Unweighted Pair-group Method Using Arithmetic Averages Research Articles

Genetic diversity of Guava varieties (Psidium guajava L.) based on morphology and ISSR molecular markers

Abstract. Hien PTT, Cham LTT, Hang VTT. 2024. Genetic diversity of Guava varieties (Psidium guajava L.) based on morphology and ISSR molecular markers. Biodiversitas 25: 1037-1045. Guava (Psidium guajava L.) is a fleshy-fruited representative belonging to the Myrtaceae family that has attracted attention due to its economic benefits. In this study, seventeen guava varieties in the Mekong Delta and Hanoi were collected and genetically validated by both morphology and DNA markers. The results revealed that fruit shape is a powerful indicator of guava discrimination. There are 12 out of 15 Inter Simple Sequence Repeats (ISSR) markers that show informative band patterns and 93 amplicons are generated with the size in length 200-2000 bp. The average polymorphic band and PIC value are 97.85% and 36%, respectively. Based on the dendrogram constructed by the Unweighted Pair Group Method using arithmetic Averages (UPGMA) method, samples were grouped into two main groups and the similarity level ranged from 55-98%. Group 2 contains Oi Tim (Purple guava), a special variety that showed high differences in terms of morphology and DNA profiles. Such information reflects the diversity of guava germplasm in Viet Nam, which is useful for conservation, and breeding strategies. The result revealed that 11 varieties expressed genetic diversity through both phenotypic traits and DNA profiles. Such information illustrates the genetic diversity among Psidium guajava, contributing to germplasm resources and plant breeding programs.

CAVE BATS (YANGOCHIROPTERA AND YINPTEROCHIROPTERA) IN GUNUNG SEWU GEOPARK: STUDY OF KARST CAVES IN WONOGIRI REGENCY

Indonesia is a country rich in biodiversity including flora and fauna. One of the high fauna diversity in Indonesia is the mammal. Some of these bats use karst caves as their roosting habitat. One of the karst areas in Indonesia is Gunung Sewu Karst. This study aims to learn more about the various bats that live in caves in the Gunung Sewu Geopark Area. This research was conducted in March-May 2023, in several caves located in the Gunung Sewu Geopark Area. Bats are caught using misnets and handnets. After they were captured, the bats were identified using Morphometry and the Shannon-Wiener index. Through another index, the Margalef index, the bat diversity in the tree cave habitats was expressed, with a discovery that there are many different species. Based on the similarity index, bats were categorized again using cluster analysis and the unweighted pair-group method using arithmetic averages (UPGMA). The entire analysis was assisted with software PAST ver 4.13. The species recorded from the three caves in this study amounted to 8 species of bats belonging to 5 genera and 5 families. Sodong Cave is a habitat with the highest level of diversity and distribution of species. All three habitats have low similarity. The existence of endemic and vulnerable species is a priority in protecting caves as their habitat and Gunung Sewu karst macroecosystems. By preserving the habitat, all biodiversity in it will be preserved.

Genetic similarity among Dendrobium species from Indonesia using RAPD markers

Abstract. Hartati S, Samanhudi, Cahyono O. 2023. Genetic similarity among Dendrobium species from Indonesia using RAPD markers. Biodiversitas 24: 5015-5021. Dendrobium genus orchids are one of the most popular commodities in the world due to their diverse range of flower sizes, shapes, and colors. To enhance plant breeding programs and genetic resources, it is necessary to obtain information on genetic similarity between orchids of the Dendrobium genus through molecular analysis techniques. Therefore, this study aimed to assess the genetic similarity among five Dendrobium species using molecular markers, specifically RAPD. The plant material used was obtained from five species of the Dendrobium spp, namely (i) Dendrobium mirbelianum; (ii) Dendrobium lamellatum, (iii) Dendrobium secundum, (iv) Dendrobium bracteosum, and (v) Dendrobium purpureum. The analysis was carried out to determine the genetic diversity of the Dendrobium orchids using RAPD markers. A total of five RAPD primers were used for amplification, namely OPD 8, OPA 7, OPA 13, OPB 12, and OPB 18. The scoring data were analyzed using NTSYS-pc (Numerical Taxonomy and Multivariate Analysis System) version 2.02 which produced data in the form of cluster dendrograms. The dendrogram that was constructed by Unweighted Pair Group Method Using Arithmetic Average (UPGMA) classified the five Dendrobium species into two main clusters. The results showed that there were two clusters, namely Cluster A consisting of D. mirbelianum, D. lamellatum, and D. secundum while Cluster B consisted of D. bracteosum and D. purpureum. Furthermore, the polymorphism of the five RAPD primers was very high, ranging from 91.6 to 100%, measuring 250-1900 bp. The coefficient of genetic similarity analyzed using the five RAPD primers ranged from 0.24 to 0.77. The species D. mirbelianum and D. lamellatum had a high coefficient of genetic similarity. Which can be discovered through parental selection in breeding program.

