AIM OF STUDY: Therefore, this study was conducted to document the ethnomedicinal knowledge of the Kenyah community. The main objectives of this study are: 1) To determine and document the diversity of medicinal plants used by the Kenyah community, 2) To determine whether the availability of modern medicine has affected Kenyah traditional medicine, and 3) To identify plants which have not been previously cited or used for previously unreported medical uses.
MATERIALS AND METHODS: We conducted repeated interviews and field surveys at the Asap-Koyan Resettlement Area, Belaga Sarawak. A total of 24 respondents from four Kenyah longhouses were interviewed in this study. Individuals possessing extensive traditional medicinal knowledge were identified via preliminary interviews or by viva voce. Translators were employed to ensure that there was no miscommunication. The results were evaluated based on the plant's total use-reports and number of respondents citing the plant. The data was also evaluated based on use-reports by ailment category.
RESULTS: Over 95% of the respondents were 40 years and older (58.21 years old ± 11.21). This was due to the younger members of the community (40 years old and below) admitting that they had almost no knowledge regarding traditional medicine, as they preferred relying on modern medicine. A total of 61 plant species were mentioned by the 24 respondents Seven plants had five or more respondents citing it, which was more than 20% of the respondents. These plants were Piper betle, Homalomena cordata, Senna alata, Annona muricata, Derris elliptica, Blumea balsamifera and Coscinium fenestratum.
CONCLUSION: Almost all of the cited plants had been previously recorded to be used in either Ayurvedic, Chinese herbal medicine, Malay traditional medicine or other Asian ethnomedicinal systems. However, there were four highly cited species that were used for treatments that were scarcely reported in past literature. These were piper betle (used by Kenyah to treat fever), Sauropus andrognus (used by Kenyah to treat fever), Derris elliptica (used by Kenyah to treat fever and influenza) and Coscinuim fenestratum (used by Kenyah to treat toxic effects from non-medical substances).
RESULTS: We show that SYMRK is essential for nodulation and endomycorrhization in Parasponia andersonii. Subsequently, it is revealed that the 5'-intron donor splice site of SYMRK intron 12 is variable and, in most dicotyledon species, doesn't contain the canonical dinucleotide 'GT' signature but the much less common motif 'GC'. Strikingly, in T. orientalis, this motif is converted into a rare non-canonical 5'-intron donor splice site 'GA'. This SYMRK allele, however, is fully functional and spreads in the T. orientalis population of Malaysian Borneo. A further investigation into the occurrence of the non-canonical GA-AG splice sites confirmed that these are extremely rare.
CONCLUSION: SYMRK functioning is highly conserved in legumes, actinorhizal plants, and Parasponia. The gene possesses a non-common 5'-intron GC donor splice site in intron 12, which is converted into a GA in T. orientalis accessions of Malaysian Borneo. The discovery of this functional GA-AG splice site in SYMRK highlights a gap in our understanding of splice donor sites.
METHODS AND RESULTS: In this study, we have developed a simplified, tissue culture-independent protocol to deliver the CRISPR/Cas9 system through in planta transformation in Malaysian rice (Oryza sativa L. subsp. indica cv. MR 219). Sprouting seeds with cut coleoptile were used as the target for the infiltration by Agrobacterium tumefaciens and we achieved 9% transformation efficiency. In brief, the dehusked seeds were surface-sterilised and imbibed, and the coleoptile was cut to expose the apical meristem. Subsequently, the cut coleoptile was inoculated with A. tumefaciens strain EHA105 harbouring CRISPR/Cas9 expression vector. The co-cultivation was conducted for five to six days in a dark room (25 ± 2 °C) followed by rooting, acclimatisation, and growing phases. Two-month-old plant leaves were then subjected to a hygromycin selection, and hygromycin-resistant plants were identified as putative transformants. Further validation through the polymerase chain reaction verified the integration of the Cas9 gene in four putative T0 lines. During the fruiting stage, it was confirmed that the Cas9 gene was still present in three randomly selected tillers from two 4-month-old transformed plants.
CONCLUSION: This protocol provides a rapid method for editing the rice genome, bypassing the need for tissue culture. This article is the first to report the delivery of the CRISPR/Cas9 system for in planta transformation in rice.