22 research outputs found
Performance Test of Vegetative Characteristics of Crossed Rice (Oryza sativa L.) Lines of Ciherang Variety X B11143D Line in Telagasari, Karawang Regency, Indonesia
Get PDFOne way to increase the genetic diversity of Ciherang as the superior variety is to cross Ciherang with the donor B11143D line as a New Plant Type (NPT) rice. This study aimed to obtain Ciherang X B11143D lines with the best vegetative characteristics in the field. The research was conducted in rice fields in Talagasari Village, Telagasari District, Karawang Regency, from May to September 2023. This experiment used a single-factor Randomized Block Design (RBD) with three replications of 22 treatments, consisting of 19 Ciherang X B11143D lines and three comparison varieties. The effect of treatment was studied using analysis of variance. The results showed that the rice lines significantly influenced the vegetative characters of Ciherang X B11143D lines in Telagasari, Karawang Regency. Based on the observed characters, several lines were selected i.e. 124.2.3, 94.3.3, and 20.4.4 lines. Those three lines chosen as backcrossing lines performed similarly compared to Ciherang as recurrent parent and inherited several important traits for rising productivity from the B11143D line as donor parent, namely the length and area of flag leaf, the total number of tillers, and the number of productive tillers, which were significantly higher than Ciherang
Introduksi Konstruk Over-Ekspresi Kandidat Gen OsWRKY76 melalui Agrobacterium tumefaciens pada Tanaman Padi Nipponbare
No full text<p>Delivering of Over-Expression Construct OsWRKY76<br />Candidate Gene in Rice cv. Nipponbare through<br />Agrobacterium tumefaciens. Aniversari Apriana, Atmitri<br />Sisharmini, Wening Enggarini, Sudarsono, Nurul.<br />Khumaida, and Kurniawan R. Trijatmiko. Plant genetic<br />improvement can be done through classical breeding or<br />genetic engineering. WRKY is a transcription factor involved<br />in regulating plant defense responses. OsWRKY76 gene is<br />located in a narrow segment of chromosome 9 which is<br />identified previously to be related to wide spectrum<br />resistance in rice. A sequence of OsWRKY76 (+1.200 bp)<br />has available in the gene bank and it makes possible to<br />isolate, clone, and construct the gene into over-expression<br />vector. The aim of this research was to assemble an overexpression<br />construct of OsWRKY76 candidate gene and<br />introduce it into rice through Agrobacterium-mediated<br />transformation. A construct of pCAMBIA-<br />1301::35S::OsWRKY76 has been successfully assembled and<br />transformed into embryogenic calli of rice cv. Nipponbare<br />using A. tumefaciens strain Agl-1 and EHA 105. A number of<br />126 independent lines has been produced, in which Agl-1<br />showed 3.8 times more efficient than EHA 105. PCR analysis<br />of randomly selected 25 independent lines showed that all<br />of them positively contained hptII gene, a selectable marker<br />used in the over-expression construct of the OsWRKY76<br />candidate gene. Based on the result, it could be concluded<br />that the over-expression construct of OsWRKY76 candidate<br />gene have been successfully introduced into the tissue of<br />Nipponbare.</p></jats:p
Introduksi Kontruk Gen CsNitr1-L dengan Ubiquitin melalui Agrobacterium tumefaciens dan Deteksi Molekulernya pada Padi Kultivar Nipponbare
No full textPupuk berbahan dasar Nirogen seperti urea dan NPK merupakan kebutuhan pkok petani padi. Untuk mengejar peningkatan hasil tanaman, pupuk yang diaplikasikan sering melebihi takaran pupuk yang dianjurkan oleh pemerintah. Hal ini memberikan dampak yang buruk bagi lingkungan sekitarnya. Berdasarkan hal tersebut perlu dilakukan upaya untuk menekan kebutuhan pupuk yaitu dengan merakit tanaman pertanian yang lebih efisien dalam penggunaan N (Nitrogen use efficiency=NUE). Gen CsNitr1-L merupakan salah satu gen terkait efisiensi penggunaan Nitrogen di tanaman. Tujuan penelitian ini adalah untuk merakit kontruks kandidat gen CsNitr1-L1 pada plasmid pCAMBIA1300-promotor Ubil dan