• Plant Disease and Agriculture
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• v.5 no.1
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• pp.34-40
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• 1999

The virus infection rates and disease symptoms of three major citrus viruses, citrus tristeza virus (CTV), satsuma dwarf virus (SDV), and citrus tatter leaf virus (CTLV), were investigated at 35 citrus orchards in Cheju Island from 1995. The infection rates of CTV, SDV, and CTLV were 69.8%, 8.6%, and 9.3%, respectively. However, depending on cultivars there were significant differences in the infection rates. The infection rates of CTV were highest in early satsuma mandarin (Citrusunshiu) with 80.9% and lowest in very early satsuma mandarin with 51.9%. In SDV, the highest was in very early stasuma mandarin with 23.1% and the lowest was in early satsuma mandarin with 6.3%. And the highest infection rate in very early satsuma mandarin with 17.9% and the lowest in tangors with 7.3% in CTLV. The symptoms of virus-infected citrus were very diverse; small and abnormal shape of fruits, abnormal leaves such as narrow boat and small spoon shapes of leaves, stem-pitting on the twig, bud-union crease and swelling of the graft part, reduction of the plant vigor and poor yields.

• The Plant Pathology Journal
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• v.33 no.3
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• pp.307-317
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• 2017

Satsuma dwarf virus (SDV) or Citrus mosaic sadwavirus (CiMV) were not consistently detected in RTPCR assay with the primer sets based on gene of Japan isolates. SDV and CiMV isolates were distinctively divided into two groups based on phylogenetic analysis of PP2 gene cloned from 22 Korean isolates, and the Korean CiMV and SDV isolates shared 95.5-96.2% and 97.1-97.7% sequence identity with Japanese isolate, respectively. We developed PP2-1 primer set based on the PP2 gene sequence of Korean isolates to simultaneously and effectively detect SDV and CiMV. And CTLV-2013 and CTV-po primer sets were newly designed for detection of Citrus tatter leaf virus (CTLV) and Citrus tristeza virus (CTV), respectively. Using these primer sets, a new multiplex PCR assay was developed as a means to simultaneously detect 4 citrus viruses, CTV, CTLV, SDV, and CiMV. The degree of detection by the multiplex PCR were consistent with those of uniplex RT-PCR for detection of each of the viruses. Therefore, the new multiplex PCR provides an efficient method for detecting 4 citrus viruses, which will help diagnose many citrus plants at the same time. We verified that 35.2% and 72.1% of 775 trees in 155 orchards were infected with SDV or CiMV (SDV/CiMV) and CTV by the multiplex-PCR assay, respectively, and CTLV was not detected in any of the trees tested.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 1999.10a
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• pp.73.2-73
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• 1999

• Journal of Plant Biotechnology
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• v.32 no.1
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• pp.45-50
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• 2005

Virus-free stocks was produced by the combination of the heat treatment of virus infected plant and shoot-tip grafting (STS). To produce virus-free stocks, the plants infected with citrus viruses were used for virus-free stock production using the modified method of STG in thermotherapy at $40^{\circ}C$ for 16 hours in the light, and at $30^{\circ}C$ for 8 hours of darkness for 4 weeks. Trifoliate orange (P. trifoliata) were used as rootstock seedling for STG. Percentages of virus-free stocks against citrus tristeza virus (CTV), satsuma dwarf virus (SDV) and citrus tatter leaf virus (CTLV) were $75.7\%,\;100.0,\%\;82.6\%$ respectively. Shoot tip size for successful STG were as small as possible. Less than $0.3\;\cal{mm}$ of shoot tips gave the hight efficiency of virus free plants but survival rates were low. And, survival rate after shoot-tip culture was analyzed and the rates were dependant on the cultivars; Yuzu cultivar showed the hight survival rate ($74.6\%$) and early satsuma mandarin (Iwasagi) was $13.3\%$ as the lowest cultivar. But citrus trees were not succeed to grown, turned brown, and died.

• Journal of Life Science
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• v.23 no.2
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• pp.267-272
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• 2013

This study was conducted to eliminate viruses from citrus-infected plants using micrografting and thermotherapy. Six citrus cultivars including a 'Setoka' hybrid were used as plant sources. The TAS-ELISA technique demonstrated that several plants were CTV positive. However, no CTV symptoms were detected in plants obtained from shoots and treated at a high temperature of $40^{\circ}C$ during the day and night and micrografted for two weeks with old trifoliate orange rootstock in vitro. Indexing of CTV, SDV, and CTLV for RT-PCR analysis of the eleven citrus seedlings, including 'Setoka', 'Samdajosang', 'Pungkwang', 'Shiranuhi', and 'Ehimekashi dai28go' was virus free following the micrografting and thermal therapy.

• 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.