Budi Setiadi Daryono

Resistance Response of Chilli (Capsicum annuum L.) F1 to Fusarium oxysporum Involves Expression of the CaChi2 Gene

Cross-breeding is a method of producing progeny with better resistance to pathogens. Resistance to pathogens usually involves pathogenesis-related (PR) proteins. Class II chitinase is an example of a defensive PR protein in plants. The class II chitinase in chilli is coded by the CaChi2 gene. In this study, we crossed susceptible with resistant chilli cultivars, analysed the F1 resistance response against pathogenic F. oxysporum, and analysed the level of CaChi2 gene expression in the F1. Data were collected using disease severity index (DSI) determination and gene expression analysis by qRT-PCR (quantitative Reverse Transcriptase Polymerase Chain Reaction). Results showed that the DSI of F1 was not significantly different from the resistant ancestor. The relative CaChi2 expression level of F1 was higher than the susceptible ancestor but not significantly different from the resistant ancestor. We concluded that the F1 can be categorised as resistant to F. oxysporum, and the CaChi2 gene is involved in the molecular defense response.

Isolation of DNA from chicken (Gallus gallus domesticus Linnaeus, 1758) feather with lysis buffer-phenol chloroform isoamyl alcohol method (PCI) and chelex method

DNA extraction material can be obtained from the body part or body of a cell-containing organism. Feather is part of the cover body that contains cell in the base. The use of feathers as a sample of DNA isolation or DNA genome sources minimizes the stress on the chicken and simplifies sampling process of many birds. This study aimed to compare the concentration and purity of the DNA isolates from lysis buffer-Phenol Chloroform Isoamyl alcohol (PCI) and chelex methods. Steps of procedure were collecting feathers from layer chicken, cutting base part of feathers, weighing feathers in 1 mg, 2 mg, 3 mg, 4 mg, and 5 mg with three repetitions, DNA isolation with buffer lysis-PCI method and with chelex method, and DNA quantification by nanodrop spectrophotometer. The average concentration produced in the lysis buffer-PCI method were 4.1 ng · µL−1, 8.0 ng · µL−1, 7.0 ng · µL−1, 9.3 ng · µL−1, and 23.3 ng · µL−1 respectively, whereas in the chelex method were 166.2 ng · µL−1, 277.8 ng · µL−1, 301.0 ng · µL−1, 318.3 ng · µL−1, and 404.3 ng · µL−1 respectively. The average purity rate produced in the lysis buffer-PCI method was 1.868, 1.617, 1.787, 1.766, and 1.826 respectively while in the chelex method were 0.723, 0.723, 0.733, 0.750, and 0.766 respectively. The conclusions of this study were the concentration of isolate by chelex method higher than buffer lysis-PCI method, the purity ratio of buffer lysis-PCI method was higher than chelex method.DNA extraction material can be obtained from the body part or body of a cell-containing organism. Feather is part of the cover body that contains cell in the base. The use of feathers as a sample of DNA isolation or DNA genome sources minimizes the stress on the chicken and simplifies sampling process of many birds. This study aimed to compare the concentration and purity of the DNA isolates from lysis buffer-Phenol Chloroform Isoamyl alcohol (PCI) and chelex methods. Steps of procedure were collecting feathers from layer chicken, cutting base part of feathers, weighing feathers in 1 mg, 2 mg, 3 mg, 4 mg, and 5 mg with three repetitions, DNA isolation with buffer lysis-PCI method and with chelex method, and DNA quantification by nanodrop spectrophotometer. The average concentration produced in the lysis buffer-PCI method were 4.1 ng · µL−1, 8.0 ng · µL−1, 7.0 ng · µL−1, 9.3 ng · µL−1, and 23.3 ng · µL−1 respectively, whereas in the chelex method were 166.2 ng · µL−1, 277.8 ng · µL−1, 301.0 ng · µL−1, 318....

Molecular characters of melon (Cucumismelo L. “Tacapa”) in response to karst critical land

Yogyakarta district has 158.600 ha critical land and spread off in three Agro Ecosystem zones. Two of them are karsts critical land. Critical lands which contain calcium carbonate in high concentration and water dehydration in upper surface give abiotic stress in wide range of plant. Melon cultivar TACAPA has superior characteristic derived from parental crossing, ♀ Action 434 and ♂ PI 371795 and potential to be developed in karsts critical land. Abscicic acid (ABA) is a phytohormone expressed by plant in abiotic stress condition. CmBG1 is a gene which regulate ABA hormone in melon. The purposes of this research were examining the molecular character of melon cultivar TACAPA in response to karsts critical land in order to study molecular characterization of CmBG1 gene. Analysis was done qualitatively by using Reverse Transcriptase-PCR (RT-PCR) and Electrophoresis, while quantitative analysis was conducted by observing absorbance score in spectrophotometer. CmBG1 gene expression is examined by using Real t...

