Kwanjai Pipatchartlearnwong

Bio: Kwanjai Pipatchartlearnwong is an academic researcher from Kasetsart University. The author has contributed to research in topics: Palm sugar & Amplified fragment length polymorphism. The author has an hindex of 2, co-authored 3 publications receiving 14 citations.

Genetic evidence of multiple invasions and a small number of founders of Asian Palmyra palm (Borassus flabellifer) in Thailand

Abstract: Borassus flabellifer or Asian Palmyra palm is an important crop for local economies in the South and Southeast Asia for its fruit and palm sugar production. Archeological and historical evidence indicated the presence of this species in Southeast Asia dating back at least 1500 years. B. flabellifer is believed to be originated in Africa, spread to South Asia and introduced into Southeast Asia through commercial routes and dissemination of cultures, however, the nature of its invasion and settlement in Thailand is unclear. Here, we analyzed genetic data of 230 B. flabellifer accessions across Thailand using 17 EST-SSR and 12 gSSR polymorphic markers. Clustering analysis revealed that the population consisted of two genetic clusters (STRUCTURE K = 2). Cluster I is found mainly in southern Thailand, while Cluster II is found mainly in the northeastern. Those found in the central are of an extensive mix between the two. These two clusters are in moderate differentiation (F ST = 0.066 and N M = 3.532) and have low genetic diversity (HO = 0.371 and 0.416; AR = 2.99 and 3.19, for the cluster I and II respectively). The minimum numbers of founders for each genetic group varies from 3 to 4 individuals, based on simulation using different allele frequency assumptions. These numbers coincide with that B. flabellifer is dioecious, and a number of seeds had to be simultaneously introduced for obtaining both male and female founders. From these data and geographical and historical evidence, we hypothesize that there were at least two different invasive events of B. flabellifer in Thailand. B. flabellifer was likely brought through the Straits of Malacca to be propagated in the southern Thailand as one of the invasive events before spreading to the central Thailand. The second event likely occurred in Khmer Empire, currently Cambodia, before spreading to the northeastern Thailand.

Towards sex identification of Asian Palmyra palm (Borassus flabellifer L.) by DNA fingerprinting, suppression subtractive hybridization and de novo transcriptome sequencing

Abstract: Background Asian Palmyra palm, the source of palm-sugar, is dioecious with a long juvenile period requiring at least 12 years to reach its maturity. To date, there is no reliable molecular marker for identifying sexes before the first bloom, limiting crop designs and utilization. We aimed to identify sex-linked markers for this palm using PCR-based DNA fingerprinting, suppression subtractive hybridization (SSH) and transcriptome sequencing. Methods DNA fingerprints were generated between males and females based on RAPD, AFLP, SCoT, modified SCoT, ILP, and SSR techniques. Large-scale cloning and screening of SSH libraries and de novo transcriptome sequencing of male and female cDNA from inflorescences were performed to identify sex-specific genes for developing sex-linked markers. Results Through extensive screening and re-testing of the DNA fingerprints (up to 1,204 primer pairs) and transcripts from SSH (>10,000 clones) and transcriptome data, however, no sex-linked marker was identified. Although de novo transcriptome sequencing of male and female inflorescences provided ∼32 million reads and 187,083 assembled transcripts, PCR analysis of selected sex-highly represented transcripts did not yield any sex-linked marker. This result may suggest the complexity and small sex-determining region of the Asian Palmyra palm. To this end, we provide the first global transcripts of male and female inflorescences of Asian Palmyra palm. Interestingly, sequence annotation revealed a large proportion of transcripts related to sucrose metabolism, which corresponds to the sucrose-rich sap produced in the inflorescences, and these transcripts will be useful for further understanding of sucrose production in sugar crop plants. Provided lists of sex-specific and differential-expressed transcripts would be beneficial to the further study of sexual development and sex-linked markers in palms and related species.

The complete chloroplast genome sequence of Asian Palmyra palm (Borassus flabellifer).

Abstract: Borassus flabellifer or Asian Palmyra palm is widely distributed in South and Southeast Asia and is horticultural and economic importance for its fruit and palm sugar production. However, its population is in rapid decline, and only a few genetic data are available. We sequenced the complete chloroplast (cp) genome of B. flabellifer to provide its genetic data for further utilization. The cp genome was obtained by Illumina sequencing and manual gap fillings providing 160,021 bp in length containing a pair of inverted repeats (IRs) with 27,256 bp. These IRs divide the genome into a large single copy region 87,444 bp and a small single copy region 18,065 bp. In total, 113 unique genes, 134 SSRs and 47 large repeats were identified. This is the first complete cp genome reported in the genus Borassus. A comparative analysis among members of the Borasseae tribe revealed that the B. flabellifer cp genome is, so far, the largest and the cp genomes of this tribe have a similar structure, gene number and gene arrangement. A phylogenetic tree reconstructed based on 74 protein-coding genes from 70 monocots demonstrates short branch lengths indicating slow evolutionary rates of cp genomes in family Arecaceae.

