Puji Lestari
Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumberdaya Genetik Pertanian

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Aktivitas Anticendawan Bacillus cereus 11UJ terhadap Rhizoctonia solani dan Pyricularia oryzae Yadi Suryadi; I Made Samudra; Tri Puji Priyatno; Dwi Ningsih Susilowati; Puji Lestari; Sutoro Sutoro
Jurnal Fitopatologi Indonesia Vol 11 No 2 (2015)
Publisher : The Indonesian Phytopathological Society (Perhimpunan Fitopatologi Indonesia)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (433.705 KB) | DOI: 10.14692/jfi.11.2.35

Abstract

Microbial secondary metabolites is an important resource for antifungal development in disease control strategy. The objective of this study was to screen Bacillus cereus 11UJ, an antagonistic rhizosphere bacteria for potential secondary metabolite production against rice fungal pathogens, i.e. Rhizoctonia solani and Pyricularia oryzae. The antagonistic effect of crude exract was evaluated using sterile filter paper discs on PDA medium. The ethyl acetate extracts of the bacterium showed a better antifungal activity to P. oryzae than those of R. solani. The inhibitory effect of the filtrate proved the potency of the isolates to produce antifungal. Analysis of pyrolysis gas chromatography-mass spectrometry showed that B. cereus 11UJ produces 3 major compounds i.e; 9,19-cyclolanostan-3-ol, acetate, (3.beta.)- (CAS) cycloartanyl acetate (13.14%); 4-(2’,2’-dimethyl-6’-methyliden-1’-cyclohexyliden)-3-methyl-2-butanone (9.72%); and stigmast-5-en-3-ol oleat (9.09%) which suggested to play an important role in the suppression of rice fungal pathogens.  
Genetic Diversity Analysis and Development of DNA Fingerprints of 20 Indonesian Local Chili Pepper Varieties Based on SSR Markers Rerenstradika Tizar Terryana; Nadia Della Savitri Ayu Ningrum; Kristianto Nugroho; Darmawan Saptadi; Helmi Kurniawan; Puji Lestari
Jurnal AgroBiogen Vol 16, No 2 (2020): December
Publisher : Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/jbio.v16n2.2020.p45-58

Abstract

Chili pepper is one of the most valuable horticultural crops, widely cultivated in Indonesia. Analysis of its genetic diversity is needed to develop successful breeding programs of local varieties. Simple sequence repeat (SSR), a robust molecular marker used for genetic diversity analysis in plant species, offers potential, reliable DNA fingerprinting method to assess genetic variation and varietal identification of chili pepper. Fifteen SSR markers were used in this study to analyze the genetic diversity and develop profiling identification of DNA fingerprint of local chili pepper varieties. Twenty local and two improved varieties of three chili pepper species, consisting of 3, 1, and 18 varieties of Capsicum frutescens, C. chinense, and C. annuum, respectively, were assessed for their SSR polymorphism. A total of 87 alleles was obtained from the polymorphism analysis with high alleles variation (2–16 alleles) with average total allele of 5.8 and average polymorphism information content (PIC) of 0.59 (0.34–0.83). Clustering and Principle Coordinate Analyses (PCoA) classified the varieties into two groups with coefficient of similarity of 0.65 indicating their high genetic variability. Most local varieties belonged to the same cluster and separated from the two improved varieties. Based on PIC values and dendrogram with selected markers, five SSR markers, i.e. EPMS441, EPMS331, EPMS335, GPMS194, and CaSSRBio1.1, were identified as SSR marker set for DNA fingerprinting purposes. SSR marker set used in this study was successful in developing the varietal identity of local chili pepper varieties, as indicated by unique code of each variety.
METODE EKSTRAKSI DNA PADA Jatropha spp. TANPA MENGGUNAKAN NITROGEN CAIR / DNA Extraction Method of Jatropha spp. without Liquid Nitrogen Kristianoto Nugroho; Rerenstradika T. Terryana; Habib Rijzaani; Puji Lestari
Jurnal Penelitian Tanaman Industri Vol 22, No 4 (2016): Desember, 2016
Publisher : Pusat Penelitian dan Pengembangan Perkebunan

