Penelitian ini bertujuan untuk mengetahui respon pertumbuhan tunas kultur mersitem apikal tanaman... more Penelitian ini bertujuan untuk mengetahui respon pertumbuhan tunas kultur mersitem apikal tanaman tebu (Saccharum officinarum) varietas NXI 1-3 secara in vitro pada media MS dengan penambahan arginin, mengetahui respon pertumbuhan tunas kultur mersitem apikal tanaman tebu (Saccharum officinarum) varietas HW-1 secara in vitro pada media MS dengan penambahan arginin., mengetahui respon pertumbuhan tunas kultur mersitem apikal tanaman tebu (Saccharum officinarum) varietas NXI 1-3 secara in vitro pada media MS dengan penambahan glutamin, dan mengetahui respon pertumbuhan tunas kultur mersitem apikal tanaman tebu (Saccharum officinarum) varietas HW-1 secara in vitro pada media MS dengan penambahan glutamin. Penelitian dilaksanakan pada Desember 2012 – Februari 2013 di Laboraturium Penelitian dan Pengembangan Usaha PT. Perkebunan Nusantara XI Surabaya. Pengamatan respon pertumbuhan tunas dilakukan 2 bulan setelah penanaman dengan parameter pengamatan jumlah tunas dan panjang tunas. Hasil ...
Sugarcane Mosaic Virus (SCMV) infection is one of the most serious problems that can result in se... more Sugarcane Mosaic Virus (SCMV) infection is one of the most serious problems that can result in severe yield loss of sugarcane. Since the symptoms of SCMV infection are similar to other biotic and abiotic stress symptoms, the development of a rapid diagnostic with high precision is required. The use of laboratory animals such as rabbits is required for antibody production in immunoassay‐based detection. However, due to its many advantages, specific chicken egg yolk immunoglobulin (IgY) has received considerable attention as an alternative antibody production in immunodiagnostics for infectious diseases. In this study, IgY antibody against SCMV recombinant coat protein (CP) was successfully obtained from chicken blood serum and tested to compare its efficacy against antibody from rabbit (IgG) using immunocapture reverse transcription‐polymerase chain reaction (IC‐RT‐PCR). The result showed that IgY and IgG could detect 0.1 g SCMV infected leaves using 1000‐times‐diluted antibodies. Th...
Sugarcane mosaic virus (SCMV, genus Potyvirus, familia Potyviridae) is a prominent pathogen of su... more Sugarcane mosaic virus (SCMV, genus Potyvirus, familia Potyviridae) is a prominent pathogen of sugarcane (Saccharum sp. hybrids). It can cause losses in susceptible varieties, in crop as well as sugar production economically. Although it has been studied in major sugar production countries, researches on the definement of SCMV from Indonesian isolate based on molecular study are very limited. This study aimed to obtain a proper recombinant antigens emanating from coat protein of SCMV from Indonesian isolate for polyclonal antibody production that can be used for immunodiagnosis purposes. A gene encodes the coat protein of SCMV (CP-SCMV) was amplified by the reverse transcription polymerase chain reaction and cloned into vector pJET1.2. The cDNA was inserted into 6X His-tag expression plasmid of pET28a(+) and overexpressed in Escherichia coli BL21(DE3) to produce recombinant protein. The highest expression had performed on 0.1M IPTG induction media for 5 hours at 37oC. SDS-PAGE analy...
Sugarcane mosaic virus (SCMV) is one among many viruses that infect sugarcane, cause yield loss, ... more Sugarcane mosaic virus (SCMV) is one among many viruses that infect sugarcane, cause yield loss, and become serious disease agents on sugarcane plantations. Since the morphological symptoms of SCMV are similar to other symptoms caused by Sugarcane streak mosaic virus (SCSMV) or nitrogen deficiency, the detection of SCMV is important through accurate diagnostic-like ELISA or RT-PCR. This research aimed to study the causative mosaic pathogen of SCMV in East Java, Indonesia, including mosaic development. The results showed that the mosaic symptom is present in all sugarcane plantations with 78% and 65% disease incidence and severity, respectively. Moreover, the detection procedure based on an amplification of cDNA of the coat protein gene sequence confirmed that SCMV was the causative agent of mosaic disease on sugarcane. Re-inoculation of healthy sugarcane plants with plant sap from a symptomatic leaf from the field showed similar mosaic or yellowish chlorotic areas on the leaf blade, and appeared on the fourth leaves upward from the inoculation leaf, in addition to showing different levels of peroxidase but not total phenol. Mosaic also correlated with the amount of total chlorophyll. Although Sucrose phosphate synthase (SPS) protein accumulation and activity were at a lower level in infected leaves, sucrose accumulation was at a higher level in the same leaves.
