus' bacterium in citrus, using various temperature treatments for time periods ranging from 2 days to 4 months. Results of quantitative polymerase chain reaction (qPCR) after treatment illustrate significant decreases in the 'Ca. L. asiaticus' bacterial titer, combined with healthy vigorous growth by all surviving trees. Repeated qPCR testing confirmed that previously infected, heat-treated plants showed no detectable levels of 'Ca. L. asiaticus', while untreated control plants remained highly infected. Continuous thermal exposure to 40 to 42°C for a minimum of 48 h was sufficient to significantly reduce titer or eliminate 'Ca. L. asiaticus' bacteria entirely in HLB-affected citrus seedlings. This method may be useful for the control of 'Ca. Liberibacter'-infected plants in nursery and greenhouse settings. [ABSTRACT FROM AUTHOR]

The citrus cultivar Carrizo is the single most important rootstock to the US citrus industry and has resistance or tolerance to a number of major citrus diseases, including citrus tristeza virus, foot rot, and Huanglongbing (HLB, citrus greening). A Carrizo genomic sequence database providing approximately 3.5× genome coverage (haploid genome size approximately 367 Mb) was populated through 454 GS FLX shotgun sequencing. Analysis of the repetitive DNA fraction indicated a total interspersed repeat fraction of 36.5%. Assembly and characterization of abundant citrus Ty3/ gypsy elements revealed a novel type of element containing open reading frames encoding a viral RNA-silencing suppressor protein ( RNA binding protein, rbp) and a plant cytokinin riboside 5′-monophosphate phosphoribohydrolase-related protein ( LONELY GUY, log). Similar gypsy elements were identified in the Populus trichocarpa genome. Gene-coding region analysis indicated that 24.4% of the nonrepetitive reads contained genic regions. The depth of genome coverage was sufficient to allow accurate assembly of constituent genes, including a putative phloem-expressed gene. The development of the Carrizo database () will contribute to characterization of agronomically significant loci and provide a publicly available genomic resource to the citrus research community. [ABSTRACT FROM AUTHOR]

Citrus Huanglongbing (HLB) is one of the most destructive diseases of citrus worldwide and is threatening the survival of the Floridian citrus industry. Currently, there is no established cure for this century-old and emerging disease. As a possible control strategy for citrus HLB, therapeutic compounds were screened using a propagation test system with 'Candidatus Liberibacter asiaticus'-infected periwinkle and citrus plants. The results demonstrated that the combination of penicillin and streptomycin (PS) was effective in eliminating or suppressing the 'Ca. L. asiaticus' bacterium and provided a therapeutically effective level of control for a much longer period of time than when administering either antibiotic separately. When treated with the PS, 'Ca. L. asiaticus'-infected periwinkle cuttings achieved 70% of regeneration rates versus <50% by other treatments. The 'Ca. L. asiaticus' bacterial titers in the infected periwinkle plants, as measured by quantitative real-time polymerase chain reaction, decreased significantly following root soaking or foliar spraying with PS. Application of the PS via trunk injection or root soaking also eliminated or suppressed the 'Ca. L. asiaticus' bacterium in the HLB-affected citrus plants. This may provide a useful tool lot the management of citrus HLB and other Liberibacter-associated diseases. [ABSTRACT FROM AUTHOR]

The citrus leaf miner, Phyllocnistis citrella Stainton (Lepidoptera: Gracillariidae), is a key pest in most citrus-growing regions worldwide. Adult citrus leafminers oviposit primarily on young elongating flush of Citrus as well as other Rutaceae and some ornamental plants. Larvae feed on the epidermal cell layer of developing leaves and injury to leaves provides a pathway for infection by the bacterium Xanthomonas citri subsp. citri (Hasse), the causal agent of Asiatic citrus canker. In this study, we quantified abundance of citrus leafminer larvae on progeny of 87 seed parent genotypes of Citrus and Citrus relatives (family Rutaceae) in the field in East--central Florida to identify those that have low abundance of leafminers. Progeny from the 87 parent genotypes varied in abundance of the leafminer. Progeny of 15 parent genotypes had a high mean abundance of more than six leafminers per flush shoot. All but one of these genotypes were in the Citrus genus. Progeny of 16 parent genotypes had zero, or nearly zero, leafminers, but none were from the Citrus genus. However, many of these 16 genotypes were from genera closely related to true citrus (subtribe Citrinae) and are sexually compatible with Citrus. Progeny of two parent genotypes in the subfamily Toddalioideae and Glycosmispentaphylla (Retz.) Corr. also had a low abundance of leafminer. Glycosmis pentaphylla also is a poor host for the Asian citrus psyllid, Diaphorina citri Kuwayama, and has biochemical resistance to the citrus weevil, Diaprepes abbreviatus (L.), so this genotype as well as others identified as poor hosts for the leafminer may prove useful in breeding programs aimed at reducing the abundance of multiple insect pests on citrus. [ABSTRACT FROM AUTHOR]