Contribution to the pollen morphology of Astragalus L. section Aegacantha Bunge (Galegeae-Fabaceae) and its systematic significance

Astragalus L. sect. Aegacantha Bunge includes 57 described species, mostly confined to Southwest Asia and Central Asia. We studied the pollen morphology of 17 species of this section present in Pakistan. Pollen morphology of sect. Aegacantha species is poorly investigated despite its systematic importance. The main aim of this study is to determine and document the pollen characters that can assist in the taxonomic identification of species in this difficult section. Pollen slides were prepared following acetolysis and examined by both light and scanning electron microscopy. Pollen size varies, with the polar axis ranging from 32.91 to 43.10 μm and the equatorial axis from 22.91 to 28.47 μm. Pollen is sub-prolate (1.15–1.33) to prolate (1.34–2.00) in shape, small to medium in size, radially symmetrical, isopolar, trizonocolporate to tricolporate, and monad. Sculpture variations (i.e. reticulate, microreticulate, perforate, microperforate, regulate, microregulate and granulate) were observed around the apertural, meridional and polar regions. Ordination analysis (principal component analysis (PCA)) revealed that the quantitative characters of pollen morphology are the most reliable characters for delimitation of species within section Aegacantha. Cluster analysis (unweighted pair group method using arithmetic average (UPGMA)) shows a partial relationship between the Aegacantha taxa clustered based on pollen characters and supports the general morphology. Further, this study shows that the pollen traits provide a baseline for phylogenetic optimisation in the investigated Aegacantha section.

Genetic diversity and differentiation of South African cactus pear cultivars (Opuntia spp.) based on simple sequence repeat (SSR) markers

Opuntia ficus-indica (L.) Mill. is one of the most recognisable agricultural crops that can withstand harsh environmental conditions. South Africa is one of the few countries that hosts a large germplasm of Opuntia cultivars, which represent an ex-situ conservation population. However, little is known about the genetic diversity in this population. Additionally, some genotypes are morphologically indistinct, and therefore, making it a challenge for novice farmers and researchers to recognise specimens in the germplasm. The current study aimed to differentiate and measure the genetic diversity in 44 cultivars that represent the South African Opuntia germplasm using eight simple sequence repeat (SSR) markers. Evidently, the cultivars comprised moderate levels of diversity (average polymorphic information content, PIC = 0.37, Nei’s unbiased gene diversity = 0.42) that discriminated 90% of the cultivars. Analysis of the cultivars with the unweighted pair-group method using arithmetic averages (UPGMA) method revealed three main clusters, whereas principal coordinate analysis (PCoA) showed indistinct clustering of cultivars based on their usage in the agricultural market. Overall, the used SSR markers were able to distinguish most cultivars in the South African Opuntia germplasm—a genetic resource that appears to hold sufficient genetic diversity to aid the conservation and breeding of novel cultivars for the agriculture market.

