Agarwood Tree Research Articles

Non-Destructive Assessment of Agarwood Resin Formation Status in Aquilaria crassna using Near-Infrared Spectroscopy

Agarwood is a unique and valuable resin synthesized within a few tree species of the tree family Thymelaeaceae primarily as a self-defence mechanism against stress causes such as fungal invasions. Instead, artificial inoculation leads to short-time harvesting practiced in intensive commercial-level cultivations. The extraction of this resin can be done only after the harvesting where accurate estimation of resin yield is crucial for avoiding immature or underdeveloped harvesting. However, the amount of resin accumulated in the trunk cannot be estimated by the outside inspection. As a result, trees are being cut down before they have reached their maximum harvest potential. Therefore, precise detection of fragrant resin formation status inside the tree has become one of the primary challenges in the agarwood production industry. Currently, the methods available are either destructive, inefficient, or inaccurate. As such, this study evaluates the possibility of the use of NIR spectroscopy as a non-destructive and quick detection method for agarwood formations status inside trees in the in-situ mode. Near-infrared (NIR) range (588–1,025 nm) spectra were acquired from resin-formed and non-formed areas from twenty tree trunks of agarwood with five different formation levels. The best agarwood formation areas of logs were assigned as class level 1 and the least was assigned as the class level 5. The spectra were acquired near the opening made for the artificial inoculation process of considered Aquilaria crassna tree trunks. The results of Soft Independent Modelling of Class Analogies showed consistent evidence of differences in the NIR spectral profiles of all levels of agarwood formations. The SIMCA discrimination between the resin developed versus not was achieved with 86% accuracy. The accuracy rates of the degree of the presence of agarwood were obtained as 80%, 100%, 85%, 90%, and 85% respectively, as assigned levels 1 to 5 in the SIMCA models. This research revealed the potential possibility of using NIR technology for the prediction of agarwood resin status inside the agarwood tree in A. crassna before harvesting the tree as a non-destructive and rapid method.
 
 Keywords: NIR spectroscopy, Agarwood, Aquilaria crassna, Non-destructive, Agarwood formation

Pharmacology and therapeutic potential of agarwood and agarwood tree leaves in periodontitis.

The main bioactive components of agarwood, derived from Aquilaria sinensis, include sesquiterpenes, 2-(2-phenethyl) chromone derivatives, aromatic compounds, and fatty acids, which typically exert anti-inflammatory, antioxidant, immune-modulating, hypoglycemic, and antitumor pharmacological effects in the form of essential oils. Agarwood tree leaves, rich in flavonoids, 2-(2-phenethyl) chromone compounds, and flavonoid compounds, also exhibit significant anti-inflammatory, antioxidant, and immune-modulating effects. These properties are particularly relevant to the treatment of periodontitis, given that inflammatory responses, oxidative stress, and immune dysregulation are key pathological mechanisms of the disease, highlighting the substantial potential of agarwood and agarwood tree leaves in this therapeutic area. However, the low solubility and poor bioavailability of essential oils present challenges that necessitate the development of improved active formulations. In this review, we will introduce the bioactive components, extraction methods, pharmacological actions, and clinical applications of agarwood and agarwood tree leaves, analyzing its prospects for the treatment of periodontitis.

Investigation of factors affecting the rapid detection of agarwood formation in <em>Aquilaria crassna</em> by near-infrared spectroscopy

