Analysis of genetic diversity and discrimination of Oil Palm DxP populations based on the origins of pisifera elite parents.

A total of 251 Dura cross Pisifera (DxP) hybrid palms from six populations descending from six parental African Pisifera origins and involving 12 progenies were analyzed with 19 selected Simple Sequence Repeats (SSR) markers. A total of 110 alleles were produced, ranging from three to eight per SSR, with a mean of 5.8 alleles per SSR locus. Of these, 68.5% were considered shared alleles by more than one population and the remaining 31.5% were population specific alleles. They generated between six and 21 haplotypes in all populations, and depending on the SSR marker, between one and 10 haplotypes within populations. Various parameters for analyzing genetic variability, differentiation and genetic structure were computed using GenAlEx, Structure and Darwin software. The obtained results confirmed microsatellites as a robust, feasible and trustful method for obtaining DNA fingerprints, tracing the source of oil palm samples. With respect to the authenticity of materials or for solving legitimacy issues, accession belonging to each population by SSR markers could be distinguished, but additional SSR should be screened for improving this process.

Isolation and Solubilisation of Inorganic Phosphate by Burkholderia spp. from the Rhizosphere of Oil Palm.

BACKGROUND AND OBJECTIVE: Phosphate-solubilising bacteria (PSB) are useful for plant growth. They inhabit different soil ecosystems such as colonizing the root environment. The aims of this study was to isolate PSB from oil palm rhizosphere and to conduct a comparative analysis of the solubility of inorganic phosphates. MATERIALS AND METHODS: Rhizospheric soil samples at 0-20 cm depth collected from the distance of 2 m away from the palm were isolated and their chemical and physical properties were analyzed. Qualitative estimation of the suspected PSB was screened by inoculating and growing them at 27°C for 10 days on NBRIP agar medium with bromophenol blue. Their abilities to solubilize AlPO4, FePO4 and Ca3(PO4)2 were examined. Phosphate solubilizing activities were tested on the NBRIP growth medium by analyzing solubilisation efficiency and soluble-P content. Genomic DNA was isolated using QIAamp® genomic DNA kit. RESULTS: A total of 15 PSB were successfully isolated from oil palm rhizosphere. During 5 days of incubation, isolate K3.1, A4 and K3.3 solubilized 53.5, 63.5 and 58.6 mg L-1 phosphate inoculated in Al3PO4, Fe3PO4 and Ca3(PO4)2, respectively. Based on the 16S rRNA gene sequence analysis, those isolates were closely related to Burkholderia arboris, Burkholderia gladioli and Burkholderia seminalis, respectively. In soil analysis, P2O5, C-organic and CEC had positive correlation with the total PSB. CONCLUSION: The existence of P promoting bacteria in oil palm rhizosphere may offer effective solution on biofertilizer agent for sustainable agriculture.

Construction of a high density linkage map in Oil Palm using SPET markers.

A high-density genetic linkage map from a controlled cross of two oil palm (Elaeis guineensis) genotypes was constructed based on Single Primer Enrichment Technology (SPET) markers. A 5K panel of hybridization probes were used for this purpose which was derived from previously developed SNP primers in oil palm. Initially, 13,384 SNPs were detected which were reduced to 13,073 SNPs after filtering for only bi-allelic SNP. Around 75% of the markers were found to be monomorphic in the progeny, reducing the markers left for linkage mapping to 3,501. Using Lep-MAP3 software, a linkage map was constructed which contained initially 2,388 markers and had a total length of 1,370 cM. In many cases several adjacent SNP were located on the same locus, due to missing recombination events between them, leading to a total of 1,054 loci on the 16 LG. Nevertheless, the marker density of 1.74 markers per cM (0.57 cM/marker) should allow the detection of QTLs in the future. This study shows that cost efficient SPET markers are suitable for linkage map construction in oil palm and probably, also in other species.

Detection of significant SNP associated with production and oil quality traits in interspecific oil palm hybrids using RARSeq.

A RARSeq based Association mapping study was performed in a population of 104 Elaeis oleifera x E. guineensis hybrids of five origins with the aim of finding functional markers associated to six productive and 19 oil quality traits. For this purpose mRNA of each genotype was isolated and double stranded cDNA was synthesized. Following digestion with two restriction enzymes and adapter ligation, a size selected pool of barcoded amplicons was produced and sequenced using Illumina MiSeq. The obtained sequences were processed with a "snakemake" pipeline, filtered and missing values were imputed. For all traits except two significant effects of the origin was observed. Genetic diversity analyses revealed high variability within origins and an excess of heterozygosity in the population. Two GLM models with Q matrix or PCA matrix as covariates and two MLM models incorporating in addition a Kinship matrix were tested for genotype-phenotype associations using GAPIT software. Using unadjusted p values (< 0.01) 78 potential associations were detected involving 25 SNP and 20 traits. When applying FDR multiple testing with p < 0.05, 25 significant associations remained involving eight SNP and six quality traits. Four SNP were located in genes with a potential relevant biological meaning.

Association Mapping Between Candidate Gene SNP and Production and Oil Quality Traits in Interspecific Oil Palm Hybrids.

Oil palm production is gaining importance in Central and South America. However, the main species Elaeis guineensis (Eg) is suffering severely from bud rod disease, restricting the potential cultivation areas. Therefore, breeding companies have started to work with interspecific Elaeis oleifera × Eg (Eo × Eg) hybrids which are tolerant to this disease. We performed association studies between candidate gene (CG) single nucleotide polymorphisms (SNP) and six production and 19 oil quality traits in 198 accessions of interspecific oil palm hybrids from five different origins. For this purpose, barcoded amplicons of initially 167 CG were produced from each genotype and sequenced with Ion Torrent. After sequence cleaning 115 SNP remained targeting 62 CG. The influence of the origins on the different traits was analyzed and a genetic diversity study was performed. Two generalized linear models (GLM) with principle component analysis (PCA) or structure (Q) matrixes as covariates and two mixed linear models (MLM) which included in addition a Kinship (K) matrix were applied for association mapping using GAPIT. False discovery rate (FDR) multiple testing corrections were applied in order to avoid Type I errors. However, with FDR adjusted p values no significant associations between SNP and traits were detected. If using unadjusted p values below 0.05, seven of the studied CG showed potential associations with production traits, while 23 CG may influence different quality traits. Under these conditions the current approach and the detected candidate genes could be exploited for selecting genotypes with superior CG alleles in Marker Assisted Selection systems.