Na lida do policial civil : estudo sobre as estratégias de formação continuada de policiais civis... more Na lida do policial civil : estudo sobre as estratégias de formação continuada de policiais civis para o atendimento aos grupos vulneráveis / Bárbara Aragão Teodoro Silva.-Belo Horizonte, 2015. 165 f., enc., il.
Event-5 is a genetically modified (GM) sugarcane that engineered by introducing the Sucrose-Phosp... more Event-5 is a genetically modified (GM) sugarcane that engineered by introducing the Sucrose-Phosphate Synthase (SPS) gene to enhance the sugar productivity. To fulfill the security requirements of genetically modified products in the safety agriculture and the enviroment, it is necessary to test the competitive ability to the primary weeds which grow in the sugarcane cultivation areas. This study aimed to determine whether the transgenic sugarcane (Event-5) potentially invasive to the weeds at the critical stage of growth, that could effect the environmental balances. This research was conducted at Screen House and Laboratory for Biosciences, State Polytechnic of Jember, designed using the replacement series method in a Randomized Complete Block Design without factorial treatment. The combination treatments were Event-5, Non-transgenic sugarcane (NT) and the weeds (Ciperus rotundus and Portulaca olecarea) densities which consists of thirty-three treatments. The competitive ability t...
Sugarcane mosaic virus (SCMV) is a plant pathogenic virus of the family Potyviridae that causes c... more Sugarcane mosaic virus (SCMV) is a plant pathogenic virus of the family Potyviridae that causes chlorosis, stunting and significantly reduced sugar productivity in sugarcane. Pathogen-derived resistance is a method used to develop SCMV-resistant sugarcane by overexpression of viral DNA. In this study, the gene encoding the coat protein (CP) of SCMV was amplified by reverse transcriptase PCR from symptomatic sugarcane leaves and used to generate transgenic sugarcane. Nucleotide sequence analysis of amplified cDNA indicated that the 998-bp-long cDNA, termed ScMVCp cDNA, codes for the CP of SCMV from the PS881 isolate. The ScMVCp cDNA was inserted into the binary vector pRI101-ON with two constructs, a full nucleotide sequence (p927) and a sequence coding for N-terminally truncated protein (p702). The constructs were then introduced into sugarcane using Agrobacterium-mediated transformation. Southern blot analysis showed a single hybridized DNA copy inserted into the genome of transgenic sugarcane lines. The inserted genes were expressed at both the RNA transcript and protein levels in the transgenic sugarcane. The highest expression was found in transgenic lines 10, 11 and 13 from the p927 construct. Artificial infection by the virus showed that p927 generated a higher resistance to virus compared with p702. This resistance was passed on to the second generation of transgenic sugarcane with 100 and 20-40% levels of resistance in the p927 and p702 transgenic lines, respectively. This report shows that the full sequence of the CP gene is required to disrupt viral assembly and packaging, thereby generating resistance to SCMV infection.
This study aims to determine the type of RNA viruses that cause mosaic disease in four sugarcane ... more This study aims to determine the type of RNA viruses that cause mosaic disease in four sugarcane varieties in Indonesia based on simultaneous detection using multiplex-RT-PCR as well as relationship between types of RNA viruses that cause mosaic disease in four varieties of sugarcane in East Java, Indonesia from the analysis of cp gene sequence. The results showed that there were two types of RNA viruses, SCMV and SCSMV, which caused mosaic disease in four sugarcane varieties at four locations in East Java, Indonesia based on RT-PCR molecular detection either singleplex-RT-PCR or multiplexRT-PCR. Molecular detection using the multiplex-RT-PCR method is considered superior to singleplexRT-PCR because it is able to diagnose several types of RNA mosaic sugarcane at once, so it is more practical and time saving. SCMV that attacked sugarcane plantations in East Java, Indonesia on two sugarcane varieties namely Ps 864 (Magetan district) and Ps 881 (Madiun district) is a SCMV cluster close...
