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Introduction: Citrus hybrids with Poncirus trifoliata L. Raf. introgression have gained interest due to their tolerance to Huanglongbing (HLB), a devastating disease for Florida citrus agriculture. While these hybrids inherit disease tolerance from P. trifoliata, they sometimes also suffer from undesirable off-flavors. Methods: A selection of thirteen genotypes were harvested over the 2020-2021 and 2021-2022 seasons. Their juices were evaluated by a trained sensory panel and were comprehensively analyzed for their chemical makeup, including soluble solids content (SSC), titratable acidity (TA), volatiles, flavonoids and limonoids. Results & discussion: Overall, along with the commercial orange cultivars 'Valencia' and 'Hamlin', the HLB-tolerant Poncirus hybrid 'US SunDragon,' and the mandarin hybrids Sugar Belle¬, FF-5-51-2, and 'US Superna' had positive citrus flavor quality. Esters, some sesquiterpenes, along with flavonoids, eriocitrin and quercetin-3-(3R-glucosylrutinoside), were positively correlated with orange flavor while b-ionone and eucalyptol were highly abundant in the mandarins. The flavonoid linarin, was more abundant in Poncirus hybrids with off-flavors than in the Poncirus hybrid 'US SunDragon', having high orange flavor. Two mandarin hybrids, FF-5-6-36 and FTP-6-32-67, were not bitter at harvest, but the juice exhibited delayed bitterness after storage at -20°C, which was associated with significant increases of limonin, nomilin, naringenin, and prunin. Interestingly, during freezer storage, a newly identified flavonoid in citrus, tricin-C-hexoside, increased dramatically across all of the genotypes. The identification of diseasetolerant hybrids with satisfactory flavor quality at juicing as well as after storage where delayed bitterness may develop, has great significance for future breeding efforts for fresh fruit or for use in stand-alone juice/juice blends. [ABSTRACT FROM AUTHOR]

Huanglongbing (HLB), or citrus greening, is the most devastating disease in citrus worldwide. Commercial citrus varieties including sweet orange (Citrus sinensis) are highly susceptible to HLB, and trifoliate orange (Poncirus trifoliata, a close Citrus relative) is widely considered resistant or highly tolerant to HLB. In this study, an intergeneric F1 population of sweet orange and trifoliate orange was genotyped by Genotyping-by-Sequencing, and high-density SNP-based genetic maps were constructed separately for trifoliate orange and sweet orange. The two genetic maps exhibited high synteny and high coverage of the citrus genome. Progenies of the F1 population and their parents were planted in a replicated field trial, exposed to intense HLB pressure for 3 years, and then evaluated for susceptibility to HLB over 2 years. The F1 population exhibited a wide range in severity of HLB foliar symptom and canopy damage. Genome-wide QTL analysis based on the phenotypic data of foliar symptom and canopy damage in 2 years identified three clusters of repeatable QTLs in trifoliate orange linkage groups LG-t6, LG-t8 and LG-t9. Co-localization of QTLs for two traits was observed within all three regions. Additionally, one cluster of QTLs in sweet orange (linkage group LG-s7) was also detected. The majority of the identified QTLs each explained 18-30% of the phenotypic variation, indicating their major role in determining HLB responses. These results show, for the first time, a quantitative genetic nature yet the presence of major loci for the HLB tolerance in trifoliate orange. The results suggest that sweet orange also contains useful genetic factor(s) for improving HLB tolerance in commercial citrus varieties. Findings from this study should be very valuable and timely to researchers worldwide as they are hastily searching for genetic solutions to the devastating HLB crisis through breeding, genetic engineering, or genome editing.

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]

Huanglongbing (HLB), also known as citrus greening, is an insidious disease in citrus and has become a threat to the sustainability of the citrus industry worldwide. In the U.S., Candidatus Liberibacter asiaticus (CLas) is the pathogen that is associated with HLB, an unculturable, phloem-limited bacteria, vectored by the Asian Citrus Psyllid (ACP, Diaphorina citri). There is no known cure nor treatment to effectively control HLB, and current control methods are primarily based on the use of insecticides and antibiotics, where effectiveness is limited and may have negative impacts on beneficial and non-target organisms. Thus, there is an urgent need for the development of effective and sustainable treatment options to reduce or eliminate CLas from infected trees. In the present study, we screened citrus-derived endophytes, their cell-free culture supernatants (CFCS), and crude plant extracts for antimicrobial activity against two culturable surrogates of CLas, Sinorhizobium meliloti and Liberibacter crescens. Candidates considered high-potential antimicrobial agents were assessed directly against CLas in vitro, using a propidium monoazide–based assay. As compared to the negative controls, statistically significant reductions of viable CLas cells were observed for each of the five bacterial CFCS. Subsequent 16S rRNA gene sequencing revealed that each of the five bacterial isolates were most closely related to Bacillus amyloliquefaciens, a species dominating the market of biological control products. As such, the aboveground endosphere of asymptomatic survivor citrus trees, grown in an organic orchard, were found to host bacterial endophytes capable of effectively disrupting CLas cell membranes. These results concur with the theory that native members of the citrus microbiome play a role in the development of HLB. Here, we identify five strains of Bacillus amyloliquefaciens demonstrating notable potential to be used as sources of novel antimicrobials for the sustainable management of HLB. [ABSTRACT FROM AUTHOR]

Citrus germplasm is conserved in genebanks at sites around the world to provide genetic resources for breeding and research programs. The value of genebank collections is particularly evident as diseases and climate change threaten citrus production areas. We provide historical, inventory, and maintenance information about national citrus collections in Russia and in the United States. The Russian Research Institute of Floriculture and Subtropical Crops (RRIFSC) in Sochi, Russia maintains a collection of 132 citrus accessions representing 50 taxa. Southern Russia is one of the most northerly citrus growing areas in the world and many accessions in the RRIFSC collection were specifically selected for cold tolerance to facilitate citrus production in this region. Tree response data taken during the frequent severe winters in Sochi has provided an understanding of the relative cold tolerance of many RRIFSC accessions. Particularly noteworthy is the array of cold-tolerant lemon accessions maintained at the RRIFSC. The Aurantioideae collections at the United States Department of Agriculture (USDA), Agricultural Research Service (ARS) National Clonal Germplasm Repository for Citrus and Dates (NCGRCD) and at the University of California Citrus Variety Collection in Riverside, California, maintain 1328 accessions of citrus cultivars and wild relatives. Because of federal and state quarantine regulations, accessions at this facility are tested for graft-transmissible pathogens and undergo therapy to eliminate known pathogens. In contrast to Sochi, desert-adapted lemon and grapefruit varieties are a major interest of the California-based collection. The collection of citrus and citrus relatives is being screened for resistance and/or tolerance genes to diseases such as huanglongbing which is currently threatening the US citrus industry. Through collaborative exchange efforts, the diversity in the two genebanks could be expanded to increase availability of desirable genetic resources to breeding and research communities throughout the world. [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]