Citrus Leaf Blotch Virus

California Pest Rating for

Citrus leaf blotch virus

Pest Rating: B

PEST RATING PROFILE

Initiating Event:

On February 26, 2018, Dr. G. Vidalakis, University of California, Director, Citrus Clonal Protection Program, informed CDFA of his detection of Citrus leaf blotch virus (CLBV) from a Bearss Lime tree at a residence in Los Angeles County. Subsequently, an official sample, which comprised a total of 4 subsamples, was collected by the CDFA from the same Bearss Lime tree and sent to the CDFA Plant Pathology Laboratory for diagnosis. On February 27, 2018, Tongyan Tian, CDFA Plant Pathologist, detected Citrus leaf blotch virus from all four subsamples using RT-qPCR and further confirmed the identity of the pathogen by conventional RT-PCR and sequencing. A temporary Q rating was assigned to the pathogen. The status, risk and consequences of introduction of CLBV to California are assessed and a pest permanent pest rating is proposed herein.

History & Status:

Background: In 1968, Dweet mottle virus (DMV) was initially detected and reported from Riverside, California, during re-indexing of a candidate Cleopatra mandarin variety (C. reticulata) on ‘Dweet’ tangor at the University of California Riverside Citrus Variety Improvement Program, the forerunner of the present Citrus Clonal Protection Program (CCPP). The candidate mandarin variety had been introduced from Florida into the Program at Riverside. The virus produced leaf chlorotic blotching symptoms that resembled, but were distinct from, symptoms produced by psorosis virus and Citrus concave gum virus. It also produced a mild exocortis reaction in Etrog citron. The parent tree did not show symptoms of damage caused by any known virus and the trunk appeared normal without any signs of stem pitting or bark discoloration, although small fruit, twig dieback and little new growth were apparent. Since the virus produced symptoms only in ‘Dweet’, it was named Dweet mottle virus (Roistacher & Blue, 1968). However, Dweet mottle virus was not reported from any commercial citrus production sites nor was it observed to produce any economic losses and was detected only once after 1963 in the CCPP indexing program (Krueger et al., 2012).

Then in 1984, at the Citrus Variety Improvement Program in Spain, Navarro and other scientists reported a new graft transmissible disease that caused a bud-union incompatibility between ‘Nagami’ kumquat and ‘Troyer’ citrange rootstock. The ‘Nagami’ kumquat had been introduced from Corsica, France. In addition to bud-union incompatibility, the presumptive virus involved caused vein clearing in certain citrus species and stem pitting in Etrog citron. However, after shoot-tip grafting, some plants produced were compatible with Troyer, but still caused stem pitting in Etrog citron, thereby, indicating the involvement of more than one virus (Navarro et al., 1984). Galipienso et al., 2000, gave further evidence of the involvement of more than one virus by demonstrating bud union crease in certain citrus species but not others when propagated on ‘Troyer’ citrange. However, chlorotic blotching in ‘Dweet’ tangor, like those induced by DMV, and stem pitting in Etrog citron were produced by all sources of the virus. In 2001-02, the causal agent in “Nagami’ kumquat was partially purified and characterized and given the candidate name, Citrus leaf blotch virus (CLBV) (Galipienso et al., 2001; Vives et al., 2001, 2002). Furthermore, these researchers detected CLBV in different citrus varieties from Japan, New South Wales (Australia), Spain, and Florida, usually associated with abnormal bud union on citrange or citrumelo. Comparison of 14 CLBV isolates from Spain, Japan, USA, France and Australia showed low genetic diversity (Vives et al., 2002). Low rates of seed transmission were demonstrated in three citrus varieties or hybrids (Guerri et al., 2004). A few years later, Vives et al., (2005) conducted partial sequence analysis to show that Dweet mottle virus from California had over 96% sequence (high) homology with citrus leaf blotch virus from Spain and therefore, suggested that DMV may be caused by CLBV. Both viruses induce mottling in ‘Dweet’ tangor and stem pitting in ‘Etrog’ citron and that, besides CLBV, a different pathogen causing bud-union crease and vein clearing may be present in ‘Nagami’ kumquat sources but not in DMV from California source. This was further demonstrated by Vives et al., (2008a) by the development of full-genome cDNA clones of CLBV that caused systemic infection in agro-inoculated herbaceous and citrus host plants and induced chlorotic blotching in ‘Dweet’ tangor and stem pitting in Etrog citron, but not vein clearing in Pineapple sweet orange or bud union crease on trifoliate rootstocks. Then in 2010, Hajeri and other researchers at the University of California, Riverside, and the USDA ARS National Clonal Germplasm Repository for Citrus and Dates (NCGRCD), Riverside, determined the complete nucleotide sequence of DMV and with phylogenetic analysis showed that DMV is an isolate of CLBV, and not a distinct species, within the genus Citrivirus.

