Inventory of Dryobalanops aromatica Gaertn. in Lae Kombih Forest Park Area, Subulussalam

Dryobalanops aromatica Gaertn. is a plant that grows in the tropical rain forests of western Indonesia which is categorized as a valuable timber tree. Dryobalanops aromatica Gaertn is a plant that is categorized as a valuable timber tree. Dryobalanops aromatica Gaertn also known as the camphor tree, lime or Sumatran camphor, this tree can grow to a height of 60 m with an average trunk diameter of 9 m. Camphor tree is a plant species belonging to the Dipterocarpaceae family which has several advantages because its tree trunk can produce a variety of high economic value commodities such as camphor, balsamic acid, resin, essential oil, beams, poles, roof construction and boards in residential buildings. The method used for the inventory of rare plants is done by using the method of vegetation analysis, the parameters in this study were the type, number, diameter and height of the Dryobalanops aromatica Gaertn. tree. Vegetation analysis was carried out by plotting 13 plots, each sample plot measuring 20 m x 20 m. The sample used is a total sample using a random sampling system method, the sampling chossen by exploring the area of the Tahura Lae Kombih area where the Dryobalanops aromatica Gaertn is located. The results shows that the plant species were found starting from the seedling, sapling, pole and tree levels. The most dominant species in this area is Dryobalanops aromatica Gaertn that has Important Value Indeks of seedling 163%, Sapling 270%, Pole 283% and Tree 216%.

First Report of Anthracnose on Cinnamomum camphora (Camphor tree) Caused by Colletotrichum fioriniae and Colletotrichum siamense in China

Camphor tree (Cinnamomum camphora) is native to east Asia, which could produce pharmaceutical metabolites, such as camphor, linalool, and so on (Chen, Tang et al. 2020). In September 2020, severe anthracnose symptoms were observed on the leaves of camphor trees in Nanchang, and estimated incidences ranged from 30% to 80%, which could inhibit leaf growth and reduce their biomass. The lesions were appeared on the leaves of annual branchlets, which the irregular dead areas appeared on leaf tips or margins (Figure 1 A and B), sometimes moving onto the shoots and small twigs. For pathogen isolation, fifteen leaves with typical symptom were randomly collected in Jiangxi Agricultural University (N28°45'38", E115°50'0.006") and the fungi were isolated from the symptomatic-asymptomatic junction and cultured on potato dextrose agar (PDA) at 25℃ in darkness. A total of 40 isolates were obtained from tissue samples, in which 32 isolates were identified as belonging to Colletotrichum spp. following the published works (Damm, Cannon et al. 2012, Damm, Cannon et al. 2012, Weir, Johnston et al. 2012). Based on the morphologies of conidia, all the 32 isolates were classified into two categories. For further precise identification, the represented isolate YK1 and YK18 were selected to analyzed using morphological characters after 7 days of incubation, and multiple genes including ITS (White, Bruns et al. 1990), ACT, GAPDH, TUB (Damm, Woudenberg et al. 2009) and RPB2 (Réblová, Gams et al. 2011). Sequences were deposited in GenBank with accession numbers from MZ229311 to MZ229326. Conidia of isolate YK1 were aseptate, primarily fusiform and measured 14.07-21.21 µm × 4.99-6.79 µm (n = 51) (Figure 1 L) and acervulus were 60.24 to 113.56 µm × 44.24 to 102.63 µm (n = 6) (Figure 1 K), while that of YK18 were one-celled, cylindric with obtuse ends (Figure 1 N) and measured 13.28-16.51 µm × 4.10-5.82 µm (n = 52) and acervulus were 73.85 to 131.70 µm × 63.93 to 105.66 µm (n = 6) (Figure 1 M). Acervulus of isolate YK1 and YK18 were produced on alfalfa stems 40 days after inoculation and dark brown to black in color. For all the genes showed greater than 99% similarity to multiple C. fioriniae and C. siamense accessions, respectively. The phylogram reconstructed from the combined dataset using W-IQ-TREE (Trifinopoulos, Nguyen et al. 2016) showed that isolate YK1 and YK18 clustered with C. fioriniae and C. siamense, respectively. Pathogenicity of both species was tested in the field by ten inoculating surface-sterilized mature leaves with puncture wound (Figure 1 C and D) and ten non-wounded young leaves with 20 µL of a conidial suspension (105 conidia ml-1) (Figure F and G). Leaves treated with sterilized water under the same conditions served as controls. After 4 to 7 days, the inoculated leaves of camphor tree developed typical dark brown to black lesions, similar to symptoms observed in the field, whereas controls remained symptomless. To fill the Koch’s postulates, C. fioriniae and C. siamense were consistently re-isolated, and confirmed morphologically and molecularly. C. siamense have been found to cause anthracnose on Cinnamomum camphora in China (Xu, 2017). To our knowledge, this is the first report of anthracnose on Cinnamomum camphora with C. fioriniae in China. In addition, this is an indication to the complexes about pathogens to anthracnose on camphor tree, which can pose serious threat to the production of Cinnamomum camphora in China.