memperoleh padi kulvitar Nipponbare hasil transformasi yang telah mengandung konstruk kandidat gen CsNitr1-L. Konstruk pCAMBIA-1300:: Ubil:: CsNitr1-L telah berhasil dirakit dan ditransformasikan ke dalam embrio muda tanaman padi kultivar Nipponbare melalui bantuan Agrobacterium tumefaciens strain LBA4404. Proses transformasi menghasilkan 146 galur independen generasi T0 Nipponbare. Analisis PCR pada galur-galur T0 menunjukan 66 diantaranya positif mengandung gen hptII dan CsNirl-L. Efisiensi transformasi yang di peroleh adalah sebesar 11,9%. Hasil analisis jumlah salinan transgen menggunakan southern Hybridization pada beberapa galur t0 positif PCR secara acak menunjukan sebanyak 4 galur T0 memiliki salinan tunggal transgen. Berdasrkan hasil tersebut, dapat disimpulkan bahwa konstruk gen CsNitr1-L telah berhasil diintroduksikan ke dalam genom tanaman padi kultivar Nipponbare dan telah diperoleh galur-galur T0 yang positif PCR mengandung gen hptII dan CsNitr1-L serta kopi tunggal transgen
Blast resistance introgression of oryzica llanos-5 on the advanced backcross population way rarem x oryzica Llanos-5
No full textUpland rice is an alternative resource for enhancing rice productivity in Indonesia. However upland rice cultivation technology is still not optimum, particularly in controlling blast pathogen (Pyricularia oryzae). The development of cultivar multiline using NIL (Near isogenic lines) population is one of the important efforts to prevent the breaking down of blast resistance in field. The advanced backcross lines were developed using the recurrent parent Way Rarem, which is high yielding but recently was destroyed by blast, and the durable blast resistant Oryzica Llanos-5, as a donor parent. The study was aimed 1) to develop NIL Way Rarem x Oryzica Llanos-5 having a higher blast resistance compared to Way Rarem; 2) to obtain molecular markers which could detect the introgression segment of blast resistance-carrying donor on the advanced backcross lines; and 3) to get information about the blast resistance inheritance of Oryzica Llanos-5. The evaluation of blast resistance demonstrated that the percentage of the blast resistant plant on BC4F1 and BC5F1 were higher than that on BC3F1 and BC4F1, respectively. The blast test using each 133 and 173 blast races on BC5F1 showed that there were several BC5F1 plants having a higher blast resistance to each both blast races compared to Way Rarem. This high blast resistance character might be derived from the introgression segments of Oryzica Llanos-5. The detection of blast resistance introgression from Oryzica Llanos-5 using SSR primers on BC3F1 population demonstrated that three markers (RM411, RM7 and RM251) on chromosome 3 and three markers (RM276, RM549 and RM585) on chromosome 6 were polymorphic markers and could identify the donor introgression based on the appearance of heterozygous allele on PCR amplification. The six SSR markers and the evaluation of blast resistance were used to select the plant on each BC4F1, BC5F1 and BC6F1 generation which was backcrossed to Way Rarem. Based on the genome wide scan in the blast resistant BC6F1 plant we found several SSR markers which could detect the donor introgression on chromosome 2, 3, 6 and 8. RM207 and RM266 were on chromosome 2, also RM404 was on chromosome 8. This is interesting information because the introgression segment on both chromosomes could not detect in earlier research by Lestari et al. (2011). Both introgression segments were located quite close to blast resistance QTL derived from Oryzica Llanos-5 alleles in RIL population Fanny x Oryzica Llanos-5 that the introgression segment on chromosome 2 was located close to Pib gene and on chromosome 8 was located close to Pi33 gene. In addition there were also introgressions segment of Oryzica Llanos-5 on chromosome 3 that could be detected by RM565 and RM514 which were similar position with blast resistance QTLs on population of cross between Fanny x Oryzica Llanos-5 varieties
Pengaruh Cekaman Aluminium terhadap Kandungan Asam Organik dalam Kalus dan Pinak Tomat (Lycopersicon esculentum Mill.)
