THE POTENTIAL USE OF SSR MARKERS TO SUPPORT THE MORPHOLOGICAL IDENTIFICATION OF INDONESIAN MUNGBEAN VARIETIES

Mungbean varieties were mainly characterized based on morphological traits. Molecular genetic approach is expected to help the breeder in identification of mungbean varieties in more detail and to protect intellectual property right. This study aimed to identify Indonesian mungbean varieties based on DNA fingerprint profile using a marker set to support morphological characters. A total of 22 Indonesian mungbean accessions were characterized based on 21 morphological traits and 55 simple sequence repeats (SSRs) primers. Of the total 22 mungbean varieties used in the present study, 16 varieties were improved varieties and remaining six varieties were local varieties originated from Java, Nusa Tenggara and Sulawesi collected in GeneBank of ICABIOGRAD. The results showed that the 21 morphological characters were not sufficient to differentiate 22 mungbean varieties, while SSR analysis revealed that eight multi-alleles markers and high polymorphic information content (PIC) values have been successfully selected for varietal identification. The selected markers enabled to differentiate each mungbean variety according to their genetic marker with the lowest distance of 0.125, demonstrating the robustness of the selected marker set as a tool to identify a specific DNA fingerprint profile as a varietal identity (ID). The genetic identity of a variety was shown by digital barcoding which represented a series of alleles produced by corresponding markers. The DNA fingerprint profile of each variety would be beneficial as reference identities of a mungbean variety.

The Potential Use of Ssr Markers to Support the Morphological Identification of Indonesian Mungbean Varieties

Mungbean varieties were mainly characterized based on morphological traits. Molecular genetic approach is expected to help the breeder in identification of mungbean varieties in more detail and to protect intellectual property right. This study aimed to identify Indonesian mungbean varieties based on DNA fingerprint profile using a marker set to support morphological characters. A total of 22 Indonesian mungbean accessions were characterized based on 21 morphological traits and 55 simple sequence repeats (SSRs) primers. Of the total 22 mungbean varieties used in the present study, 16 varieties were improved varieties and remaining six varieties were local varieties originated from Java, Nusa Tenggara and Sulawesi collected in GeneBank of ICABIOGRAD. The results showed that the 21 morphological characters were not sufficient to differentiate 22 mungbean varieties, while SSR analysis revealed that eight multi-alleles markers and high polymorphic information content (PIC) values have been successfully selected for varietal identification. The selected markers enabled to differentiate each mungbean variety according to their genetic marker with the lowest distance of 0.125, demonstrating the robustness of the selected marker set as a tool to identify a specific DNA fingerprint profile as a varietal identity (ID). The genetic identity of a variety was shown by digital barcoding which represented a series of alleles produced by corresponding markers. The DNA fingerprint profile of each variety would be beneficial as reference identities of a mungbean variety

Gene Duplication to Reveal Adaptation Clue of Plant to Environmental Stress: A Case Study of NBS-LRR Genes in Soybean

Gene duplication to reveal adaptation clue of plant to environmental stress: A case study of NBS-LRR genes in soybean. Puji Lestari, Suk-Ha Lee, I Made Tasma, and Asadi. Adaptive strategies of plant to stress are fine-tuned by adjusting several activities including molecular mechanism which involve duplicated genes responsive to environmental changes. Genes responsive to the environmental stresses which are retained after small scale duplication are part of plant genome duplication. However, less information of duplicated genes could be adaptive to environmental changes in plant. This review presents an overview of duplication events in plant genomes which impact to gene duplication in relation to environmental changes, gene duplication as an adaptation mechanism, a case of duplicated nucleotide binding site-leucine-rich repeat (NBS-LRR) genes in soybean, and the gene duplication implementation for plant breeding in Indonesia. Notably, genome duplication events generate gene duplication and contribute to adaptive evolution against environmental changes. Generalization of plants to adapt the stressful conditions also probably improves our understanding of gene duplication as a mechanism of adaptation. Several recently duplicated NBS-LRR genes in soybean retain disease resistance QTL and the differential expression convince their contribution to biotic stress resistance in soybean. Proposed models of NBS-LRR genes duplication process may help to understand these genes response to the environmental changes. The duplication of genes resistant to pest/disease particularly NBS-LRR provides important information to select breeding parents and develop molecular markers related to desease resistance to genetically improve soybean in Indonesia. Overall, it may therefore be possible to enhance breeding which targets on genes tolerance/resistance to abiotic/biotic stress, and provide a molecular basis for crop-stress protection strategy and more improved soybean varieties specified for harsh environment

Activation of CD147 with Cyclophilin A Induces the Expression of IFITM1 through ERK and PI3K in THP-1 Cells

CD147, as a receptor for Cyclophilins, is a multifunctional transmembrane glycoprotein. In order to identify genes that are induced by activation of CD147, THP-1 cells were stimulated with Cyclophilin A and differentially expressed genes were detected using PCR-based analysis. Interferon-induced transmembrane 1 (IFITM1) was detected to be induced and it was confirmed by RT-PCR and Western blot analysis. CD147-induced expression of IFITM1 was blocked by inhibitors of ERK, PI3K, or NF-κB, but not by inhibitors of p38, JNK, or PKC. IFITM1 appears to mediate inflammatory activation of THP-1 cells since cross-linking of IFITM1 with specific monoclonal antibody against it induced the expression of proinflammatory mediators such as IL-8 and MMP-9. These data indicate that IFITM1 is one of the pro-inflammatory mediators that are induced by signaling initiated by the activation of CD147 in macrophages and activation of ERK, PI3K, and NF-κB is required for the expression of IFITM1