Agarwood is a highly valued fragrant resin produced inside a few tree species belonging to the family Thymalaeaceae as a self-defense response to plant stress. The amount of resin developed inside the tree cannot be estimated by outside inspection. Consequently, harvesting trees before they reach their potential yield is a severe drawback to the Agarwood industry. Therefore, developing effective techniques for detecting Agarwood resin status inside the tree species has become a critically important task for the agarwood industry to increase productivity. The present study evaluates the factors affecting Near-Infrared Spectroscopy (NIRS) models when predicting agarwood formation inside A. crassna trunks using NIR spectroscopy. The research used 110 wood specimens obtained from well-grown Agarwood trees in a commercial plantation in Nawimana GS Division, Matara District, Sri Lanka. NIR meter FQA-NIR Gun (588-1100nm) with a custom-made probe was used to acquire NIR reflectance spectra without outside light interference. SIMCA models were built to identify the agar resin-developed wood log areas from the normal wood areas in the tree trunk. SIMCA prediction models were built to investigate three influencing factors, namely present or absent outside tree bark, surface roughness and wood thickness agarwood prediction. Better prediction results were obtained from the bark-removed samples (at the accuracy rates of 97%) to the bark present (85%), smooth wood surfaces (98%) to the rough surface (90%) and 2mm thickness (98%) to the other thickness. The most effective wavelength for the separation of Agarwood present and absent samples was located at 978 nm of NIR. The study has demonstrated the potential possibility of using NIR spectroscopy to identify the agarwood formation in A. crassna in non-destructive and rapid mode.

FORMULATION AND STABILITY OF STERILE GEL OF AGARWOOD LEAVES EXTRACT (Gyrinops Versteeg (Gilg.) Domke) FOR DIABETIC WOUNDS

Agarwood leaves contain high levels of secondary metabolites due to increased metabolic processes in agarwood trees infected with fungi. Through this metabolic process, leaves contain secondary metabolites, flavonoids, and tannins. This causes agarwood leaves to have potential as an anti-inflammatory agent for treating diabetic wounds, aimed to formulate a sterile gel preparation of agarwood leaf extract and evaluate its quality, including sterility tests and stability tests (cycling tests) including organoleptic tests, homogeneity, spreadability, adhesive power, viscosity, and pH. A sterile gel formula was prepared with an extract concentration of 4% and variations of Carbopol 940 (gelling agent), namely 0.75%, 1.0%, and 1.25%. The results showed that F1, F2, and F3 were sterile and free of bacterial and fungal growth. The stability test before and after the cycling test in organoleptic tests F1, F2, and F3 were brown, had a distinctive odor of gaharu leaves, and had a soft texture. The homogeneity tests F1, F2, and F3 Homogeneous. The pH values before the cycling test were F1 5.63, F2 4.55, and F3 4.57, whereas after the cycling test, they were F1 5.05, F2 4.87, and F3 4.63. Adhesion time before cycling test F1 02.97 seconds, F2 03.15 seconds, F3 04.86 seconds while after cycling test F1 02.64 seconds, F2 03.04 seconds, F3 03.08 seconds. Spreadability test before cycling test F1 5.4 cm, F2 4.6 cm, F3 3.7 cm, while after cycling test F1 5.0 cm, F2 4.7 cm, F3 3.5 cm ...

Taxonomy, phylogeny, and successful cultivation of Marasmiellus scandens (Basidiomycota) associated with Aquilaria sinensis (agarwood tree) in China

Endophytic fungi are isolated from all parts of plants and several have been reported from Aquilaria sinensis and most of them are ascomycetous species. In this study, an endophytic Marasmiellus species collected from Aquilaria sinensis in Guangxi Zhuang Autonomous Region, China was isolated. Phylogenetic analyses of the Internal Transcribed Spacer (nrITS) gene region and morphological analyses confirmed the endophytic fungus as Marasmiellus scandens. In addition, fruiting bodies of M. scandens were successfully obtained in sawdust grow bags. A full description, illustrations, cultivation results, and a phylogenetic tree to show the placement of M. scandens are provided. Marasmiellus scandens is reported here as a new country and host record. In addition, this is the first report of the M. scandens cultivation in its endophytic form.

First Report of Infestation of the Agar Defoliator <i>Heortia vitessoides</i> (Moore) on the Agar Wood Tree <i>Aquilaria malaccensis</i> Benth. in South India

Massive infestation of the agar defoliator Heortia vitessoides (Moore) (Lepidoptera, Pyraloidea, Crambidae) was observed in Thiruvananthapuram and Kollam districts in Kerala on agar wood tree Aquilaria malaccensis Benth. (Thymelaeaceae), during February – December, 2022. Of the 119 trees observed in Thiruvananthapuram and Kollam districts, 115 were infested and six of them died due to the pest attack.