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/littri.v22n4.2016.159-166

Abstract

Physic nut is one of potential plant which produces biofuel. It is necessary to do an intensive breeding program either conventionally or molecularly based to develop new varieties of physic nut. All this time the physic nut DNA extraction methods always used liquid nitrogen to destroy the plant tissue. Liquid nitrogen is not always available, especially in remote areas. Physic nut has high latex content which makes extraction process more difficult. The aim of this study was to find the technique of DNA extraction which can give a good result without using liquid nitrogen. This research was conducted in Molecular Biology Laboratory, ICABIOGRAD Bogor from November to Desember 2015. There were five Jatropha species from Thailand which used in this research e.g. Jatropha curcas, Jatropha podagrica, Jatropha gossypifolia, Jatropha multifida, and Baliospermum solanifolium. The extraction method used modification CTAB by addition of PVP, sodium metabisulphite, sucrose and ascorbic acid. The young leaves were used as the part to be extracted. The results showed that the modified method could produce a good quality and quantity of DNA. The banding pattern of DNA amplification clearly visible under UV light. This method can reduce the dependency of liquid nitrogen. AbstrakEkstraksi DNA merupakan salah satu ta hap penting dalam kegiatan pemuliaan berbasis molekuler. Jarak merupakan salah satu tanaman dengan kandungan getah cukup tinggi sehingga proses ekstraksi DNA cukup sulit dilakukan. Selama ini metode ekstraksi yang dikembangkan umumnya menggunakan nitrogen cair untuk menghancurkan jaringan tanaman. Nitrogen cair merupakan senyawa yang cukup sulit untuk didistribusikan ke laboratorium yang letaknya jauh dari kota. Tujuan penelitian ini adalah untuk memperoleh teknik ekstraksi DNA jarak yang mampu menghasilkan DNA dengan kualitas dan kuantitas yang baik tanpa menggunakan nitrogen cair. Penelitian ini dilaksanakan di Laboratorium Biologi Molekuler, Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumberdaya Genetik Pertanian (BB Biogen), Bogor pada bulan November hingga Desember 2015. Terdapat lima spesies jarak asal Thailand yang digunakan pada penelitian ini yaitu Jatropha curcas, Jatropha podagrica, Jatropha gossypifolia, Jatropha multifida, dan Baliospermum solanifolium. Ekstraksi DNA dilakukan melalui modifikasi buffer CTAB melalui penambahan senyawa PVP, natrium metabisulfit, sukrosa dan asam askorbat. Bagian tanaman yang digunakan berupa daun yang masih muda. Hasil penelitian menunjukkan DNA jarak yang diekstraksi dengan metode ini mempunyai kualitas dan kuantitas yang baik serta mampu diamplifikasi dengan baik. Metode ini mampu mengurangi ketergantungan terhadap nitrogen cair yang ketersediaannya terbatas di laboratorium yang lokasinya jauh dari kota.Kata kunci: Jatropha spp., ekstraksi, DNA, nitrogen cair
EVALUASI KANDUNGAN PROTEIN DAN SIFAT PASTA BERAS JAPONICA DENGAN MARKA DNA Puji Lestari; Hee Jong Koh
BERITA BIOLOGI Vol 12, No 2 (2013)
Publisher : Research Center for Biology-Indonesian Institute of Sciences

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/beritabiologi.v12i2.526

Abstract

An evaluation of protein content and pasting properties of rice using molecular markers approach is still limited. This study was aimed to evaluate protein content and pasting properties of japonica rice using DNA marker in complement to conventional tools. This study was further analysis based on the data of protein content and pasting properties of 22 japonica varieties and genotyping data using 30 DNA markers. Multiple regression analysis was performed to formulate equations to estimate the protein content and pasting properties. As a result, the protein content and Rapid Visco Analyzer (RVA) pasting properties values of 22 varieties demonstrated in the range of japonica rice with high variation. Correlation between protein content and RVA pasta properties showed differential profiles. Several type of DNA markers tested in this study showed allele variation among varieties. To estimate the protein content and several parameters of pasting properties of japonica rice, six marker sets in the form of model equations were successfully developed. High resolutions of equations with signifikant R2 in range of 0.95 - 0.99 showed their potency to predict rice physicochemical properties. These developed marker sets are an initial step which needs further validation in order to be able for evaluation of thephysicochemical properties in early generation of breeding and germplasm collection of japonica rice in the future.
KERAGAMAN GENETIK 24 VARIETAS PADI SAWAH DAN PADI GOGO (Oryza sativa L.) INDONESIA BERDASARKAN MARKA SSR Kristianto Nugroho; Slamet Slamet; Puji Lestari
Scripta Biologica Vol 4, No 1 (2017)
Publisher : Fakultas Biologi | Universitas Jenderal Soedirman