No full text<p class="p1">The purpose of this research was to evaluate the effects of Al stress on citric, malic and oxalic acid content of <em>L. esculentum </em>cv. Intan callus and plantlet, also aluminum content of <em>L. esculentum </em>plantlet. Callus was induced from cotyledone of <em>L. </em><em>esculentum </em>on Murashige &amp; Skoog (MS) media containing 10<span class="s1">-7 </span>M NAA and 10<span class="s1">-6 </span>kinetin. The callus was then transferred step wisely at 3 weeks interval to media containing 220, 275, 330, 385, 440, 550, 825, and 1100 μM AlCl<span class="s1">3</span>. The callus cultures on the control media and media with the addition of 550 μM AlCl<span class="s1">3 </span>were able to regenerate and produce shoots after 8 passages of subculture. The shoots from media with the addition of 550 μM AlCl<span class="s1">3 </span>were transferred into the media with addition of 825 μM AlCl<span class="s1">3</span>, then to the media with 1100 μM AlCl<span class="s1">3</span>. The High Pressure Liquid Chromatography (HPLC) analysis showed that Al stress callus and plantlets contained malic acid, but no citric and oxalic acid. The content of malic acid in callus decreased with increasing AlCl<span class="s1">3 </span>concentration from 0 to 385 μM. On the other hand, the content of malic acid in callus increased with increasing AlCl<span class="s1">3 </span>concentration from 440 μM to 1100 μM. Similarly, the content of malic acid in root increased with increasing concentration of AlCl<span class="s1">3 </span>from 550 μM to 1100 μM. The result of Neutron Activation Analysis showed that Al content in root decreased as the amount of AlCl<span class="s1">3 </span>increased in the media. These results suggested that <em>L. esculentum </em>callus and plantlet respond to the Al stress by producing higher amount of malic acid.</p></jats:p
Keragaman Jumlah Salinan Transgen Galur T0 Padi Kultivar Nipponbare Berdasarkan Analisis qPCR dengan Penanda Gen hptII
No full text<p><strong>Keragaman Jumlah Salinan </strong><strong>Transgen</strong><strong> </strong><strong><em>CsNitr1-L</em></strong><strong> pada Galur Transforman T0 Padi Kultivar Nipponbare. </strong>Perakitan tanaman transgenik dengan menggunakan bantuan <em>Agrobacterium tumefaciens</em> menghasilkan penyisipan transgen yang berbeda, baik dalam jumlah salinan maupun letak transgen dalam genom tanaman. Penelitian ini bertujuan untuk menganalisis keberadaan kimera dan jumlah salinan pada tiap anakan dalam satu rumpun dan beberapa rumpun padi Nipponbare transforman T0 yang berasal dari kalus yang sama. Gen <em>CsNitr1-L</em> pada plasmid biner pCAMBIA1300 ditranformasikan ke dalam genom tanaman padi kultivar Nipponbare dengan menggunakan <em>A. tumefaciens </em>strain LBA 4404. Analisis molekuler dilakukan terhadap tiga anakan dari masing-masing empat rumpun tanaman Nipponbare transgenik generasi T0 (event 1, 2, 3, dan 4) dan empat kelompok rumpun tanaman T0 yang berasal dari kalus yang sama. Dari masing-masing kelompok tersebut diambil 3 rumpun T0. Hasil analisis qPCR menunjukkan bahwa anakan-anakan yang berasal dari satu rumpun tanaman T0 memiliki jumlah salinan transgen yang seragam. Selain itu, hasil analisis qPCR juga mengungkap bahwa tidak semua tanaman yang berasal dari kalus yang sama memiliki jumlah salinan transgen yang seragam. Implikasi dari hasil ini bahwa setiap rumpun padi T0 yang tumbuh dari kalus hasil transformasi perlu dipisah saat aklimatisasi supaya benih T1 yang dihasilkan seragam. <strong></strong></p></jats:p
Identification of A Major Quantitative Trait Locus for Grain Weight In Rice Using Microsatellite Marker