Direct imaging of single UvrD helicase dynamics on long single-stranded DNA

Fluorescence imaging of single-protein dynamics on DNA has been largely limited to double-stranded DNA or short single-stranded DNA. We have developed a hybrid approach for observing single proteins moving on laterally stretched kilobase-sized ssDNA. Here we probed the single-stranded DNA translocase activity of Escherichia coli UvrD by single fluorophore tracking, while monitoring DNA unwinding activity with optical tweezers to capture the entire sequence of protein binding, single-stranded DNA translocation and multiple pathways of unwinding initiation. The results directly demonstrate that the UvrD monomer is a highly processive single-stranded DNA translocase that is stopped by a double-stranded DNA, whereas two monomers are required to unwind DNA to a detectable degree. The single-stranded DNA translocation rate does not depend on the force applied and displays a remarkable homogeneity, whereas the unwinding rate shows significant heterogeneity. These findings demonstrate that UvrD assembly state regulates its DNA helicase activity with functional implications for its stepping mechanism, and also reveal a previously unappreciated complexity in the active species during unwinding

Nodulation and Growth of a Supernodulating Soybean Mutant SS2-2 Symbiotically Associated with Bradyrhizobium Japonicum

Mutan kedelai penghasil nodulsuper menunjukkan kelemahan dalam kontrol autoregulasi pada nodulasi dan perbedaan fenotip dibandingkan dengan tipe liarnya. Studi untuk mengevaluasi karakter pertumbuhan dan nodulasi dari kedelai penghasil nodulsuper dalam asosiasinya dengan Bradyrhizobium japonicum dilakukan dalam penelitian ini. Tiga genotip kedelai, yaitu mutan kedelai penghasil nodulsuper SS2-2, tipe liarnya Sinpaldalkong 2 dan kedelai kontrol Jangyeobkong diinokulasi dengan B. japonicum USDA 110, kemudian ditumbuhkan di rumah kaca dalam kondisi terkontrol. Karakter nodulasi, fiksasi nitrogen (Acetylene Reduction Activity/ARA), pertumbuhan tanaman, dan hasil biji ditentukan untuk mengevaluasi asosiasi simbiotik antara B. japonicum dan kedelai nodulsuper. Kedelai yang diinokulasi dengan B. japonicum menunjukkan peningkatan jumlah dan berat kering nodul serta berat kering total tanaman dibandingkan dengan tanpa inokulasi. Tanaman SS2-2 yang diinokulasi menunjukkan jumlah nodul sekitar 20 kali lipat lebih tinggi daripada tipe liarnya. Inokulasi B. japonicum ternyata juga meningkatkan fiksasi nitrogen seiring dengan perkembangan nodul. Tanaman S2-2 lebih pendek dan menghasilkan fiksasi nitrogen (ARA) lebih tinggi, tetapi spesifik ARA dan hasil biji lebih rendah dibandingkan dengan tipe liarnya. Berdasarkan hasil evaluasi terhadap nodulasi dan pertumbuhannya, interaksi Rhizobium dan kedelai penghasil nodulsuper SS2-2 mempunyai respon asosiasi simbiotik lebih rendah dibandingkan kedelai penghasil nodul normal (kedelai yang tidak mendapat perlakuan mutasi)

Strut Support with Tricortical Iliac Allografts in Unstable Proximal Humerus Fractures: Surgical Indication and New Definition of Poor Medial Column Support

Background The execution of fibular allograft augmentation in unstable proximal humerus fractures (PHFs) was technically demanding. In this study, the authors evaluated the clinical and radiographic outcomes after tricortical iliac allograft (TIA) augmentation in PHFs. Methods We retrospectively assessed 38 PHF patients treated with locking-plate fixation and TIA augmentation. Insertion of a TIA was indicated when an unstable PHF showed a large cavitary defect and poor medial column support after open reduction, regardless of the presence of medial cortical comminution in preoperative images. Radiographic imaging parameters (humeral head height, HHH; humeral neck-shaft angle, HNSA; head mediolateral offset, HMLO; and status of the union), Constant score, and range of motion were evaluated. Patients were grouped according to whether the medial column support after open reduction was poor or not (groups A and B, respectively); clinical outcomes were compared for all parameters. Results All fractures healed radiologically (average duration to complete union, 5.8 months). At final evaluation, the average Constant score was 73 points and the mean active forward flexion was 148°. Based on the Paavolainen assessment method, 33 patients had good results and 5 patients showed fair results. The mean loss of reduction was 1.32 mm in HHH and 5.02% in HMLO. None of the parameters evaluated showed a statistically significant difference between the two groups (poor and not poor medial column support). Conclusions In unstable PHFs, TIA augmentation can provide good clinical and radiological results when there are poor medial column support and a large cavitary defect after open reduction