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Agroforensic, a New Emerging Study using Molecular Marker Technique
(Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian, 2022-01-19) Listanto...[at al], Edy; Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian
Abstract. Agroforensic will identify and solve the problem when one case such as genetic diversity of crops should be declared, and in other situation need some confirmation like as detection of genetic modified organism (GMO) on food product or agrobusiness. Identification of DNA fingerprinting in forensics can be presented as DNA pattern profile containing repetitive and varied sequences among the investigated individuals. Several PCR-based technologies have been used in such genetic analysis. This study was conducted based on SSR analysis and GMO detection using a total of 13 SSR markers and 2 specific primers, respectively. The SSR samples consisted of 4 samples of sticky rice, 3 samples of brown rice, 8 samples of white rice, and 2 samples of corn, while GMO detection was conducted on 3 samples of corn flour. The aims of this study were to determine the genetic validity between the tested samples compared to the local control rice based on SSR analysis, to confirm the similarity of corn samples based on SSR analysis, and to identify the GMO content of corn flour sample based on qualitative PCR. The results of SSR analysis on sticky rice, brown rice, and white rice showed that their DNA band patterns were different from the local controls. This result was also supported by NTSys analysis. The results of SSR analysis on corn samples indicated that there were different DNA banding pattern between the control and sample corn based on six SSR primers. The results of GMO detection showed that three samples of corn flour were negative. Agroforensic can be performed using SSR method based on PCR technique.
The Application of Gamma Ray Irradiation to Increase Triterpenoid Compounds in Embryogenic Calli of Centella asiatica L. Urban
(Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian, 2022-01-19) Roostika ...[at al], Ika; Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian
Abstract. Centella asiatica L. Urban has bioactive compounds from triterpenoid group. Raw materials that are not standardized are one of the obstacles in supplying the needs of the pharmaceutical industry. The in vitro culture technology may solve this problem. The purpose of this study was to determine the effect of gamma ray irradiation on growth and triterpenoid content of embryogenic calli of C. asiatica. The three accessions tested were CASI 016, CASI 002, and released varieties Castina 1. Gamma ray irradiation was carried out at doses of 0, 1, 2, 3, 4, and 5 Krad. The results showed that the in vitro response of each accession was genotype dependent. Castina 1 contained higher triterpenoid compounds than the other accessions. However, the gamma ray irradiation treatment up to 5 Krad has not been able to increase the content of these compounds. The treatment could only increase the content of asiaticoside and asiatic acid of accession CASI 016 at the dose of 5 Krad. There was an increase of asiaticoside and madecasoside content of accession CASI 002. It is expected that the increase of gamma ray irradiation dose will increase the triterpenoid compounds content in embryogenic calli of C. asiatica.
Application of Phosphate Solubilizing Microbes to Promote the Effectiveness of Rock Phosphate on Cacao Seedling Growth in Acid Soil
(Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian, 2022-01-19) Sasmita ...[at al], Kurnia Dewi; Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian
Abstract. The effectiveness of rock phosphates (RP) on plant growth is often lower than that of chemically synthesized fertilizer SP-36. Phosphate solubilizing microbes (PSM) are potentially used to enhance RP dissolution and agronomic effectiveness for cacao plant. This study was carried out to assess the ability of several PSM to dissolve insoluble phosphate, produce phytohormones, and increase the effectiveness of RP in cacao seedling at acid soil. Five phosphate solubilizing bacteria (PSB) and one phosphate solubilizing fungi (PSF) were evaluated for their ability to solubilize insoluble phosphate in liquid medium (added Ca3[PO4]2, RP, and AlPO4) and to produce phytohormones. Six isolates were also evaluated in cacao nursery consisting of nine treatments (without P fertilizer; SP-36 fertilizer; only RP; RP with inoculation of five PSB and one PSF, respectively). The result showed that the highest P solubilization in liquid medium with Ca3(PO4)2, RP, and AlPO4 was obtained by Aspergillus niger F-E1, Burkholderia ambifaria B-SS1.2, and Pseudomonas fluorescens B-SS13.5, respectively. All isolates could produce the IAA, GA3, trans-zeatin, and kinetin. Whereas B. ambifaria B-SS1.2, B. cepacia B-SS19.7, and A. niger F-E1 increased shoot dry weight and relative agronomic effectiveness (RAE) of RP. Notably, the B. ambifaria B-SS1.2 increased N, P, and K uptake in shoot of cacao seedlings. The utilization of these microbes potentially promote the effectiveness of RP for cacao plant growth in acid soil.
