► We tested citrus rootstock varieties for tolerance to Ca. L. asiaticus (Las). ► Percentage of Las-positive plants and Las genome numbers varied between varieties. ► Few HLB disease symptoms were ...observed in most rootstock varieties. ► Carrizo citrange, US-897 and US-942 were classified as tolerant to Las. ► US-802, US-812, and Volkamer lemon were classified as moderately tolerant.
Candidatus Liberibacter asiaticus (Las) is the suspected causal agent of Huanglongbing (HLB) in Florida and other citrus producing countries around the world. Although little resistance to HLB is found within commercial citrus varieties, tolerance has been reported for some varieties that are commonly used as rootstocks. This study assesses the response of eight different rootstock varieties, which include the Citrus×Poncirus trifoliata hybrids Carrizo citrange, US-802, US-812, US-897, and US-942, Benecke trifoliate orange, Volkamer lemon, and Cleopatra mandarin, to Las under controlled conditions in the greenhouse. The percentage of plants detected positive for Las did not exceed 7% in the genotypes Benecke, Carrizo, US-802, US-942, and Volkamer during the first 6 months after graft-inoculation (mai). Pruning resulted in much increased numbers of plants with detectable Las levels and induced disease symptoms in previously asymptomatic plants. At 12 mai, percentages of Las-positive plants were 91–96% for US-802, US-812, Cleopatra, and Volkamer, and 70–78% for US-897 and US-942. Lowest percentages (44–52%) were observed for Benecke and Carrizo. The number of Las genomes per g of leaf tissue in Las-positive plants was considerably lower in Benecke and US-897 seedlings with 4.5×105 and 1.6×106 compared with Cleopatra (1.0×107), while numbers for the other genotypes ranged from 3.1–4.9×106. At 12 mai, foliar disease symptoms, though variable in extent, were prominent in most genotypes, except US-897. Shoot masses were significantly reduced in Cleopatra, Benecke, US-802, and Volkamer seedlings in response to Las at this time. Continued observation until 18 mai found no significant growth reductions in Benecke seedlings. Our results suggest classifying Carrizo, US-897, and US-942 as tolerant, US-802, US-812, and Volkamer lemon as moderately tolerant, and Cleopatra mandarin as susceptible to Las. Despite irregular growth, low rates of infection and low Las numbers indicate some resistance of Benecke to Las. Additional greenhouse experiments and field observations confirmed findings for US-802, US-897, US-942, and Cleopatra, although results for US-802 were more variable.
Key message
Plant biostimulants did not improve citrus health and productivity in an HLB-endemic environment after two years, but fulvic acids increased concentrations of some nutrients and ...influenced the rhizosphere microbial community.
Fibrous root loss is one of the main factors associated with citrus tree decline due to Huanglongbing (HLB) disease in Florida, and there is increased interest to use soil amendments such as plant biostimulants to improve root health and crop production. The effect of individual and combined applications of seaweed extracts, humic substances, and microbial inoculants on root and tree health, tree nutritional status, and productivity was investigated in an HLB-endemic environment. Experiments were conducted in two differently managed commercial citrus orchards. The effect of the treatments on the composition and structure of the citrus rhizosphere bacterial and fungal community was examined and linked to variations in root macro- and micronutrient concentrations. Overall, treatments had no effect on disease index ratings, total and specific root lengths, root physiological responses, and fruit quality and yield after two years of application. Only the individual applications of fulvic acids resulted in statistically significant increases in root and leaf macro- and micronutrient concentrations compared to the control treatment, an effect that differed between trials. These increases in root macro- and micronutrient concentrations were linked to variations in the relative abundance of specific bacterial and fungal taxa in the citrus rhizosphere. Our results suggest that longer-term and higher application rates of fulvic acids may be necessary to translate rhizosphere effects into measurable effects on tree health and productivity in HLB-endemic conditions. We show that factors such as different management practices and soil and environmental conditions may determine efficacy of biostimulants on root and tree health.
