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Characterization of Begomovirus The Causal Agent of Leaf Curl Disease on Cucumbers

Cucumber - Cucumis sativus L. is an important vegetable crops in Indonesia. Productivity of cucumber affected by inefficient and ineffective cultural practices in addition to the problems with pests and diseases. One of important disease on cucumber that has the potential to effect the production is leaf curl disease caused by Begomovirus. 


Leaf curl in Cucumber - Leaf curl diseases caused by Begomovirus has been reported on several crops in Indonesia and become a serious concern effecting yield loss. Tobacco plantation in East Java suffered 30% damage from infection of Tobacco leaf curl begomovirus in 1984. Since early 2000 infection of Pepper yellow leaf curl begomovirus on chillipepper has been reported in West and Central Java and disease management for this yellow disease on chillipepper has not been achieved successfully. Tomato yellow leaf curl begomovirus was also reported to cause epidemic on tomato plants in West and Central Java. 

Recently, leaf curl disease was observed in cucumber plants especially in Klaten, central Java. Similar disease was reported earlier since 1996, and virus identification showed that nucleotide sequence of the virus has high similarity with Tomato leaf curl New Delhi begomovirus (TLCV-New Delhi). Infection of Begomovirus on Cucurbitaceae has actually became problems in several countries, for instance Squash leaf curl begomovirus caused leaf curl disease on squash (Cucurbita maxima) and Watermelon curly mottle begomovirus caused yellow leaf and dwarfing on melon (C. melo) in United States. 

Insect Vector - Begomovirus is only transmitted by whitefly (Bemisia tabaci) as its insect vector and whitefly is known to have a broad host range and capable to colonize a number of plants. Identification and characterization of Begomovirus causing leaf curl disease of cucumber in Java has not been done despite its importance. Studies on host range and transmission of Begomovirus is important in order to develop disease management strategy. The aims of this research is to detect viruses infecting cucumber plants, to identify Begomovirus infected cucumber, to study transmission efficiency of Begomovirus by vector B. tabaci, and to determine the host range. 

Samples from Field - Field observation and samples collection was conducted in West Java (Bogor, Subang), Central Java (Tegal, Sukoharjo), and Yogyakarta (Sleman). Leaves showing typical symptoms of leaf curling, mosaic, blistering and yellowing were collected for further identification. Infection of Begomovirus, SqMV, ZyMV, and CMV was detected from all samples, but no infection of TRSV nor WMV was detected. Mix infection was found, for instance samples from Bogor and Sleman (Kalasan dan Ngemplak) were infected by 4 viruses (Begomovirus, SqMV, ZyMV, dan CMV) and showed more severe symptoms i.e. leaf curling, mosaic, yellowing and blistering. 

Identification - Nucleotide sequences obtained from sequencing of DNA fragments as the amplicon using polymerase chain reaction (PCR) method ranging from 1474 to 1633 bp for TEGAL, KALASAN, NGEMPLAK, SUKOHARJO, and BOGOR isolates. Begomovirus genome amplified using universal primer pAL1v1978/pAR1c715 covers part of replication gene, coat protein gene and common region which is the region of the genome commonly used for identification of Begomovirus. Sequence homology among the five isolates ranging from 96.1% to 99.3%, indicated close relationship. BOGOR isolate showed different symptoms from other isolates and its relationship is not as close with the other isolates (TEGAL, KALASAN, NGEMPLAK, and SUKOHARJO). All isolates showed high homology, i.e. 95.7% to 98.6%, to Tomato leaf curl New Delhi virus-[Cucumber:Indonesia] (AB613825) from Klaten, Central Java. Phyllogenetic analysis showed that all TLCV isolates infecting cucumber has close relationship with other isolates infecting other Cucurbitaceae plants (Cucurbita moschata and Luffa acutangula) and Solanaceae plants (Capsicum annuum and Solanum lycopersicum). 

Transmission - One insect vector (B. tabaci) was able to transmit TLCV and causing up to 60% disease incidence. The more whiteflies used in the transmission, the more disease incidence gained. Transmission using 10 and 20 whiteflies resulted on the highest disease incidence (86.67%). First symptom was observed 8 and 9 days after inoculation and symptoms were developed earlier when more whiteflies was used in the transmission. The shortest incubation period (8 days) was obtained in transmission using 10 whiteflies. Therefore, using 10 whiteflies was considered as the most efficient transmission for TLCV on cucumber. 

Spread - Host range of TLCV isolates from cucumber involves Cucurbitaceae (five varieties of Cucumis sativus, C. melo, Citrullus lanatus, Cucurbita pepo, Luffa acutangula) and Solanaceae (Lycopersicon esculentum, Solanum melongena, Nicotiana tabacum). The shortest incubation period (9 days) was obtained on tomato. The highest disease incidence (91.67%) was occurred on cucumber var. Sabana, whereas the lowest disease incidence was occurred on tobacco (16.67%). Although incubation period and disease incidence on Cucurbitaceae and Solanaceae was not obviously different, but symptom severity was more observed on cucumber than other Cucurbitaceae and Solanaceae. It seems that interaction between isolate of Begomovirus and variety of plant may influence disease incidence. 

Knowledge about host range of TLCV and its transmission is very important to control the disease. Infected plant will serve as virus inoculum in the field and insect vector will cause wide disease incidence and spread.

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