Backcrossing of Soybean Lines Containing Aluminium Tolerance Gene into Superior Soybean Variety, Biosoy
(Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian, 2022-01-19) Pardal ...[at al], Saptowo J.; Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian
Abstract. Two new superior varieties of soybeans, Biosoy 1 and Biosoy 2 have the advantage of high productivity and bigger seeds than favorite local variety Grobogan. These varieties have been planted in several provinces in Java and showed a good performance. However, these varieties are not so adaptive in acid soil with high aluminum content. Therefore, to expand the planting area of Biosoy varieties outside Java, which is generally an acidic land with high Al content, it is necessary to improve its tolerance to Al stress. Two transgenic soybean lines (GM2 and GM5) carrying the Al tolerance gene (MaMt2) have shown tolerance to acidic soils pH 3.7–4.8 based on evaluation in the Biosafety Containment. Those transgenic lines have been crossed with the Biosoy variety. F1 soybean plants carrying the MaMt2 gene were then backcrossed to the Biosoy variety to produce BC1F1 seeds. The aim of this research was to do molecular confirmation of MaMt2 gene insertion in the BC1F1 soybean plants and backcrossing the positive BC1F1 to Biosoy 1 variety. Materials used for the research were seven plants of BC1F1-GM2, six plants of BC1F1-GM5, and Biosoy 1 variety. The research works included PCR analysis of BC1F1 plants, backcrossing of BC1F1-MaMt2 plants with Biosoy 1, and harvesting of BC2F1 seeds. The PCR analysis of 13 samples of BC1F1 plant showed that all samples were positive carrying the MaMt2 gene (100%). BC2F1 seeds have been produced from the second backcross. These BC2F1 soybean seeds will be used as backcross material in the next studies.
Biofertilizer Increases Nutrient Use Efficiency (NUE) of Nitrogen, Phosphorus, and Potassium at Leaves Level of Artemisia annua L.
(Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian, 2022-01-19) Rahman ...[at al], Wiguna; Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian
Abstract. An efficient cultivation procedure is required to support the production of artemisinin from Artemisia annua L. As a secondary plant metabolite, artemisinin production is influenced by environmental factors such as the availability of nutrients. This research aimed to evaluate nutrient use efficiency (NUE) of N, P, and K and biofertilizer effectiveness in A. annua cultivation under treatments of various rates of nutrients provided in relation to artemisinin production. Leaves dry mass, leaves nutrients concentration (N, P, and K), artemisinin content, and artemisinin yield were measured to analyse NUE. Four indices of NUE, i.e. partial factor productivity (PFP), apparent crop recovery efficiency (RE), physiological efficiency (PE), and agronomic efficiency (AE), were used for evaluation. The result showed that the biofertilizer increases the artemisinin content, artemisinin yield, and NUE. Two of the four indices, PFP and AE, show the significant role of biofertilizer on NUE. The NUE level of plants with biofertilizer treatment increased 2.2 and 4.5 times than plants without biofertilizer. Therefore, it is recommended to use biofertilizer in A. annua cultivation for artemisinin production.
Cells Density Affects Cell Production of Citrus limonia in Flask and Air-Lift Bioreactor Cultures and Limonin Farming
(Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian, 2022-01-19) Agisimanto ...[at al], Dita; Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian
Abstract. Japansche citroen (JC) is popular citrus rootstock in Indonesia due to well-adapted to biotic and abioticstresses. JC is naturally rich of secondary metabolites. Therefore, cells production of JC through tissue culture is potential for continuous metabolites farming in local pharmaceutical industries. The objectives of study were to to determine the best initial cells density (ICD) in flask and air-lift bioreactor (ALB) cultures for mass cells proliferation and to determine the limonin content in cells. Callus was induced from nucellus and habituated callus were collected to initiate cell suspensions. Selected cells were transferred to flask and ALB cultures at the density of 2, 4, and 6 mg/ml for 90 days of culturing. An increasing ICD affected cell growth in flask and ALB cultures. Cells in flask culture grew slower and were produced lower in quantity than in ALB. At 4 mg/ml, growth rate (GR) of cells in flask culture at day 30 reached maximum of 4.12 mg/day, 87-folds lower than in ALB that reached 360 mg/day. Limonin at 120.725 μg/g dried cells (DW) were harvested from ALB within 30 days, while total phenolic was 9.883±0.471 mg GAE/g DW and total flavonoid was 0.144±0.011 mg QE/g DW. This study confirmed the advantages of cell culture as a perpetual source for production of biomass content metabolites that amenable lower operational cost compared to natural production for local industries.
