Secondary metabolite profiling of four host plants leaves of wild silk moth Attacus atlas L.

Secondary metabolites may affect insect herbivores’ host plant preferences. Attacus atlas L. larvae are known have a wider variety of host plants compared with other members of the Attacus genus. This research compared the metabolic profiles of four A. atlas host plants: keben (Barringtonia asiatica (L.) Kurz), dadap (Erythrina lithosperma Miq.), gempol (Nauclea orientalis L.), and soursop (Annona muricata L.). Leaves were collected from Sawit Sari Research Station, Yogyakarta. Terpenoid was extracted by macerating the leaves in ethyl acetate and subjecting them to GC-MS analysis, while alkaloid, tannin, and flavonoid were extracted through percolation. Total alkaloids, tannins, and flavonoids were measured using spectrophotometric analysis. Multivariate data analysis using PAST ver. 3.0 was performed on the GC-MS data. Based on the PCA scatter plot of the GC-MS data, keben leaves were clustered separately from the other three leaves by PC1. Dadap and gempol leaves were clustered together due to the phytol content while caryophyllene was detected only in soursop leaves. Neophytadiene was detected in all of the leaves, suggesting that this terpenoid may serve as a signal to locate the host plants. Keben leaves contained the lowest alkaloids and highest tannins and flavonoids compared with the other leaves. These secondary metabolites may determine the host plant suitability for culturing the A. atlas

Optimization of qRT-PCR Annealing Temperature of WRKY45 Gene for Detection of Resistance Genes Against Xanthomonas oryzae pv. oryzae on Black Rice Cempo Ireng

Plant pathogens constrain the development of black rice farming. One of these pathogens is Xanthomonas oryzae pv. oryzae (Xoo), causing a bacterial leaf blight disease. The disease disrupts crop growth and reducing yields. Cempo ireng is a local pigmented rice cultivar from Yogyakarta, Indonesia, which is reported for its high resistance to Xoo. One of the rice resistance mechanisms to Xoo infection is a molecular defense employing plant resistance genes such as WRKY45. Comparing the expression of resistance-related genes of the resistant cultivar to the susceptible ones is needed to elucidate the resistance mechanism of the black rice to Xoo. For this purpose, the expression of WRKY45 gene at the level of mRNA can be performed using qRT-PCR. The success of qPCR analysis is greatly influenced by the accuracy of the annealing primer temperature of the corresponding gene. This study aimed to optimize the primer's annealing temperature for WRKY45 gene. The optimization was done by a temperature gradient PCR. Determination of the optimal annealing temperature was selected based on the profile of the amplification curve, melt curve, melt temperature and the Ct value obtained. The annealing temperature gradient used in this study was ranging from 52°C to 60°C. The results showed that the best annealing temperature for WRKY45 gene primers is 58.3°C based on the amplification curve, melt curve, melt peak and Ct value of 29.21

1H NMR Fingerprinting of Medicinal Herbs Contain Chemical Drug Material Allopurinol

A study to differentiate the pure medicinal herbs from the mix medicinal herbs with chemical drug material has been done. For this purpose, we conducted fingerprinting of commercial medicinal herbs and chemical drug material allopurinol using 1H-NMR followed with chemometrics analysis. Nine commercial traditional herbal medicines claimed for rheumatic were used as samples as well as allopurinol as the chemical drug standard. Extraction of samples was done by ultrasonicator for 15 min in methanol-d4 containing 0.01% TMSP as an internal standard. Each type of herbal medicine was prepared in three replicates. The phytochemical analysis was done by 500 MHz JEOL NMR. The chemometrics analysis was done using SIMCA software following the 1H NMR spectra processing with MNOVA software. All spectra showed no contamination with allopurinol. The specific signals of allopurinol at aromatic regions were confirmed not present when the spectra were stacked together. Hence, the result of OPLS-DA analysis convinced that the herbs were clearly separated the medicinal herbs into 3 classes. Jamu 1 is separated from others showed very high intensity of several signals which may indicate an addition of chemical medicines but not allopurinol. The clear separation of other two groups may corresponds to the similarity of ingredients. These results also showed that most of traditional medicines which produced by small industries, the traditional medicines contain no active pharmaceutical ingredients (allopurinol) indicating a high safety of Indonesia traditional medicines

Antioxidant Activity of Banana Kepok Kuning (Musa paradisiaca L.) and Cavendish (Musa acuminata Colla, AAA) Peel Extracts and the Potential as Chicken Meat Preservative

