Plastid DNA Barcoding and RtActin cDNA Fragment Isolation of Reutealis Trisperma: A Promising Bioresource for Biodiesel Production.

Reutealis trisperma belonging to the family Euphorbiaceae is currently used for biodiesel production, and rapid development in plant-based biofuel production has led to its increasing demand. However, massive utilization of bio-industrial plants has led to conservation issues. Moreover, genetic information on R trisperma is still limited, which is crucial for developmental, physiological, and molecular studies. Studying gene expression is essential to explain plant physiological processes. Nonetheless, this technique requires sensitive and precise measurement of messenger RNA (mRNA). In addition, the presence of internal control genes is important to avoid bias. Therefore, collecting and preserving genetic data for R trisperma is indispensable. In this study, we aimed to evaluate the application of plastid loci, rbcL, and matK, to the DNA barcode of R trisperma for use in conservation programs. In addition, we isolated and cloned the RtActin (RtACT) gene fragment for use in gene expression studies. Sequence information was analyzed in silico by comparison with other Euphorbiaceae plants. For actin fragment isolation, reverse-transcription polymerase chain reaction was used. Molecular cloning of RtActin was performed using the pTA2 plasmid before sequencing. We successfully isolated and cloned 592 and 840 bp of RtrbcL and RtmatK fragment genes, respectively. The RtrbcL barcoding marker, rather than the RtmatK plastidial marker, provided discriminative molecular phylogenetic data for R Trisperma. We also isolated 986 bp of RtACT gene fragments. Our phylogenetic analysis demonstrated that R trisperma is closely related to the Vernicia fordii Actin gene (97% identity). Our results suggest that RtrbcL could be further developed and used as a barcoding marker for R trisperma. Moreover, the RtACT gene could be further investigated for use in gene expression studies of plant.

In Silico Analysis of Partial Fatty Acid Desaturase 2 cDNA From Reutealis trisperma (Blanco) Airy Shaw.

Reutealis trisperma oil is a new source for biodiesel production. The predominant fatty acids in this plant are stearic acid (9%), palmitic acid (10%), oleic acid (12%), linoleic acid (19%), and α-eleostearic acid (51%). The presence of polyunsaturated fatty acids (PUFAs), linoleic acid, and α-eleostearic acid decreases the oxidation stability of R. trisperma biodiesel. Although several studies have suggested that the fatty acid desaturase 2 (FAD2) enzyme is involved in the regulation of fatty acid desaturation, little is known about the genetic information of FAD2 in R. trisperma. The objectives of this study were to isolate, characterize, and determine the relationship between the R. trisperma FAD2 fragment and other Euphorbiaceae plants. cDNA fragments were isolated using reverse transcription polymerase chain reaction (PCR). The DNA sequence obtained by sequencing was used for further analysis. In silico analysis identified the fragment identity, subcellular localization, and phylogenetic construction of the R. trisperma FAD2 cDNA fragment and Euphorbiaceae. The results showed that a 923-bp partial sequence of R. trisperma FAD2 was successfully isolated. Based on in silico analysis, FAD2 was predicted to encode 260 amino acids, had a domain similarity with Omega-6 fatty acid desaturase, and was located in the endoplasmic reticulum membrane. The R. trisperma FAD2 fragment was more closely related to Vernicia fordii (HM755946.1).

An ethnobotanical study of medicinal plants used by the Tengger tribe in Ngadisari village, Indonesia.

The people of Tengger, Indonesia have used plants as traditional medicine for a long time. However, this local knowledge has not been well documented until recently. Our study aims to understand the utilization of plants in traditional medicine by the people of Tengger, who inhabit the Ngadisari village, Sukapura District, Probolinggo Regency, Indonesia. We conducted semi-structured and structured interviews with a total of 52 informants that represented 10% of the total family units in the village. The parameters observed in this study include species use value (SUV), family use value (FUV), plant part use (PPU), and the relative frequency of citation that was calculated based on fidelity level (FL). We successfully identified 30 species belonging to 28 genera and 20 families that have been used as a traditional medicine to treat 20 diseases. We clustered all the diseases into seven distinct categories. Among the recorded plant families, Poaceae and Zingiberaceae were the most abundant. Plant species within those families were used to treat internal medical diseases, respiratory-nose, ear, oral/dental, and throat problems. The plant species with the highest SUV was Foeniculum vulgare Mill. (1.01), whereas the Aloaceae family (0.86) had the highest FUV. Acorus calamus L. (80%) had the highest FL percentage. The leaves were identified as the most used plant part and decoction was the dominant mode of a medicinal preparation. Out of the plants and their uses documented in our study, 26.7% of the medicinal plants and 71.8% of the uses were novel. In conclusion, the diversity of medicinal plant uses in the Ngadisari village could contribute to the development of new plant-based drugs and improve the collective revenue of the local society.

Transcription profile data of phorbol esters biosynthetic genes during developmental stages in Jatropha curcas.

Jatropha curcas is currently known as an alternative source for biodiesel production. Beside its high free fatty acid content, J. curcas also contains typical diterpenoid-toxic compounds of Euphorbiaceae plant namely phorbol esters. This article present the transcription profile data of genes involved in the biosynthesis of phorbol esters at different developmental stages of leaves, fruit, and seed in Jatropha curcas. Transcriptional profiles were analyzed using reverse transcription-polymerase chain reaction (RT-PCR). We used two genes including GGPPS (Geranylgeranyl diphospate synthase), which is responsible for the formation of common diterpenoid precursor (GGPP) and CS (Casbene Synthase), which functions in the synthesis of casbene. Meanwhile, J. curcas Actin (ACT) was used as internal standard. We demonstrated dynamic of GGPPS and CS expression among different stage of development of leaves, fruit and seed in Jatropha.