Studi Pembuatan Antiserum Poliklonal Untuk Deteksi Cepat Virus Mosaik Mentimun Pada Krisan

Virus mosaik mentimun merupakan salah satu patogen penting pada berbagai tanaman hortikultura, termasuk tanaman krisan. Untuk mengetahui secara dini infeksi virus pada tanaman, maka perlu dikembangkan metode deteksi cepat. Penelitian ini bertujuan mendapatkan antiserum poliklonal untuk deteksi cepat virus mosaik mentimun pada krisan. Penelitian dilaksanakan di Laboratorium Virologi Balai Penelitian Tanaman Hias, Segunung dari bulan April sampai Desember 2000. Antiserum terhadap CMV pada tanaman krisan telah dihasilkan dengan cara penyuntikan virus murni CMV pada kelinci dengan konsentrasi setiap penyuntikan sebesar 1 mg/ml. Antiserum yang diuji terdiri dari enam periode pengambilan darah. Pengujian menggunakan metode ELISA tidak langsung. Hasil pengujian menunjukkan bahwa dari enam kali pengambilan darah ternyata bereaksi positif, yaitu dengan adanya konsentrasi antibodi dalam darah meningkat. Antiserum juga dapat digunakan untuk mendeteksi langsung terhadap ekstrak daun krisan yang terinfeksi CMV. Kepekaan antiserum tertinggi pada pengambilan darah ke empat dan ke enam dengan konsentrasi 1/100 dan 1/500 terhadap pengenceran sampel 1/10 dan 1/100. Study on developing of polyclonal antiserum for rapid detection of cucumber mosaic virus on chrysanthemum. Cucumber mosaic virus is one the major pathogens on some horticulture crops, including chrysanthemum. A rapid detection method should be developed to support the evaluation of initial infection of the virus in plants. The objective of this experiment was to obtain polyclonal antiserum to CMV on chrysanthemum for rapid detection. The experiment was conducted in Virology Laboratory of Indonesian Ornamental Plants Institute in Segunung from April to December 2000. A polyclonal CMV antiserum had been produced by injections of purified CMV into rabbits with concentration 1 mg/ml each injections. The antiserum from six bleeding periods were tested. An indirect ELISA method was used to determine the sensitivity of the antiserum. Results indicated that six bleeding periods had positive reaction, with concentration of the virus antibodies increased gradually from the first to the sixth bleedings. The antiserum can also be directly used to detect CMV from infected chrysanthemum plants. The highest antiserum sensitivity were the fourth and sixth bleedings with concentration 1/100 and 1/500 to sample dilution 1/10 and 1/100

Molecular Studies on Bromovirus Capsid Protein II. Functional Analysis of the Amino-Terminal Arginine-Rich Motif and Its Role in Encapsidation, Movement, and Pathology

AbstractThe N-terminal region of the brome mosaic bromovirus (BMV) coat protein (CP) contains an arginine-rich motif that is conserved among plant and nonplant viruses and implicated in binding the RNA during encapsidation. To elucidate the functional significance of this conserved motif in the BMV CP, a series of deletions encompassing the arginine-rich motif was introduced into a biologically active clone of BMV RNA3, and their effect on replication, encapsidation, and infection in plants was examined. Analysis of infection phenotypes elicited onChenopodium quinoarevealed the importance of the first 19 N-proximal amino acids of BMV CP in encapsidation and pathogenicity. Inoculation ofC. quinoawith three viable variants of BMV RNA3 lacking the first 11, 14, and 18 N-terminal amino acids of the CP resulted in the development of necrotic local lesions and restricted the spread of infection to inoculated leaves. Progeny analysis from symptomatic leaves revealed that, in each case, virus accumulation was severely affected by the introduced mutations and each truncated CP differed in its ability to package genomic RNA. In contrast to these observations inC. quinoa,none of the CP variants was able to establish either local or systemic infections in barley plants. The intrinsic role played by the N-terminal arginine-rich motif of BMV CP in packaging viral RNAs and the interactions between the host and the truncated CPs in modulating symptom expression and movement are discussed

Kepatogenan Satelit Rna Yang Berasosiasi Dengan Cucumber Mosaic Virus (Cmv-satrna) Pada Tanaman Cabai

Pathogenicity of RNA satellite associated with cucumber mosaic virus (CMV-satRNA) on hot pepper plant. The objective of this experiment was to determine the pathogenicity of RNA satellite associated with CMV (CMV-satRNA) on hot pepper plant and effects of mix infection of CMV-satRNA and other viruses naturally infecting hot pepper, PVY (potato virus Y) and TMV (tobacco mosaic virus). Two green house experiments were conducted in Gedong Meneng Bandar Lampung during June—November 2004. The treatments of the the first experiment were arranged in a completely randomized design with five replications. The treatments were hot pepper plant inoculated with CMV-satRNA, severe strain CMV-G, and control uninoculated plant. The second treatment was arranged in a complete block randomized design in a factorial experiment with four replications. The first factor was single inoculation of CMV-satRNA, PVY, TMV, double inoculatoin of CMV-satRNA and PVY, CMV-satRNA and TMV, PVY and TMV, and control uninoculated plant. The second factor was two hot varieties, Taro and Laris. The results of the first experiment showed that infection of CMV-satRNA did not significantly reduce the yield of hot pepper plant compared to severe strain CMV-G. The result of the second experiment indicated that mix infection of CMV-satRNA and TMV or CMV-satRNA and PVY did not induced a synergetic reaction on hot pepper plants

