Genetic Similarity Index Research Articles

English

In the present study, genetic variability in 10 commercial chilli pepper varieties viz. Gemini, G-334, Agnirekha, Pusa jwala, Mangala, Black diamond, Sindura, Pusa jyothi, Badiga-2 and Teja (branch), cultivated in the local area of Andhra Pradesh  State, India, using random amplified polymorphic DNA (RAPD) markers was examined. Out of thee RAPD markers used for screening 10 chilli pepper genotypes, DNA amplification was observed only with OPAB02; this primer produced monomorphic band  in Gemini,G-334,  Black diamond and Sindura and polymorphic bands in Agnirekha, Pusa jwala, Mangala, Pusa jyothi, Badiga-2 and Teja (Branch). Genetic variabilty in terms of  DNA pattern produced in the above varieties could be used as a marker to distinguish between them. Dendrogram generated by OPAB02 primer showed that  the 10 chilli pepper varieties could be grouped into four clusters. Average genetic similarity index revealed 100% similarity between varieties of first cluster, 50% similarity between varieties of second and fourth clusters and 32% genetic similarity between  varieties of third cluster. UPGMA cluster analysis will be useful in chilli pepper  breeding programmes and germplasm conservation. Since the commercial value of chilli pepper is based on pungency level, future studies are aimed at molecular marker based pungency phenotyping. Key words:  Capsicum annuum, Randomly Amplified Polymorphic DNA, Dendrogram, Polymerase  Chain Reaction.

Polymorphic microsatellite differences among four cultured populations of two selected tilapia strains

AbstractGenetic diversity plays a vital role for maintainingeconomically important traits in fish populations.The goal of this study was to assess the geneticvariability among four tilapia populations in China.Two populations named XA (Oreochromis aureus,30 samples) and AN (Egyptian Oreochromis niloticus,24 samples) were gathered from Freshwater Fish-eries Research Center (FFRC), and the other twonamed AL (O. aureus, 30 samples) and AJ (Egyp-tian O. niloticus, 30 samples) were from GuangxiFisheries Research Institute (GFRI). GFRI intro-duced AL and AJ from XA and AN, and selectedthem for generations. We want to know if AL andAJ have genetic changes after selection and gener-ations. Fifteen polymorphic microsatellite markerswere used. Our results indicated that the four tila-pia populations showed moderate genetic variabil-ity. The polymorphism information content (PIC)of the four populations was from 0.28 to 0.39; A e was from 1.67 to 2.15; the H e was from 0.35 to0.46; the genetic similarity indexes were from 0.94to 0.69 and also the clustering result showed thatthese four populations were highly related. Therewere no significant changes in genetic variationbetween late-introduced population and early-introduced population of O. aureus. Our resultshelped to understand the genetic variability of tila-pia populations in China and for future marker-assisted selection for important economic traits.Keywords: microsatellite, Nile tilapia, Blue tila-pia, heterozygosity, genetic diversity, selection,loss of genetic variationIntroductionTilapia belongs to Perciformes Cichlida and it isnative to Africa and Middle East. Tilapia (Oreochr-omis mossambicus) was introduced to China in theearly 1950s. Later, many species and strains (Or-eochromis niloticus, Oreochromis aureus, etc.) wereintroduced to China too (Li 2001; Li & Li 2001;Ye 2008). After their introduction to China, tila-pia has been undergone artificial selection forvarious economically important traits, such asfast growth, resistance to environmental stressorsand high percentage of males. Since the expandedproduction of tilapia started, China has been thelargest supplier of tilapia in the world till now(Li, Li, Tang & Cai 2009).Fresh Water Fisheries Research Center (FFRC) isthe most famous genetic and breeding resourcesCenter of Tilapia in China well known as the bestquality of purebred Tilapia parents. And now,FFRC was the head of China Agriculture Techni-que System-tilapia (CARS). The Blue tilapia(O. aureus, XA) was introduced from Auburn Uni-versity by FFRC in 1983, and they have selectedand bred a new Blue tilapia strain, named “Xia’ao1”. This strain is widely used for cross-breedingwith Nile tilapia to get all male tilapia fry inChina. The Nile tilapia samples (AN) used in thisstudy were introduced from Central laboratory forAquaculture Research, Abbassa, Ministry of Agri-culture and Land Reclamation of Egypt by FFRC in1999. Guangxi Fisheries Research Institute (GFRI)was the most famous tilapia fry productive unit inChina as the high male rate of cross-bred tilapiafry. GFRI introduced two strains of tilapia: XA(O. aureus) and AN (Egyptian O. niloticus) from

