• Journal of Microbiology and Biotechnology
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• v.29 no.6
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• pp.984-988
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• 2019

Blue mold in citrus is caused by Penicillium italicum. In this study, the P. italicum-specific primers were developed for rapid detection based on the conserved genes RPB1 and RPB2 among Penicillium genomes. The two primer pairs RPB1-a and RPB1-b proved to be specific to detect P. italicum. The PCR assay among 39 fungal isolates and the colonial, pathogenic morphologies and molecular methods validated the specificity and reliability of these two primer pairs. This report provided a method and P. italicum-specific primers, which might greatly contribute to citrus postharvest industry.

• Horticultural Science & Technology
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• v.33 no.3
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• pp.390-395
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• 2015

The synthesis of chimeras is a breeding approach for horticultural crops. In our breeding program, a new diploid citrus chimera, named 'Hongrou Taoye' (Citrus sinensis [L.] Osbeck + Citrus unshiu Marc.), was found arising at the junction where a 'Taoye' sweet orange (C. sinensis) scion was grafted onto Satsuma mandarin (C. unshiu). As an artificial chimera, its fruit traits derived from the L1 cell layer, with juice color and carotenoid complement, in which ${\beta}$-cryptoxanthin accumulated predominantly, similar to those of Satsuma mandarin. By contrast, traits originating from the L2/L3 cell layer, including pollen, seed, and rind aroma characteristics, were the same as those of 'Taoye' sweet orange (the scion). SSR and cpSSR analyses showed that both nuclear and chloroplast genomes of the chimera were a combination of both donor parents. 'Hongrou Taoye' thus combined the valuable traits of both donor plants, and therefore has good potential in citrus fresh market.

• Journal of Plant Biotechnology
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• v.43 no.3
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• pp.261-271
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• 2016

Citrus is an economically important fruit crop widely growing worldwide. However, citrus production largely depends on natural hybrid selection and bud sport mutation. Unique botanical features including long juvenility, polyembryony, and QTL that controls major agronomic traits can hinder the development of superior variety by conventional breeding. Diverse factors including drastic changes of citrus production environment due to global warming and changes in market trends require systematic molecular breeding program for early selection of elite candidates with target traits, sustainable production of high quality fruits, cultivar diversification, and cost-effective breeding. Since the construction of the first genetic linkage map using isozymes, citrus scientists have constructed linkage maps using various DNA-based markers and developed molecular markers related to biotic and abiotic stresses, polyembryony, fruit coloration, seedlessness, male sterility, acidless, morphology, fruit quality, seed number, yield, early fruit setting traits, and QTL mapping on genetic maps. Genes closely related to CTV resistance and flesh color have been cloned. SSR markers for identifying zygotic and nucellar individuals will contribute to cost-effective breeding. The two high quality citrus reference genomes recently released are being efficiently used for genomics-based molecular breeding such as construction of reference linkage/physical maps and comparative genome mapping. In the near future, the development of DNA molecular markers tightly linked to various agronomic traits and the cloning of useful and/or variant genes will be accelerated through comparative genome analysis using citrus core collection and genome-wide approaches such as genotyping-by-sequencing and genome wide association study.

• The Plant Pathology Journal
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• v.33 no.1
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• pp.21-33
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• 2017

Citrus blast caused by bacterium Pseudomonas syringae is a very important disease of citrus occuring in many areas of the world, but with few data about genetic structure of the pathogen involved. Considering the above fact, this study reports genetic characterization of 43 P. syringae isolates obtained from plant tissue displaying citrus blast symptoms on mandarin (Citrus reticulata) in Montenegro, using multilocus sequence analysis of gyrB, rpoD, and gap1 gene sequences. Gene sequences from a collection of 54 reference pathotype strains of P. syringae from the Plant Associated and Environmental Microbes Database (PAMDB) was used to establish a genetic relationship with our isolates obtained from mandarin. Phylogenetic analyses of gyrB, rpoD, and gap1 gene sequences showed that P. syringae pv. syringae causes citrus blast in mandarin in Montenegro, and belongs to genomospecies 1. Genetic homogeneity of isolates suggested that the Montenegrian population might be clonal which indicates a possible common source of infection. These findings may assist in further epidemiological studies of this pathogen and for determining mandarin breeding strategies for P. syringae control.