The study of brome mosaic virus and arabidopsis

Includes bibliographical references.The purpose of this project was to provide me the opportunity to work in a lab with a professor to learn about the virus, Brome Mosaic Virus, and the plant, Arabidopsis. Work was to be conducted under the guidance of both the lab coordinator and a graduate student in the lab when needed. Additional understanding was to be gained in the propagation of the virus, purification of virus particles, the extraction of virion RNA and electrophoresis of DNA from the plant Arabidopsis. DNA gels were to be run after the PCR reaction had been completed on extracted DNA from the Arabidopsis plant. The work performed would allow for the other researchers to continue their own work and the experience and research done combined with study of papers and other resources was to culminate in the production of this thesis.B.S. (Bachelor of Science

Inter- and Intramolecular recombinations in the Cucumber Mosaic Virus genome related to adaptation to Alstroemeria

In four distinct alstroemeria-infecting cucumber mosaic virus (CMV) isolates, additional sequences of various lengths were present in the 3' nontranslated regions of their RNAs 2 and 3, apparently the result of intra- and intermolecular recombination events. Competition experiments revealed that these recombined RNA 2 and 3 segments increased the biological fitness of CMV in alstroemeri

Uji Kepekaan Antiserum Poliklonal Untuk Deteksi Cepat CMV Dengan Metode ELISA Tidak Langsung Pada Tanaman Anthurium

. Rahardjo, I.B., E. Diningsih, and Y. Sulyo. 2008. Sensitivity Test of Cucumber Mosaic Virus Polyclonal Antisera for Rapid Detection of CMV with Indirect ELISA on Anthurium. Cucumber mosaic virus is one of the major pathogens on some horticulture crops including Anthurium. A rapid detection method should be developed to support the evaluation of initial infection of the virus in crops. A serological method commonly used for rapid detection of plant viruses is ELISA (enzyme-linked immunosorbent assay). The objective of the experiment was to determine the optimum concentration of CMV antiserum for rapid detection on anthurium. The experiment was done in Virological Laboratory of Indonesia Ornament Crops Research Institute in Segunung (1.100 m asl.), from April to July 2005. A polyclonal CMV antiserum had been produced by injections of purified CMV into a rabbit with the concentration of 1 mg/ml each injection from previous research. Antiserum concentration was measured with the spectrophotometer at 280 nm wave length. The sensitivity tests of the antiserum was carried out with indirect ELISA method. The results showed that the optimum concentration of the antiserum and the optimum sample dilution for detection of CMV infected anthurium were 1/1,000 and 1/5, 1/1,000 and 1/10 respectively

Uji Kepekaan Gamma-Glob U Lin an Ti Serum Poliklonal Cu Cum Ber Mo Saic VI Rus Untuk Deteksi Cepat CMV Dengan Metode Elisa Tidak Langsung Pada Tanaman Tapak Dara