Characteristic of Indigenous Leuconostoc mesenteroides EN 17-11 Protease and Its Stability during Storage at Cold and Freezing Temperatures
(Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian, 2022-01-19) Khusniati ...[at al], Tatik; Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian
Abstract. Protease as protein hydrolyzer is an imported enzyme used in the food and pharmaceutical industries in Indonesia. To reduce imports, indigenous lactic acid bacteria can be used as a source of protease. This study aimed to determine the characteristics of indigenous Leuconostoc mesenteroides EN 17-11 protease and its stability during storage at cold and freezing temperatures. Protease activity was detected using the modified Horikoshi method. The protease stability after storage was tested at 0, 7, 14, 21, and 28 days. Relative protease activity with a value ≥50% indicates protease activity in stable conditions. All treatments were statistically analyzed using ANOVA. The results showed that the optimum protease activity of L. mesenteroides EN 17-11 was reached at pH 6.0 with activity of 0.9970 U/ml and temperature of 40°C with activity of 1.1166 U/ml (p<0.05). The protease stability at incubation time of 60 min was reached at pH 5.0–7.5 with a relative activity of 55.29–100.00%, and a temperature of 20–50°C with a relative activity of 51.92–100.00% (p<0.05). The protease stability at storage for 28 days with a relative activity of ≥50% at freezing temperature was higher than at cold temperature. The highest protease stability was reached at freezing temperature for 7 days with a relative activity of 77.31% (p<0.05). It was concluded that the protease stability of indigenous L. mesenteroides EN 17-11 was better at freezing than at cold temperature.
Characterization and Interrelationships of the Number of Vessel Bundles with Yield Components in Various Genotypes of Soybean (Glycine max [L.] Merrill)
(Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian, 2022-01-19) Karyawati, Anna S., Dyah P. Fitrawantio; Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian
Abstract. Soybean is the third most important source of vegetable protein and agricultural commodity in Indonesia. Physiological results of soybean crop are determined by the relationship source strength and sink capacity. Source is associated with assimilation of CO2, and sink is associated with a photo-assimilate allocated to storage or plant growth. Physiological characters of the number of vessels there is the xylem-phloem tissue cannot be separated but has different functions, so affect growth and yield of crops. Therefore, to increase productivity of soybeans it is necessary to examined the relationship between number of vessels and soybeans yield. The purpose of this study was to assess the diversity of vessels number and correlation of vessel number and yield in 36 genotypes of soybean. Research was conducted in the Agro Techno Park, University of Brawijaya, Jatikerto, Kromengan District, Malang Regency, East Java Province from March to June 2020. The research used randomized block design with three replications. The observation of vessel number was conducted in Plant Breeding Laboratory, Faculty of Agriculture, University of Brawijaya. Data was analyzed using Scott-Knott's test facilitated by SASM-Agri application and the correlation analysis by OPSTAT application. The results showed that the number of vessels of soybean plants in the range of 10.5–17.5. Genotypes that have a large number of vessel bundles are genotypes Anjasmoro × Argopuro, Anjasmoro × UB2, Tanggamus × UB1, Tanggamus × UB2, Argopuro × Tanggamus, Argopuro × Grobogan, Argopuro × UB1, Argopuro × UB2, Grobogan × UB2, UB1 × Tanggamus, UB1 × UB2, Anjasmoro, Argopuro, and Tanggamus. Moreover, it was discovered that the number of vessels was positively correlated with plant heigh (r = 0.333) and negatively correlated with the number of empty pods (r = -0.365). Although, number pod empty pods had no significant effect on the various soybean genotypes.