Diversity of the Cave-Dwelling Bat (Chiroptera) in the Ngobaran Coastal Area, Karst of Gunung Sewu

Bats (Chiroptera) are divided into two suborders (Yinpterochiroptera and Yangochiroptera). More than 50% of species of bats use caves as their roosting sites. Thus, they play a crucial role in the cave ecosystem. For that reason, they also exist as keystone species in the karst area. Gunung Sewu is one of the karst areas in Indonesia that best exemplifies tropical karst. Furthermore, Gunung Sewu is still at risk of habitat loss despite being designated as a Geopark. This study aimed to understand the diversity of bats that live in caves in the karst region of Gunung Sewu, specifically in four caves near Ngobaran Beach between April and June 2020. A harp net and misnet placed at the cave’s entrance were used to capture bats for data collection. After they were captured, the bats were identified using Morphometry and the Shannon-Wiener index. Through another index, Margalef index, the bat diversity in the four cave habitats was expressed, with a discovery that there are many different species. Based on the Jaccard similarity index, bats were categorized again using cluster analysis and the unweighted pair-group method using arithmetic averages (UPGMA). A total of nine species across five families were identified. The diversity of existing species variety was also recognized by analyzing the composition of the four cavern inhabitants. The four cave ecosystems’ bat diversity was divided into three categories: moderate variety, low similarity, and high species diversity. Except for Cekelan 1 Cave and Gebyog Cave (P=0.015), other variations did not demonstrate a meaningful difference (P0.05). This demonstrates how different each ecosystem is. Therefore, they could be classified as potentially spoiled habitats, demanding additional conservation efforts.

The Relationship of Plant Types Familia Araceae on The Mountain Sidole Track Central Sulawesi Based on Morphological Character

The Araceae family plant in the Mount Sidole has a diversity and variety of life forms. The purpose of this study is to describe the family relationship of Araceae family based on morphological characters. Sampling using the path method (belt transect) with free collection techniques. The size of the track used is 10 m wide and ± 15 km long along the Sidole Mountain climbing track at an altitude of 400-1780 m above sea level. Morphological data measurement refers to 37 meristic and qualitative characters. Processing data using the File Editor Program (PFE) and analyzed with the Multivariate Statistical Package (MVSP) program. Determination of similarity index with the Simple Matching Coefficient method and kinship between types using the Unweighted Pair-Group Method Using Arithmetic Average (UPGMA). The results of specimen collection are 26 species. Based on the dendrogram of Familia Araceae divided into three groups. Group I are 9 species of hemiepipitic groups which cluster at a coefficient of 76%, namely Amydrium medium, Pothos scandens, Epipendrum sp., Scindapsus treubii, S. pictus, Rhaphidophora sp., R. sylvestris, R. lobbii and R. puberula. Group II are 8 species of geophytes grouping at 77% coefficient, namely Colocasia oresbia, C. esculenta, Xanthosoma violaceumm, X. sagittifolium, Apoballis mutata, Alocasia sp., A. longiloba and A. Macrorrhizos. Group III are 9 species of geophytes grouping at 82% coefficient, namely Spathiphyllum commutatum, Schismatoglottis inculta, S. lancifolia, S. wallichii, S. asperata, S. cadieri, S. calyptrata and S. calyptrata ssp flagelliforme and Aglaonema nitidum.

Paleoecology of a Lower Cretaceous (Barremian) fish community of the Morro do Chaves Formation, Sergipe-Alagoas Basin, NE Brazil

Opportunities to examine the structure of paleocommunities are extremely rare, due to the incompleteness of the fossil record. Regarding the Morro do Chaves Formation, there are many studies about the fauna and flora since 1870. However, paleoecological data are relatively scarce. According to the most recent study, the palynological records of the Morro do Chaves Formation were related to a disturbed paleoenvironment, caused by marine incursions in the northern sector of the Sergipe-Alagoas Basin during Lower Cretaceous. Thus, this paper aims to analyze the paleoecological relationships of the paleocommunity of fishes in the Morro do Chaves Formation, as the possible responses to the paleoenvironmental disturbances. From the basic matrix of abundance data for four fish orders found in 11 pelitic levels, similarity matrices were computed, using the Bray-Curtis distance coefficient. Cluster analysis by the unweighted pair-group method using arithmetic averages (UPGMA) was performed on these matrices, for grouping the levels. The advantages of multivariate analysis in paleoecological studies are also discussed. In addition, the Morro do Chaves Formation is composed of active piscivorous predators, benthophagous and planktiphagous fishes. The aulopiform fishes probably acted as top-down regulators in the paleocommunity, controlling the local fish populations. Mawsonia and “Lepidotes” are usually recorded in similar outcrops and inferred to be associated fauna. While Mawsonia likely possesses a sit-wait strategy, “Lepidotes” would be a relatively more active predator.