Culture dependent bacteria from the gut of Neurozerra conferta Walker, the insect pest of agarwood trees possess enzymatic functions potentially associated with its aroma

Culture dependent bacteria from the gut of Neurozerra conferta Walker, the insect pest of agarwood trees possess enzymatic functions potentially associated with its aroma

Effects of Trichoderma Harzianum and Fusarium Solani Inoculant on the Resin Content of Agarwood (Gyrinops Versteegii (Gilg.) Domke)

This study aimed to determine which fungal inoculants, Trichoderma harzianum or Fusarium solani in a solid or liquid form, produced the highest resin content in agarwood trees. This experimental study used a randomized completely block design (RCBD) with seven treatments in triplicates: solid Trichoderma harzianum inoculants in pellets(A) and capsules (B), liquid T. harzianum inoculants (C), solid Fusarium solani inoculants in pellets (D) and capsules (E), liquid F. solani inoculants (F), and control or without inoculants (G). Parameters observed were the color and aroma of the infected agarwood and the resin content of the harvested sapwood. Quantitative data obtained from observations/measurements were analyzed statistically with analysis of variance and a subsequent LSD test at a 5% significance level following a significant result. The results showed that treatments using different fungal inoculants had no significant effects on the resin content of agarwood. However, inoculants in pellets and capsules tended to increase the resin content. The resin content of agarwood ranged from 15.97% to 21.53%, and the highest level was obtained from treatment A (solid T. harzianum inoculants in a pellet form)

Propagation of Gaharu Plant Gyrinops Versteegii Species Provenant Beringin Throught in Vitro Culture

Grynops verstegii is a non-timber forest product commodity that has a very high economic value. People know this tree as the agarwood tree. In Lombok, the Beringin provenance is one of the leading provenances, so it is very important to conserve its germplasm. This study aims to find alternatives for gaharu development through seed production using in vitro culture techniques. This research was carried out in July 2021 at the Tissue Culture Unit of the Immunobiology Laboratory, University of Mataram. In this experiment there were 8 media treatments with different concentrations of BAP (Ssitokini) and NAA (auxin). The results showed that callus growth occurred on MS 1, MS 3, MS 4, MS 6, and MS 7 media. However, the best growth was obtained in MS 1 treatment with a combination of 1.0 ppm BAP and 0.5 ppm NAA. While most of the samples experienced browning and contamination.

Microencapsulation of agarwood (Aquilaria crassna) essential oil by spray drying

Agarwood essential oil is the essential oil extracted from the heartwood of agarwood trees (Aquilaria sp.) after inoculation or natural infection of one of several fungal species. The essential oil is proved to carry antioxidant, antimicrobial and anti-stress activities. Microencapsulation of essential oil included emulsification of oil into maltodextrin solution and spray drying of emulsion at different levels of inlet temperature and compressed air pressure. Microencapsulation process was qualified based on following parameters including process’s microencapsulation yield, moisture content of powder, powder particle size, encapsulated oil’s composition, oil recovery and drying yield. Microencapsulation yield was affected by inlet temperature whereas drying yield changed as air pressure varied from 1.5 to 2.5 kgf/cm2. Other parameters were impacted by both variables. Besides, oil composition changed at an acceptable degree after microencapsulation process, in which there were an increase in percentage and the presence of some components but the main components still remained in the encapsulated oil. Finally, the drying air temperature of 150 °C and air pressure of 2.5 kgf/cm2 were the best parameters to obtain both microencapsulation yield of 89.12% and drying yield of 79.00%.