Hydrogel Containing Borassus flabellifer L. Male Flower Extract for Antioxidant, Antimicrobial, and Anti-Inflammatory Activity

Abstract: Borassus flabellifer L. is a plant in Arecaceae family, widely distributed and cultivated in tropical Asian countries. The purpose of this study was to identify the bioactive compounds of B. flabellifer L. male flower ethanolic extract and investigate the antioxidant, anti-inflammatory, and antibacterial activities against Cutibacterium acnes. Total phenolic compounds and total flavonoids in B. flabellifer L. male flower ethanolic extract were determined by the Folin–Ciocalteu method and aluminum chloride colorimetric assay, respectively. Active substances in the extract and their quantities were analyzed by liquid chromatography and mass spectrometry (LC–MS/MS). The antioxidant evaluation was carried out using DPPH, ABTS free radical scavenging assays, and FRAP assay. C. acnes inhibitory activity was performed by the broth microdilution method. Anti-inflammatory activity was determined by the protein denaturation assay. In addition, gel containing different amounts of B. flabellifer L. male flower extract was formulated. The physical stability of the gel was observed by measuring viscosity and pH after six heating and cooling cycles, as well as 1-month storage at 4, 30, and 45 °C. The total phenolic content in the extract was 268.30 ± 12.84 mg gallic acid equivalent/g crude dry extract. The total flavonoid contents in the extract were 1886.38 ± 55.86 mg quercetin equivalent/g extract and 2884.88 ± 128.98 mg EGCG equivalent/g extract, respectively. The LC–MS/MS analysis revealed the presence of gallic acid, coumarin, and quercetin and the concentrations of quercetin, coumarin, and gallic acid in B. flabellifer male flower ethanolic extract were 0.912, 0.021, and 1.610 µg/mL, respectively. DPPH and ABTS antioxidant assays indicated that the B. flabellifer L. male flower extract had IC50 values of 31.54 ± 0.43 and 164.5 ± 14.3 µg/mL, respectively. FRAP assay revealed that the B. flabellifer male flower extract had high ferric ion reducing power. The extract was able to inhibit C. acnes bacteria with a minimum inhibitory concentration (MIC) of 250 mg/mL. At 250 and 500 µg/mL, the extract demonstrated the highest anti-inflammatory activity. The gel containing 31.25% w/w and 62.5% w/w showed good physical stability after six heating and cooling cycles, as well as 1-month storage.

Transferability, development of simple sequence repeat (SSR) markers and application to the analysis of genetic diversity and population structure of the African fan palm ( Borassus aethiopum Mart.) in Benin

Abstract: In Sub-Saharan Africa, Borassus aethiopum Mart. (African fan palm) is an important non-timber forest product-providing palm that faces multiple anthropogenic threats to its genetic diversity. However, this species is so far under-studied, which prevents its sustainable development as a resource. The present work is a first attempt at characterizing the genetic diversity and population structure of B. aethiopum across nine collection sites spanning the three climatic regions of Benin, West Africa, through the use of microsatellite markers. During a first phase we relied on the reported transferability of primers developed in other palm species. We find that, in disagreement with previously published results, only 22.5% of the markers tested enable amplification of B. aethiopum DNA and polymorphism detection is very low. In a second phase, we generated a B. aethiopum-specific genomic dataset through high-throughput sequencing and used it for the de novo detection of microsatellite loci. Among the primer pairs targeting these, 11 detected polymorphisms and were further used for analyzing genetic diversity. Across the nine sites, expected heterozygosity (He) ranges from 0.263 to 0.451 with an overall average of 0.354, showing a low genetic diversity. Analysis of molecular variance (AMOVA) shows that within-site variation accounts for 53% of the genetic variation. Accordingly, the low number of migrants and positive values of the fixation index (F) in sites from both the Central (Sudano-Guinean) and the Southern (Guinean) climatic regions suggest limited gene flow between sites. The global correlation between genetic and geographic distances is weak; however, our clustering analyses indicate that B. aethiopum palms from Save (Center) are genetically more similar to those from the North than to samples from other Central sites. In the light of our results, we discuss the use of inter-species transfer vs. de novo development of microsatellite markers in genetic diversity analyses targeting under-studied species, and suggest future applications for our molecular resources. We propose that, while prominent short-range pollen and seed dispersal in Benin explain most of our results, gene flux between the Central and Northern regions, as a result of animal and/or human migrations, might underlie the Save discrepancy.