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DNA profiling of X-chromosomal short tandem repeats (X-STR) has exceptional value in criminal investigations, especially for complex kinship and incest cases. In this study, Investigator® Argus X-12 Quality Sensor (QS) kits were successfully used to characterize 12 X-STR loci in 199 unrelated healthy Kedayan individuals living in Sabah and Sarawak, Malaysia. The LG1 haplogroup (DXS8378 - DXS10135 - DXS10148) has the largest HD (0.9799) as compared with all other closely linked haplotype groups examined (LG2; DXS7132-DXS10074-DXS10079, LG3; DXS10103-DXS10101-HPRTB and LG4; DXS10134-DXS7423-DXS10146). Data from statistical analysis showed that high combined of PD M , PD F , MEC_ Krüger , MEC_ Kishida , MEC_ Desmarais , and MEC_ Desmarais_duo values (0.999999994405922, 0.99999999999999, 0.999990463834938, 0.999999975914808, 0.999999975985006, and 0.999996491927194, respectively) in the Kedayan. In a two-dimensional scaling (MDS) plot and dendrogram constructed using allele frequencies at the 12 X-STR loci, Kedayan appear to be most closely related to their other Austronesian populations including the Malays and Filipinos as compared with other reference population groups. Findings from the present study thus demonstrate high genetic variability across the 12 tested X-STR loci and can be used for population studies and forensic applications.

In this study, 268 samples for unrelated males belonging to the five major human subpopulation groups in Ghana (Akan, Ewe, Mole-Dagbon, Ga-Dangme and Guang) were genetically characterised for 23 Y chromosome short tandem repeat (STR) loci using the Powerplex® Y23 STR kit. A total of 263 complete haplotypes were recorded of which 258 were unique. The haplotype diversity, discriminating capacity and match probability for the pooled population data were 0.9998, 0.9627 and 0.0039, respectively. The pairwise genetic distance (R ST ) for the Ghanaian datasets and other reference populations deposited in the Y-STR Haplotype Reference Database (YHRD) were estimated and mapped using multidimensional scaling (MDS) plot. The Guang and Ewe were significantly different from the Akan, Mole-Dagbon and Ga-Dangme. However, the five Ghanaian datasets were all plotted close together with other African populations in the MDS data mapping.

Genetic polymorphisms at 23 Y chromosome short tandem repeat (STRs) loci included in the Powerplex® Y23 PCR kit were successfully scored in 128 unrelated Kedayan individuals living in Sabah, East Malaysia. Complete haplotypes were recorded for all individuals and included 92 different types with 72 being unique to single male subjects. Three important forensic statistics were calculated from these data; haplotype diversity = 0.993, discriminating capacity = 0.719, and match probability = 0.015. The Kedayan appear to be most closely related to Malays and Filipinos in a multidimensional scaling plot and are separated from other mainland Asia populations including Thais and Hakka Han. These new data for Kedayan have been deposited in the YHRD database (accession number: YA004621). Our statistical analyses showed the reliability of Y-STR loci for geographically extended use in forensic casework and for studying human population history.

Short repetitive regions in autosomal and Y chromosomes known as short tandem repeats (STRs) are currently used for DNA profiling in crime investigations. However, DNA profiling requires a sufficient quality and quantity of DNA template, which is often not obtained from trace evidence or degraded biological samples collected at the scene of a crime. Here, we assessed autosomal and male DNA components extracted from crime scene and mock casework samples using the Casework Direct Kit, Custom and compared the results against those obtained by extraction of matching samples using well-established Maxwell 16 System DNA IQ Casework Pro Kit. The quantity and quality of extracted DNA obtained using both Casework Direct Kit, Custom and Maxwell 16 System DNA IQ Casework Pro Kit were analyzed using PowerQuant Systems followed by autosomal and Y-chromosome STR profiling using GlobalFiler Express PCR Amplification Kit and PowerPlex Y23 System, respectively. Our results showed that the Casework Direct Kit and Maxwell 16 DNA IQ Casework Pro Kit have more or less equal capacity to extract inhibitor free DNA, but that the latter produces slightly better quality and more DNA template and subsequently higher numbers of STR allele calls for autosomal and Y-STR analyses. Nonetheless, the Casework Direct Kit, Custom is the quicker and cheaper option for extraction of good, clean DNA from high content material and might best be used for extraction of reference samples. Such reference DNA samples typically come from buccal swabs or freshly drawn blood. So, in general, they can confidently be expected to have a high nucleic acid content and to be inhibitor-free.