Sugarcane mosaic virus (SCMV) is a mosaic disease that has spread over sugarcane plantations in I... more Sugarcane mosaic virus (SCMV) is a mosaic disease that has spread over sugarcane plantations in Indonesia. The important step to overcome the disease is to detect the pathogen as early as possible. Detection of the pathogen can be achieved using the immunodiagnostic method by employing a specific antibody against the viral coat protein. The objective of this research was to produce a polyclonal antibody using the recombinant coat protein of SCMV, and to test its sensitivity for detection of SCMV in the symptomatic plant. The gene encoding of the coat protein was cloned using the RT-PCR Kit and total RNA isolated from symptomatic sugarcane leaves cultivar PS-881. Nucleotide sequences analysis of the cloned cDNA indicated that the cDNA contained 998 nucleotides and named SCMVCp-cDNA. The cDNA was then inserted into a His-tag expression plasmid of pET28a and overexpressed in Escherichia coli BL21 (DE3) to produce a recombinant protein. The recombinant fused protein SCMVCp was strongly expressed in an insoluble fraction, with a molecular size of around 44 kDa, without the addition of an IPTG inducer. Purification of the recombinant protein using an affinity Ni-NTA resin, followed by SDS-PAGE separation, resulted in a high purity of the protein and used as an antigen to raise the polyclonal antibody in a rabbit. The sensitivity of the antiserum determined by western blot analysis showed that the antiserum was able to detect the recombinant protein at a concentration of 10 ng. The western blot analysis also detected a clear single band of 36.7 kDa of the SCMV coat protein in symptomatic sugarcane leaves and not in healthy leaves. Interestingly, when the sample proteins were prepared using low-speed centrifugation, the corresponding coat protein was detected in a soluble fraction by western blot analysis. Thus, the antiserum was successfully used for indirect-ELISA analysis using the soluble protein fraction. The results provide an easy method to detect and diagnose SCMV infection using the immunodiagnostic.
Penelitian ini bertujuan untuk mengetahui respon pertumbuhan tunas kultur mersitem apikal tanaman... more Penelitian ini bertujuan untuk mengetahui respon pertumbuhan tunas kultur mersitem apikal tanaman tebu (Saccharum officinarum) varietas NXI 1-3 secara in vitro pada media MS dengan penambahan arginin, mengetahui respon pertumbuhan tunas kultur mersitem apikal tanaman tebu (Saccharum officinarum) varietas HW-1 secara in vitro pada media MS dengan penambahan arginin., mengetahui respon pertumbuhan tunas kultur mersitem apikal tanaman tebu (Saccharum officinarum) varietas NXI 1-3 secara in vitro pada media MS dengan penambahan glutamin, dan mengetahui respon pertumbuhan tunas kultur mersitem apikal tanaman tebu (Saccharum officinarum) varietas HW-1 secara in vitro pada media MS dengan penambahan glutamin. Penelitian dilaksanakan pada Desember 2012 – Februari 2013 di Laboraturium Penelitian dan Pengembangan Usaha PT. Perkebunan Nusantara XI Surabaya. Pengamatan respon pertumbuhan tunas dilakukan 2 bulan setelah penanaman dengan parameter pengamatan jumlah tunas dan panjang tunas. Hasil ...
Sugarcane Mosaic Virus (SCMV) infection is one of the most serious problems that can result in se... more Sugarcane Mosaic Virus (SCMV) infection is one of the most serious problems that can result in severe yield loss of sugarcane. Since the symptoms of SCMV infection are similar to other biotic and abiotic stress symptoms, the development of a rapid diagnostic with high precision is required. The use of laboratory animals such as rabbits is required for antibody production in immunoassay‐based detection. However, due to its many advantages, specific chicken egg yolk immunoglobulin (IgY) has received considerable attention as an alternative antibody production in immunodiagnostics for infectious diseases. In this study, IgY antibody against SCMV recombinant coat protein (CP) was successfully obtained from chicken blood serum and tested to compare its efficacy against antibody from rabbit (IgG) using immunocapture reverse transcription‐polymerase chain reaction (IC‐RT‐PCR). The result showed that IgY and IgG could detect 0.1 g SCMV infected leaves using 1000‐times‐diluted antibodies. Th...