Mixed Modeling in Genetic Divergence Study of Elite Popcorn Hybrids (Zea mays var. everta)

Popcorn is a food highly appreciated throughout the world, generating billions of dollars annually just in the North American market alone. Even in the face of the historical superiority of American hybrids, which occupy almost 100% of the grain production fields in Brazil, our researchers have been working to develop cultivars that combine important traits for the crop, such as tolerance to leaf diseases and high rates of grain yield and popping expansion. This work investigated the degree of genetic divergence among 40 diallel hybrids of popcorn, 16 parents and 6 elite controls using mixed models to discriminate superior materials to be adopted in the Brazilian agribusiness. Based on the individual Best Linear Unbiased Prediction (BLUP) of each of the 15 variables analyzed, the Unweighted Pair-Group Method using Arithmetic Averages (UPGMA) and Tocher clusters were performed, and the study of Pearson correlation was carried out. The results pointed out that there is genetic variability among the genotypes evaluated and that the best candidates for composing superior genotypes are in the combination between the P10×L77 parents and/or between the P7 and L88 lines. Linear correlations showed that earlier flowering and taller genotypes exhibited an association with materials more tolerant to Exserohilum turcicum intensity.

Microsatellite analysis in some watermelon (Citrullus lanatus) genotypes

Conservation of genetic resources is essential for the continuation of future crop production. Watermelon (Citrul-lus lanatus), a member of Cucurbitaceae, is widely distributed in tropical and subtropical regions. The aim of this study was to reveal the genetic relationships with the help of microsatellite markers in a watermelon collection free of unnecessary repetitions, and to determine the success of SSR (Simple Sequence Repeats) primers developed in cucurbits. In this study, 96 watermelon genotypes with good agronomic characteristics were used among the geno types collected from different regions of Turkey and purified up to the S4-S6 (self-pollination) stage. In the study, 33 SSR primer pairs were used to determine the genetic relationship between watermelon genotypes. In the study, a total of 67 bands were obtained with SSR primers. As a result of UPGMA (Unweighted Pair Group Method Using Arithmetic Averages) analysis, genotypes showed similarity at the level of 0.84-1.00. The number of alleles detected per primer varied between 1 and 6. In terms of the total number of alleles obtained, CMCT44 (5 units) and Cgb4767 (6 units) loci produced the most alleles. Primers with high polymorphism rate and allele excess were determined, and the possibilities for use in genetic stability analyses, variety differentiation and other genetic analyses were determined.

Evaluation of genetic divergence of coffee genotypes using the volatile compounds and sensory attributes profile.

The quality of the coffee beverage is related to the chemical, physical, and sensory attributes of the coffee beans that vary with the geographic location of the crop, genetic factors, and post-harvest processing. So, the objective of this study was to evaluate the genetic divergence of 27 genotypes of Coffea canephora using the volatile compounds and sensory attributes profile to select genotypes that produce a coffee beverage with high sensory quality. This genetic diversity was estimated from the Euclidean distance matrix using non-standard data and the Unweighted Pair-Group Method Using Arithmetic Averages (UPGMA). The 2-furyl-methanol, 4-ethenyl-2-methoxyphenol, furfural, 5-methylfurfural, methylpyrazine, and 2,6-dimethylpyrazine were predominating volatile compounds in the genotypes. The sensory attributes had a positive Pearson's correlation with the total score. The volatile compounds had a different relative contribution to the genetic divergence between the genotypes of C. canephora. The 4-ethenyl-2-methoxyphenol, 2-furyl-methanol, and furfural were volatile compounds that most contributed to the formation of the groups in the UPGMA dendrogram. The relative contribution of sensory attributes to dissimilarity among genotypes was 6.42% to 20.20%. Therefore, this study verified the relative contribution of volatile compounds, in specially 4-ethenyl-2-methoxyphenol, 2-furyl-methanol, and furfural, and sensory attributes (flavor, mouthfeel, and bitterness/sweetness) to the genetic divergence between the genotypes of the three clonal varieties. Thus, this work points out compounds that positively contribute to the sensory quality of the Conilon coffee beverage.