Clonal woody crop germplasm collections often originate and are grown in distinct geographical locations. Because the degree of cold-hardiness is known to be a factor in the successful use of dormant bud cryopreservation for Malus, it was suggested that material from relatively warmer climates would not respond to cryopreservation as well as material from colder environments. To test this hypothesis, the effect of growing provenance on cryosurvival of dormant buds from three Malus (apple) cultivars grown in three locations (Geneva, NY; Davis, CA; and Corvallis, OR) was tested in 3 consecutive years. Dormant winter buds were harvested at the three locations, cryopreserved, and bud viability was tested by grafting. The collective 3-year mean viability for cryopreserved dormant apple buds for the three locations ranged from 63% to 81% of the buds surviving with the highest survival from the Corvallis site; however, the Geneva twigs were exposed to the lowest preharvest temperature. These results suggest that the temperature at the growing location may not hinder application of the dormant bud cryopreservation method with Malus to the extent previously speculated. [ABSTRACT FROM AUTHOR]

Citrus huanglongbing is one of the most destructive diseases of citrus worldwide. The disease is associated with three different species of 'Candidatus Liberibacter', of which 'Ca. L. asiaticus' is the most widely distributed. An optimized system using 'Ca. L. asiaticus'-infected periwinkle cuttings was developed to screen chemical compounds effective for controlling the bacterial population while simultaneously assessing their phytotoxicity. The optimal regeneration conditions were determined to be the use of vermiculite as a growth medium for the cuttings, and a fertilization routine using half-strength Murashige and Tucker medium supplemented with both naphthalene acetic acid (4 µg/ml) and indole-3-butyric acid (4 µg/ml). This system allowed a plant regeneration rate of 60.6% for 'Ca. L. asiaticus'-infected cuttings in contrast to the <1% regeneration rate with water alone. Two chemical agents, penicillin G sodium and 2,2-dibromo-3-nitrilopropionamide (DBNPA), were found to be effective at eliminating or suppressing the 'Ca. L. asiaticus' bacterium in this periwinkle regeneration system. When treated with penicillin G sodium at 50 µg/ml, all plants regenerated from 'Ca. L. asiaticus'-infected cuttings were 'Ca. L. asiaticus' negative as determined by both nested polymerase chain reaction (PCR) and quantitative real-time PCR. In addition, DBNPA was also able to significantly reduce the percentage of 'Ca. L. asiaticus'-positive plants and the titer of the 'Ca. L. asiaticus' bacterium at 200 µl/liter. [ABSTRACT FROM AUTHOR]

Genic microsatellites or simple sequence repeat markers derived from expressed sequence tags (ESTs), referred to as EST–SSRs, are inexpensive to develop, represent transcribed genes, and often have assigned putative function. The large apple ( Malus × domestica) EST database (over 300,000 sequences) provides a valuable resource for developing well-characterized DNA molecular markers. In this study, we have investigated the level of transferability of 68 apple EST–SSRs in 50 individual members of the Rosaceae family, representing three genera and 14 species. These representatives included pear ( Pyrus communis), apricot ( Prunus armeniaca), European plum ( P. domestica), Japanese plum ( P. salicina), almond ( P. dulcis), peach ( P. persica), sour cherry ( P. cerasus), sweet cherry ( P. avium), strawberry ( Fragaria vesca, F. moschata, F. virginiana, F. nipponica, and F. pentaphylla), and rose ( Rosa hybrida). All 68 primer pairs gave an amplification product when tested on eight apple cultivars, and for most, the genomic DNA-derived amplification product matched the expected size based on EST ( in silico) data. When tested across members of the Rosaceae, 75% of these primer pairs produced amplification products. Transferability of apple EST–SSRs across the Rosaceae ranged from 25% in apricot to 59% in the closely related pear. Besides pear, the highest transferability of these apple EST–SSRs, at the genus level, was observed for strawberry and peach/almond, 49 and 38%, respectively. Three markers amplified in at least one genotype within all tested species, while eight additional markers amplified in all species, except for cherry. These 11 markers are deemed good candidates for a widely transferable Rosaceae marker set provided their level of polymorphism is adequate. Overall, these findings suggest that transferability of apple EST–SSRs across Rosaceae is varied, yet valuable, thereby providing additional markers for comparative mapping and for carrying out evolutionary studies. [ABSTRACT FROM AUTHOR]

