Quick detection of citrus greening by PCR method with specific and universal primers

Huanglungbin or Greening is caused by ploem restricted unculturable bacterium, identified as Liberobacter. In general, Liberobacter found in Indonesia is Liberobacter asiaticum. The aim of this research was to develop the effective primers for Polymerase Chain Reaction (PCR) method in order to detect citrus greening quickly. Primers used in this research were specific Primers (011, 012c, 0A1) and universal Primers (fDl/rPI). The specific Primers (011, 012c) were more effective and specific in detecting citrus greening than those of universal Primers (fDlirP1). In addition, the advantage of specific primers was that it could be used to amplify DNA greening of 1160 by only, but could not be used to amplify DNA of other sistemic pathogens. Keywords: Huanglungbin (HLB) â liberobacter â polymerase chain reaction â quick detection â specific and universal primer

SEED HEALTH EVALUATION IN THE PROCESS OF FREE-VIRUS CITRUS SEED PRODUCTION ON KAMPAR REGENCY, RIAU PROVINCE OF INDONESIA

Kampar is one of the largest citrus seed production centers in Indonesia which has a wide distribution range. In anticipating and preventing the spread of dangerous systemic diseases and the presence of vector pests, a survey was conducted to evaluate the health of the seeds and the application of technology to the process of citrus production in Kampar, Riau, Indonesia. The survey was conducted in November 2014 in Sungai Pinang, Tambang, Kampar Regency and at Padang Marpoyan Seed Center, Riau, Indonesia. The sample was selected by random sampling method on scattered seeds in captivity, Foundation Block (FB), Budwood Multiplication Block (BMB) plant in the protected screen house and in farmer land. The results of the survey showed that seed growers that have not used patches from the BMB mother tree are already available on the farmer's land of 1200 stems since 2014 while BMB in BBI is available since 2012. The condition of the parent FB and BMB trees managed by the Agriculture Service in Padang Marpoyan Seed Center is sufficiently good and protected in a screen house. The results of the seed health examination showed that the FB and BMB plants were free from Huanglongbing (HLB) and Citrus Tristeza Virus (CTV) diseases. Whereas, the field of BMB plants on the farmer's land with no screen house protection and Parent Tree showed that the plant's population in the field were infected with CTV by 66.7%. The seed plants spread to 100% small seed grower were infected with CTV. In the medium seed grower cluster, CTV infection occurred as much as 87.5%. In large seed grower, the indexing results showed a positive mark and contained Liberibacter asiaticum pathogens that cause HLB

Exploration of Trichoderma spp. and fungal pathogen that causes a strawberry anthracnose and examination of in vitro antagonistic activity

Strawberry (Fragaria Vesca L.) is a kind of fruit that has high economic value. The obstacles that often arise in strawberry cultivation are pests and diseases. One of the alternatives to control the anthracnose is a biological agent known as Trichoderma spp. Trichoderma spp is antagonistic to the pathogen. The objective of this research is to explore the Trichoderma spp. as an antagonistic agent to the fungal pathogen that causes anthracnose disease in strawberry. The Trichoderma spp. was isolated by pour plate method while the pathogen was isolated by a direct plating method. The antagonistic activities of Trichoderma spp. were tested in vitro towards fungal pathogen by dual culture assay. The pathogen and antagonistic agent were paired by using three methods of pairing. The fungal pathogen has a similarity to the members of the Genus Colletotrichum and the two isolated antagonists have a similarity to the member of the Genus Trichoderma. The two Trichoderma spp. (TKL1 and TKL2) have significantly inhibited a radial growth of the pathogen. An introduction of the antagonists before the pathogen has given the best growth inhibition of the pathogen. The inhibitory effect of the two Trichoderma spp. is not significantly differed from each other at p>0,05. The microscopic examination showed that the most common mode of the action was mycoparasitism. The Trichoderma hyphae grew alongside and coiled compactly around the hyphae of the fungal pathogen isolates

THE EFFECT OF SINGLE AND DUAL INFECTIONS OF CITRUS TRISTEZA VIRUS AND CITRUS VEIN ENATION VIRUS ON CITRUS SPECIES

Viral diseases caused by CTV and CVEV induce dwarfing on plants and severe damage and yield loss to the citrus crop. The virus transmitted through infected plant material and by the same vectors, aphid. Problems cited in the field are complex of symptoms of the two diseases that are difficult to distinguish. This study aimed to determine the effect of single and dual infections of CTV and CVEV on Purut (Citrus hystrix) and Lime (Citrus aurantifolia). The research was conducted at the screen house and laboratory of Balitjestro, Batu and University of Brawijaya in October 2014 to April 2015. CTV and CVEV isolates used were from Balitjestro’s collection. Treatments conducted were single inoculation for CTV and CVEV, 3 combinations of CTV and CVEV dual inoculation. Each treatment was carried out both on Purut and Lime. A total of 6 treatments and 5 replications were arranged in a Randomized Complete Design; each replication comprised of two plants. The results showed that the symptoms on Purut appeared faster and more severe than those on Lime. Both single and dual infections of CTV and CVEV affected the incubation period of CTV on vein clearing symptoms, cupping and vein enation. Symptom of vein clearing was found the fastest at dual infections treatment of CTV followed by CVEV. Cupping symptom was seen the fastest in the treatment of dual infections of CVEV followed by CTV, and symptom of vein enation was found only in a single infection of CVEV. The highest disease intensity and infection of vein clearing and cupping were observed on dual infections of CVEV followed by CTV while the highest infections of vein clearing and stem pitting were obtained from simultaneous dual infection of CTV and CVEV. The results of serologic testing by DAS ELISA on three observations (44 days after inoculation (dai), 86 dai, and 142 dai) showed that a dual-infection showed greater absorbance value than single infection treatment did. Plants growths on dual infections treatment were slower than those on single infection

Study of entomophatogenic fungus to control vector insect of citrus tristeza virus on citrus

Citrus Tristeza Virus (CTV) disease is a silent killer, which threatens to decrease productivity, quality and even death of citrus plants and the erosion of genetic resources. Spreading in the field very quickly by the intermediate insect vector pest, aphid (Toxoptera citricida, T. Aurantii and A. Gosypii). The microbes studied for potential biopesticide candidates are: Beauveria bassiana and Hirsutella citriformis, and Metarhizium anisopliae (Metch) Sorokin previously reported to control Diaphorina citri pests resulting effectiveness of > 25% and was able to suppress yield loss up to 10%. The objectives of the study examined the effectiveness of entomopathogen in controlling the pest of CTV vector, Toxoptera citricida, in the laboratory and screen house, to findout the physiological, biochemical and molecular physiology of entomopathogen. The results showed that the best entomopathogen suspension concentration was B.bassiana 106 followed by H. citriformis 106 and M. anisopliae 106. Entomopatogen B. bassiana and H. citriformis effectively controled the CTV vector pest in the laboratory. In the semi-field experiments at the screen house, the most effective result was H.citriformis 106 and the combination of H.citriformis 106 + B.bassiana 106, killing up to 50% and 100% on day 7th H.citriformis had the most physiological character, was able to develop optimally at a temperature of 20-400C and humidity between 60-80%. The biochemical character of the entomopathogenic fungus B.bassiana contained cellulase enzyme and phosphate solvent and IAA hormone, at most compared to the others. H.citriformis had not been found to contain enzymes and hormones. The molecular biochemical characterization of entomopathogenic fungi using FS1 and NS2 primers more clearly distinguished isolates and entomopathogenic species