Vi rus mosaik ketimun merupakan salah satu patogen penting pada berbagai tanaman hortikultura, termasuk tanamantapak dara. Untuk dapat mengetahui secara dini infeksi vi rus pada tanaman, perlu dikembangkan metode deteksicepat. Salah satu metode serologi yang pal ing banyak digunakan dewasa ini untuk deteksi vi rus secara cepat adalahen zyme-linked immunosorbent as say (ELISA). Penelitian ini bertujuan memperoleh gamma-glob u lin murni dari an ti -serum poliklonal cucumber mosaic virus (CMV) dan mengetahui konsentrasi optimalnya untuk deteksi cepat.Penelitian ini dilakukan di Laboratorium Virologi, Balai Penelitian Tanaman Hias, Segunung, dari bulan Januarisampai Desember 2001. An ti serum diproduksi dengan cara penyuntikan vi rus murni CMV secara bertahap padakelinci dengan konsentrasi setiap penyuntikan sebesar 1 mg/ml yang dilakukan pada penelitian sebelumnya.Pemurnian gamma-glob u lin mengikuti metode Clark & Adam. Konsentrasi gamma-glob u lin diukur denganspektrofotometer pada panjang gelombang 280 nm. Pengujian kepekaan gamma-glob u lin terhadap an ti gen dilakukandengan metode ELISA tidak langsung. Hasil pengujian menunjukkan konsentrasi gamma-glob u lin sebesar 1 mg/mldengan uji ELISA tidak langsung, konsentrasi gamma-glob u lin yang op ti mal untuk deteksi CMV adalah sebesar 1mg/ml dengan pengenceran enzim con ju gated goat antirabbit dan sampel, masing-masing sebesar 1/25.000 dan 1/10atau konsentrasi gamma-glob u lin sebesar 1mg/ml dengan pengenceran enzim con ju gated goat antirabbit dan sampel,masing-masing sebesar 1/10.000 dan 1/100.AB STRACT. Rahardjo, I.B., Y. Sulyo, and E. Diningsih. 2004. Sen si tiv ity test of gamma-glob u lin of cu cum bermo saic vi rus polyclonal an ti serum for rapid de tec tion of CMV with in di rect ELISA on Vinca sp. Cu cum bermo saic vi rus is one of the ma jor patho gens on some hor ti cul ture crops in clud ing Vinca sp. A rapid de tec tion methodshould be de vel oped to sup port the eval u a tion of ini tial in fec tion of the vi rus on crops. A serological method com -monly used for rapid de tec tion of plant vi ruses is ELISA. The ob jec tives were to pro duce pu ri fied gamma-glob u lin ofCMV an ti serum and to de ter mine the op ti mum con cen tra tion for rapid de tec tion of the vi rus. The ex per i ment was donein Virological Lab o ra tory of In do ne sian Or na men tal Crops Re search In sti tute in Segunung, from Jan u ary to De cem -ber 2001. A polyclonal CMV an ti serum had been pro duced by in jec tions of pu ri fied CMV into rab bit with the con cen -tra tion of 1 mg/ml each in jec tion from pre vi ous re search. Clark & Adam method for gamma-glob u lin pu ri fi ca tion wasfol lowed. Gamma-glob u lin con cen tra tion was mea sured with spectrophotometer on wave length of 280 nm. The testof gamma-glob u lin sen si tiv ity was car ried out with in di rect ELISA method. The re sults showed that the gamma-globulincon cen tra tion ob tained in this study was 1 mg/ml. The op ti mum con cen tra tion of the gamma-glob u lin for CMVde tec tion with in di rect ELISA was 1 mg/ml with 1/25,000 and 1/10 of en zyme con ju gated goat antirabbit di lu tion andsam ples, re spec tively, or the op ti mum con cen tra tion of the gamma-glob u lin was 1 mg/ml with 1/10,000 and 1/100 ofen zyme con ju gated goat antirabbit di lu tion and sam ples, re spec tively

Toxicity of Bacillus thuringiensis-Derived Pesticidal Proteins Cry1Ab and Cry1Ba against Asian Citrus Psyllid, Diaphorina citri (Hemiptera)

The Asian citrus psyllid (ACP), Diaphorina citri Kuwayama (Hemiptera), is an important pest of citriculture. The ACP vectors a bacterium that causes huanglongbing (HLB), a devastating and incurable disease of citrus. The bacterium Bacillus thuringiensis (Bt) produces multiple toxins with activity against a diverse range of insects. In efforts to provide additional control methods for the ACP vector of HLB, we identified pesticidal proteins derived from Bt for toxicity against ACP. The trypsin proteolytic profiles of strain-derived toxins were characterized. Strain IBL-00200, one of six strains with toxins shown to have basal activity against ACP was selected for liquid chromatography-mass spectrometry (LC-MS/MS) identification of the individual Cry toxins expressed. Toxicity assays with individual toxins derived from IBL-00200 were then performed. The activated form of the Cry toxins Cry1Ab and Cry1Ba were toxic to ACP with LC50 values of approximately 120 µg/mL. Disruption of the midgut epithelium was associated with the toxicity of both the IBL-00200-derived toxin mixture, and with Cry1Ba. With further optimization of the efficacy of Cry1Ab and Cry1Ba, these toxins may have practical utility against ACP. Bt toxins with activity against ACP may provide an additional tool for management of ACP and the associated HLB disease, thereby providing a more sustainable and environmentally benign approach than repeated application of broad-spectrum insecticides

Penggunaan Pelacak Nonradioaktif (Digoxigenin-dna Probe) Untuk Mendeteksi Peanut Stripe Virus

The use of nonradioactive probe (Digoxigenin-DNA) for detection of peanut stripe virus. The objective of this experiment was to develop the nonradioactive-labeled probe to detect peanut stripe virus (PStV) in peanut leaves and seeds. Digoxigenin labeled cDNA (dig-DNA probe) was synthesized from recombinant plasmid (pHS1.23) using polymerase chain reaction (PCR). The probe containing 1.195 bp (base pair) corresponding to 3' termini, included part of NIb (nuclear inclusion body) gene, coat protein gene, and 3' untranslated region of PStV genome was used to detect the existence of PStV in peanut leaves and seeds of infected peanut plants

Report on mixed monolayer films of a saturated and an unsaturated polar lipid

Study of mixed monolayer films of stearic acid-oleic acid and DPPC-POPCPreprin