• Citrus x Poncirus hybrids have poor eating quality due to the Poncirus parent • In this study, three such hybrids had composition analysis similar to citrus fruit • Two hybrids had volatile composition closer to that of Poncirus trifoliata fruit • One hybrid had a flavonoid aglycone profile similar to that of pure Poncirus • A pattern of inheritance of Poncirus chemical traits could be seen Citrus breeders have made numerous hybridizations that included Poncirus trifoliata L.Raf. (P. trifoliata), a relative of the genus Citrus , in their pedigree to produce hybrids that are cold- and disease-resistant. Initial hybrids of Citrus × P. trifoliata typically produce fruit that have unacceptable flavor, but more advanced hybrids with greater proportions of Citrus in their pedigrees have various degrees of off-flavor and taste. This study reports the volatile constituents, as well as nonvolatile limonoids and flavonoids, in the juice of six Citrus hybrids containing P. trifoliata in their pedigrees, compared to two hybrids derived solely from Citrus. Secondary metabolite profiles reflected the complex genetic backgrounds of those hybrids. Hybrids 6_23−20 and US 119 had juice composition most similar to that of P. trifoliata , with more volatiles, and in particular, more esters and sesquiterpenes with higher amounts in total. Furthermore, hesperidin concentration in US 119 was low (10 μg g−1) in comparison with other hybrids (125−326 μg g−1) while concentrations of narirutin and isosakuranetin-7-O-rutinoside were unusually high (135−211 μg g−1 and 304−182 μg g−1, respectively) in comparison with the other hybrids (5−60 μg g−1 and 10−75 μg g−1, respectively), which is in agreement with the composition of pure P. trifoliata. Juice of P. trifoliata derived hybrids 5−18-24, 5−18-31, 1−76-100, and 1−77-105 presented secondary metabolite compositions closer to those of pure Citrus hybrids. Based on volatiles, limonoids, and flavonoid profiles, we conclude that it is possible to generate advanced Citrus hybrids including P. trifoliata in their pedigrees that approach the quality of common Citrus types. [ABSTRACT FROM AUTHOR]

• A large variation in total oil content was found among14 avocado genotypes. • Oleate, palmitate and linoleate were the major fatty acid components in all genotypes. • Acetaldehyde, C-6 aldehydes and mono/sesquiterpenes were the major VOCs. • '35707' and 'PA-6206' were rich in total oil content and unsaturated fatty acids. Worldwide avocado consumption is growing due to potential health benefits. Most research has been focused on 'Hass', which does not perform well in Florida. Fatty acids and volatile organic compounds (VOCs) were evaluated in 14 avocado genotypes grown in East-Central Florida conditions and compared to 'Hass'. Two saturated and five unsaturated fatty acids were detected. Total oil content (TOC) was 11–25%, with 58.2–71.5% unsaturated fatty acids (UFA). 'FL Hass' contained 20% TOC, near the average for genotypes tested, but with a low UFA of 61.9%. 'PA-6206' (a 'Hass' x 'Bacon' seedling) and '35707' (a 'Catalina' seedling) had higher TOC and UFA than other genotypes, while the mostly West Indian types such as 'Simmonds', 'Day', 'Pflume' and 'Miguel' had low TOC. Detected VOCs were acetaldehyde, hexanal, (E)-2-hexenal, limonene, α-cubebene, α-copaene, and β -caryophyllene. Most genotypes contained most of the VOCs. 'Monroe' lacked C6 aldehydes, and 'Pflume', 'Bernecker-43', 'Lula' and '35707' lacked some or all sesquiterpenes. [ABSTRACT FROM AUTHOR]