Bananas are a type of fruit that is consumed, with the peel being discarded as waste. However, the peel contains phytochemical compounds, including phenolic and flavonoids, which act as natural antioxidants. Therefore, this study aimed to determine the antioxidant activity of banana peel of Kepok Kuning (KCE) (Musa paradisiaca L.) and Cavendish (CCE) (Musa acuminata Colla, AAA) extracts, as well as eluviate their preservative potency. The sample was extracted using 80% methanol, and the antioxidant activities of KCE and CCE were evaluated using DPPH and FRAP. The extracts and the control antioxidant butylated hydroxyl toluene (BHT) were tested for their preservatives potency on chicken meat at concentrations of 5%, 10%, and 15%, respectively. After 8 days of incubation in the refrigerator, the color change was recorded and analyzed using ImageJ, while free fatty acid contents were determined by titration. The results showed that 15% KCE and 15% CCE exhibited higher antioxidant activities than 0.1% BHT. There was no significant difference in the L (lightness) and b (yellowness) values between the treatments, but variations were evident in the a (redness) values. FFA values were significantly different between treatments, with untreated control registering the highest value at 0.41%, and the KCE 15% and CCE 15% concentrations showing the lowest values at 0.12%. Considering these results, both extracts could be developed as preservatives for chicken meat

Callus Induction from Cotyledon of Gyrinops versteegii (Gilg.) Domke

Gyrinops versteegii is an endemic plant in eastern Indonesia that produced agarwood with high quality and economic value. This plant has been threatened by overexploitation which leads on decreasing in the natural population. This research aimed to induce in vitro callus formation to support the sustainable utilization of G. versteegii, which in the end, may support plant productivity. Callus induction was investigated using cotyledon and cultured on MS medium supplemented with several combination of plant growth regulators. The observations were done for the number of explants turned into callus and the duration for the first callus formation. The results showed that combination of 3 mg/L NAA + 0.5 mg/L BAP was recorded as the best combination for callus induction (63.63%). Callus with friable structure and bright color are obtained within nine days of incubation and showed the characteristic of embryonic callus. This result is expected to give a significant opportunity to conserve the natural population of G. versteegii

Metabolite profiling of agarwood (Gyrinops versteegii (Gilg.) Domke) leaves from difference growth locations using Thin Layer Chromatography

Gyrinops versteegii (Gilg) Domke is an agarwood-producing plant originating from Eastern Indonesia. Leaves have antioxidant and anticancer activity due to their secondary metabolites. However, secondary metabolites are strongly influenced by the environment. This study aims to compare the terpenoid, phenolic, and flavonoid profiles of agarwood leaves grown in three locations, namely Bogor, Mataram, and Sambas using Thin Layer Chromatography (TLC), in order to obtain the most potential agarwood G. versteegii for further development as herbal medicine. Agarwood leaf powder was extracted using Soxhlet with 80% methanol as solvent, followed by liquid-liquid fractionation using chloroform and aquadest. The crude extract and the obtained fraction were subjected to metabolite profiling using TLC. Racing data factors from TLC were analyzed using hierarchical cluster analysis (HCA). The results showed that the Mataram sample produced higher yields for both extracts and fractions. TLC detected that the chloroform fraction contained more terpenoids, followed by phenolics and flavonoids, while the aqueous fraction contained the same number of positive bands for terpenoids and phenolics. In terms of origin, the Bogor sample showed six terpenoid positive bands higher than the other samples. A similar pattern was observed in the phenolics, while the flavonoids were the same for all samples. HCA resulted in Mataram being clustered separately from the Bogor and Sambas samples. Agarwood leaves from Mataram cannot be used to replace agarwood leaves from Bogor and Sambas as herbal ingredients

Color Stability of Phycoerythrin Crude Extract (PECE) from Rhodomonas Salina Toward Physicochemical Factors

Rhodomonas salina produces Cr-phycoerythrin545 as its designated phycoerythrin (PE) with an absorption maximum at 545 nm and a shoulder 564 nm. PE has potential to be applied as colorants, pharmaceutical agents, and fluorescent dye tags. The stability of the PE color is influenced by the physicochemical factors of the solution. This study aimed to analyze the color stability of PECE against chemical (ethanol and pH) and physical (light and temperature) factors. PECE was prepared from freeze-dried biomass of R. salina and was extracted in phosphate buffer solution (pH = 6.0) using a freeze-thaw method in -25 oC (2 hours) and 4 oC (24 hours). The resulting extract was concentrated and dried in a freeze-dryer. Analyses were conducted using UV-visible and fluorescence spectrophotometer. PECE showed color stability against light of white fluorescent lamp exposure up to 8 hours, temperature exposure up to 40 oC, ethanol solution up to concentration of 20 % (v/v), and pH range 3.9-8.42. Results from this study can be useful for extraction, purification, and future application of Cr-PE545