• Journal of Plant Biotechnology
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• v.42 no.4
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• pp.326-335
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• 2015

Citrus is an economically important fruit tree with the largest amount of fruit production in the world. It provides important nutrition such as vitamin C and other health-promoting compounds including its unique flavonoids for human health. However, it is classified into the most difficult crops to develop new cultivars through conventional breeding approaches due to its long juvenility and some unique reproductive biological features such as gamete sterility, nucellar embryony, and high level of heterozygosity. Due to global warming and changes in consumer trends, establishing a systematic and efficient breeding programs is highly required for sustainable production of high quality fruits and diversification of cultivars. Recently, reference genome sequences of sweet orange and clementine mandarin have been released. Based on the reference whole-genome sequences, comparative genomics, reference-guided resequencing, and genotyping-by-sequencing for various citrus cultivars and crosses could be performed for the advance of functional genomics and development of traits-related molecular markers. In addition, a full understanding of gene function and gene co-expression networks can be provided through combined analysis of various transcriptome data. Analytic information on whole-genome and transcriptome will provide massive data on polymorphic molecular markers such as SNP, INDEL, and SSR, suggesting that it is possible to construct integrated maps and high-density genetic maps as well as physical maps. In the near future, integrated maps will be useful for map-based precise cloning of genes that are specific to citrus with major agronomic traits to facilitate rapid and efficient marker-assisted selection.

• Korean Journal of Breeding Science
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• v.40 no.2
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• pp.184-187
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• 2008

A new early maturing satsuma mandarin (Citrus unshiu Marc.) 'Haryejosaeng' was developed as a nucellar seedling selection of 'Tachima Wase' (C. unshiu Marc.) that was crossed with C. natsudaidai Hayata at the National Institute of Subtropical Agriculture in Jeju island in 1992. The 'Jegam ga No. 2', a first selection from the seedlings fruited in 2001 was finally named "Haryejosaeng" through field evaluation trials at three locations in Jeju island from 2003 to 2004. 'Haryejosaeng' produces seedless fruit maturing in early November, and has higher soluble solids and lower acidity than 'Miyagawa Wase', the leading early-maturing satsuma mandarin cultivar in Jeju island. Fruit weight is moderate at about 80~90 g and its shape is compressed-oblate globose with a light orange color. The rind thickness of about 2 mm provides easy peeling. The flesh shows light orange colored and contains 10 to 11 Brix and 1 to 1.1% acidity when mature. Tree shows vigorous growth and spreading thornless twigs. Alternate bearing is similar to 'Miyagawa Wase'. 'Haryejosaeng' is susceptible to citrus scab disease and melanose, but resistant to citrus canker.

• Journal of Life Science
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• v.21 no.10
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• pp.1452-1459
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• 2011

Citrus is a common term and genus (Citrus) of flowering plants in the rue family, Rutaceae. Citrus is believed to have originated in the part of Southeast Asia bordered by Northeastern India, Myanmar (Burma) and the Yunnan province of China. The taxonomy and systematics of the genus are complex and the precise number of natural species is unclear, as many of the named species are clonally propagated hybrids, and there is genetic evidence that even some wild, true-breeding species are of a hybrid origin. One of the most popular sequences for phylogenetic inference at the generic and infrageneric levels in plants is the chloroplast trnL-trnF region. We evaluated the seven taxa with the trnL-trnF region to estimate phenotypic relationships within the genera Citrus, Poncirus, and Fortunella in Korea. Alignment of the DNA sequences required the addition of numerous gaps. Sequence variation within Citrus was mostly due to insertion/deletion. Within the genus Citrus, C. lomonia and C. sinensis were relatives and sistered with C. aurantium in the four phylogenetic analyses (MP, ML, ME, and NJ). However, some external nodes were poorly supported.

• Horticultural Science & Technology
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• v.35 no.1
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• pp.121-130
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• 2017