Six new hybrid rootstocks, ‘US-1279’, ‘US-1281’, ‘US-1282’, ‘US-1283’, ‘US-1284’, and ‘US-1516’, were released from the U.S. Department of Agriculture (USDA) citrus breeding program to provide ...improved tree tolerance to huanglongbing (HLB), the most destructive disease facing the citrus industry in the United States and many other parts of the world. Five of these new rootstocks were released based on field performance in trials with the rootstocks propagated by stem cuttings, rather than the traditional propagation using nucellar seedlings. In this study, we evaluated the fruit, seed, and seedling characteristics of these new rootstocks, along with four other USDA rootstocks of commercial importance. The study included a determination of the percentage of true-to-type and off-type seedlings by both plant morphology and simple sequence repeat (SSR) markers. All 10 rootstocks produced an acceptable number of seeds and good seedling emergence from those seeds. The rootstocks ‘Swingle’, ‘US-802’, ‘US-812’, ‘US-1283’, ‘US-1284’, and ‘US-1516’ had a high percentage of true-to-type seedlings and correspondingly good potential to be propagated by seeds. However, no true-to-type plants were observed among seedlings from the rootstocks ‘US-1279’, ‘US-1281’, and ‘US-1282’, indicating that economical seed propagation will be impossible for these cultivars. The 10 SSR marker sets used in this study were observed to easily differentiate the 10 rootstocks studied, and readily distinguished true-to-type and off-type seedlings among progeny from all 10 rootstock clones. This study presents information of significant value for commercial nurseries involved in propagation of citrus rootstocks, and those involved in citrus rootstock breeding and development around the world. We propose the use of these 10 SSR marker sets as readily applicable for accurate identification of most citrus rootstock cultivars and their true-to-type seedlings.
Citrus crops have a long history of cultivation as grafted trees on selected rootstock cultivars, but all current rootstocks have significant limitations and traditional methods of rootstock breeding ...take at least 2–3 decades to develop and field test new rootstocks. Citrus production in the United States, and other parts of the world, is impaired by a wide range of biotic and abiotic problems, with especially severe damage caused by the disease huanglongbing (HLB) associated with
Candidatus
Liberibacter asiaticus. All major commercial citrus scion cultivars are damaged by HLB, but tree tolerance is significantly improved by some rootstocks. To overcome these challenges, the USDA citrus breeding program has implemented a multi-pronged strategy for rootstock breeding that expands the diversity of germplasm utilized in rootstock breeding, significantly increases the number of new hybrids evaluated concurrently, and greatly reduces the time from cross to potential cultivar release. We describe the key components and methodologies of this new strategy, termed “SuperSour,” along with reference to the historical favorite rootstock sour orange (
Citrus aurantium
), and previous methods employed in citrus rootstock breeding. Rootstock propagation by cuttings and tissue culture is one key to the new strategy, and by avoiding the need for nucellar seeds, eliminates the 6- to 15-year delay in testing while waiting for new hybrids to fruit. In addition, avoiding selection of parents and progeny based on nucellar polyembryony vastly expands the potential genepool for use in rootstock improvement. Fifteen new field trials with more than 350 new hybrid rootstocks have been established under the SuperSour strategy in the last 8 years. Detailed multi-year performance data from the trials will be used to identify superior rootstocks for commercial release, and to map important traits and develop molecular markers for the next generation of rootstock development. Results from two of these multi-year replicated field trials with sweet orange scion are presented to illustrate performance of 97 new hybrid rootstocks relative to four commercial rootstocks. Through the first 7 years in the field with endemic HLB, many of the new SuperSour hybrid rootstocks exhibit greatly superior fruit yield, yield efficiency, canopy health, and fruit quality, as compared with the standard rootstocks included in the trials.