In California, the seed transmissibility of citrus leaf blotch virus caused concern to the citrus nursery industry. Consequently, Kreuger et al. (2012) reported that all citrus trees at CCPP and NCGRCD were tested for the presence of the virus using RT-PCR with local DMV positives and a CLBV positive from Florida as positive controls. The virus was not detected in the tested trees. Furthermore, they failed to detect it during surveys of field trees exhibiting bud union abnormalities for the presence of specific pathogens and therefore, while the overall status of CLBV in California is presently unknown, they believe that this virus if present at all, is only at a low incidence. This is because the close identity of CLBV and DMV has likely prevented CLBV from becoming introduced into California. All introductions of new citrus germplasm are indexed into ‘Dweet’ tangor as well as other indicator species at CCPP and NCGRCD. Reaction of CLBV in ‘Dweet’ tangor would enable detection of this virus, even if the actual identity of the virus was not known at the time of indexing. Detection of positives or even misidentifications would have been eliminated by thermal therapy or shoot-tip grafting before release (Kreuger et al., 2005, 2012).

Citrus leaf blotch virus has been reported in China, Corsica (France), Cuba, Italy, Japan, New South Wales (Australia), New Zealand, Spain, Florida, Arkansas, Oregon, and California (USA). In Arkansas and Oregon, the virus was found in peony plants showing stunting and gnarled irregularities, however, since the virus was found in both symptomatic and asymptomatic material, it could not be associated with the disease and its role in peony health is currently unknown. Nonetheless, CLBV may easily move between propagation cycles via mechanical and seed transmission of clonally propagated peony plants (Gress et al., 2017).

Citrus leaf blotch virus not only causes symptomless infection in most citrus but also, is unevenly distributed within an infected plant, thereby presenting a possible challenge for its detection. In greenhouse studies, Vives et al. (2002) detected CLBV consistently in young leaves of infected ‘Nagami’ kumquat, ‘Owari’ Satsuma, Navelina and Navel oranges, however, detection in old leaves of other citrus species (Eureka lemon, Marsh grapefruit and Nules Clementine) was not consistent, particularly in Pineapple sweet orange. Detection of the virus in field trees was even less consistent, and not detected in neighbor trees showing similar symptoms possibly due to low titer or uneven distribution of the virus in the plant.

Hosts: Citrus spp., including C. sinensis, C. limon, C. unshiu, C. paradisi, Poncirus trifoliata, P. trifoliata x C. sinensis (Harper et al., 2008), C. medica (Etrog citrus), C. reticulata x C. sinensis (‘Dweet’ tangor) (Roistacher & Blue, 1968), Fortunella margarita (kumquat “Nagami’) (Navarro et al., 1984), Prunus avium cv. Red-lamp (sweet cherry) (Wang et al., 2016), Actinidia sp. (kiwifruit) (Zhu et al., 2016), Paeonia lactiflora (peony) (Gress et al., 2017). Experimental hosts include Nicotiana cavicola (Guardo et al., 2009), N. occidentalis and N. benthamiana (Vives et al., 2008b).