Gas chromatography analysis of the microwave-aided extracted agarwood oil from physically induced Aquilaria malaccensis trees in Northern Thailand

Agarwood (Aquilaria malaccensis) oil is one of the essential bioactive materials in the scent/ perfume industry for the middle east and gulf countries. The built-in functionality and availability of bioactive components of oil make agarwood more precious. In this study, the physically induced infected agarwood tree chops were extracted by microwave assisted Soxhlet extraction. Moreover, the extracted oil samples were analyzed against control not infected trees using Gas chromatography-mass spectroscopy (GC-MS) by GCMS (Agilent GC 6890zN MS 5973). The microwave assisted Soxhlet extraction revealed a higher amount of oil extraction. In the physically induced samples, 24 signal peaks were recorded employing GC-MS. The most prominent peak area recorded was -5-hydroxy-3-methyl-1-indanone detected at minute 41.89 with a peak area of 17.64%, followed by -methyl 4,4,7-trimethyl 4.7-dihydro indan -6- carboxylate at 23.17 min, occupying an area of 15.54%; followed by 3-carene recorded at 46.61 minutes with an area. According to this research, the microwave-assisted Soxhlet extraction method outperformed and can be utilized in the agarwood industry for enhanced oil production.

Growth and Coppicing Ability of the Critically Endangered Agarwood (Aquilaria Malaccensis Lam.) Tree in Monoculture and Polyculture in North East India

ABSTRACT Agarwood (Aquilaria malaccensis Lam.) (Thymelaeaceae) is valued in many cultures because of its distinctive fragrance and its use in incense, perfumery, and traditional medicine. Large-scale harvesting from natural populations caused rapid depletion of the species in the wild, and the species is now listed as “Critically Endangered” and almost extinct in the wild. The promotion of this species in agroforestry systems may pave the way for its conservation. The present study aimed to quantify the growth and coppicing ability of agarwood under monoculture and polyculture stands. Monoculture stands aged more than 20 years, and stem girth (1.37 m above ground) of 60–70 cm had the highest growth increment of 3.73 cm yr−1. Monoculture stands (>10 years old) also recorded higher number of coppicing stocks than polyculture stands. The highest coppice growth increment (4.07 cm yr−1) was recorded in the 70–80 cm stem girth in monoculture stands. Based on the results, we recommend the promotion of agarwood trees with good coppice management in forestry programs to conserve the species while also meeting the economic needs of rural communities.

Diversity and Biosynthetic Activities of Agarwood Associated Fungi

Agarwood is a fragrant dark resin produced in plants belonging to the family Thyme-laeaceae and which has a high economic value. The unique fragrance and medicinal applications intensify the value of agarwood. The wild populations of agarwood trees are highly threatened by high economic demand. Therefore, it is worthwhile to develop an artificial agarwood induction technology for the countries that rely on agarwood from the natural habitat of the plants. Fungal induction of agarwood has been shown to be an efficient method. Interestingly, most of the fungi known from agarwood are endophytic. In this paper, we supplement and update the bioactivity of fungi associated with agarwood and their ability to induce agarwood formation. According to the existing literature, 59 endophytic fungal strains of 16 genera induce agarwood production, most of which belong to Fusarium (28 identified strains). Hence, Fusarium is a good candidate for further studies on fungal induced agarwood production.

Identification of bacterial species from healthy wood of Aquilaria crassna using colony PCR

Agarwood is one of the most expensive, sacred and valuable woods used in the biotechnology industry. This natural raw material is in high demand as a commodity and can be used to manufacture a variety of products. Isolating a microbe from its environment is crucial because several bacterial strains can produce novel compounds for biotechnological applications. Exploration of bacterial strains, such as bacterial endophytes, has been associated with bioactive compounds of the agarwood plant. The purpose of this study was to isolate and identify bacteria from Agarwood trees in the Kaeng Hang Maeo Sub-district of Chanthaburi Province, Thailand by using molecular biology techniques. A colony PCR technique without prior DNA extraction for rapid and simple detection of bacteria was performed. After the DNA sequence analysis, the results showed that most populations in the normal layer are belonged to Pantoea dispersa (99%). Through analysis of phylogenetic tree by neighbor-joining method, the sequences of the 16s rRNA gene were analyzed to understand evolutionary relationships of bacteria found in agarwood. In conclusion, good amplification of the 16s rRNA sequence (1.5 kb) was detected with the specificity of the target sequences of the 16s RNA. In addition, the isolated bacterial strains are the most common species in agarwood, however, there has been no previous research on isolating these bacterial strains in Aquilaria crassna.