Most of the commercially important citrus scion cultivars are susceptible to Huanglongbing (HLB), which is the most devastating disease the citrus industry has ever faced. Because the rootstock can ...influence the performance of the scion in various ways, including disease and pest tolerance, use of superior rootstocks can assist citrus growers with minimizing the negative effects of HLB. The objective of this study was to assess rootstock effects on the horticultural performance and early production potential of ‘Hamlin’ sweet orange ( Citrus sinensis ) trees in commercial field settings under HLB-endemic conditions. Two field trials were conducted in different locations in Central and Southeast Florida. The trials were established in 2015 and included 32 diverse diploid and tetraploid rootstock cultivars and advanced selections. One trial was performed in Highlands County, FL, on a poorly drained flatwoods-type site. Another trial was performed in Polk County, FL, on a well-drained sandy Central Florida Ridge site. Horticultural traits including tree height, canopy volume, trunk diameter, canopy health, leaf nutrient content, yield, and fruit quality were assessed during the 2018–19 and 2019–20 production years. Significant differences were found among trees on different rootstocks for most of the measured traits, particularly tree vigor and productivity, but rootstock effects also varied by location. Rootstocks that induced large tree sizes, such as the diploid mandarin × trifoliate orange hybrids ‘X-639’, ‘C-54’, ‘C-57’, and ‘C-146’, also induced higher yield, but with lower yield efficiency. Most of the tetraploid rootstocks significantly reduced tree size, among which ‘Changsha+Benton’, ‘Green-3’, ‘Amb+Czo’, ‘UFR-3’, and ‘UFR-5’ induced high yield efficiency. Therefore, these rootstocks have the potential to be used in high-density plantings. However, trees on some of these small size-inducing rootstocks had a higher mortality rate and were more vulnerable to tropical force winds. This study provides important information for the selection of rootstocks with the greatest production potential in an HLB-endemic environment, especially during the early years of production.
The devastating citrus disease huanglongbing (HLB) associated with the phloem-limited bacteria Candidatus Liberibacter asiaticus (CLas) has caused a more than 70% reduction in citrus production since ...its discovery in Florida in 2005. Most citrus scion cultivars are sensitive to HLB, whereas some cultivars used as rootstocks are tolerant. Using such tolerant rootstocks can help trees to cope better with the disease’s impact. Evaluating rootstock effects on a grafted scion in the field takes many years, but shorter-term evaluation is imperative to aid in rootstock selection for an HLB-endemic production environment. In this study, we investigated grafted healthy and CLas-infected citrus trees under controlled greenhouse conditions. The objectives were to identify traits suitable for assessing grafted tree tolerance in advance of longer-term field studies and aiding in the selection of superior rootstock cultivars. We assessed 10 commercially important rootstocks grafted with ‘Valencia’ sweet orange scion and with known field performance. At 6, 9, 15, and 21 months after graft inoculation (mai), leaf CLas titers were determined and canopy health was evaluated. Plants were destructively sampled at 21 mai to assess plant biomasses and other physiological and horticultural variables. There was little influence of the rootstock cultivar on CLas titers. Surprisingly, few HLB foliar disease symptoms and no differences in soluble and nonsoluble carbohydrate concentrations were measured in infected compared with healthy plants, despite high CLas titers and significant reductions in plant biomasses. Most trees on rootstocks with trifoliate orange parentage were less damaged by HLB than other rootstocks, although results did not always agree with reported field performance. Among the different variables measured, leaf size appeared to be most predictive for grafted tree assessment of HLB sensitivity. The results of this study provide a better understanding of the strengths and weaknesses of assessing rootstock influence on grafted tree performance in a controlled greenhouse environment. Although such studies provide valuable information for cultivar tolerance to HLB, other rootstock traits will ultimately contribute to field survival and productivity in an HLB endemic production environment.
Huanglongbing (HLB) is a devastating disease of citrus that is found in most citrus production areas around the world. The bacterium associated with HLB resides in and damages the phloem, restricting ...the movement of photosynthates throughout the plant and leading to tree decline. Considerable root loss can be observed in affected trees even when few disease symptoms are visible aboveground. Root traits can substantially influence tree performance and use of superior rootstocks is one strategy to manage tree health and reduce production losses in a disease-endemic environment. Citrus rootstocks are typically propagated by seed, but due to the increased demand for some of the best-performing cultivars, propagation by other methods is being used to overcome seed shortages. In this research, differences in root architecture and root growth of six different rootstocks propagated by seed, cuttings, and tissue culture, and their influence on the grafted ‘Valencia’ ( Citrus sinensis ) scion were investigated. A field trial was established in southwest Florida in 2017. Trees were evaluated for their performance during the first 2 years after planting and a subset of trees was excavated for detailed analysis of root architectures and biomass distribution. Significant differences among propagation methods were found for the rootstock trunk diameter and the lateral (structural) root length, which were largest in seed-propagated rootstocks. Most of the other horticultural and root architectural traits were not significantly influenced by the rootstock-propagation method; however, many of the measured variables were significantly influenced by the rootstock cultivar regardless of the propagation method. The results showed that rootstocks propagated by cuttings and tissue culture were similar to seed-propagated rootstocks in their influence on the grafted tree during the early years of growth in the field.