Nine commercial varieties of pineapples in Malaysia (Josapine, Maspine, MD2, Sarawak, Gandul, N36, Moris, Crystal Honey and Yankee) were collected from various places in Peninsular Malaysia and analysed for cultivar identifications using nine simple sequence repeat (SSR) markers. A total of 27 alleles have been observed which ranged from 2 to 5 with an average of 3 alleles per locus. The polymorphic information content (PIC) value ranged from 0.3426 (Acom_82.8) to 0.6561 (Acom_67.2) with a mean of 0.4524 while the heterozygosity value ranged from 0.1097 (TsuAC021) to 0.8010 (TsuAC039) with a mean of 0.5481. The pairwise Nei's genetic distances had also been calculated and the value ranged from 0.0562 (Gandul and Josapine) to 0.6383 (MD2 and Yankee) with an average value of 0.3169. The above data emphasised a moderate level of polymorphisms among the nine varieties. A dendrogram was constructed by using the unweighted pair group method with arithmetic mean (UPGMA) which showed all the nine successfully differentiated pineapple commercial varieties. A principal coordinate analysis (PCoA) was also had been generated which revealed an agreement with the dendrogram output. Therefore, these nine SSR markers can be used to identify the nine selected commercial varieties to ensure pure planting materials. [ABSTRACT FROM AUTHOR]

The progress in next-generation sequencing (NGS) has transformed the discovery and development of microsatellite markers, particularly for understudied species. In this study, one of the NGS platforms adopted was restriction site-associated DNA sequencing (RAD-seq) to identify and develop microsatellite markers for taro. A total of 8,490,604 clean reads were generated and assembled into 222,445 contigs with only 3,280 contigs found to contain a total of 3454 repeats. From this total, only 1,790 repeats were successfully designed as a flanking primer, and 14 microsatellites were selected at random for cultivar identification of Malaysian taro. The analysis of 14 novel simple sequence repeat (SSR) markers only showed that twelve SSR markers were polymorphic with the remaining two SSR markers, CesSSR9 and CesSSR11, monomorphic. The analysis of polymorphic SSR markers revealed the presence of 50 alleles ranging between two (CesSSR5) and seven (CesSSR13) having an average of 4.17 alleles per locus. The values of the polymorphic information content ranged from 0.05 (CesSSR5) to 0.59 (CesSSR7) with an average of 0.33 per locus. The unweighted pair group method with arithmetic mean (UPGMA) dendrogram showed that cultivar MINYAK was unable to be differentiated from cultivar PUTIH. As such, these newly developed microsatellite markers will provide a tool for molecular identification in taro and provide necessary information for future biodiversity conservation and gene bank management of taro. [ABSTRACT FROM AUTHOR]

Bacterial leaf blight (BLB) and blast are among the most destructive diseases of rice. In order to develop new varieties that can resist various races of the pathogens Xanthomonas oryzae pv. oryzae (Xoo) and Magnaporthe oryzae, which cause these two diseases, respectively, it is important to identify suitable markers and a marker-aided selection strategy. Seventeen foreground microsatellites associated with resistance against BLB and blast were screened for polymorphism for possible adoption in a breeding program for resistant cultivar development. A total of 472 background simple sequence repeats (SSRs) were also screened for polymorphism in order to recover the recurrent parent genome. All 72 F1 progenies were also analysed with the 17 foreground and 472 background markers in comparison to the alleles of their parents. The markers were validated using 218 BC2F2 plants. The result revealed six tightly linked polymorphic foreground markers (Xa21FR, pTA248, Xa13prom, MP, RM21 and RM13) associated with resistance against Xoo pathotype P7.7 and two microsatellites (RM6836 and RM8225) associated with resistance against blast pathotype P7.2. A total of 79 polymorphic background markers were identified for recovering the recurrent parent genome. The result also showed that the markers had goodness of fit (blast resistance) to 1:2:1 ratio for single gene model in BC2F2 progenies. These genetically confirmed microsatellite markers were recommended for use in marker-assisted backcross breeding and other marker-assisted selection of BLB and blast resistance genes in order to develop varieties with durable and broad spectrum resistance against the two major rice diseases. [ABSTRACT FROM AUTHOR]

