Molecular Biology Reports

BACKGRAUNDHomeobox (HOX) transcript antisense RNA (HOTAIR) and HOX genes are reported to be more expressed in various cancers in humans in recent studies. The role of HOTAIR and HOXD genes in acute myeloid leukemia (AML) and chronic myeloid leukemia (CML) is not well known. MATERIALS AND METHODSIn this study, expression levels of HOXD8, HOXD9 and HOXD11 from HOXD gene family and HOTAIR were determined from peripheral blood samples of 30 AML and 30 CML patients and 20 healthy volunteers by quantitative Real Time PCR. RESULTSWe determined that the expression levels of HOXD9 and HOXD11 in the AML patients were significantly lower than the control group (p<0.001 and p=0.002, respectively). There was no significant difference in the expression levels of HOTAIR and HOXD8 when compared to the control group. In the CML patients there was a significant increase in the expression level of HOTAIR when compared to the control group (p=0.002). The expression levels of HOXD9 and HOXD11 were found to be significantly lower than the control group (p<0.001). CONCLUSIONOur study showed that HOTAIR may not be a biomarker in the diagnosis and is not significantly correlated with the clinicopathological prognostic characteristics of AML. Additionally; it can be said that HOTAIR is oncogenic by suppressing the expression of HOXD9 and HOXD11 but not HOXD8 in CML patients. The expression profiles of HOTAIR may be a potential biomarker in the diagnosis of CML patients in predicting and monitoring drug resistance.

BackgroundLymphocyte enhancer factor-1 (LEF-1) is a member of the LEF-1/TCF family of transcription factors that are critically involved in canonical Wnt/{beta}-catenin Signaling to regulate B Lymphocyte proliferation and survival. Alteration of LEF1 expression and function leads to leukemogenesis as well as other several neoplasms. Aimsto identify mutations in exons two and three of the LEF1 among B-CLL Sudanese patients. Also, to functionally analyze the detected SNPs using different in silico tools. Materials and methodsImmuno-phenotype for the detection of B cells CD5 and CD19 markers was performed on 128 B-CLL Sudanese patients by using a flow cytometry technique. DNA extraction, conventional PCR, and Sanger sequencing were applied to the LEF1 gene. Also, we performed a mutational analysis for identified SNPs using bioinformatics tools. ResultsA positive CD5 & CD19 expression was found in B-CLL patients. No mutation was observed in exon two. While four mutations were observed in exon three; two of them were not reported in previous studies. Interestingly, splicing analysis predicted that these mutations could lead to splicing defects in LEF1 pre-mRNA due to their potential effects on splicing regulatory elements (i.e. ESE). Conclusionthe two mutations Pro134Pro and Ile135Asn (novel mutation) were detected in all enrolled CLL patients and they could be used as diagnostic and/or prognostic markers for CLL. Therefore, further in vitro and in vivo functional studies with a large sample size are required to verify the splicing effect of the detected mutations in LEF1 pre-mRNA.

TP53 is a tumour suppressor gene. Its inactivation plays a significant role in the molecular pathology of cancers. TP53 germline mutations increase the risk of developing multiple primary cancers. However, the role of alterations in TP53 germline DNA in head and neck cancers (HNCs) is not well-established. HNCs comprise one of the most frequent cancers in South Asia. The present discovery study reports the investigation of germline variations in the TP53 gene in a cohort of 30 HNC patients from Karachi, Pakistan. Blood samples were collected and genomic DNA was extracted from white blood cells. TP53 has 11 exons, where exon 1 is not transcribed. After quality control of DNA, amplification of seven selected exons along with their splice sites, two intronic regions (introns 2-3 and 3-4), and 3UTR were carried out. Sanger sequencing was carried out in order to identify germline variations. Comparison with wild type sequence revealed rs1642785 G>C (intron 2-3) variation in 63.2%, PIN3 duplication (rs17878362) in intron 3-4 in 94.7%, and rs1042522 G>C in exon 4 (p.R72P) in 66.6% of the cases. In 3UTR, 13.4% of the analyzed cases carried either one of two variants, i.e., 17:7669567_8delCA or 17:7669560C>G. The latter variations are reported for the first time in literature. In conclusion, we report three highly frequent germline variations and two newly discovered variations in 3UTR of TP53 germline DNA in HNC patients from Pakistan. These results shall contribute to delineating the genetic component of HNCs with potential translational implications.