Sugarcane mosaic virus (SCMV, genus Potyvirus, familia Potyviridae) is a prominent pathogen of su... more Sugarcane mosaic virus (SCMV, genus Potyvirus, familia Potyviridae) is a prominent pathogen of sugarcane (Saccharum sp. hybrids). It can cause losses in susceptible varieties, in crop as well as sugar production economically. Although it has been studied in major sugar production countries, researches on the definement of SCMV from Indonesian isolate based on molecular study are very limited. This study aimed to obtain a proper recombinant antigens emanating from coat protein of SCMV from Indonesian isolate for polyclonal antibody production that can be used for immunodiagnosis purposes. A gene encodes the coat protein of SCMV (CP-SCMV) was amplified by the reverse transcription polymerase chain reaction and cloned into vector pJET1.2. The cDNA was inserted into 6X His-tag expression plasmid of pET28a(+) and overexpressed in Escherichia coli BL21(DE3) to produce recombinant protein. The highest expression had performed on 0.1M IPTG induction media for 5 hours at 37oC. SDS-PAGE analy...
Sugarcane mosaic virus (SCMV) is one among many viruses that infect sugarcane, cause yield loss, ... more Sugarcane mosaic virus (SCMV) is one among many viruses that infect sugarcane, cause yield loss, and become serious disease agents on sugarcane plantations. Since the morphological symptoms of SCMV are similar to other symptoms caused by Sugarcane streak mosaic virus (SCSMV) or nitrogen deficiency, the detection of SCMV is important through accurate diagnostic-like ELISA or RT-PCR. This research aimed to study the causative mosaic pathogen of SCMV in East Java, Indonesia, including mosaic development. The results showed that the mosaic symptom is present in all sugarcane plantations with 78% and 65% disease incidence and severity, respectively. Moreover, the detection procedure based on an amplification of cDNA of the coat protein gene sequence confirmed that SCMV was the causative agent of mosaic disease on sugarcane. Re-inoculation of healthy sugarcane plants with plant sap from a symptomatic leaf from the field showed similar mosaic or yellowish chlorotic areas on the leaf blade, and appeared on the fourth leaves upward from the inoculation leaf, in addition to showing different levels of peroxidase but not total phenol. Mosaic also correlated with the amount of total chlorophyll. Although Sucrose phosphate synthase (SPS) protein accumulation and activity were at a lower level in infected leaves, sucrose accumulation was at a higher level in the same leaves.
Na lida do policial civil : estudo sobre as estratégias de formação continuada de policiais civis... more Na lida do policial civil : estudo sobre as estratégias de formação continuada de policiais civis para o atendimento aos grupos vulneráveis / Bárbara Aragão Teodoro Silva.-Belo Horizonte, 2015. 165 f., enc., il.
Event-5 is a genetically modified (GM) sugarcane that engineered by introducing the Sucrose-Phosp... more Event-5 is a genetically modified (GM) sugarcane that engineered by introducing the Sucrose-Phosphate Synthase (SPS) gene to enhance the sugar productivity. To fulfill the security requirements of genetically modified products in the safety agriculture and the enviroment, it is necessary to test the competitive ability to the primary weeds which grow in the sugarcane cultivation areas. This study aimed to determine whether the transgenic sugarcane (Event-5) potentially invasive to the weeds at the critical stage of growth, that could effect the environmental balances. This research was conducted at Screen House and Laboratory for Biosciences, State Polytechnic of Jember, designed using the replacement series method in a Randomized Complete Block Design without factorial treatment. The combination treatments were Event-5, Non-transgenic sugarcane (NT) and the weeds (Ciperus rotundus and Portulaca olecarea) densities which consists of thirty-three treatments. The competitive ability t...