Characterization of Genomic Variation on Three Indonesian Oil Palm Genotypes Analyzed Using Next-Generation Sequencing HiSeq
(Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian, 2022-01-19) Tasma ...[at al], I Made; Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian
Abstract. Oil palm is an important estate crop and Indonesia is the largest palm oil producer in the world. Since the average yield is relatively low, a more efficient breeding program that utilizes the latest technological developments in DNA analysis is necessary. The objectives of this study were to (1) characterize genomic variations of three Indonesian oil palm genotypes based on whole genome sequencing using next-generation sequencing (NGS) HiSeq; (2) provide genomic data resources for future oil palm breeding program. Whole genome sequencing was performed on three Indonesian oil palm genotypes (Dura, Pisifera, and Elaeis oleifera from Brazil). Sequence data were aligned with that of the Eg5 genome reference sequences. The average sequence coverage depth for all loci was 34 and more than 95% of the genome was sequenced at least ten times. Comparison of the sequences revealed 3,735,311 sequence changes, an average of one sequence change per 197 bases. Among these changes, 3,371,088 were SNPs and 364,233 were insertions and deletions (indels). DNA variations found within exons total 329,087. Of these, 214,068 SNPs variations were capable of causing modification in the amino acid composition of the encoded proteins (nonsynonymous mutations). A genomic database was constructed to manage DNA variations resulted from this study. The SNPs collected from this study should be useful for constructing high density oil palm SNP chip to expedite future national oil palm breeding program.
Characterization of Japansche Citroen Rootstock Somaclones and In Vitro Selection for Aluminium Tolerance
(Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian, 2022-01-19) Sukmadjaja ..[at al], Deden; Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian
Abstract. Japansche citroen (JC) is a type of citrus which is widely used as rootstock in citrus cultivation. In order to expand citrus cropping on marginal acid land, a JC citrus clones that are tolerant of acid soil conditions are needed. Therefore, it is necessary to assemble the citrus genotype by increasing genetic diversity to obtain tolerant traits for aluminum and low pH. In this study, mutation induction experiments with gamma ray irradiation were carried out, followed by in vitro selection methods. The aims of this research was to increase genetic variation of JC citrus through induced mutation by gamma ray irradiation and to obtain candidates of JC citrus somaclones that are tolerant to Al at low pH. The research was conducted at the Tissue Culture Laboratory, Indonesian Center for Agricultural Biotechnology and Genetic Resources Research and Development (ICABIOGRAD), Bogor. The plant material used was JC citrus plantlet from immature embryos treated with gamma ray irradiation at doses of 0, 4.5, 5, and 5.5 Gy. Evaluation was conducted to morphological characters, followed by in vitro selection for tolerance to Al at low pH by using selecting agent of aluminium trichloride hexahydrate. The results showed that morphological variations found in JC citrus somaclones after being irradiated with gamma rays, including the formation of the winged leaves (unifoliolatus) that are not found in the control JC. There were 32 selected somaclones to be tolerant to Al at low pH.
Characterization of Plant Architecture and Yield Trait of Castor (Ricinus communis L.) Germplasm Suitable for Mechanical Harvesting
(Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian, 2022-01-19) Anggraenia, Tantri Dyah Ayu, Rully Dyah Purwati; Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian
Abstract. Characterization of castor (Ricinus communis L.) germplasm on plant architecture and yield trait is important for breeding program of dwarf variety with high yield, suitable for mechanical harvesting. This study aimed to conduct complete characterization of castor germplasm on plant stature and seed and oil yield trait. The materials used in this study were 29 castor accessions. The accessions were planted and observed for plant architecture and yield traits. The results showed that the germplasm collections had variation on plant height, i.e. normal, semidwarf, and dwarf, with internode length on the main stem ranged from 3.79 to 9.64 cm and number of branches from 3 to 15. The length of fruit raceme of main stem varied from 21 to 95 cm and the seed weight per raceme ranged from 6.85 to 124.83 g. Oil content ranged from 24.23 to 50.37%. There were positive correlation between (p-value = 0.05) the yield trait and its component.However, only fruit number per raceme that had direct effect. Castor accessions 18A and 70B had characters dwarf, low number of branch, long fruit raceme, high seed yield, and oil content. These accessions could be utilized as genetic material potential for new castor plant variety suitable for mechanical harvesting.