BackgroundChronic Myeloid Leukemia (CML) is initiated in the bone marrow due to the chromosomal translocation t(9;22), resulting in the fusion oncogene BCR-ABL. Tyrosine kinase inhibitors (TKIs) targeting BCR-ABL have transformed fatal CML into an almost curable disease. However, TKIs lose efficacy during disease progression, and the mechanism of CML progression remains to be fully understood. Additionally, common molecular biomarkers for CML progression are lacking. Our studies previously detected ANKRD36 (c.1183_1184 delGC and c.1187_1188 dupTT) associated exclusively with advanced phase CML. However, clinical validation of this finding was pending. Therefore, this study aimed to clinically validate mutated ANKRD36 as a novel biomarker of CML progression. Materials and MethodsThe study enrolled 124 patients in all phases of CML, recruited from Mayo Hospital and Hameed Latif Hospital in Lahore, Punjab, between January 2020 and September 2023. All response criteria were adopted from the European LeukemiaNet guideline 2020. Informed consent was obtained from all study subjects. The study was approved by scientific and ethical review committees of all participating centers. Sanger sequencing was employed to detect ANKRD36 mutations in CML patients in accelerated phase (AP) (n=11) and blast crisis (BC) (n=10), with chronic-phase CML (CP-CML) patients as controls (n=103). Samples were processed using Big Dye Terminator Cycle Sequencing Ready Reaction kits and sequenced using ABI Prism 3730 Genetic Analyzer, and sequencing using forward and reverse primers for ANKRD36. Data was analyzed using SPSS version 26. ResultsDuring our study, 17% of CML patients progressed to advanced phases AP-CML n= 11 (8.9%) and BC-CML n=10 (8.1%). The chronic- and advanced-phase patients showed significant difference with respect to male-to-female ratio, hemoglobin level, WBC count, and platelet count. Sanger sequencing detected ANKRD36 mutations c.1183_1184 delGC and c.1187_1185 dupTT exclusively in all AP- and BC-CML patients but in none of the CP-CML patients. Nevertheless, mutations status was not associated with male-to-female ratio, hemoglobin level, WBC count, and platelet count, which makes ANKRD32 as an independent predictor of early and terminal disease progression in CML. ConclusionsThe study confirms ANKRD36 as a novel genomic biomarker for early and late CML progression. Further prospective studies should be carried out in this regard. ANKRD36, although fully uncharacterized in humans, shows the highest expression in bone marrow, particularly myeloid cells and correlation with high WBC count. Functional integrated genomic studies are recommended to further explore the role of ANKRD36 in the biology and pathogenesis of CML.

BackgroundAcute myeloid leukemia (AML) is a heterogeneous malignancy caused by the proliferation of neoplastic myeloid progenitor cells. Mutations in ten-eleven translocation methylcytosine dioxygenase 2 (TET2), a key regulator of DNA demethylation, are common in AML and play a role in its progression and response to treatment. MethodsWe performed a phylogenetic analysis of 25 TET2 gene sequences (12 AML and 13 normal) from Pakistani individuals using multiple sequence alignment, phylogenetic tree construction, and distance matrix evaluation. We also identified population-specific expression patterns and mutational hotspots. ResultsAML samples showed significant deletions in the catalytic domain (positions 693- 876) and lower sequence conservation compared to controls. These deletions may impair TET2 enzymatic activity and disrupt the epigenetic regulation. The Phylogenetic tree also showed two significant clades supported by high bootstrap values, differentiating AML from control individuals in the evolutionary direction. ConclusionOur findings reveal distinct TET2 mutational hotspots associated with acute myeloid leukemia (AML) in the Pakistani population. These results have implications for targeted epigenetic therapy and motivate large-scale, cross-population research to unravel the global effects of TET2 variations.

