Nurmalasari Darsono

Dr. Nurmalasari Darsono


Research Focus & Metrics

Academic Career Length

10Years (2017-Present)

(Calculated once Scilit publication data is synced: current year − year of first publication + 1)

Representative Publications

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24 July 2017 in Agronomy
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Journal Article
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HAHardian Susilo Addy
Hardian Susilo Addy
NDNurmalasari Darsono
Nurmalasari Darsono
AWAgus Heri Setyo Wahyudi
Agus Heri Setyo Wahyudi
ASAhmil Sholeh
Ahmil Sholeh
CACahya Anugrah
Cahya Anugrah
BSBambang Sugiharto
Bambang Sugiharto
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.

31 August 2018 in Molecular Biology Reports
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Journal Article
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RARetnosari Apriasti
Retnosari Apriasti
SWSuvia Widyaningrum
Suvia Widyaningrum
WHWeny N. Hidayati
Weny N. Hidayati
WSWidhi Dyah Sawitri
Widhi Dyah Sawitri
NDNurmalasari Darsono
Nurmalasari Darsono
BSBambang Sugiharto
Bambang Sugiharto

13 June 2018 in Agronomy
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Journal Article
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NDNurmalasari Darsono
Nurmalasari Darsono
NANovita Niswatun Azizah
Novita Niswatun Azizah
KPKiky Mey Putranty
Kiky Mey Putranty
NANatalia Tri Astuti
Natalia Tri Astuti
HAHardian Susilo Addy
Hardian Susilo Addy
BSBambang Sugiharto
Bambang Sugiharto
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.

30 April 2024 in Preprints.org
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Preprint
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INIntan Ria Neliana
Intan Ria Neliana
BSBambang Sugiharto
Bambang Sugiharto
WSWardatus Soleha
Wardatus Soleha
SSSuherman Suherman
Suherman Suherman
NDNurmalasari Darsono
Nurmalasari Darsono
WSWidhi Sawitri
Widhi Sawitri
Sugarcane mosaic virus (SCMV) and sugarcane streak mosaic virus (SCSMV) are the primary viruses responsible for mosaic diseases in sugarcane. In this study, the mosaic viruses SCMV and SCSMV with the involvement of sorghum mosaic virus (SrMV) were surveyed from eight commercial sugarcane cultivars grown in fields at four locations in East Java, Indonesia. Sugarcane leaves with mosaic symptoms were assessed for disease incidence and severity, and symptomatic leaves were collected for analysis. The incidence and severity of mosaic symptoms were the highest in the PS881 and NX04 cultivars grown in Kediri compared to the other cultivars. RT-PCR analysis showed that SCSMV was present in all sugarcane cultivars and locations. Interestingly, double mixed infections with SCMV were found in PS881, PS882, and Cening, whereas triple mixed infections with SrMV were found in the PS881 cultivar. Phylogenetic analysis grouped SCSMV, SCMV, and SrMV together with isolates from Thailand (KP987832.1), Florida, USA (MT725538.1), and Agentina (EU189037.1). The expression of ascorbate peroxidase gene (Apx) was increased in the virus-infected sugarcane cultivars and prominently elevated in the triple mixed infection of PS881 cultivars. However, the catalase gene (Cat) was slightly upregulated in the mixed infection group. The chlorophyll content was significantly reduced in the leaves of PS58, PS881, PS882, NX04, and HW Merah, in parallel with the expression of the photosystem I gene (PsaA). The expression of photosynthetic genes, such as ribulose-1,5 bisphosphate carboxylase small sub-unit (RbcS), phosphoenolpyruvate carboxylase (Pepc), sucrose-phosphate synthase (Sps1), and PsaA, was significantly reduced and almost depleted in the triple mixed infection of PS881, except for Pepc expression, which remained approximately half of that in the control healthy sugarcane. The decrease in photosynthetic activity was confirmed by the decrease in ribulose-1,5 bisphosphate carboxylase (Rubisco) and PEPC proteins, as detected by immunoblot analysis. The decrease in photosynthetic parameters indicated that the mosaic viral infection substantially reduced the growth and yield of sugarcane.