Genetic diversity evaluation in wild Muntingia calabura L. based on Random Amplified Polymorphic DNA (RAPD) markers

This present research was aimed to contribute fundamental knowledge about genetic diversity in wild Muntingia calabura. Random Amplified Polymorphic DNA (RAPD); a method that has been widely used to study genetic relationships in fruit crops. In this research investigation, eight accessions were collected, four accessions from Bangkok, Thailand and four accessions from Medan, North Sumatera, Indonesia. DNA fingerprinting of the eight accessions was performed using twelve RAPD markers (OPA-01, OPA-02, OPA-07, OPA-11, OPC-08, OPC-09, RAPD-01, RAPD-02, RAPD-03, RAPD-04, RAPD-05, RAPD-06). The obtained results revealed that all twelve primers produced polymorphic bands. UPGMA (unweighted pair-group method using arithmetic averages) method was used to determine the genetic distance. Based on dendrogram analysis, the eight accessions of M. calabura formed two main clusters. The range of genetic distance based on RAPD scores was 0.50–0.85. The RAPD technology is suitable for anonymous genomes due to its adaptability to situations with limited quantities of DNA and higher efficiency, and low cost. Researchers hope that present work is impactful and useful for genetic breeding programs in the future.

Genetic diversity in Coffea canephora genotypes for leaf nutrient concentration


 This study analyzed the genetic diversity in Coffea canephora genotypes by univariate and multivariate statistical analysis, based on concentrations of macro- and micronutrients in coffee leaves in the stages of pre-flowering and grain filling. The experiment was arranged in randomized blocks with three replications, in a 42x2 factorial design, in which factor one represented the evaluated genotypes and factor two the periods of leaf sampling, i.e., pre-flowering and grain filling. The data of leaf nutrient concentrations were subjected to analysis of variance by the F test (p <0.01), and genetic parameters were estimated. For the study of genetic diversity, the genotypes were grouped by the hierarchical unweighted pair-group method using arithmetic averages (UPGMA). The relative importance of a trait to predict genetic diversity was also studied. There is genetic divergence for leaf nutrient concentration in C. canephora genotypes. With a maximum limit of 60% of dissimilarity between genotypes, four groups were also formed by UPGMA. For the 42 evaluated genotypes, leaf S concentration was the most important trait for genetic diversity; this genotypic variability should be investigated to enhance the efficiency of nutritional diagnosis.
 Highlights
 
 There is genetic variability for leaf nutrient concentration in Coffea canephora
 The leaf concentration of nutrients in the phenological stage of pre-anthesis is different from the leaf concentration of nutrients in the coffee bean filling period.
 Leaf concentration of sulfur was the characteristic that contributed most to the genetic diversity among the 42 genotypes, followed by the concentration of copper, boron and magnesium.
 Genotypic variability for leaf nutrient concentration should be considered for better nutritional management.

Floral morphology of robusta coffee genotypes

AbstractKnowledge about the diversity in the morphological characteristics of robusta coffee (Coffea canephora Pierre ex Froehner) flowers is fundamental to understanding reproduction processes, interpret pollination‐related mechanisms, and contribute to breeding studies. The objective of this study was to analyze and characterize morphological variability of flower traits among robusta coffee cultivar Conilon genotypes of high agronomic performance. The experiment was arranged in a randomized block, split‐plot design. The main plot corresponded to six Conilon coffee genotypes and the subplots to 2 yr of evaluation (2018 and 2019). Eight floral traits were measured and analyzed through analysis of variance by the F test (<.01), Tocher's optimization method and hierarchical Unweighted Pair Group Method using Arithmetic Averages (UPGMA) clustering. The relative importance of each character was also estimated to predict genetic diversity, and the degree of trait interrelation was assessed by Spearman's correlation. Genotype Pirata had outstanding floral morphological structure and was grouped separately by Tocher and UPGMA clustering. Style length was the major contributor to genetic diversity, followed by the lengths of the corolla tube and stigmatic lobes. The flower trait characteristics were positively correlated, and corolla tube length and style length were most strongly correlated. These results contribute to other studies related to the interpretation of the mechanisms related to pollination and reproduction of this species, and also outline diversity of genetic resources of interest to plant‐breeding programs.