Symptoms: Citrus leaf blotch virus causes symptomless infection in most citrus species and cultivars (Vives et al., 2008a). However, CLBV (and the isolate, DMV) induce chlorotic blotching or mottling in ‘Dweet’ tangor and stem pitting ‘Etrog’ citron. Although CLBV does not induce bud union crease on trifoliate rootstock (Vives et al., 2008a), it has been found to be usually associated with abnormal bud union on citrange or citrumelo rootstock. A different pathogen or interaction of CLBV with a different pathogen is likely the cause of bud union crease and vein clearing symptoms (Vives et al., 2005).

Damage Potential: Citrus leaf blotch virus causes chlorotic leaf blotching in ‘Dweet’ tangor and stem pitting in Etrog citron. Although it does not induce bud union crease in several citrus species it is usually associated with bud union crease symptoms in citrange and citrumelo rootstocks and therefore, an interaction between CLBV and other agent(s) cannot be ruled out. There are no reports of yield losses due to CLBV and the virus can cause symptomless infections in most citrus species and cultivars. In California, CLBV (aka DMV) is a regulated pathogen and its distribution is unknown or at best likely to be of low incidence. CLBV (aka DMV) was not reported from any commercial citrus production sites in California nor was it observed to produce any economic losses (Krueger et al., 2012). However, in certain scion-rootstock combinations using ‘Dweet’ tangor and Etrog citron rootstocks there may be a potential for disease development due to CLBV.

Transmission: Citrus leaf blotch virus is transmitted in citrus by grafting and seed. CLBV dispersal occurs primarily by propagation of infected buds. Low rates of seed transmission in at least three citrus species and hybrid, ‘Troyer’ citrange (Citrus sinensis x Poncirus trifoliata), ‘Nagami’ kumquat (Fortunella margarita) and sour orange (C. aurantium), has been demonstrated (Guerri et al., 2004). Also, CLBV has been mechanically transmitted to Nicotiana cavicola (Guardo et al., 2009), by sap inoculation to N. occidentalis and N. benthamiana (Vives et al., 2008b), and transmitted from citrus to citrus by contaminated knife blades (Roistacher et al., 1980). The virus is not transmitted by vectors (Galipienso et al., 2000).

Worldwide Distribution: Asia: China, Japan; Europe: Italy, Spain; North America: USA, Cuba; Oceania: New South Wales (Australia), New Zealand (Cao et al., 2017; Gress et al., 2017; Guardo et al., 2007; Harper et al., 2008; Hernández-Rodríguez, 2016; Navarro et al., 1984; Roistacher & Blue, 1968; Vives et al., 2002; Wang et al., 2016).

Official Control: Citrus leaf blotch virus is on the ‘Harmful Organism’ list for Uruguay (USDA PCIT, 2018). CLBV (aka DMV) is a regulated pathogen in California’s mandatory Citrus Nursery Stock Pest Cleanliness Program (CCR, Title 3, Division 4, Chapter 4, Subchapter 6, Section 3701).

California Distribution: The distribution in California is unknown. If at all present, it is likely to be only at a low incidence (Kreuger et al., 2005, 2012. See: ‘Background’).

California Interceptions: No official interceptions have been reported.

The risk Citrus leaf blotch virus would pose to California is evaluated below.

Consequences of Introduction:

1) Climate/Host Interaction: Although the distribution of Citrus leaf blotch virus in California, is presently unknown and is likely to be only at a low incidence (Kreuger et al., 2012), if not regulated, it may be possible for the pathogen to have a widespread establishment in symptomatic and non-symptomatic infected citrus varieties in commercial citrus-growing regions of the State.

Evaluate if the pest would have suitable hosts and climate to establish in California. Score: 3

– Low (1) Not likely to establish in California; or likely to establish in very limited areas.

– Medium (2) may be able to establish in a larger but limited part of California.

– High (3) likely to establish a widespread distribution in California.