Biospecimens have become an integral part of life science research, pharma and drug development. There is increased demand for biospecimens from patients with assured quality of collection, processing, and storage. Establishing a biobank with high-quality biospecimens is an expensive aspect due to the requirement of costly freezers and liquid nitrogen facilities. Many studies have explored various storage reagents for biological samples. Still, the need of the hour is the one that can preserve biological samples at ambient room temperature and suitable for diagnosis and research. InstaPRESERVE is an eco/user-friendly solution with long-term sustainable alternative for freezer or liquid nitrogen systems for biobanking. Blood and saliva specimens were collected and stored in InstaPRESERVE at room temperature and conventional storage conditions. The genomics and proteomics analysis of the DNA, RNA, and protein extracted from the -80{degrees}C and InstaPRESERVE stored specimens and fresh specimens indicated that the specimens were well-conserved at room temperature compared to frozen specimens. Innovative technologies and approaches are being evaluated in biobanks for sample integrity, reducing collection, processing, and storage costs, increasing speed and throughput, and reducing energy expenditure. Preserving the biospecimens in InstaPRESERVE at room temperature eliminates expensive storage systems, cold-chain during transportation, and pre-processing requirements. The qualitative and quantitative results of DNA, RNA, and protein ensured stabilized storage for a longer period. The use of InstaPRESERVE as a specimen storage medium is emerging as an alternative in biobanking for specimen storage at room temperature. The existing and proposed biobanks may consider employing InstaPRESERVE as a reliable resource for biospecimens storage.

The use of molecular techniques to deal with plant molecular breeding requires the extraction of genomic DNA in good quantity and quality, which can be influenced by method of extraction and source of DNA (plant species, plant part or tissues). However, this research focuses on plant tissue source and tried to describe the quality and quantity of DNA isolated from different tissues of barley crop. CTAB protocol was used for the isolation of DNA and both the quality and quantity of this DNA was validated through gel electrophoresis, Qubit quantification and PCR amplifications. The result showed that DNA could be successfully extracted from all tissues of plants and the yield of DNA obtained was variable ranging from 179ng l-1 in stem to 750ng l-1 in young leave. Band intensity of genomic and PCR amplified DNA was good for DNA isolated from young and matured leave. Faint band was observed in the PCR amplification for DNA isolated from stem but no to unreliable amplification was obtained for seeds and roots, respectively. Thus, for any molecular technique in barley crop research, the best tissue for DNA isolation using modified CTAB, is young or matured leave and alternatively stem can be used as DNA source.

Aibika (Abelmoschus manihot) is a tropical leafy vegetable with great potential in the prevention of malnutrition. Its high fibre and micronutrient content makes it a valuable complement to staple foods such as rice and starchy crops consumed in South Asia and the Pacific. The present study is the first to report the development of a set of 21 nuclear single sequence repeat (SSR) markers of A. manihot from genomic sequence data obtained using NGS technology. The DNA library was prepared from a pool of four aibika accessions and sequenced using an Illumina MiSeq system. A total of 1,295,217 pair-end reads were generated. Raw data are available in the European Nucleotide Archive (accession number PRJEB88210; https://www.ebi.ac.uk/ena/browser/view/PRJEB88210). The sequences were assembled using the ABySS software. In total 651,320 contigs were generated. Using MISA Perl script and Primer3 software, we identified 8014 SSR motifs, of which 4637 had a suitable primer design. The contigs were blasted with NCBI-BLAST on okra reference genome. After selection based on motif type, repeat length, amplicon size and occurrence blast, we retained 91 candidate SSR loci, which were tested on 23 aibika accessions. Finally, we validated 21 high quality SSR loci by genotyping 45 accessions from three Pacific countries. The number of alleles per locus ranged from 3 to 21 with an average of 7.81 alleles/locus. The 21 selected SSRs were found to be useful in discriminating between accessions and revealing the diversity structure of A. manihot. They will help to optimize genebanks management and breeding programmes, and guide future collection activities.