The genome of the camphor tree and the genetic and climatic relevance of the top-geoherbalism in this medicinal plant

Camphor tree (Cinnamomum camphora (L.) J. Presl), a species in the magnoliid family Lauraceae, is known for its rich volatile oils and is used as a medical cardiotonic and as a scent in many perfumed hygiene products. Here, we present a high-quality chromosome-scale genome of C. camphora with a scaffold N50 of 64.34 Mb and an assembled genome size of 755.41 Mb. Phylogenetic inference revealed that the magnoliids are a sister group to the clade of eudicots and monocots. Comparative genomic analyses identified two rounds of ancient whole-genome duplication (WGD). Tandem duplicated genes exhibited a higher evolutionary rate, a more recent evolutionary history and a more clustered distribution on chromosomes, contributing to the production of secondary metabolites, especially monoterpenes and sesquiterpenes, which are the principal essential oil components. Three-dimensional analyses of the volatile metabolites, gene expression and climate data of samples with the same genotype grown in different locations showed that low temperature and low precipitation during the cold season modulate the expression of genes in the terpenoid biosynthesis pathways, especially TPS genes, which facilitates the accumulation of volatile compounds. Our study lays a theoretical foundation for policy-making regarding the agroforestry applications of camphor tree.

First Report of Leaf Spot Disease on Cinnamomum camphora (Camphor tree) Caused by Epicoccum poaceicola in China

As an important industrial, pharmaceutical and evergreen shade tree (Singh and Jawaid 2012), the camphor tree (Cinnamomum camphora) has been coppiced in Jiangxi Province, China. From 2017 to 2020, we noticed many camphor trees with leaf spots, with an incidence estimated at 50 to 75%, which could severely inhibit leaf growth and reduce their biomass. A dark-green circle with a watery spot appeared on the infected leaves at the initial stage, and necrosis with forming shot-spots surrounded by yellow halos occurred (Figure 1 A). Five leaves with typical symptoms were sampled and washed with tap water for ca. 15 min. Isolation and morphological analysis were performed following the method of Bao et al. (2019). Among 61 fungal isolates, 49 showed the same culture characters. Colonies on PDA were villose and regular, the reverse was scarlet at the edge of the colony, which was ca. 8.75 cm after 7 days of inoculation (Figure 1 I). Chlamydospores were aseptate, dark brown, smooth, in chains or solitary, ellipsoidal to ovoid, 4.8–9.6 × 4.8–11.1 μm (Figure 1 J). The pycnidia were produced on PDA and varied from 47.4 to 85.8 µm (mean 60.2 µm) × 38.6 to 66.8 μm (mean 49.7 μm) (n = 17) (Figure 1 K). Conidia were hyaline, unicellular, elliptical to ovoid, 4.3-6.4 µm (mean 5.1 µm) × 2.3-3.3 µm (mean 2.8 µm) (n = 52) (Figure 1 L). Pathogenicity tests of isolate XW-9 was carried out in the field. Ten leaves were wounded with a sterilized insect needle and inoculated with mycelium plugs (7-mm diameter). Non-colonized PDA plugs served as the negative controlIn addition, conidial suspensions (105 conidia/mL) of isolate XW-9 were sprayed on surface-sterilized leaves with a further ten leaves being sprayed with sterile water as the control. Symptoms described in this study appeared in 100% of the mycelium-inoculated leaves and more than 80% of the conidium-inoculated leaves after 7 days post-inoculation (Figure 1 B-E). No symptoms were seen in the controls (Figure 1 C). Three days after inoculation, brown spots resembling those observed in the field developed on the inoculated leaves, and some lesions turned into shot holes on the infected leaves (Figure 1 G & H). However, no symptoms were observed on the controls (Figure 1 F). The fungus was re-isolated from the margins of the leaf spots and labelled P-XW-9A. The gene regions for ITS, LSU, tub2, RPB2 and ACT of isolates XW-9 and P-XW-9A were amplified and sequenced. The sequences of rDNA-ITS, LSU, tub2, RPB2 and ACT of XW-9 were GenBank MW142397, MW130844, MW165322, MW446945 and MW165324, respectively and those of P-XW-9A were GenBank MW142398, MW130845, MW165323, MW446946 and MW165325, respectively (Lumbsch, et al. 2000; Aveskamp, et al. 2009; Hou et al. 2020). Phylogenetic analysis using concatenated sequences of ITS, LSU, RPB2, and tub2 showed that isolates XW-9 and P-XW-9A formed a single clade with the reference strain of E. poaceicola CBS 987.95 (Figure 2). Thus, XW-9 was identified as E. poaceicola based on its morphological and molecular characteristics. Significantly, the recovered isolate P-XW-9A also aligned with E. poaceicola fulfilling the criteria for Koch's Postulates. E. poaceicola was only reported as a fungal pathogen of Phyllostachys viridis in China (Liu et al. 2020). To our knowledge, this is the first report of leaf spot disease on camphor trees caused by E. poaceicola in China and our findings will be useful for its management.