Bafa Gölü'nün (Türkiye) ekolojik ve trofik durumunun fitoplankton bazlı değerlendirilmesi

Phytoplankton groups are one of the major quality element to be used in the evaluation of the trophic and ecological state of freshwater ecosystems according to the EU Water Framework Directive. This research was made to assess the trophic and ecological status of Lake Bafa in Turkey, on the basis of phytoplankton communities. Büyük Menderes River is one of the most important factor that carries pollutants to Lake Bafa. The eight sampling station were assigned to evaluate the ecological and trophic state of the lake. Phytoplankton species were collected monthly for 2 years study period. Most commonly used phtoplankton indices Q index and Carlson’s Trophic State Index (TSI), and different versions of diversity indices were used to estimate trophic and ecological state of the lake. Similarities between the sampling stations were clustured by using the unweighted pair group method using arithmetic average (UPGMA), based on phytoplankton communities. Correlations between the applied indices were determined by using Pearson Correlation. After the identification of collected phytoplanktons, total of 63 taxa which belong to classis of Cyanophyceae (11.2%), Bacillariophyceae (49.2%), Chlorophyceae (23.8%), Xanthophyceae (1.5%), Euglenophyceae (11.2%) and Dinophyceae (3.1%) were detected. The 1st and 2nd stations were the most similar stations to each other (88%) according to phytoplankton communities. Secchi disc depth (SD) and TP played an important role in the distribution of phytoplankton species in Lake Bafa. The highest significant positive correlation was determined between Q and TSI (r = 0.987, p˂0.01). Considering the TDI values in the phytoplankton composition of the lake, it can be said that although the productivity status of the studied lake is still “mesotrophic”, it has a tendency towards “eutrophic” state. According to the Q values, the first five stations reflect the moderate ecological state, while the 6th, 7th and 8th stations represent the poor ecological state.

Effects of heavy metals on bacterial community structure in the rhizosphere of Salsola collina and bulk soil in the Jinchuan mining area

It is important to understand the changes in microbial properties of plant rhizospheres to elucidate the interaction between heavy metals and plant communities. In this study, high throughput sequencing method was used to analyze the effects of heavy metals on rhizosphere microbial community structure of the autochthonous plant Salsola collina in the Jinchuan mining area. The operational taxonomic units (OTU) number and species diversity, and richness of the bacterial community in the rhizosphere of Salsola collina were higher than those of the bulk soil. Proteobacteria, Actinobacteria, and Firmicutes were the major bacterial phyla, and Salsola collina, Kaistobacter, Citrobacter, Acinetobacter and Pseudomonas were the main genera. Principal component analysis (PCA) and unweighted pair group method using arithmetic average (UPGMA) cluster analysis showed that there were significant differences in bacterial community structure between site and between rhizosphere and bulk soil. The moisture content, total nitrogen, phosphorus, magnesium, nickel, chromium, lead, and zinc contents, and pH had significant effects on species richness of the bacterial community, with total phosphorus content having a significant effect on species diversity. The finding of this study could be used as an important guideline for bioremediation of heavy metals in mining area.

RAPD, ISSR, and SCoT markers based genetic stability assessment of micropropagated Dendrobium fimbriatum Lindl. var. oculatum Hk. f.- an important endangered orchid

Dendrobium fimbriatum is an ornamental and medicinal orchid listed in the Red data book of IUCN. Phytohormones' effect on the in vitro regeneration of the orchid was studied using Mitra medium supplemented with different growth regulators. KN produced effective shoot formation when present alone or in combination with IBA or NAA. The shooting was gradually increased when KN concentration was increased from 0.8 to 4.8 mg L-1, but the opposite response was observed with BAP at higher concentration (4.8 mg L-1). IBA either in combination with BAP or KN promoted effective root development and multiplication. Micropropagated orchids grown in the basal medium devoid of any phytohormone showed 100% monomorphism, while low genetic polymorphism of 1.52% (RAPD-Random Amplification of Polymorphic DNA), 1.19% (ISSR-Inter Simple Sequence Repeat) and 3.97% (SCoT-Start Codon Targeted) was exhibited among the regenerants propagated in the hormone enriched medium. UPGMA (Unweighted pair group method using arithmetic averages) dendrograms showed the grouping of mother plant (MP) with the in vitro regenerants. The principal coordinate analysis (PCoA) further confirmed the clustering patterns as determined by the cluster analysis. The study reported for the first time the successful in vitro propagation of Dendrobium fimbriatum and their genetic stability assessment using molecular markers.