2) Known Pest Host Range: The natural host range is limited primarily to Citrus Other hosts include sweet cherry and kiwifruit reported from China and peony reported from Arkansas and Oregon. Experimental hosts include, Nicotiana cavicola, N. occidentalis and N. benthamiana.

Evaluate the host range of the pest.

Score: 1

– Low (1) has a very limited host range.

– Medium (2) has a moderate host range.

– High (3) has a wide host range.

3) Pest Dispersal Potential: Citrus leaf blotch virus has high reproduction within its plant host, although unevenly distributed within infected plants. It is transmitted by grafting, seed, and mechanically. Its ability for long distance spread through infected seed render it a high rating for dispersal.

Evaluate the natural and artificial dispersal potential of the pest.

Score: 3

– Low (1) does not have high reproductive or dispersal potential.

– Medium (2) has either high reproductive or dispersal potential.

– High (3) has both high reproduction and dispersal potential.

4) Economic Impact: Citrus leaf blotch virus is a regulated pathogen under California’s mandatory Citrus Nursery Stock Pest Cleanliness Program. Under this program any citrus stock found positive for the pathogen would be eliminated before release for commercial planting. This pathogen causes chlorotic leaf blotching in ‘Dweet’ tangor and stem pitting in Etrog citron. Although it does not induce bud union crease in several citrus species, it is usually associated with bud union crease symptoms in citrange and citrumelo rootstocks and therefore, an interaction between CLBV and other agent(s) cannot be ruled out. There are no reports of yield losses due to CLBV and the virus can cause symptomless infections in most citrus species and cultivars. Researchers have stated that CLBV has not been reported from commercial citrus production sites in California nor was it observed to cause any economic losses. If citrus stock were not regulated, it is likely that in certain scion-rootstock combinations using ‘Dweet’ tangor and Etrog citron rootstocks there may be a potential for disease development due to CLBV. In such a case, it is estimated that CLBV could lower crop yield and value and trigger the loss of markets.

Evaluate the economic impact of the pest to California using the criteria below.

Economic Impact: A, B, C

A. The pest could lower crop yield.

B. The pest could lower crop value (includes increasing crop production costs).

C. The pest could trigger the loss of markets (includes quarantines).

D. The pest could negatively change normal cultural practices.

E. The pest can vector, or is vectored, by another pestiferous organism.

F. The organism is injurious or poisonous to agriculturally important animals.

G. The organism can interfere with the delivery or supply of water for agricultural uses.

Economic Impact Score: 3

– Low (1) causes 0 or 1 of these impacts.

– Medium (2) causes 2 of these impacts.

– High (3) causes 3 or more of these impacts.

5) Environmental Impact: No environmental impact is expected, however, if not regulated, CLBV may impact home/urban plantings of citrus host plants.

Evaluate the environmental impact of the pest on California using the criteria below.

Environmental Impact: E

A. The pest could have a significant environmental impact such as lowering biodiversity, disrupting natural communities, or changing ecosystem processes.

B. The pest could directly affect threatened or endangered species.

C. The pest could impact threatened or endangered species by disrupting critical habitats.

D. The pest could trigger additional official or private treatment programs.

E. The pest significantly impacts cultural practices, home/urban gardening or ornamental plantings.

Environmental Impact. Score: 2

– Low (1) causes none of the above to occur.

– Medium (2) causes one of the above to occur.

– High (3) causes two or more of the above to occur.

Consequences of Introduction to California for Citrus leaf blotch virus: 12

Add up the total score and include it here. (Score)

-Low = 5-8 points

–Medium = 9-12 points

-High = 13-15 points

6) Post Entry Distribution and Survey Information: Evaluate the known distribution in California. Only official records identified by a taxonomic expert and supported by voucher specimens deposited in natural history collections should be considered. Pest incursions that have been eradicated, are under eradication, or have been delimited with no further detections should not be included. (Score)

Evaluation is (0). While the distribution of CLBV in California is currently not known, there is no evidence that it is established within the State.