No full textRice is one of the major staple foods in the world, especially in Asia. Improving yield potential of superior cultivars is important to meeting the demand for rice production, which is increasing due to human population increase, climate change, and degradation of agricultural resources, such as land and water. In this study, a BC3F2 population developed from an intraspecific cross between Ciherang and a new plant type line (B11143D) was used in a quantitative trait locus (QTL) analysis. Ciherang is a high yielding rice cultivar with good grain quality which has been planted in 37% of the irrigated rice area in Indonesia. The objective of this study was to identify QTL(s) for yield components on chromosome 12, which can be used to improve the elite cultivar Ciherang or other popular cultivars through marker-assisted breeding. A total of two hundred BC3F2 lines were evaluated in the greenhouse during this study. The population was observed for eight agronomic traits including days to heading (dth), plant height (ph), flag leaf length (fll), panicles per plant (ppl), panicle length (pl), grains per panicle (gpp), 1000-grain weight (gw), and yield (yld). Four simple sequence repeats (SSR) markers (RM3472, RM28048, RM28195, and RM1986) were used for targeted mapping on chromosome 12. Linkage analysis identified a QTL for 1000-grain weight located on chromosome 12 at position 53.5 cM–73 cM
Identification of A Major Quantitative Trait Locus for Grain Weight In Rice Using Microsatellite Marker
Get PDFRice is one of the major staple foods in the world, especially in Asia. Improving yield potential of superior cultivars is important to meeting the demand for rice production, which is increasing due to human population increase, climate change, and degradation of agricultural resources, such as land and water. In this study, a BC3F2 population developed from an intraspecific cross between Ciherang and a new plant type line (B11143D) was used in a quantitative trait locus (QTL) analysis. Ciherang is a high yielding rice cultivar with good grain quality which has been planted in 37% of the irrigated rice area in Indonesia. The objective of this study was to identify QTL(s) for yield components on chromosome 12, which can be used to improve the elite cultivar Ciherang or other popular cultivars through marker-assisted breeding. A total of two hundred BC3F2 lines were evaluated in the greenhouse during this study. The population was observed for eight agronomic traits including days to heading (dth), plant height (ph), flag leaf length (fll), panicles per plant (ppl), panicle length (pl), grains per panicle (gpp), 1000-grain weight (gw), and yield (yld). Four simple sequence repeats (SSR) markers (RM3472, RM28048, RM28195, and RM1986) were used for targeted mapping on chromosome 12. Linkage analysis identified a QTL for 1000-grain weight located on chromosome 12 at position 53.5 cM–73 cM
Isolasi, Identifikasi, dan Karakterisasi Cendawan Blas Pyricularia oryzae Hasil Rejuvenasi
No full text<p>Isolation, Identificaton, and Charaterization of Blast Fungus Pyricularia oryzae Derived from Rejuvenation. Puji Lestari, Wawan, Tri P. Priyatno, Wening Enggarini, Reflinur, and Yadi Suryadi. Blast disease caused by Pyricularia oryzae (Po) is one of the important diseases on rice crop in Indonesia. This study was aimed at obtaining blast fungus pathogen rejuvenated from preserved seed origin, agar medium and rice leaf tissue. In addition, we identified blast isolates for long-term storage collection, and characterized isolates towards their specific locus of genes virulence. The results revealed that 