Plant growth‐promoting activity of endophytic bacteria from sweet sorghum (Sorghum bicolor (L.) Moench)

Application of high levels of chemical fertilizers for optimal growth of sweet sorghum causes environmental degradation. Plant growth‐promoting bacteria have biotechnological importance because they can improve the growth and health of important agronomic plants. This study aimed to isolate, characterize, and identify endophytic bacteria associated with sweet sorghum (cv. KCS105), and also to study the inoculation effects of selected isolates on sorghum growth. In this study, 35 isolates were evaluated for their ability to support plant growth. The results showed that seven isolates were diazotrophic, six were capable of dissolving phosphate, six produced IAA and could detect ACC‐deaminase activity, and three inhibited the growth of pathogenic fungi. Nine isolates exhibiting mechanisms for promoting plant growth from the Alphaproteobacteria (Devosia), Firmicutes (Bacillus, Paenibacillus, Staphylococcus), and Actinobacteria (Microbacterium, Brachybacterium) phyla were identified. In addition, the Paenibacillus sp. BB7, Bacillus sp. PIB1B, and Bacillus sp. PLB1B isolates showed increasing effects on plant growth in greenhouse tests. Endophytic bacterial isolates which display plant growth‐promoting features can potentially be employed as biofertilizer agents. They may also address environmental damage problems resulting from the use of chemical fertilizers and pesticides

Toxicity of Ocimum basilicum L. leaf extract against Spodoptera exigua Hübner (Lepidoptera: Noctuidae)

The beet armyworm Spodoptera exigua Hübner (Lepidoptera: Noctuidae) is one of the most significant shallot pests worldwide, which affects agricultural production by approximately 100% in Indonesia. The growing awareness of sustainable agriculture encourages the search for natural alternatives to synthetic pesticides that promote resistance and contaminate the environment. Therefore, this study aims to evaluate the efficacy of basil (Ocimum basilicum L) leaf extract against the 2nd and 3rd instar larvae of S. exigua. The extracts were prepared using maceration and hydrodistillation and the pathogenicity was carried out against the 2nd and 3rd instar of S. exigua using three replicates with several doses of extract ranging from 0- 2.0%. The extracts were applied to the larvae using contact and topical methods. The results showed that maceration extraction yields a greater extract with different types compared to those of the hydrodistillation method. Maceration extract of basil leaves with the contact application method to 2nd instar showed better results than the topical application with the highest mortality rate of 75% (F=24.464; P< 0.001) and LC50 of 0.007%. This indicated that basil leaf extract with the contact application method has great potential to be developed as a botanical insecticide to control S. exigua in the field as part of Integrated Pest Management (IPM)

A bibliometric analysis of botanical insecticides for Lepidopteran insects over the period 1985-2022

Lepidoptera is an economically important insect pest that attacks many commercially important crops. Synthetic insecticides are currently most extensively utilized to control this pest. The use of massive doses of synthetic pesticides has resulted in resistance and resurgence. Therefore, developing an environmentally acceptable technique for controlling insect pest populations, such as using natural plant components as insecticides, can assist in mitigating the negative impacts of synthetic pesticides. Plant pesticides or botanical insecticides are becoming increasingly popular. This bibliometric research aimed to demonstrate the growth, awareness, importance, international cooperation or collaboration and knowledge gaps in developing synthetic botanical insect pesticides in the future. Information used in this research was culled from 138 scientific papers (Scopus database) and analyzed using VOSviewer 1.6.17. Descriptive statistics was used to identify the rate of botanical insecticide development in controlling these insect pests by analyzing the contributing documents by year, country and bibliometric analysis of country and keyword co-occurrence. The research started in 1985 and exponential growth occurred after 2006. In addition, the trend peaked in 2020 and is currently being maintained. The scientific papers were distributed from 41 countries, with Brazil being the most productive. The bibliographical network shows the relatedness of information about keywords between countries. The results obtained can help recognize existing knowledge gaps that need to be addressed and considered in developing botanical insecticides to control this lepidopteran pest