Traditional foliar spray and soil drench applications of crop protection compounds have been ineffective at managing huanglongbing (HLB) in citrus. Trunk injection is a technique that delivers crop ...protection compounds directly into the tree vasculature, which optimizes compound availability while minimizing drift, runoff, and damage to non-target organisms. Five-year-old HLB-affected ‘Valencia’ sweet orange (Citrus sinensis) trees were injected with the insecticide imidacloprid or the antibacterial oxytetracycline in October 2020 and April 2021. Trees were monitored for tree health, psyllid colonization, bacterial titers, fruit quality, fruit drop, and yield for two production seasons. Imidacloprid injection caused 63% mortality of psyllid adults within one week and reduced progeny survival by 80%, though the efficacy waned within two months. Injection with oxytetracycline significantly improved tree health, reduced bacterial titers, and reduced preharvest fruit drop by more than 3-fold with corresponding increases in yield. Residue dynamics varied by injected compound and tissue analyzed. These results suggest trunk injection could be an effective delivery method for existing or novel therapeutics targeting either the insect vector or the pathogen causing HLB.
While the rootstock genotype (belowground part of a plant) can impact rhizosphere microbial communities, few studies have examined the relationships between rootstock genotype-based recruitment of ...active rhizosphere bacterial communities and the availability of root nutrients for plant uptake. Rootstocks are developed to provide resistance to disease or tolerance of abiotic stresses, and compost application is a common practice to also control biotic and abiotic stresses in crops. In this field study, we examined: (i) the effect of four citrus rootstocks and/or compost application on the abundance, diversity, composition, and predicted functionality of active rhizosphere bacterial communities, and (ii) the relationships between active rhizosphere bacterial communities and root nutrient concentrations, with identification of bacterial taxa significantly correlated with changes in root nutrients in the rhizosphere.
The rootstock genotype determined differences in the diversity of active rhizosphere bacterial communities and also impacted how compost altered the abundance, diversity, composition, and predicted functions of these active communities. Variations in the active bacterial rhizobiome were strongly linked to root nutrient cycling, and these interactions were root-nutrient- and rootstock-specific. Direct positive relationships between enriched taxa in treated soils and specific root nutrients were detected, and potentially important taxa for root nutrient uptake were identified. Significant differences in specific predicted functions were related to soil nutrient cycling (carbon, nitrogen, and tryptophan metabolisms) in the active bacterial rhizobiome among rootstocks, particularly in soils treated with compost.
This study illustrates that interactions between citrus rootstocks and compost can influence active rhizosphere bacterial communities, which impact root nutrient concentrations. In particular, the response of the rhizobiome bacterial abundance, diversity, and community composition to compost was determined by the rootstock. Specific bacterial taxa therefore appear to be driving changes in root nutrient concentrations in the active rhizobiome of different citrus rootstocks. Several potential functions of active bacterial rhizobiomes recruited by different citrus rootstocks did not appear to be redundant but rather rootstock-specific. Together, these findings have important agronomic implications as they indicate the potential for agricultural production systems to maximize benefits from rhizobiomes through the choice of selected rootstocks and the application of compost. Video Abstract.
Trunk injection is a targeted delivery of pesticides, insecticides, nutrients, or other plant protection materials into the stem or trunk of woody plants as an alternative to spraying or soil ...drenching. Trunk injection has historically been used for disease and pest management of high-value forest tree species or ornamental plants when aerial applications are problematic due to spatial problems and health-related concerns. An interest in using the injection technique for protection of agricultural crops in commercial production systems has emerged more recently, where foliar applications and soil drenches have proven ineffective or pose environmental hazards. This review provides an overview of the basic principles of trunk injection and the plant physiological implications, its current use in commercial agriculture and other plant systems, and associated risks.
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