This study was carried out to investigate a method for producing cultured virus - free ovules for breeding high - quality Citrus cultivars. Ovules from the immature fruits of three citrus cultivars native to Jeju (Dongjeongkyool, Cheongkyool, and Jikak) and two cultivars of Citrus unshiu Marc. (Miyagawa wase and Haryejosaeng) that were thought to be infected with Citrus tristeza virus (CTV) were cultured on MS2 medium (Murashige - Skoog [MS] basal medium containing $500mg{\cdot}L^{-1}$ malt extract, $50g{\cdot}L^{-1}$ sucrose, $1.0 mg{\cdot}L^{-1}$ kinetin, and $8g{\cdot}L^{-1}$ agar). After four weeks of culture, 10, 21, 13, 5, and 7 somatic embryos and 2, 4, 2, 4, and 5 white callus cells (surrounding green somatic embryos) were obtained from Dongjeongkyool, Cheongkyool, Jikak, Miyagawa wase, and Haryejosaeng, respectively. After six weeks of culture, somatic embryos were obtained from cultured cells grown on MT basal medium supplemented with malt extract ($500mg{\cdot}L^{-1}$), lactose ($70g{\cdot}L^{-1}$), and agar ($16g{\cdot}L^{-1}$). Over 60% of the somatic embryos from citrus cultivars native to Jeju developed into normal plants on MS basal medium supplemented with malt extract ($500mg{\cdot}L^{-1}$), sucrose ($50g{\cdot}L^{-1}$), and agar ($8g{\cdot}L^{-1}$) after 10 weeks of culture. Normal plants were regenerated from two Citrus unshiu Marc. cultivars on MT basal medium supplemented with sorbitol (1.0 M), galactose (1.0 M), $GA_3$ ($1.0mg{\cdot}L^{-1}$), and Gelrite ($3g{\cdot}L^{-1}$). The absence of virus in plants generated from cultured ovules was confirmed by RT - PCR and antigen - antibody reactions. Therefore, virus - free Citrus cells can be obtained for breeding high - quality citrus cultivars using the biotechnological technique evaluated in this study.

• Journal of Life Science
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• v.21 no.12
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• pp.1659-1665
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• 2011

In Citrus, an $F_1$ segregation population of 150 plants was constructed from a cross between 'Kiyomi' (C. unshiu ${\times}$ C. sinensis) carrying the male sterility trait and 'Jinkyool' (C. sunki). Sequence-related amplification polymorphism (SRAP) combined with bulked segregant analysis was used to develop markers linked to male fertility. In the $F_1$ population, 66 out of 150 seedlings had aborted anthers and the ratio of male sterile plants to fertile plants in the progenies matched the expected Mendelian segregation ratio of 1:1 ($x^2$ =2.16 at p=0.05). From the profiling of the 197 SRAP primer sets, three SRAP primer sets (F4/R27, F39/R60, and F15/R37) that were closely linked to the target trait were identified and successfully converted into a sequence characterized amplified region (SCAR) marker for selection of male fertility in citrus. The SCAR marker, using the pMS 33U/pMS 1462L primer set specifically, produced a single 1.4-Kb fragment that was linked to male fertility. Our results suggested that this SCAR marker can be useful for marker-assisted selection of male sterile individuals in breeding $F_1$ progenies in Citrus.

• Journal of Life Science
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• v.22 no.7
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• pp.871-879
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• 2012

Polyploids are a potentially important germplasm source in seedless citrus breeding program. Seedlessness is one of the most promising traits of commercial mandarin breeds that mandarin triploid hybrids possess permanently. The formation of new constant triploid hybrids can be recovered through diploid species hybridization from the fusion of divalent gametes at low frequencyor intra-and inter-ploidy crosses. However, extensive breeding work based on small $F_1$ hybrid seeds developed is impossible without a very effective aseptic methodology and ploidy event. In this study, in vitro embryo culture was employed to recover natural hybrids from monoembryonic diploid, open-pollinated mandarin. Flow cytometry was used to determine ploidy level. A total of 10,289 seeds were extracted from 792 fruits having approximately 13 seeds per fruit. Average frequency of small seeds developed was 7.1%, while the average frequency of small seeds per fruit were: 8.9% for 'Clementine' 10.2% for 'Harehime' 2.6% for 'Kamja' 3.1% for 'Pyunkyool' 2.8% for 'Sadookam' and 7.0% for 'Wilking' mandarin. Average size of a perfect seed was $49.52{\pm}0.07mm^2$ ('Clementine') while the small seed measured $7.95{\pm}0.04mm^2$ ('Clementine'), which was about 1/6 smaller than the perfect seed. In total, 731 small seeds were obtained and all of them contained only one embryo per seed. The efficiency of 'Clementine' was 14 times higher than 'Wilking' and more than 109 times higher than 'Pyunkyool'. The basic information on spontaneous polyploidy provides for the hybridization of constant triploids and increases the efficiency of conventional cross.