22.7% isolates stored in agar media produced a typical mycelium of Po. The specificity of total genomic DNA banding pattern of isolates which were analyzed using specific primers encoding virulence genes of Cut1, Erg2, and Pwl2, showed six haplotypes consist of B-001 (1 isolate), C-011 (1 isolate), D-111 (8 isolate), F-110 (1 isolate), G-100 (3 isolate), and H-101 (2 isolate). None of haplotypes A-000 and E-010 were found among the isolates studied. In regard to its pathogenicity, the majority of Po fungal isolates had Cut1, Pwl2, and Erg2 genes. Among the total isolates of the Po fungus, the greatest proportion of genes were Pwl2 (87.5%) followed by Cut1 (75%) and Erg2 (62.4%) genes.</p><p> </p><p><strong>Abstrak</strong></p><p>Penyakit blas yang disebabkan oleh cendawan Pyricularia oryzae (Po) merupakan salah satu penyakit penting pada pertanaman padi di Indonesia. Penelitian ini bertujuan untuk mendapatkan cendawan patogen blas yang berasal dari hasil rejuvenasi simpanan benih, media agar, dan jaringan daun padi. Selain itu, identifikasi koleksi isolat blas yang ditujukan untuk penyimpanan jangka panjang dan karakterisasi kespesifikan tiap isolat terhadap lokus spesifik gen virulensi. Hasil pengujian menunjukkan 22,7% isolat yang tersimpan dalam media agar masih menunjukkan tipikal miselium Po. Kespesifikan pola pita DNA genomik total isolat yang dianalisis berdasarkan primer penyandi gen spesifik virulensi Cut1, Erg2, dan Pwl2, diperoleh sebanyak enam haplotipe meliputi B-001 (1 isolat), C-011 (1 isolat), D-111 (8 isolat), F-110 (1 isolat), G-100 (3 isolat), dan H-101 (2 isolat). Tidak ditemukan haplotipe A-000 dan E-010 pada isolat Po yang dianalisis. Dalam hubungannnya dengan patogenisitas, mayoritas cendawan Po mempunyai gen Cut1, Pwl2, dan Erg2. Di antara total isolat cendawan Po, gen yang paling besar ditemukan proporsinya adalah gen Pwl2 (87,5%) diikuti Cut1 (75%) dan Erg2 (62,4%).</p></jats:p
Detection and quantification of splicing variants of Hd3a gene in oil palm
Get PDFAlternative splicing is a complex process that contributes to the generation of diverse mRNA and protein isoforms, including in oil palm (Elaeis guineensis). Despite their importance, many functions of alternative splicing genes remain poorly characterized. This study aims to investigate splicing variants of gene encoding Heading date 3a in E. guineensis (EgHd3a) using the GenBank database and ClustalW algorithm. To ensure the data accuracy and reliability of design isoform‐ specific primers, special emphasis is given to primer design techniques and validation using polymerase chain reaction (PCR) and quantitative real‐time (qRT)‐PCR analysis. The designed primers demonstrated high specificity and discrimination between mRNA specimens. Nucleotide variations at the 3’‐end influenced the specificity of primers with the addition of GC composition. Furthermore, qRT‐PCR analysis revealed a strong correlation between Ct values and gene concentration for the isoforms which indicates a reliable amplification of EgHd3a. Although two isoforms, Hd3a‐X2 and Hd3a‐X3, showed slightly higher than acceptable PCR efficiency values, caution is advised to prevent non‐specific amplification. Despite the challenge posed by the limitation of primer positioning due to alternative splicing, the chosen primer proved optimal for analysis. This study highlights the importance of considering alternative splicing in gene quantification experiments and provides insights into the critical steps, methods, and quality control measures necessary for accurately detecting alternative splicing events, contributing to understanding this complex biological process