Huntingtons disease (HD), a rare neurodegenerative disorder, is inherited in an autosomal dominant manner, and caused by a pathological trinucleotide expansion at exon1 of the HTT locus. Previous studies have described the haplogroups at the HTT locus that can explain the differences in prevalence of HD. We have selected three informative SNPs (rs762855, rs3856973 and rs4690073) to study these haplogroups in an Indian sample. Our results show that the genotype frequencies are significantly different between cases and controls for these SNPs. More than 90% of both cases and controls belong to Haplogroup A which is the predominant European haplogroup.

BACKGROUNDResearch on peripheral leukocyte gene expression in human health and disease is growing rapidly. However, how to process sample efficiently, simply and stably, and how to reflect human physiological state preferably remains critical issues in large cohort studies.\n\nMETHODSWe used RNA-seq to explore the differences of gene expression profiles among whole blood (WB) and three groups of leukocytes from buffy coat (BC) extraction, red blood cell (RBC) lysis and peripheral blood mononuclear cell (PBMC) isolation.\n\nRESULTSThe residual globin mRNA in leukocytes from RBC lysis (1.00% {+/-}1.23%) and PBMC isolation (0.06% {+/-} 0.03%) was much less than that in leukocytes from BC extraction (17.48% {+/-} 6.95%) and WB (24.46% {+/-} 6.43%), resulting in higher transcriptome mapping rates and larger numbers of detected genes. The expression of 616 genes associated with leukocyte function was slightly higher in leukocytes from RBC lysis than that from BC extraction and WB, but barely detected in leukocytes from PBMC isolation.\n\nCONCLUSIONSWe suggest that sample processing based on RBC lysis could allow better applications of gene expression profiling of peripheral leukocytes in large cohort studies.

Huntingtons disease (HD) is an autosomal dominant neurodegenerative disorder with profound phenotypic characters. HD is at present incurable and there are several trials going on to find a cure. HD is caused when there is a mutation in the Huntingtin gene which is found to be associated with axonal transport. Diagnosis is based on the signs and symptoms of the patients but by that time the psychomotor problems have already reached the level from where reversing the disease is impossible. Blood based biomarkers can be used for the diagnosis of the disease at an early stage. In this study several gene expression study data were analyzed and there were 329 Differentially Expressed Genes (DEGs) in all the three chosen datasets. Protein protein interaction network was created using STRING and CytoHubba plug-in was used to identify top ten hub genes which are CXCL8, PSMC6, UBE2D1, UBE2D1, CD27, UBE2D3, SF3B1, CASP3, EIF4E, BIRC2 and PTEN. Online software Enrichr was used for Gene Ontology and KEGG pathway enrichment analysis to find out the biological process, molecular function, cellular component and the pathways that were enriched in HD. This study finds out that those genes which were present in all the three datasets namely FNDC3A, BCLAF1 and ALCAM were not the hub genes. So further studies are required for identifying a potential biomarker of HD.