Diversidad genética de una colección de hongos entomopatógenos de Ecuador utilizando un enfoque AFLP modificado

Se determinó la diversidad genética de 41 cepas ecuatorianas de hongos entomopatógenos y un aislamiento de los EEUU, de una colección mantenida por el INIAP (Instituto Nacional Autónomo de Investigaciones Agropecuarias) utilizando un protocolo modificado de AFLP (Amplified Fragment Length Polymorphism). Se encontraron índices de similitud genéticos inferiores a 0.50 y 121 bandas polimórficas. El análisis AMOVA reveló que la variación entre grupos y dentro de grupos contribuían en cantidades similares (59% y 41% respectivamente) a la variación genética total detectada. El dendograma construido a partir del coeficiente genético de Jaccard muestra doce grupos, de los cuales siete contienen aislamientos agrupados por género. De estos siete grupos, cinco contienen aislamientos agrupados en base al huésped de donde se recolectaron. Los valores de Bootstrap muestran doce relaciones filogenéticas confiables con valores de confianza mayores al 70%. El análisis de componentes principales (PCA) produjo seis grupos; cuatro de ellos contienen aislamientos asociados por género. Estos resultados sugieren la existencia de una diversidad genética considerable dentro de la colección de hongos entomopatógenos del INIAP, y una tendencia de agrupamiento relacionada con el huésped de donde fueron aislados. No se encontraron regiones genómicas comunes dentro de las cepas más virulentas de hongos entomopatógenos. La elevada diversidad genética dentro de esta colección representa una fuente potencial de genotipos con actividad bioinsecticida potente.

Posidonia oceanica genetic and biometry mapping through high-resolution satellite spectral vegetation indices and sea-truth calibration

In the framework of Posidonia oceanica (PO) preservation activities, a small-scale restoration pilot project was implemented in 2005 at a Santa Marinella site to replace the loss of this important species of seagrass in this zone of the central Tyrrhenian coast via an innovative transplantation approach. In this context, taking into account the recent advances in the fields of high-resolution (HR) satellite/airborne remote-sensing and genetics laboratory analysis techniques, we propose this integrated methodology for monitoring changes in transplanted meadows in regard to perspective to provide support in the assessment of the entire local PO and seagrass population dynamic. According to specific information requirements in terms of radiometric and spectral/spatial resolution, the multispectral data currently available from the QuickBird polar satellite’s four-band (red, green, blue visible and near-infrared) HR sensor were exploited for methodology implementation using a practical ‘image-based’ approach to account for atmospheric and water column turbidity typical of this mid-coastal Mediterranean region. First, the extents and types of seagrass cover were suitably mapped, and then also the distributions of specific vegetation parameters related to PO dynamics and health were assessed by exploiting the remotely sensed satellite-derived radiance signals and point sea-truth calibration measurements of the bio-genetic parameters. In particular, we implemented maps of leaf area index, genetic similarity, and density Giraud indices corresponding to distributions of PO patches using multivariate and data-mining models (artificial neural network) based on appropriately preprocessed radiometric and auxiliary (bathymetry) input variables.

A fusant of Sphingomonas sp. GY2B and Pseudomonas sp. GP3A with high capacity of degrading phenanthrene

A fusant strain F14 with high biodegradation capability of phenanthrene was obtained by protoplast fusion between Sphingomonas sp. GY2B (GenBank DQ139343) and Pseudomonas sp. GP3A (GenBank EU233280). F14 was screened and identified from 39 random fusants by antibiotic tests, scanning electron microscope (SEM) and randomly amplified polymorphic DNA (RAPD). The result of SEM analysis demonstrated that the cell shape of fusant F14 different from parental strains. RAPD analysis of 5 primers generated a total of 70 bands. The genetic similarity indices between F14 and parental strains GY2B and GP3A were 27.9 and 34.6%, respectively. F14 could rapidly degrade phenanthrene within 24h, and the degradation efficiency was much better than GY2B and GP3A. GC-MS analysis of metabolites of phenanthrene degradation indicated F14 had a different degradation pathway from GY2B. Furthermore, the fusant strain F14 had a wider adaptation of temperatures (25-36°C) and pH values (6.5-9.0) than GY2B. The present study indicated that fusant strain F14 could be an effective and environment-friendly bacterial strain for PAHs bioremediation.