The Influence of Protein Level on Feed with the Ovarian Follicle Development of Laying Hens Gallus gallus

Laying chicken attracts interest from farmers because of its high egg production. Feed quality and quantity determine chicken egg production level. The purpose of this study is to determine the influence of protein level on feed into the development of follicles in laying hens. This research used 20 individuals of day-old chick (DOC) Lohmann Brown, divided into four groups with five individuals in each group. In the first treatment, chickens are reared for 7 weeks and being fed with 21% protein level. In the second treatment, chickens are reared for 7 weeks and being fed with a protein level of 14%. In the third treatment, chickens are reared for 8 weeks and being fed with 21% protein level and for 8 weeks with 14% protein level at the fourth treatment. Body weight was measured every week. In the seventh and eighth week, each group was dissected for the organ harvesting and tissues processing. Ovaries were prepared and weighted to observe development of the follicle using hematoxylin-eosin staining and immunohistochemistry (IHC). Follicular morphology can be classified and quantified serially on each slide. Data were analyzed by T-test to see its significance (P < 0.05). The results of the study explained that the treatment with 21% protein level on feeding increases primordial and secondary follicle development in the seventh and eighth week significantly compared with 14% protein level. At the age of 8 weeks, there are an increasing number of follicle developments significantly on 21% protein level group compared with 14% protein level group.

Detection of Resistance Against Begomovirus Using a SCAR Marker in Melon (Cucumis melo L. cv. Hikapel)

Begomovirus is the biggest genus of the Geminiviridae family, which triggers melon crop failure in Indonesia. Begomovirus infection is characterized by the emergence of chlorosis spots within the leaves, as well as curly and dried leaves, yellowish leaf-stalks, and stunted plants. But melon with resistance to Begomovirus genetically has not been found until now. This study was conducted to identify resistance against Begomovirus in melon using a SCAR marker. The melon variety Hikapel was used in this study. A total of 58 melon plants were observed visually to score the Begomovirus symptoms. Begomovirus infection was then detected using CPA5 and CPA2 markers. If Begomovirus particles are amplified at 770 bp, this result is identical to Squash leaf curl Philippine virus (SLCPV). Detection of Begomovirus resistance was conducted in nine samples using a PCR method. The results showed that the virus particle was amplified at 770 bp using CP primers, and melon DNA was amplified at 1198 bp using SCAR. This revealed that Hikapel melon had resistance to SLCPV. There have been no previous reports about melon with genetically proven resistance to Begomovirus.

Ethnobotany of Bamboo in Sangirese, North Celebes

Advances in technology have encouraged the growth of a variety of biodiversity processing methods. These conditions encourage efforts to use natural resources more broadly. Ethnobotany can be used to document the local knowledge of the plants and their utilization. This study is aimed to describe the various forms of bamboo utilization by Sangirese. The method used was survey with purposive sampling. The data were analyzed descriptively based on exploration results and interviews with the people. The results showed there were 11 species of bamboo that are directly related to the life of local community in Sangirese, consisting of seven species of Bambusa , three species of Schizostachyum , and each one species of Dendrocalamus and Gigantochloa . Local community use bamboo for a variety of daily necessities; as building materials, handicraft, furniture, ceremonies, musical instruments, transportation, medicine, ornamental plants, as well as food. These exploration results are expected to complement the data of bamboo ethnobotany in Indonesia. Applicatively, this research is expected to be a reference in the national strategy of bamboo germplasm resource conservation.

The occurrence and identification of powdery mildew on melon in Java, Indonesia

Powdery mildew is a serious disease and causes considerable decline in production and fruit quality of melon (Cucumis melo L.) grown in Java, Indonesia. The fungal species that cause powdery mildew in Indonesia have never been reported. Therefore, the lack of knowledge about the biology of these fungi will inhibit in the handling of the eradication of powdery mildew further. A survey was conducted in 58 locations of melon cultivation in Central Java, East Java and West Java to study the occurrence of powdery mildew in Java. Powdery mildew species involved in the disease development were identified morphologically based on the anamorph characters such as the mycelium/hyphae, germ tube, appressoria, conidia and fibrosin bodies. In order to confirm the identification, the complete ITS (internal transcribed spacer) region of rDNA of powdery mildew species was amplified using ITS 1 and ITS 4 primers, sequenced directly and compared with sequences from the GenBank. Twelve isolates of powdery mildew species were...