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (986.786 KB) | DOI: 10.20884/1.sb.2017.4.1.350

Abstract

The use of molecular marker is an efficient approach to analyze the genetic diversity and it can be used widely in biological studies. The characterization of rice germplasms by using molecular markers technique is more precise because it is not influenced by environmental factors. The purpose of this study was to analyze the genetic diversity of 24 varieties of lowland and upland rice by using 15 SSR markers. The results showed as many as 86 alleles were detected in 24 rice varieties, with the average number of alleles per marker was 5.73 and the range of alleles per locus was 2-10. The average of major allele frequency was 43% with the lowest score was 26% on RM6997 and RM536 markers and the highest score was 65% on RM60 marker. A total of 14 SSR markers were able to discriminate heterozygous alleles within a range between 0.17 (RM105) to 1.00 (RM201, RM263, RM416, RM518 and RM223). The value of gene diversity ranged from 0.48 (RM60) to 0.81 (RM536) with an average of 0.70. The value of PIC (Polymorphic Information Content) ranged from 0.38 (RM105) to 0.78 (RM536) with an average of 0.65. The phylogenetic analysis showed that 24 rice varieties separate into two main clusters in the coefficient of 0.63. The first cluster consists of 12 lowland varieties and the second cluster consists of 12 upland varieties. The genetic diversity data in this study were expected could be a valuable information in the rice plant breeding activities in the future.
METODE EKSTRAKSI DNA CABAI (Capsicum annuum L.) MENGGUNAKAN MODIFIKASI BUFFER CTAB (CETHYL TRIMETHYL AMMONIUM BROMIDE) TANPA NITROGEN CAIR Kristianto Nugroho; Rerenstradika T Terryana; Puji Lestari
Scripta Biologica Vol 4, No 2 (2017)
Publisher : Fakultas Biologi | Universitas Jenderal Soedirman

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (747.778 KB) | DOI: 10.20884/1.sb.2017.4.2.423

Abstract

Chili pepper is an agricultural commodity having high economic value. The production and supply of chili pepper frequently did not match the increased demand; it caused the market price fluctuated. It is important to create new varieties of chili pepper with high production trait to overcome the scarcity. Therefore the plant breeding activities for chili pepper should be done intensively in both conventional and molecular-based to obtain varieties of chili pepper with expected qualities. In molecular breeding, DNA extraction is the crucial steps of the process. If extracted DNA has an excellent quality and quantity,  the next processes normally could be completed with the high-quality result. To date, most methods of DNA extraction used liquid nitrogen to destroy the tough carbohydrates of plant tissue. Liquid nitrogen is nitrogen gas in a fluid state which quite difficult to be distributed to the remote laboratory wit no available storage facility. This study aimed to obtain a modified DNA extraction method, in particular for chili pepper, which capable to produce DNA with high quality and quantity without using liquid nitrogen. The sample used consisted of eight F2 plants including their hybrid-parental of the Kencana and the 0207. This research applied modified Doyle and Doyle method for extraction. Modification of extraction buffer is done through the addition of the 1% (w/v) PVP (Polyvinylpyrrolidone) and 0.2% (v/v) β-mercaptoethanol. The results showed that the DNA extracted using this method has good quality and quantity, capable of being amplified by using SSR (Simple Sequence Repeat) primer and could be digested by restriction enzyme EcoRI. Besides, this method can reduce dependence on the use of liquid nitrogen, in particular for remote laboratories with no available storage facility.