Sugarcane mosaic virus (SCMV) is a plant pathogenic virus of the family Potyviridae that causes c... more Sugarcane mosaic virus (SCMV) is a plant pathogenic virus of the family Potyviridae that causes chlorosis, stunting and significantly reduced sugar productivity in sugarcane. Pathogen-derived resistance is a method used to develop SCMV-resistant sugarcane by overexpression of viral DNA. In this study, the gene encoding the coat protein (CP) of SCMV was amplified by reverse transcriptase PCR from symptomatic sugarcane leaves and used to generate transgenic sugarcane. Nucleotide sequence analysis of amplified cDNA indicated that the 998-bp-long cDNA, termed ScMVCp cDNA, codes for the CP of SCMV from the PS881 isolate. The ScMVCp cDNA was inserted into the binary vector pRI101-ON with two constructs, a full nucleotide sequence (p927) and a sequence coding for N-terminally truncated protein (p702). The constructs were then introduced into sugarcane using Agrobacterium-mediated transformation. Southern blot analysis showed a single hybridized DNA copy inserted into the genome of transgenic sugarcane lines. The inserted genes were expressed at both the RNA transcript and protein levels in the transgenic sugarcane. The highest expression was found in transgenic lines 10, 11 and 13 from the p927 construct. Artificial infection by the virus showed that p927 generated a higher resistance to virus compared with p702. This resistance was passed on to the second generation of transgenic sugarcane with 100 and 20-40% levels of resistance in the p927 and p702 transgenic lines, respectively. This report shows that the full sequence of the CP gene is required to disrupt viral assembly and packaging, thereby generating resistance to SCMV infection.
This study aims to determine the type of RNA viruses that cause mosaic disease in four sugarcane ... more This study aims to determine the type of RNA viruses that cause mosaic disease in four sugarcane varieties in Indonesia based on simultaneous detection using multiplex-RT-PCR as well as relationship between types of RNA viruses that cause mosaic disease in four varieties of sugarcane in East Java, Indonesia from the analysis of cp gene sequence. The results showed that there were two types of RNA viruses, SCMV and SCSMV, which caused mosaic disease in four sugarcane varieties at four locations in East Java, Indonesia based on RT-PCR molecular detection either singleplex-RT-PCR or multiplexRT-PCR. Molecular detection using the multiplex-RT-PCR method is considered superior to singleplexRT-PCR because it is able to diagnose several types of RNA mosaic sugarcane at once, so it is more practical and time saving. SCMV that attacked sugarcane plantations in East Java, Indonesia on two sugarcane varieties namely Ps 864 (Magetan district) and Ps 881 (Madiun district) is a SCMV cluster close...
Sugarcane mosaic virus (SCMV) is a mosaic disease that has spread over sugarcane plantations in I... more Sugarcane mosaic virus (SCMV) is a mosaic disease that has spread over sugarcane plantations in Indonesia. The important step to overcome the disease is to detect the pathogen as early as possible. Detection of the pathogen can be achieved using the immunodiagnostic method by employing a specific antibody against the viral coat protein. The objective of this research was to produce a polyclonal antibody using the recombinant coat protein of SCMV, and to test its sensitivity for detection of SCMV in the symptomatic plant. The gene encoding of the coat protein was cloned using the RT-PCR Kit and total RNA isolated from symptomatic sugarcane leaves cultivar PS-881. Nucleotide sequences analysis of the cloned cDNA indicated that the cDNA contained 998 nucleotides and named SCMVCp-cDNA. The cDNA was then inserted into a His-tag expression plasmid of pET28a and overexpressed in Escherichia coli BL21 (DE3) to produce a recombinant protein. The recombinant fused protein SCMVCp was strongly expressed in an insoluble fraction, with a molecular size of around 44 kDa, without the addition of an IPTG inducer. Purification of the recombinant protein using an affinity Ni-NTA resin, followed by SDS-PAGE separation, resulted in a high purity of the protein and used as an antigen to raise the polyclonal antibody in a rabbit. The sensitivity of the antiserum determined by western blot analysis showed that the antiserum was able to detect the recombinant protein at a concentration of 10 ng. The western blot analysis also detected a clear single band of 36.7 kDa of the SCMV coat protein in symptomatic sugarcane leaves and not in healthy leaves. Interestingly, when the sample proteins were prepared using low-speed centrifugation, the corresponding coat protein was detected in a soluble fraction by western blot analysis. Thus, the antiserum was successfully used for indirect-ELISA analysis using the soluble protein fraction. The results provide an easy method to detect and diagnose SCMV infection using the immunodiagnostic.
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