Assessments of genetic diversity have been claimed to be significantly efficient in utilising and managing resources of genetic for breeding programme. In this study, variations in genetic were observed in 65 pineapple accessions gathered from germplasm available at Malaysian Agriculture Research and Development Institute (MARDI) located in Pontian, Johor via 15 markers of simple sequence repeat (SSR). The results showed that 59 alleles appeared to range from 2.0 to 6.0 alleles with a mean of 3.9 alleles per locus, thus displaying polymorphism for all samples at a moderate level. Furthermore, the values of polymorphic information content (PIC) had been found to range between 0.104 (TsuAC035) and 0.697 (Acom_9.9), thus averaging at the value of 0.433. In addition, the expected and the observed heterozygosity of each locus seemed to vary within the ranges of 0.033 to 0.712, and from 0.033 to 0.885, along with the average values of 0.437 and 0.511, respectively. The population structure analysis via method of delta K (ΔK), along with mean of L (K) method, revealed that individuals from the germplasm could be divided into two major clusters based on genetics (K = 2), namely Group 1 and Group 2. As such, five accessions (Yankee, SRK Chalok, SCK Giant India, SC KEW5 India and SC1 Thailand) were clustered in Group 1, while the rest were clustered in Group 2. These outcomes were also supported by the dendrogram, which had been generated through the technique of unweighted pair group with arithmetic mean (UPGMA). These analyses appear to be helpful amongst breeders to maintain and to manage their collections of germplasm. Besides, the data gathered in this study can be useful for breeders to exploit the area of genetic diversity in estimating the level of heterosis. [ABSTRACT FROM AUTHOR]

Objectives: This study includes the direct sequencing of cornulin (CRNN) gene to elucidate the possible mechanism of CRNN downregulation and explore the genetic imbalances at 1q21.3 across oral squamous cell carcinoma (OSCC) samples. Materials and methods: In mutation screening of CRNN gene, gDNA from OSCC tissues were extracted, amplified, and followed by direct sequencing. OSCC samples were also subjected to fragment analysis on CRNN gene to investigate its microsatellite instability (MSI) and loss of heterozygosity (LOH). Immunohistochemistry was performed to validate CRNN downregulation in OSCC samples. Results: No pathogenic mutation was found in CRNN gene, while high frequency of allelic imbalances was found at 1q21.3 region. MSI was found more frequent (25.3 %) than LOH (9.3 %). Approximately 22.6 % of cases had high MSI which reflects higher probability of inactivation of DNA mismatch repair genes. MSI showed significant association with no betel quid chewing ( p = 0.003) and tongue subsite ( p = 0.026). LOH was associated with ethnicity ( p = 0.008) and advanced staging ( p = 0.039). The LOH at 1q21.3 was identified to be as an independent prognostic marker in OSCC (HRR = 7.15 (95 % CI, 1.41-36.25), p = 0.018). Downregulation of CRNN was found among MSI-positive OSCCs and was associated with poor prognosis ( p = 0.044). Conclusion: This study showed a significant correlation between LOH/MSI at 1q21.3 with clinical outcomes and that downregulation of CRNN gene could be considered as a prognostic marker of OSCC. Clinical relevance: Insights of the downregulation mode of CRNN gene lays the basis of drug development on this gene as well as revealing its prognostic value. [ABSTRACT FROM AUTHOR]

The most vital aspect of marker-assisted backcross breeding is the recurrent parent genome recovery. This enables the selection of only parents with recovered recipient/recurrent parent genome in addition to the targeted genes. The recurrent parent genome recovery (RPGR) ensures that non-desirable genomic segments are removed while the gene of interest is sustained in the recombined progenies without further segregations. This work was aimed at quantifying the RPGR of backcross populations with introgression of bacterial leaf blight resistance genes. Putra-1, a Malaysian elite variety, high yielding with inherent resistance to blast but susceptible to bacterial leaf blight (BLB), was crossed with IRBB60 which is resistant to BLB disease. The IRBB60 has four Xoo resistance genes—Xa4, xa5, xa13 and Xa21. Tightly linked polymorphic functional and SSR markers were used for foreground selection at every stage of backcrossing to select progenies with introgressed target genes. Background selection was done to quantify the percentage of RPGR in the selected lines using 79 confirmed polymorphic microsatellites. Result obtained showed that the percentage of RPGR was 80.11% at BC1F1, 95.30% at BC2F1 and 95.9% at BC2F2. The introgression of Xa4, xa5, xa13 and Xa21 Xoo resistance genes were faster through the adopted marker-assisted backcross breeding compared to what could be obtained through conventional breeding. All the 16 selected lines displayed resistance to BLB with three lines showing high resistance to the disease. The blast resistance contained in the genetic background of Putra-1 was also sustained in all the selected lines. The newly developed lines were recommended as new rice varieties for commercial cultivation. [ABSTRACT FROM AUTHOR]