Information of the Passiflora genome is still very limited. Understand the evolutionary relationship between different species of Passiflora, and develop a large number of SSR markers to provide a basis for the genetic improvement of Passiflora. Applying restriction site associated DNA sequencing (RAD-Seq) technology, we studied the phylogeny, simple sequence repeat (SSR) and marker transferability of 10 accessions of 6 species of Passiflora. Taking the partial assembly sequence of accessions P4 as the reference genome, we constructed the phylogenetic tree using the detected 46,451 high-quality single nucleotide polymorphisms (SNPs), showing that P6, P7, P8 and P9 were a single one while P5 and P10 were clustered together, and P1, P2, P3 and P4 were closer in genetic relationship. Using P8 as the reference genome, a total of 12,452 high-quality SNPs were used to construct phylogenetic tree. P3, P4, P7, P8, P9 and P10 were all single branch while P1 and P2 were clustered together, and P5 and P6 were clustered into one branch. A principal component analysis (PCA) revealed a similar population structure, which four cultivated passion fruits forming a tight cluster. A total of 2,614 SSRs were identified in the genome of 10 Passiflora accessions. The core motifs were AT, GA, AAG etc., 2-6 bases, 4-16 repeats, and 2,515 pairs of SSR primer were successfully developed. Tthe SSR transferability in cultivated passion fruits is the best. These results will contribute to the study of genomics and molecular genetics in passion fruit.

Thrombophilia is defined as the willingness of blood to clot easily in a situation of imbalances between fibrinolysis and coagulation. It is classified as inherited and acquired thrombophilia. Several studies revealed that the inherited thrombophilia is strongly associated with single nucleotide polymorphisms (SNP) or deletions on certain genes, such as FV Leiden (rs6025), MTHFR1 (rs1801133), MTHFR2 (rs1801131), Serpine-1 (rs1799768), and Factor II (rs1799963). This study aims to develop an SNP detection panel based on Kompetitive Allele Specific Primer-polymerase chain reaction (KASP-PCR) technique. Results revealed that 86.5% of susceptible patients (n = 111) contain at least one mutation; while seven of them harbor three thrombophilia-associated mutations simultaneously. A clear allelic discrimination was observed for all tested samples. The prevalence of each mutation among different countries and ethnic groups are in line with the findings of this study. Rather than expensive and time-consuming approaches, the current assay enables the cost-effective advantage of the KASP-PCR, which reduces the diagnostic cost with a fast and convenient way. After clinical validation and approval, it can be used in hospitals, research centers, and diagnostic laboratories to determine the genetic susceptibility of individuals to thrombosis and for research purposes.

With its rising fatality rates, oral cancer is one of the most concerning public health issues. To reduce disease-related mortality and morbidity, advancements in screening and detection are critical. Finding specific biomarkers is one of the most successful approaches for screening, diagnosing, and staging this dreadful disease. In this study differentially expressed genes associated with oral cancer were analyzed using RNASeq to find the potential biomarkers. Functional enrichment of upregulated genes found that 253 genes were present in the plasma membrane. Three clusters were formed using KMean Clustering from the PPI networks, and highly connected hub genes were identified from each cluster. Eventually, expression and survival analyses of hub genes were performed using The Cancer Genome Atlas (TCGA) database targeting Head and Neck Squamous Cell Carcinoma. Among those genes, expression levels of eight genes SLC2A1, ITGA6, LAMC2, COL1A2, COL1A1, TNC, THY1, and CD276 have significantly changed in Head and Neck Squamous cell carcinoma. There are reports that suggest these genes were significantly dysregulated in Oral Squamous cell carcinoma and can be explored further as potential biomarkers for margin clearance.