Construction and Introduction of OsAER1::LeAlaAT Cassette to Improve the Nitrogen Use Efficiency in Rice cv. Mekongga
(Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian, 2022-01-19) Sisharmini ...[at al], Atmitri; Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian
Abstract. Nitrogen use efficiency (NUE) in rice can be improved by developing a variety that absorbs and use nitrogen (N) more efficiently. We have successfully isolated the Lycopersicon esculentum alanine aminotransferase (LeAlaAT) gene from tomato, which is responsible for improved efficiency of N use in plants, and Oryza sativa alkenal reductase (OsAER1) promoter from rice cv. Awan Kuning. These gene and promoter can be used to develop NUE rice through the genetic engineering approach. The purposes of this study were to construct an OsAER1::LeAlaAT expression plasmid cassette, to introduce the constructed plasmid cassette into genome of rice cv. Mekongga, and to test the transgenic rice lines at several levels of N concentration. The cassette was constructed by replacing the β-glucuronidase (GUS) gene derived from the pCAMBIA1300_OsAER1::GUS cassette with LeAlaAT gene. The recombinant plasmid was then transformed into Agrobacterium tumefaciens vector strain LBA4404 and introduced into rice cv. Mekongga. The plant transformation technique was carried out at with immature embryos as explants. Molecular analysis was performed using PCR to evaluate the presence of targeted gene construct introgression in the rice lines and the efficacy test of NUE was carried out hydroponically to determine the performance of lines at several levels of N concentrations. The results showed that the OsAER1 promoter and LeAlaAT gene were successfully constructed in pCAMBIA1300 vectors and introduced to Agrobacterium vector. Rice transformation cv. Mekongga resulted in 50 transgenic rice lines (T0). Hydroponic rapid screening of NUE in 50 T1 transgenic rice lines using Yoshida solution with ¼ strength of N, resulted in five selected rice lines based on superior biomass dry weight compared to control, namely M40, M41, M45, M50, and M54 lines. The T1 generation of rice lines was tested hydroponically to evaluate the response and plant performance at several N concentrations treatment. Compared to the control, the transgenic lines showed the best performance at ¼ strength of N concentration. Further NUE testing is required to determine the performance of selected rice lines in greenhouses and confined field trials.
Current Status of Tidal Swamp Rice Varieties and Its Improvement for Fe Toxicity Tolerance and Biofortification
(Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian, 2022-01-19) Sabran ...[at al], Muhamad; Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian
Abstract. Tidal swampland is now getting high priority in Indonesia agricultural development, in particular for rice production. Productivity of rice in swampland, both in tidal swamp and monotonous swamp is low compared to that in other agroecosystem due to adverse constraints, such iron toxicity, salinity, acidity, and flooding, as well as other constraints, such as socio-economic, culture, and infrastructures. Effort to increase rice productivity has been done through varietal improvement. Thirty-five swamp rice varieties have been released in the last twenty years. However, the adoption rates of those varieties are low and some varieties are rarely found in the field. Advances in biotechnologyprovide new tools for effective crop improvement in tidal swampland. Several rice varieties have been developed through molecular breeding approach. It is also possible to select rice lines using specific markers corresponding to OsIRT and OsFer2 genes that play a role in Fe2+ transport regulation and deposit in rice grain, respectively. This molecular breeding approach will not only improve rice tolerance to iron toxicity in marginal lands such as tidal swampland, but also iron biofortification of rice to mitigate iron deficiency in population that consume rice as staple food.
Cytological and Molecular Identifications of Seedless Tangerine Derived from Endosperm Culture
(Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian, 2022-01-19) Martasari ...[at al], Chaireni; Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian
Abstract. Endosperm cultures have been established on tangerine citrus cv. Siam Madu. To verify the ploidy levels of the regenerant derived-plants, both cytological and molecular assessments were applied. The research was conducted from March until June 2017 at the Laboratory of Plant Breeding and Genetic Resources of the Indonesian Citrus and Subtropical Fruits Research Institute (ICSFRI). A total of 25 tangerine citrus accessions served as the evaluated materials after they were grafted into JC rootstock for 16 weeks. The results showed that the stomatal density, length, and width were varied among the tested accessions. The lowest stomatal density was observed in SM42. While, SM11 showed the highest stomatal density with smallest value in stomatal size trait. The highest ratio of stomatal length and width was detected in SM56. Based on the cytological observation, 13 accessions were observed to have triploid, 11 accessions were diploids, and one accession had haploid chromosome structures. Molecular characterization using SSR primers (TTTA41, TAA15, and CAT01) resulted that most of tested accessions had the same number of amplified alleles, except for SM18, SM47, SM49, SM51, and SM54 which showed polymorphic bands compared to the controls.
Design and In Vitro Test of sgRNA for the CRISPR/Cas9 Plasmid Construct of the SQS Gene of Artemisia annua L.
(Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian, 2022-01-19) Koerniati, Sri; Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian
Abstract. Artemisinin is the main compound of the dihydroartemisinic acid (DHA)-based antimalarial drug. Its synthesis might be impeded by squalene synthase (SQS) gene. This study aimed to design and synthesis sgRNA of SQS gene of Artemisia annua L., to amplify DNA fragment of the target gene, and to set up in vitro test to finding out sgRNA function. The CRISPR/Cas9 system is applied as it provides a more precise modification of the gene sequences. This preliminary study started with designing single guide RNA (sgRNA) and investigating their functions in directing the Cas9 protein to cut the target. The methods included designing sgRNA and primers using available tools online, synthesizing sgRNA, amplifying SQS DNA target, and setting up in vitro testing using kits and according to manufacturer instructions. The experiment resulted in two sgRNAs and two SQS DNA fragments (targets) that were incubated with the Cas9 protein at 37°C for 3 h in the in vitro test reaction. Both sgRNAs functioned since both 1.6 and 1.4 kbp DNA fragments became two smaller DNA fragments. These indicate that the CRISPR/Cas9 vector construct to target the SQS gene in the A. annua plant can use these two sgRNAs.
The Diversity of Morpho-Agronomic Characters and Identification of Early Maturity Cassava (Manihot esculenta Crantz.) Germplasm
(Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian, 2022-01-19) Wahyuni ...[at al], Tinuk Sri; Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian
Abstract. The breeding of cassava superior varieties requires the collection of germplasm support that accommodates various gene sources and has a wide variety of characters. This research was conducted to analyze morpho-agronomic characters of 100 cassava accessions, to examine its relationship to tuber yield, and to identify accessions with early maturity and high yield potential. The field experiment was carried out at the Muneng Experimental Station of ILETRI in 2019. The characters observed included 31 characters of leaves, stems, yields, and tuber yield components at 7 and 10 MAP. The results showed that most of the characters varied widely, except for petiole length, central leaf lobe length, stem diameter, distance between leaf scar, and number of marketable tuber at 7 MAP, and weight of marketable, unmarketable, and average tuber at 7 and 10 MAP. Path analysis showed that the number of total tubers per plant and the average tuber weight at 7 MAP had a large direct effect and a positive equal effect on the total. Meanwhile, the other 11 characters had less direct effect than the indirect effect through these two important characters. Tuber yield of 100 accessions harvested at 7 MAP was equivalent to 6.8–50.6 t/ha, with average of 28.8 t/ha. Meanwhile, it was between 8.0–68.4 t/ha at 10 MAP, with average of 38.1 t/ha. With K-mean cluster analysis, eight accessions were obtained having fresh tuber yield ranged of 39.6–50.6 t/ha and 46.20–67.20 t/ha at 7 and 10 MAP, respectively. The eight selected accessions were MLG 10361, MLG 10328, MLG 10292, MLG 10371, MLG 10353, MLG 10352, MLG 10360, and MLG 10308. Accessions identified with early maturity or high yield potential need further evaluation with repeated trials and evaluation of various other advantages.
DNA Barcoding of Vatica bantamensis, a Critically Endangered Tree Endemic to Banten, Indonesia
(Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian, 2022-01-19) Hariri ...at al, Muhammad Rifqi; Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian
Abstract. Vatica bantamensis is a member of the family Dipterocarpaceae endemic to Ujung Kulon National Parks in Banten Province, Indonesia. It is classified as a critically endangered species by IUCN Red List. Under this category, the species is considered to be facing an extremely high risk of extinction in the wild. The objectives of this study were to provide easy and accurate identification of V. bantamensis for the conservation and sustainable use of the species. In the present study, we conducted DNA barcoding of V. bantamensis using an rbcL marker. We used V. rassak as a comparison species since both species are morphologically closely related. A total of 606‒608 bp nucleotides have been successfully amplified and sequenced. The dendrogram constructed using the neighbor-joining method with Tamura 3-parameter showed that V. bantamensis used in this study were clustered together with V. rassak. These two species were separated clearly against other Vatica species and two outgroup species, namely Dryobalanops lanceolata and Shorea pinanga, obtained from the NCBI database. The rbcL sequence can discriminate V. bantamensis from V. rassak in six different sites (136, 507, 522, 565, 566, and 567). This study demonstrated that the rbcL sequence could authenticate the samples of V. bantamensis and support the correct identification of this endangered plant species.