Agronomical use as baby leaf salad of Silene vulgaris based on morphological, biochemical and molecular traits

In this study, the genetic variability of 14 accessions of bladder campion (Silene vulgaris), collected from different parts of Spain, were evaluated by molecular, morphological, agronomical and biochemical analysis. Molecular analysis was carried out using the ISSR technique with 10 primers, producing a total of 125 DNA bands, 80 of which were polymorphic among the accessions. A dendrogram drawn on the basis of a similarity matrix using the UPGMA algorithm revealed that the 70 samples of bladder campion plants could be classified into two major clusters at a Jaccard's genetic similarity index of 0.25, confirming that genetic variation was very high among and within accessions. An analysis of the morphological, agronomical and biochemical traits was a less efficient way of grouping the accessions due to the high inter-population variability observed for the analyzed traits used. Some accessions showed a very high level of nutritional compounds (glutathione, total phenols, and antioxidant capacity) and morphological characteristics, which, taken together, makes them attractive candidates for consumption as a ready-to-eat product. However, high concentrations of some anti-nutritional compounds (nitrates and oxalates) were found in some accessions. The results show sufficient variation exists in the desired traits within and among accessions in the germplasm to pursue a breeding programme to develop S. vulgaris cultivars as a high quality, minimally processed product.

Reduced genetic diversity in endemic Brazilian Lymania spp (Bromeliaceae) populations and implications for conservation

We analyzed the genetic diversity of populations of two sympatric species of Lymania (Bromeliaceae), both endemic to the Atlantic rainforest of southern Bahia (Brazil). Lymania azurea has a restricted occurrence, while Lymania smithii has a wider distribution. Our aim was to provide genetic data to contribute to the design of more efficient conservation strategies for these bromeliads, possibly justifying inclusion in the official Brazilian list of Endangered Species. Up to now, L. azurea has been classified by the Brazilian Ministry of the Environment as "data deficient". We sampled four populations of L. azurea throughout its distribution area in southern Bahia and two populations of L. smithii in the same region. Genotyping was performed with 48 random amplified polymorphic DNA markers. Based on the Jaccard genetic similarity index, L. smithii has greater diversity than L. azurea. An analysis of molecular variation showed greater genetic variance within than between populations for both species. L. azurea was found to have 20% inbreeding, probably due to population fragmentation, with L. smithii showing only 10%. When we analyzed pairs of populations of L. azurea within a conservation unit, we found low population structure (ФST = 0.098), apparently due to a large degree of gene flow between them. In disturbed areas, we found a higher ФST (0.372). We found low genetic variability for L. azurea, probably as a consequence of habitat fragmentation, supporting the need for its inclusion in the Brazilian list of endangered flora.

Estimation of Genetic Diversity in Rice (<i>Oryza sativa</i> L.) Genotypes using Simple Sequence Repeats

Simple Sequence Repeats (SSRs) are co-dominant DNA markers and have a wide range of applications in the field of genetics. In the present investigation, 24 rice genotypes, belonging to Pakistan and imported from IRRI, Taiwan were examined for the determination of genetic diversity using twenty four SSRs. Our results showed that the primers used had polymorphism when applied to the tested genotypes. The distinguishing factor with respect to both locations were primers designed from chromosome 2,3,4,5,7 and 8. A total of 76 loci were produced by twenty four SSRs among 24 rice genotypes with an average of 3.17 loci per genotype. All markers produced polymorphic fragments among most of the genotypes. A total of 995 alleles were detected with an average of 13.87 alleles per locus. Among all the primers, RM-1 produced highest number of bands, “120” followed by RM-154 who developed 108 bands, RM-124 produced 65 bands shown and lowest number of bands (8) produced by primer RM-489. The similarity matrix clearly revealed that most closely genotypes were SRS-62 and SRS-505, securing the similarity value of 0.9234. It mean that these genotypes are 92.34 % similar, likewise genotypes Bas-370 and Bas-Pak, Bas-370 and S.S.Shaheen had the score of 0.8966 and 0.8840 respectively. The most dissimilar genotypes are IRRI-12 and EF-1-2-51-2002, scoring the value of 0.5601; it means they are 56.15% dissimilar. Similarity coefficient ranged from 0.54 to 0.9234. The average genetic similarity index was calculated as 0.73. Dendrogram divided twenty four rice genotypes into four main clusters and each cluster was subdivided into two furthers groups. Among all the genotypes, IRRI-4 showed distinct behavior in dendrogram, proving it to be useful for breeding program.