Mini-review Teknologi Produksi Teh Herbal Gaharu

Agarwood tea is a prospective herbal tea product in the Indonesian marketplace. This product was made from agarwood trees, especially Aquilaria and Gyrinops genera. Agarwood leaves were the primary raw material of agarwood tea products. The leaves processing method into herbal tea products has adopted the conventional tea production method. This method has an essential contribution to maintaining the quality of agarwood tea products. This mini-review emphasizes the importance of systematic procedure and its effect on agarwood tea production technology. Agarwood tea production started with the standardization of raw material preparation. Raw material selection and sterilization were the essential procedure at this stage. Agarwood tea production after raw material preparation involves three steps: drying, chopping, and oxidizing. Dried leaves could produce agarwood tea with more robust characteristics than fresh leaves. Chopping the leaves is essential for agarwood tea product packaging. Oxidizing the leaves could produce agarwood tea with a better tannin concentration. It could be concluded that leaf preparation, drying, chopping, and oxidizing the leaves is an essential processing technology for agarwood leaves to produce a good quality tea.

Agarwood Tree Characteristics based on Different Growing Habitat and Ecophysiological Attributes in the Papuan Tropical Forests

Understanding tropical forest characteristics, including forest derivative products such as agarwood, is pivotal. This study focused on perceiving an ideal habitat of the lowland tropical forest ecosystem of West Papua for agarwood-producing tree species. In further, this study identified a close association of biotic and abiotic characteristics to underpin agarwood-producing tree growth. Six sites in three different locations were established to ascertain forest composition and the associated insects. Soil samples were collected to analyze their physical and chemical properties using the Kjeldahl, the Walky and Black, and Atomic Absorption Spectrophotometry (AAS). In addition, temperature, relative humidity, and light intensity were measured. The results noticed two agarwood producing species, Gyrinops caudate and Aquilaria sp., found at an altitude of 400–402 masl in Manokwari, 200–300 masl in Teluk Wondama, and 167.7 masl in Teluk Bintuni. There were no significant differences in temperature, relative humidity, and light intensity. The soil cation exchange capacity was moderate in Manokwari, but it was low in Teluk Wondama and Teluk Bintuni. Total C-organic content in three research sites was classified as high. However, total N content in the three research sites was in the moderate category. The ideal habitat for agarwood in the West Papua forest is from the lowlands to the highlands. Environmental conditions that support the growth of agarwood are moderate temperature, high humidity, and moderate light intensity. Soil fertility also affected the natural growth of agarwood trees. Keywords: abiotic, agarwood association, lowland tropical forest, soil characteristics

BIO-INDUKSI RANTING CABANG GAHARU (GYRINOPS VERSTEEGII) DI PERKEBUNAN GAHARU DESA PEJARING LOMBOK TIMUR

Bio-induction on the branch of agarwood tree (Gyrinops versteegii) is a good alternative method to increase resin productivity. The aim of this research is to applied bio-induction on the branch of G. versteegii with different diameters. Different branch diameter was the treatment on this research including branch 1 (diameter 70 mm ± 5 mm), branch 2 (85 mm ± mm), branch 3 (100 mm ± mm). Fusarium solani isolate of Institute for Technology Research and Development of Non-Timber Forest Product was used as a bio-induction agent. This isolate was cultured on bean sprout broth media for 28 days. Bio-induction was carried using standard procedure developed by Forest Research and Development Agency. Observation of the result conducted 3 months after bio-induction. There were 3 parameters of bio-induction result including resin area production, discoloration in form of browning on branch, and fragrance level of resin. Branch 2 has the largest resin area production. However, branch 3 has the highest score of browning and fragrance level. This result indicated a correlation between browning on branch and fragrance level quality. It could be concluded that bio-induction on larger branch diameter could produce higher productivity of resin, especially on browning and fragrance level parameter

Agarwood formation in Aquilaria beccariana and Aquilaria microcarpa in response to inoculation of newly isolated fungi from Brunei Darussalam