Genome-Wide Identification and Functional Characterization of the Trans-Isopentenyl Diphosphate Synthases Gene Family in Cinnamomum camphora

Trans-isopentenyl diphosphate synthases (TIDSs) genes are known to be important determinants for terpene diversity and the accumulation of terpenoids. The essential oil of Cinnamomum camphora, which is rich in monoterpenes, sesquiterpenes, and other aromatic compounds, has a wide range of pharmacological activities and has therefore attracted considerable interest. However, the TIDS gene family, and its relationship to the camphor tree (C. camphora L. Presl.), has not yet been characterized. In this study, we identified 10 TIDS genes in the genome of the C. camphora borneol chemotype that were unevenly distributed on chromosomes. Synteny analysis revealed that the TIDS gene family in this species likely expanded through segmental duplication events. Furthermore, cis-element analyses demonstrated that C. camphora TIDS (CcTIDS) genes can respond to multiple abiotic stresses. Finally, functional characterization of eight putative short-chain TIDS proteins revealed that CcTIDS3 and CcTIDS9 exhibit farnesyl diphosphate synthase (FPPS) activity, while CcTIDS1 and CcTIDS2 encode geranylgeranyl diphosphate synthases (GGPPS). Although, CcTIDS8 and CcTIDS10 were found to be catalytically inactive alone, they were able to bind to each other to form a heterodimeric functional geranyl diphosphate synthase (GPPS) in vitro, and this interaction was confirmed using a yeast two-hybrid assay. Furthermore, transcriptome analysis revealed that the CcTIDS3, CcTIDS8, CcTIDS9, and CcTIDS10 genes were found to be more active in C. camphora roots as compared to stems and leaves, which were verified by quantitative real-time PCR (qRT-PCR). These novel results provide a foundation for further exploration of the role of the TIDS gene family in camphor trees, and also provide a potential mechanism by which the production of camphor tree essential oil could be increased for pharmacological purposes through metabolic engineering.

Camphor tree, Cinnamomum camphora (L.); Ethnobotany and pharmacological updates

This review paper highlights the recent updates of the fragrant camphor tree (Cinnamomum camphora) and camphor oil is used as a medicine for controlling many human diseases, relief of pain, inflammation and irritation in the body and skin. It can also be very effective in treating and preventing some serious, life threatening diseases. Recently medicinal plants (sweet worm wood; Artemisia annua) containing camphor essential oil has been tested against corona virus (SARS-CoV-2) disease (Covid-19). Commercially, camphor is very important with many biological properties and is used as a topical ointment since camphor is a very toxic substance and oral consumption of camphor should be avoided.

Autotrophic and heterotrophic contributions to soil respiration in a subtropical camphor tree forest

Understanding the contributions of autotrophic respiration (Ra) and heterotrophic respiration (Rh) to total soil respiration (Rs) is necessary for accurate prediction of global carbon balance and net ecosystem production under environmental change. In this research, annual Rs and Rh and estimated were investigated by using a root trenching experiment in a Camphor tree (Cinnamomum camphora) forest in subtropical China for two years to qualify the relative contribution of Ra and Rh components to Rs, and to determine the environmental factors that control the seasonal changes in Ra, Rh and Rs. The results showed that annual mean Rs was 405 ± 219 gC m-2 year-1 in the studied forests, of which Rh and Ra were 240 ± 120 gC m-2 year-1 and 164 ±102 gC m-2 year-1, respectively. The contribution of Rh to Rs averaged 58.1%, ranging from 45 to 81%. The seasonal changes in Rs and Rh were highly correlated with soil temperature, but not to soil water content. Our results suggest microbial community and activity make a primary contribution to carbon flux released from soil to atmosphere in the studied forest ecosystems.