Acute promyelocytic leukemia is a distinct subtype of acute myeloid leukemia characterized by the t(15;17) translocation, leading to the PML (Promyelocytic leukemia protein)-RARA (Retinoic Acid Receptor Alpha) fusion protein. Although PML-RARA fusion is common, there are 20 more fusion events also reported in APL. All -trans retinoic acid (ATRA) is a standard drug for APL, leading to significant improvement in patient outcomes; nevertheless, a small fraction of patients still experience relapse, and some patients exhibit resistance to the drug. Long non-coding RNAs (LncRNAs) are recognized as promising biomarkers for cancer diagnosis, prognosis, and treatment response. In this study, we used ATRA-Resistant (AP1060) and ATRA -Sensitive (NB4), both treated and untreated cell line transcriptomic data retrieved from the NCBI Gene Expression Omnibus(GEO) database to perform transcriptomic analysis with bioinformatic tools. We utilized the LncRAnalyzer pipeline to predict the lncRNAs, followed by differential expression analysis using DESeq2. Weighted Gene Co-expression Network Analysis (WGCNA) was employed to construct lncRNA co-expression modules associated with ATRA resistance. BEDTools is used to identify cis-acting target genes of lncRNAs.LncRNA -miRNA sponging identified by miRanda algorithm. The identified miRNAs reveal their significant role in APL and other leukemia subtypes. The results of the study show that the identified lncRNAs from the miRNA-LncRNA network are promising biomarkers for ATRA resistance.

BackgroundHomologous recombination (HR) pathway is a DNA double-stranded breaks repair pathway well-known for its high level of accuracy. Low HR pathway efficiency clinically known as homologous recombination deficiency (HRD) was identified in some cancers such as breast and ovarian cancers and studies have reported the sensitivity of HRD cancer cells to DNA repair inhibitors such as Olaparib. However, current techniques including immunofluorescence-based technique are qualitative-based, hence lack sensitivity to determine the functionality of HR pathway. Additionally, some of the techniques including gene expression arrays require expression study of wide range genes involve in HR pathway, which is not cost-effective. The aim of the study is to optimise a PCR-based assay (Norgens Homologous Recombination kit) that can be employed to quantitate HR efficiency in cells, which accurately reflects the functional status of HR pathway. Methods and FindingsThe kit has two test plasmids (dl-1 and dl-2) with partial deletions in the LacZ gene and the plasmids are generated from modification of pUC19. HR-proficient (HeLa and AsPC-1) and HR-deficient (CAPAN-1 cells) cancer cell lines were transfected with the two plasmids to generate functional LacZ gene (i.e. recombinant product). The recombinant product was quantified by real-time PCR. Although recombinant product was generated in all the cell lines, our real-time PCR demonstrated a high quantity of recombinant product in HeLa cell line whilst low quantity in CAPAN-1 and AsPC-1 cell lines. The quantity of recombinant product generated and quantified reflects HR pathway efficiency. ConclusionOverall, the results have provided some evidence that the PCR-based kit can be suitably employed for quantification of HR efficiency provided appropriate transfection method and reagent are used. However, further study is required to confirm HR efficiency status of AsPC-1 cells to ascertain the low HR efficiency detected by the kit in these cells.

Sandalwood (Santalum album L.) is highly valued aromatic tropical tree. It is known for its high quality heartwood and oil. In this study, 39 genic and genomic SSR markers were used to analyze the genetic diversity and population structure of 177 S. album accessions from 14 populations of three states in India. High genetic diversity was observed in terms of number of alleles 127, expected heterozygosity (He) ranged from 0.63-0.87 and the average PIC was 0.85. The selected population had relatively high genetic diversity with Shannons information index (I) >1.0. 0.02 mean coefficient of genetic differentiation (FST) and 10.55 gene flow were observed. AMOVA revealed that 92% of the variation observed within individuals. Based on cluster and Structure result, individuals were not clustered as per their geographical origin. Furthermore the clusters were clearly distinguished by principal component analysis analysis and the result revealed that PC1 reflected the moderate contribution in genetic variation (6%) followed by PC2 (5.5%). From this study, high genetic diversity and genetic differentiation was found in S. album populations. The genetic diversity information of S. album populations can be used for selection of superior genotypes and germplasm conservation to promote the tree improvement of S. album populations.