–Not established (0) Pest never detected in California, or known only from incursions.

-Low (-1) Pest has a localized distribution in California, or is established in one suitable climate/host area (region).

-Medium (-2) Pest is widespread in California but not fully established in the endangered area, or pest established in two contiguous suitable climate/host areas.

-High (-3) Pest has fully established in the endangered area, or pest is reported in more than two contiguous or non-contiguous suitable climate/host areas.

Final Score:

7) The final score is the consequences of introduction score minus the post entry distribution and survey information score: (Score)

Final Score: Score of Consequences of Introduction – Score of Post Entry Distribution and Survey Information = 12

Uncertainty:

The in-state distribution of CLBV is not currently known. Also, the impact of infection related to crop damage and losses is not known.

Conclusion and Rating Justification:

Based on the evidence provided above the proposed rating for Citrus leaf blotch virus is B.

References:

Cao, M. J., Y. -Q. Yu, X. Tian, F. Y. Y. and, R. H. Li and C. Y. Zhou. 2017. First report of Citrus leaf blotch in lemon in China. Plant Disease 101: 8. https://doi.org/10.1094/PDIS-10-16-1500-PDN

Galipienso, L., L. Navarro, J. F. Ballester-Olmos, J. Pina, P. Moreno, and J. Guerri. 2000. Host range and symptomatology of a graft transmissible pathogen causing bud union crease of citrus on trifoliate rootstocks. Plant Pathology 49: 308–314.

Galipienso, L., M. C. Vives, P. Moreno, R. G. Milne, L. Navarro and J. Guerri. 2001. Partial characterization of Citrus leaf blotch virus, a new virus from Nagami kumquat. Archives of Virology 146: 357–368.

Gress, J. C., S. Smith, and I. E. Tzanetakis. 2017. First report of Citrus leaf blotch virus in peony in the U.S.A. Plant Disease 101: 637. https://doi.org/10.1094/PDIS-08-16-1218-PDN

Guardo, M., G Sorrentino, T. Marletta and A. Carusa. 2007. First report of Citrus leaf blotch on kumquat in Italy. Plant Disease 91: 104.

Guardo, M., O. Potere, M. A. Castellano, V. Savino and A. Caruso. 2009. A new herbaceous host for Citrus leaf blotch virus. Journal of Plant Pathology 91: 485-488.

Guardo, M., G. Sorrentino and A. Caruso. 2015. Characterization and incidence of Citrus leaf blotch virus in Southern Italy. 12^th International Citrus Congress – International Society of Citriculture. Acta Horticulturae 1065: 825-83.

Hajeri, S., C. Ramadugu, M. Keremane, G. Vidalakis and R. Lee. 2010. Nucleotide sequence and genome organization of Dweet mottle virus and its relationship to members of the family Betaflexiviridae. Arch Virol 15: 1523-1527. DOI 10.1007/s00705-010-0758-1

Harper, S. J., K. M. Chooi and M. N. Pearson. 2008. First report of Citrus leaf blotch virus in New Zealand. Plant Disease 92: 1470. https://doi.org/10.1094/PDIS-92-10-1470C

Hernàndez-Rodríguez, L., J. M. Pérez-Castro, G. García-García, P. Luis Ramos-González, V. Zamora-Rodríguez, Xenia Ferriol-Marchena, Inés Peña-Bárzaga and L. Batista-Le Riverend. 2016. Citrus leaf blotch in Cuba: first report and partial molecular characterization. Tropical Plant Pathology 41: 147. https://doi.org/10.1007/s40858-016-0078-4

Krueger, R. R., J. A. Bash and R. F. Lee. 2005. Phytosanitary status of California citrus. International Organization of Citrus Virologists Conference Proceedings (1957-20), 16 (16): 468-472. https://escholarship.org/uc/item/3667q9qn

Krueger, R. R., J. A. Bash and R. F. Lee. 2012. Dweet mottle virus and Citrus leaf blotch virus. http://ucanr.edu/blogs/blogcore/postdetail.cfm?postnum=7112

Navarro, L., J. A. Pina, J. F. Ballester-Olmos, P. Moreno and M. Cambra. 1984. A new graft transmissible disease found in Nagami kumquat. In: Timmer L. W., and J. A. Dodds (eds) Proceedings of the 9th Conference of the International Organization of Citrus Virologists, IOCV, Riverside, pp 234–240.