Background: Most bacteria are not culturable, but can be identified through molecular methods such as metagenomics studies. Due to specific metabolic requirements and symbiotic relationships, these bacteria cannot survive on typical laboratory media. Many economically and medically important bacteria are unculturable; including phloem-limited plant pathogens like Candidatus Liberibacter asiaticus (CLas). CLas is the most impactful pathogen on citrus production, is vectored by the Asian citrus psyllid (ACP, Diaphorina citri), and lacks an effective treatment or resistant cultivars. Research into CLas pathogenicity and therapy has been hindered by the lack of persistent pure cultures. Work to date has been mostly limited to in planta studies that are time and resource intensive. Results: We developed and optimized an in vitro protocol to quickly test the effectiveness of potential therapeutic agents against CLas. The assay uses intact bacterial cells contained in homogenized tissue from CLas-infected ACP and a propidium monoazide (PMA) assay to measure antimicrobial activity. The applicability of PMA was evaluated; with the ability to differentiate between intact and disrupted CLas cells confirmed using multiple bactericidal treatments. We identified light activation conditions to prevent PCR interference and identified a suitable positive control for nearly complete CLas disruption (0.1% Triton-X 100). Isolation buffer components were optimized with 72 mM salt mixture, 1 mM phosphate buffer and 1% glycerol serving to minimize unwanted interactions with treatment and PMA chemistries and to maximize recovery of intact CLas cells. The mature protocol was used to compare a panel of peptides already under study for potential CLas targeting bactericidal activity and identify which were most effective. Conclusion: This psyllid homogenate assay allows for a quick assessment of potential CLas-disrupting peptides. Comparison within a uniform isolate largely eliminates experimental error arising from variation in CLas titer between and within individual host organisms. Use of an intact vs. disrupted assay permits direct assessment of potential therapeutic compounds without generating pure cultures or conducting extensive in planta or field studies. [ABSTRACT FROM AUTHOR]

Background: Citrus Huanglongbing (HLB) is a bacterial disease with high economic significance. The associated agent Candidatus Liberibacter asiaticus is a fastidious, phloem-limited, intracellular bacterium that is transmitted by an insect vector the Asian citrus psyllid (ACP). The genome of Ca. L. asiaticus contains protein secretion machinery that suggests host cell modulation capacity of this bacterium. Results: A total of 28 candidate effectors, an important class of secreted proteins, were predicted from the Ca. L. asiaticus genome. Sequence specific primers were designed for reverse transcription (RT) and quantitative PCR (qPCR), and expression was validated for 20 of the effector candidates in infected citrus with multiple genetic background. Using detached leaf inoculation, the mRNA of effectors was detected from 6 h to 7 days post ACP exposure. It was observed that higher bacterial titers were associated with a larger number of effectors showing amplification across all samples. The effectors' expression were compared in citrus hosts with various levels of HLB tolerance, including susceptible Duncan grapefruit and Washington navel orange, tolerant citron and Cleopatra mandarin, and resistant Pomeroy trifoliate and Carrizo citrange. Across all genotypes relatively high expression was observed for CLIBASIA_03695, CLIBASIA_00460, CLIBASIA_00420, CLIBASIA_04580, CLIBASIA_05320, CLIBASIA_04425, CLIBASIA_00525 and CLIBASIA_05315 in either a host-specific or -nonspecific manners. The two genotypes in each HLB-response group also show effector-expression profiles that seem to be different. In a companion study, the expression of effectors was compared between leaves and roots of own-rooted citrus that had been Ca. L. asiaticus-infected for more than a year. Results indicated relatively high expression of CLIBASIA_03875, CLIBASIA_04800 and CLIBASIA_05640 in all leaf and some root tissues of citron, Duncan and Cleopatra. Conclusion: This temporal and spatial expression analysis of Ca. L. asiaticus effectors identified candidates possibly critical for early bacterial colonization, host tolerance suppression and long-term survival which are all worthy of further investigation. [ABSTRACT FROM AUTHOR]