Regulatory elements of stx2 gene and the expression level of Shiga-like toxin 2 in Escherichia coli O157:H7

BACKGROUND/PURPOSE Shiga-like toxin (Stx) is an important factor in the pathogenesis of Escherichia coli O157:H7 infection and is responsible for some severe complications. Stx2 is usually associated with hemolytic uremic syndrome in humans. Its expression is regulated by elements located upstream of the stx2 gene, including stx2-promoter sequence, ribosome binding site, and the antiterminator q gene. The present study aimed to find the correlation between regulatory elements and the expression level of Stx2 in two local isolates of E. coli O157:H7. METHODS Two local E. coli O157:H7 strains SM-25(1) and KL-48(2), originating from human and cattle feces, respectively, and an E. coli reference strain, ATCC 43894, were investigated. The complete stx2 gene covering the sequences of promoter, ribosome binding site, and open reading frame and q gene of each strain was analyzed. The magnitude of Stx2 production was detected with a reverse passive latex agglutination method and Stx mediated cellular damage was determined with the Vero cell assay. RESULTS A comparison of the complete stx2 gene contained stx2-promoter, ribosome binding site, and q genes of two local strains KL-48(2) and SM25(1), and the E. coli ATCC 43894 showed that the amino acid sequences were identical. Both local isolates were Stx negative in the reverse passive latex agglutination test and nontoxic in the Vero cell assay. CONCLUSION The expression level of Shiga-like toxin of the two local isolates of E. coli O157:H7 did not only depend on the regulatory elements of the stx2 gene.

Genetic diversity of local durian (Durio zibethinus Murr.) cultivars of South Kalimantan’s province based on RAPD markers

South Kalimantan on the Borneo Island is one of the center origin and diversity of durian (Durio spp.) germplasm in Indonesia. This study was conducted to investigate the genetic diversity and relationships of local durian (Durio zibethinus Murr.) cultivars from South Kalimantan’s Province by RAPD markers. Eleven plant samples of local durian cultivars trees and five selected RAPD primers, namely OPA-01, OPA-02, OPA-07, OPA-16, OPA-18, and OPA-19 were used. DNA samples were extracted from leaf tissues according to the protocol of Nucleon Phytopure (Amersham Bioscience). Amplifications were performed on a PCR Thermal Cycler (Applied Biosystem 9700). Variation of the PCR products of each primer was scored based on the presence (1) and absence (0) of bands on the images taken from the gels. The NTSys-PC software was used to compute Dice’s coefficients of similarity, and the dendrogram was constructed using UPGMA method. Results showed that 11 local durian cultivars of South Kalimantan’s Province have high di...

Effect of Biofertilizer Addition on Nitrous Oxide Emission

Application of nitrogen fixing biofertilizer, such as Azotobacter, has a potential for reducing nitrous oxide (N2O) emission. The aim of this study was to examine the effect of nitrogen fixing biofertilizer addition to common practices of urea and fresh cattle manure usages for maize (Zea mays L.) growing on N2O emission. The field experiment was conducted at Gunung Kidul, Yogyakarta, Indonesia. The treatments were addition of fresh cattle manure (M), fresh cattle manure added with nitrogen fixing biofertilizer (MB), urea (U), urea added with nitrogen fixing biofertilizer (UB), and control (no N fertilizer added). Nitrogen contents of the added urea and fresh cattle manure were adjusted to be equal. Urea and fresh cattle manure were given three times throughout the experiment period, i.e. 12, 30, and 48 days after planting (DAP). Urea was given at a rate of 44, 29, and 15 kg.ha -1 , respectively while fresh cattle manure was given at a rate of 6000, 4000, and 2000 kg.ha -1 , respectively. The emitted N2O was collected using a closed-chamber method at 24, 42, 60, and 72 DAP and were determined using Gas Chromatograph. Soil properties including available N (NH4 + -N and NO3 - -N) and organic C contents were also analyzed. On the harvesting time, the harvest index and the grain yield were determined. Biofertilizer addition influence decomposition process of cattle manure and urea that led to mineralization and nitrification of residual organic matter and hence to cause soil NH4 + N in the order concentration of M treatment > MB > U > UB > C, and soil NO3 - -N of MB treatment > M > U > UB > C. Reduction of NO3 - N was resulted in the highest N2O emission of M > U > MB > UB > C (P UB > U > M> C treatments. Available mineral N and soil organic C contents strongly affected N2O emission (P < 0.01). The results suggested that biofertilizer addition to common agricultural practices reduce N2O emission and simultaneusly increased grain yield, and harvest index of maize.