Roistacher, C. N., and R. L. Blue. 1968. A psorosis-like virus causing symptoms only on ‘Dweet’ tangor. International Organization of Citrus Virologists Conference Proceedings (1957-2010), 4(4): 13-18.

Roistacher, C. N., E. M. Nauer and R. C. Wagner. 1980. Transmissibility of cachexia, Dweet mottle, psorosis and infectious variegation viruses on knife blades and its prevention. Proceedings of the 8^th Conference of the International Organization of Citrus Virologists, IOCV, Riverside 1980: 225-229.

USDA PCIT. 2018. USDA Phytosanitary Certificate Issuance & Tracking System. Retrieved March 15, 2018. 3:25:54 pm CDT. https://pcit.aphis.usda.gov/PExD/faces/ReportHarmOrgs.jsp.

Vives, M. C., L. Galipienso, L. Navarro, P. Moreno and J. Guerri. 2001. The nucleotide sequence and genomic organization of Citrus leaf blotch virus: Candidate type species for a new virus genus. Virology 287: 225-233.

Vives, M. C., L. Galipienso, L. Navarro, P. Moreno and J. Guerri. 2002. Citrus leaf blotch virus: a new citrus virus associated with bud union crease on trifoliate rootstocks. International Organization of Citrus Virologists Conference Proceedings (1957-2010), 15 (15): 205-212.

Vives, M. C., L. Rubio, L. Galipienso, L. Navarro, P. Moreno and J. Guerri. 2002. Low genetic variation between isolates of Citrus leaf blotch virus from different host species and different geographical origins. Journal of General Virology 83: 2587–2591.

Vives M. C., J. A. Pina, J. Juarez, L. Navarro, P. Moreno and J. Guerri. 2005. Dweet mottle disease is probably caused by Citrus leaf blotch virus. 16th Conference of the International Organization of Citrus Virologists Conference Proceedings (1957-2010), 15 (16): 251-256.

Vives, M. C., S. Martin, S. Ambros, A. Renovell, L. Navarro, J. A. Pina, P. Moreno, J. and J. Guerri. 2008a. Development of a full-genome cDNA clone of Citrus leaf blotch virus and infection of citrus plants. Molecular Plant Pathology 9:787–797.

Vives, M. C., P. Moreno, L. Navarro and J. Guerri. 2008b. Citrus leaf blotch virus. In: Rao, G. P., A. Myrta and K. Ling (eds). Characterization, Diagnosis and Management of Plant Viruses, vol. 2. Pp. 55-67. Studium Press, Houston, TX, USA.

Wang, J., D. Zhu, Y. Tan, X. Zong, H. Wei and Q. Liu. 2016. First report of Citrus leaf blotch virus in sweet cherry. Plant Disease 100:1027.

Zhu, Chen-xi, Wang, Guo-ping, Zheng, Ya-zhou, Yang, Zuo-kun, Wang, Li-ping, Xu, Wen-xing and N. Hong. 2016. RT-PCR detection and sequence analysis of coat protein gene of Citrus leaf blotch virus infecting kiwifruit trees. Acta Phytopathologica Sinica, 46 (1): 11.

Responsible Party:

John J. Chitambar, Primary Plant Pathologist/Nematologist, California Department of Food and Agriculture, 3294 Meadowview Road, Sacramento, CA 95832. Phone: 916-262-1110, plant.health[@]cdfa.ca.gov.

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