The assessment of genetic diversity is essential for the conservation and breeding purposes. This study aimed to assess and evaluate the genetic diversity of 116 accessions of mango (Mangifera indica) germplasm using microsatellite markers. The DNA was extracted from young fresh leaf before genotyping using microsatellites to determine the allele size. The analysis of 20 polymorphic microsatellite markers revealed a total of 122 alleles ranging from two (MiIIHR10, MiIIHR21, and MiIIHR25) to 11 alleles per locus (MiIIHR28 and MiIIHR30) with an average of 6.1. The mean polymorphic information content (PIC) value was 0.4585 which ranged between 0.0081 (MiIIHR10) to 0.9573 (MiIIHR28). The UPGMA dendrogram indicated that the accessions were divided into two major clusters, which were divided into several sub-clusters based on their genetic distance matrix values. Some accessions were highly similar to each other, probably due to the duplication of collected accessions or insufficient microsatellite markers to differentiate them. The analysis of the population structure of the individuals also showed two subpopulations, suggesting that the accessions could be separated into two groups, which supported the generated dendrogram. The findings of this study facilitate improved conservation management of the germplasm and help to find strategies for future breeding activities.
Clonal propagation is widely used for Acacia breeding and commercial planting. When a large number of clones are handled, problems with mixings are commonly confronted. Detection of admixture in Acacia clones based on morphology particularly at seedling stage is not feasible. However, molecular markers are commonly used to test the genetic fidelity of planting materials. This paper reports the detection of mislabelling in Acacia clonal progenies using a sequence tagged microsatellite (STMS) genetic marker system. Progenies from two mapping populations were clonally propagated and field planted for phenotypic and genotypic evaluation at three locations in Malaysia: (a) Forest Research Institute Malaysia field station at Segamat, Johor, (b) Borneo Tree Seeds and Seedlings Supplies Sdn, Bhd. (BTS) field trial site at Bintulu, Sarawak and (c) Asiaprima RCF field trial site at Lancang, Pahang. During field planting mislabelling was reported at Segamat, Johor and similar was suspected for Bintulu, Sarawak. Screening revealed mislabelling events in both populations. A total of 18.52% mislabelling incidences were detected from both sites, of which 17.39% of mislabelling was detected for fibre length cross and 20% for wood density cross. The assignment test efficiently reestablished the mislabelled ramets to the respective clones. Future studies should be focused on the utilization of a higher number of markers, e.g. SSR or SNPs to increase a discrimination power. A high number of SNPs can be generated within a short period of time compared to SSR, but SNPs could be cost inhibitory. Multiplexing microsatellite combinations along with sample bulking will further reduce the processing time when screening large populations. The use of assignment test would efficiently assign mislabelled individuals to the respective clones. It is concluded that checking for mislabelling is imperative for future breeding and for analyses such as QTL mapping where a correlation between genotypic and phenotypic data is determined.
Mango, or Mangifera indica as it is scientifically known, is a fruit crop that has significant importance to the global economy. This study characterised 10 commercially potential mango progenies using a total of 10 SSR markers. The analysis of the 10 SSR markers revealed the presence of 34 alleles. Each locus had 2–6 alleles equal to an average of 3.40 alleles per locus. The data also showed 0.115 (MiIIHR21) to 0.692 (MiIIHR28) polymorphic information content (PIC), equal to an average of 0.431. Each locus had heterozygosity and gene diversity values ranging from 0.044 (M1) to 0.957 (MiIIHR29) and from 0.122 (MiIIHR21) to 0.730 (MiIIHR29), respectively, averaging 0.480 and 0.476 each. The range of the pairwise genetic distance was 0.000 (ND014«ND019) to 0.611 (ND020«SB003). The dendrogram analysis showed that each mango progeny was successfully differentiated using the 10 SSR markers except for ND014 and ND019. The allele combination using these 10 SSR markers could be used to develop a DNA profile for fingerprinting purposes for each progeny except for ND014 and ND019, which can only be differentiated with the addition of an extra marker. The developed DNA fingerprinting profile could be used for cultivar identification and authentication. Additionally, breeders could also use the developed DNA profiles for plant variety protection (PVP) purposes.
More than 200 different cultivars of durian exist worldwide but Durio zibethinus or Musang King (MK) is the most premium and prized durian fruit among the recommended varieties. Early identification of this premium variety is critical to protect from non-authentic MK durian cultivars. However, the MK variety's morphological traits are nearly identical to other varieties. Currently, the identification of durian varieties is mostly performed via evaluation of leaf shape, fruit shape, aroma, taste and seed shape and this requires trained personnel for the morphology observation. To enable the rapid identification of the MK variety, PCR amplification of ten durian varieties using six gene candidates from the chloroplast genome was first performed to obtain DNA probes that were specific to the MK durian variety. PCR amplification of ten durian varieties using primers designed confirmed that the nadhA gene sequence showed an obvious difference in the MK variety from other durian varieties. The unique sequence of MK was used as a DNA probe to develop an electrochemical biosensor for the direct identification of the MK durian variety. The electrochemical biosensor was based on the hybridization response of the immobilized DNA probe with the target DNA from the MK variety and was monitored via differential pulse voltammetry technique. Under optimal conditions, the DNA electrochemical biosensor showed a low detection limit at 10% of MK genomic DNA concentration with a wide linear calibration range of 0.05–1.5 µM (R2 = 0.9891) and RSD value of 3.77% (n = 3). The results of the developed DNA biosensor provide high promise for the development of portable sensors employed in the determination of MK variety in the field.
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