IntroductionGastric adenocarcinoma (GA) is responsible for thousands of deaths annually and significantly affects patients quality of life. To increase knowledge about this disease, several studies have focused on the transcriptional control mechanism by analyzing the role of transcription factors (TFs) in GA. MethodsThis study identified differential expression (DE) of TFs in GA samples from 74 patients using NGS, biostatistical analysis in R, and comparison with 68 normal tissue samples in the genetic databases of the National Center for Biotechnology Information (NCBI). Results1,564 human transcription factors were DE. Of these, 87 were selected using the established cutoff value of AUC [&ge;] 0.95, and 25 were analyzed for their influence on GA carcinogenesis. Based on their role in biological processes, nine TFs were found to be underexpressed due to their growth suppression and cell differentiation properties, notably the TFs IRF3 and CXXC1. Sixteen TFs were overexpressed, associated with drug resistance, disease promotion, metastasis, and poor prognosis in GA, such as AHR, NCOA3, RRB1, and STAT3, respectively. ConclusionThis study integrated systematic bioinformatics and NGS, which revealed novel TFs with important oncogenic and suppressive functions in GA. Therefore, it provides new insights into genes with potential biomarkers for prognostic prediction in GA, which may offer important implications for clinical practice and may effectively target treatment, monitoring, or disease recurrence.

In Brazil, it is estimated that at least 10% of the native flora, which corresponds to about 4 or 5 thousand plant species, are used for food purposes. Among these species, Pereskia aculeata Mill, known as ora-pro-nobis and belonging to the Cactaceae family, stands out. It is considered a Unconventional Food Plant (UFP) and a vegetable of traditional use, playing an important role in the diet and economy of rural and urban families. Due to their high nutrient content, they have both economic and social significance, being a target of interest for scientific research. Despite the potential for human use of this species, there is a lack of knowledge about its diversity and genomic structure, resulting in few genetic and molecular studies that can contribute to the conservation and improvement of this species in general. It is essential to acquire information about its genomic structure, as well as to carry out studies related to the comparison, selection and optimization of protocols for obtaining quality DNA and in sufficient quantity, in order to guarantee the success of molecular and evolutionary analyses. The aim of this research was to evaluate qualitatively and quantitatively methods of isolation of genomic DNA from Pereskia aculeata Mill. Significant variations were observed in the DNA extracted and in the purity observed between the different protocols tested, with the isolation by the protocol described by Sambrook et al. (1989) being the most efficient in terms of concentration and purity, followed by the modified method proposed in this work. This work is a pioneer in comparing methods for isolating DNA from Pereskia aculeata Mill. in terms of quality and quantity. In addition, it was found that there is still a scarcity of molecular studies on this species, both worldwide and in Brazil, which indicates the existence of a promising gap for the development and realization of scientific research in this field.

Kiwifruit (Actinidia spp.) is a significant plantation crop belonging to family Actinidiaceae, having deciduous, dioecious, and scrambling vines with chromosome number 2n=58. Dioecy in kiwifruit forms the basis for several breeding programs. The present study was carried out for diversity analysis in kiwifruit genotypes using RAPD markers. 7 kiwifruit genotypes (2 males viz. Allision & Tomuri and 5 females viz. Hayward, Bruno, Allision, Monty & Abbott) were analysed for molecular polymorphism using 94 RAPD primers, out of which 23 primers amplified the genomic DNA in all the genotypes. RAPD data was analysed using NTSYS-pc software and dendrogram construction was done using UPGMA method. Two separate clusters of male and female genotypes were formed. Similarity matrix indices showed maximum similarity between Tomuri (M) and Allision (M) with a similarity coefficient of 0.719 while Abbott (F) and Allision (M) were found to have least similarity having a similarity coefficient of 0.521. Four RAPD primers amplified unique amplicons in Monty, Hayward, Bruno, Allision (M) and Abbott and two primers amplified unique amplicons in Allision (M) & Tomuri (M) along with the male and female plants of Allision genotype respectively. Therefore, these primers can help in distinguishing the genotypes of kiwifruit and can also be validated as putative markers for the sex identification in kiwifruit.