Assessment of Genetic Variation in Selected Members of Family Apocynaceae using SDS-PAGE

Genetic variation of any plant species is very interesting in reducing genetic vulnerability as well as stabilizing production. In this regard, a study was undertaken to analyze the genetic variation among selected members of family Apocynaceae by using sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). In this study, a total of 53 protein bands were analyzed, out of them all bands were polymorphic with a total of 100% polymorphism. The electrophoresis of the proteins revealed protein bands in the range of 151 kD to less than 11 kD molecular weight. The similarity index calculated on the basis of presence and absence of protein bands ranged from 0.04 to 0.200. A dendrogram was constructed based on UPGMA (unweighted pair group method using arithmetic averages) clustering method revealed three clusters. Cluster I contained three species namely Thevetia peruviana, Catharanthus roseus and Nerium indicum, in which Thevetia peruviana and Catharanthus roseus were more close than Nerium indicum, while cluster II included only one species namely Rauvolfia serpentina. Carissa carandus emerged as the most primitive species forming an out group (cluster III). Thus, this study revealed that the SDS-PAGE method plays a key role in the study of protein based variation among selected plant species.

ASSESSMENT OF GENETIC RELATIONSHIP AMONG FIVE MANDARIN CULTIVARS USING RAPD-PCR MARKER

Genetic relationship among five mandarin cultivars was assessed using Randomly Amplified Polymorphic DNA (RAPD) markers. Five primers were selected after screening of many primers which generated 119 total bands. Clustering pattern analysis, the Unweighted Pair-group Method using Arithmetic averages (UPGMA) dendrogram based on Nei's genetic distance grouped five mandarin cultivars into two main clusters where S. mandarin and Ch. Mandarin formed one cluster and the rest three cultivars grouped together into two sub clusters, where C. mandarin and Sat. Mandarin formed one sub cluster and the other sub cluster included only B. mandarin cultivar. Inter-species similarity index (Dice and Jaccard) and the UPGMA dendrogram based on Nei's genetic distance revealed that S. mandarin and Sat. mandarin as the most closely related species among the five citrus cultivars under study.

THE USE OF GENETIC DISTANCE AND GROUPING METHODS TO PREDICT Eucalyptus Pellita F. MUELL GENITORS FOR HYBRIDIZATION

The objective of this study was to use quantitative traits to estimate the genetic distance among E. pellita provenances and progenies, to inform possible hybridization strategies in a species improvement program. A provenance and progeny test with 118 progenies from seven provenances was evaluated. The following quantitative traits were measured at seven years of age: diameter at breast height (DBH); height; and individual volume. The data were submitted to REML/BLUP analysis to obtain the predicted genetic value (BLUP). From this, the Mahalanobis (D²) genetic distance was estimated for provenances and progenies, which were then grouped by Tocher’s method, the unweighted pair group method using arithmetic averages (UPGMA), and principal component analysis (PCA). In total, 29 divergent groups were obtained among progenies based on Tocher’s method, which showed greater reliability according to the cophenetic correlation coefficient than UPGMA. The opposite was found between provenances, where the results for UPGMA demonstrated greater clustering reliability. Based on principal component analysis (PCA), the M. Ray and Tully provenances were the most similar, while Connl. A and Orchard were the most divergent. Height was the most important trait in estimating genetic distance. The results obtained offer important insights for breeding programs; with this information, crosses can be designed between contrasting individuals among and within provenances to obtain E. pellita hybrids, validating the possible heterotic groups identified through the genetic distance and grouping methods.