The Effect of Coating Application Using Chitosan Enzymatic Depolymerization on Anthracnose Disease Suppression in Mango (Mangifera indica L.) cv. ’Arumanis’
(Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian, 2022-01-19) Suryadi ...[at al], Yadi; Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian
Abstract. Mango cv. ’Arumanis’ is more prone to postharvest disease. An alternative anthracnose disease control is necessary to reduce use of synthetic pesticides. The research was aimed to develop bioformulation against Colletotrichum spp. Seventeen mango phyllosphere isolates belonging to Gram-positive (7 isolates) and Gram-negative (10 isolates) bacteria were obtained from Bogor and Pasuruan. The FAM 1 isolate produced the highest chitinase (14.7±1.11 ppm). Chitosan (CS) depolymerized with chitinase (D3K, BK2P of Bogor isolates) and mixed with tripolyphosphate (TPP) at a concentration ratio of 5:1 (v/v) could inhibit the growth of Colletotrichum spp. compared with control under in vitro test, however, the level of inhibition was lower (<50%). Further formulations of CS (depolymerized with chitinase of Pasuruan isolates (EGM 1, FAM 1, and BMH) mixed with tripolyphosphate (TPP) using concentration ratio of 5:2 and 5:3 (v/v) was effective to suppress the fungal growth colonies in vitro. The application with a formula CS (FAM 1 isolate) and TPP mixture at ratio 5:2 showed as the highest anthracnose disease suppression in vivo. Candidate formulation to reduce Colletotrichum spp. infections have been obtained in the laboratory. However, in vivo test of various mango cultivars in the field still needs to be done.
The Effect of FeSO4 Concentration on the Callus Growth of Two Chili (Capsicum annuum L.) Varieties
(Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian, 2022-01-19) Yunita ...[at al], Rossa; Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian
Abstract. In vitro selection is a method that can produce varieties that are tolerant to abiotic stresses such as Fe poisoning. FeSO4 is a compound that can be used as an in vitro selection agent for resistance to Fe toxicity. Each genotype has a different response to FeSO4. For this reason, it is necessary to determine the optimal concentration for each genotype to be selected so that the selection results are more accurate. This study aimed to observe the response of callus growth to Fe toxicity and to obtain the lethal concentration of 50 (LC50) FeSO4 value for callus of two chili varieties Prima Agrihorti and Rabani Agrihorti. This study used a complete factorial design with the first factor was variety and the second factor was the concentration of FeSO4 (0, 54, 108, 162, and 215 mg/l). The number of repetitions was 10 times. The observed parameters were the percentage of browning callus, callus diameter, and callus performance. To determine the LC50 value, the curve analysis program was used. The results showed the interaction between varieties and the concentration of FeSO4 has not significant effect, but when the FeSO4 concentration increase, the growth was inhibited, shown as smaller diameter and the callus turned to brown. The LC50 FeSO4 value in the variety Prima Agrihorti was 130.341 mg/l, while that of Rabani Agrihorti was 115.511 mg/l. These values can be used for in vitro selection of callus of chili varieties Prima Agrihorti and Rabani Agrihorti.
Effect of Nitrogen Fixation and Phosphate Solubilizing Bacteria on Growth and Yield of Lowland Rice in Different Soil Type
(Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian, 2022-01-19) Ikhwani ...[at al], Ikhwani; Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian
Abstract. Efficiency of nitrogen and phosphate fertilization in soil for rice cultivation could be improved using microorganism biofertilizer. This study aimed to investigate the effect of nitrogen fixation and phosphate solubilizing bacteria on growth and yield trait of lowland rice in alluvial and latosol soils. The experiment was conducted at greenhouse arranged in factorial randomize block design with two factors (soil type and bacterial isolates) and three replications. The first factor was two soil type consisted of alluvial and latosol soils. The results showed that alluvial soil type had the best results on growth and yield more than latosol soil. The filled grain on alluvial soil had number/plant highest of 1,755.5 more than latosol soil. The latosol isolate on alluvial soil had highest for plant height (9.50 cm), 3.33 of tiller number, and 2.0 of panicle number per plant more than alluvial isolate. The interaction of soil type and isolate effects showed no significant response for all traits at generative stage. In term of yield component trait, latosol isolate had better response for grain compared with alluvial isolate, both in alluvial and latosol soils. This finding had potential to latosol isolate further in alluvial and latosol soils.

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