Abstract. Mohammad YH, Shivanand P, Metali F, Taha H, Matussin NBA, Abdul-Halim AMA, Mohaimin AZ. 2021. Agarwood formation in Aquilaria beccariana and Aquilaria microcarpa in response to inoculation of newly isolated fungi from Brunei Darussalam. Biodiversitas 22: 4131-4138. Aquilaria is an important agarwood-producing tree, but naturally occurring agarwood is rare. Artificial induction method is the most commonly used to produce agarwood in a short time. Out of the 21 species of Aquilaria, Brunei Darussalam houses two species that produce high quality agarwood, namely Aquilaria beccariana and A. microcarpa. The aims of this study are to artificially induce agarwood formation using eight different induction treatments: six fungal inoculant treatments with two control treatments, and to isolate and identify fungal strains from naturally infected agarwood tree using DNA barcoding. Agarwood formation was observed at four different time intervals i.e. 1, 3, 6 and 9 months of post-inoculation. Results exhibited that the area and the total length of discoloration zone were significantly greater at 6 and 9 months of incubation period. Induction treatment did not significantly affect the area and the total length of discoloration. FTIR analysis revealed that two aromatic compounds (C – H and C – C stretching) were detected at the wavelength ranges of 3000 – 2800 cm-1, and 1500 – 1400 cm-1 respectively, and OH group was detected at 3600 – 3400 cm-1. Nine fungal strains from Aspergillus, Fusarium, Mucor and Trichosporon were isolated and identified from a naturally infected agarwood, and the last two fungi (Mucor and Trichosporon) were never been previously reported.

Pemberdayaan Masyarakat pada Budi Daya dan Pengembangan Produk Pohon Gaharu (Aquilaria sp.) di Kabupaten Tolitoli, Sulawesi Tengah

Tolitoli District has a potential for agarwood trees in the forests, but it has not been optimally utilized and preserved by the local communities. Therefore, efforts are needed to increase understanding and community involvement in cultivation and product development of agarwood trees. The objectives of this activity are: to explore agarwood tree species in the forest and sources of local inoculums using a purposive sampling method for agarwood production, to assess the suitability of agro-climates for agarwood tree cultivation, to conduct face-to-face training and practice for agarwood tree breeding and bio-induction by injection, and to identify socio-economic conditions to support agarwood tree cultivation for community empowerment through product development and cultivation in Kabupaten Tolitoli. The project was implementated by exploring and identifying the existence of natural agarwood trees and agarwood farmers, training, and mentoring. The results of exploration and identification showed that Kabupaten Tolitoli has natural resources of agarwood trees in the forest and local inoculums for the bio-induction of agarwood formation. Also, the local community, including the local government, practitioners, farmers, and educational institutions, have great interest and desire for product development and cultivation of agarwood trees. Thus, it can be concluded that community empowerment through product development and cultivation of agarwood trees in Kabupaten Tolitoli can be implemented.

The effect of medium formulation and BAP concentration of growth and development of Aquilaria malaccensis Lam. shoot in vitro

Tissue culture of agarwood tree (Aquilaria malaccensis Lam) was done to produce a large number of transplants in the laboratory. The compositions of medium culture are commonly modified to obtain the best growth of explants to regenerate into plantlets. Our study aimed to evaluate and compare different medium formulations supplemented with BAP on agarwood shoot culture growth. In this study we used MS, NN, B5, AR and WP medium containing 0 (control treatment), 0.25, 0.5, 1.0 and 2.0 mg/L BAP. The shoot tips were cultured for 16 weeks in the treatment media. Height of shoots, number of leaves, number of adventive shoots, and number of roots was recorded every week to determine the growth. The results showed that after 16 weeks of culture, the best medium for shoot height was NN medium with BAP fortification (3.54cm) for number of leaves and adventive shoots proliferation was Gamborg B5 medium fortified with 0.5 mg/L BAP (21.63 leaves and 8,38 shoots), while NN medium without BAP was best for rooting.