Molecular genetic diversity within Myrmeleontidae family

Antlions are insects which feed on ants, insect which dig a pit and lies in wait for ants and other insects. Twelve species of Myrmeleontidae family as antlions and many specimens were identified in different locations in Fars province in Iran. To unveil the genetic similarity between these species, their DNA was extracted by modified CTAB method and with the use of seventeen 10-nucleotides primers of random amplified polymorphic DNA (RAPD); the genetic analysis of them was investigated. After PCR, agarose 1.5 % was used for electrophoresis. The obtained electrophoresis bands had base pairs range between 150 and 1,000 bp. The maximum of polymorphic bands belonged to OPH5, N13, and the minimum of polymorphic bands belonged to OPA7 primers. Different genetic similarity indices were found between eight species of antlions. Possibility of use of RAPD marker together with morphological studies for classification and identification of antlions is discussed.

Molecular and ampelographic characterization of some grape hybrids (Vitis vinifera L.)

23 hybrids were selected as potential cultivar candidates due to their superior characteristics and were created at the Ataturk Horticultural Central Research Institute (AHCRI) or the TekirdaAŸ Viticulture Research Institute (TVRI). In this study, the ampelographic and molecular characteristics of 23 grapevine hybrids created in two different breeding studies were determined. Ampelographic data were collected during two vegetation periods in Yalova, Turkey. Total of 68 ampelographic characteristics were identified for 23 hybrids. Polymerasechain reaction (PCR) - based microsatellite technique was used to analyse 23 hybrids and 2 standard varieties. The genetic relationships were defined with 20 simple sequence repeat (SSR) markers, and genetic similarity indices, principal component analysis graphs and phylogenetic relationship dendrograms were produced depending on the hybrid and SSR marker size of the allele which ranged from 88 to 294 bp. Additionally, ampelographic and molecular scores were evaluated together. After evaluation of molecular and ampelographic data together, similarity ratio was increased and ampelographic characterisation was dominated by molecular characterization. It may be related using more ampelographic characters in the study. Variety candidates which were obtained from different Institutes were characterized and taken under protection with this study. The information in the obtained database can be used for cultivar identification, parentage analysis, and legal protection. Also, some ampelographic characterization results will be used in determining the priority of registration. Key words: Ampelography, characterization, hybrids, simple sequence repeat (SSR), Vitis vinifera L.

Molecular characterization of genetic diversity in some durum wheat (Triticum durum Desf.) in Palestine

Random amplified polymorphic DNA (RAPD) analysis was conducted to estimate the genetic diversity and relatedness among 10 wheat cultivars and landraces. Five out of 70 decamer random primers showed distinctly polymorphic bands when used to amplify genomic DNA. The primers yielded a total of 157 RAPD markers of which 150 were considered as polymorphic. Each primer produced 18 to 37 polymorphic bands with an average of 30 markers per primer. DNA fragments size ranged from 350 to 1910 bp. The mean similarity indices ranged from 0.05 to 0.68 with an average of 0.29. The unweighted pair group method with arithmetic mean (UPGMA) dendrogram based on Jaccard genetic similarity index, grouped the 10 landraces and cultivars into four main clusters. Despite the clustering of most landraces in the same cluster, high genetic variation was observed within cluster-I in which the genetic distance ranged from 73% between Hiteyah8 and White Dibeyah to 32% between Black Hiteyah and Yellow Hiteyah. The result of this study confirms high genetic diversity within these cultivars and landraces, and therefore, underlined the significant values of these cultivars and landraces as gene pool for durum wheat genetic improvement. Thus, RAPD offer a potentially simple, rapid and reliable method to evaluate genetic variation and relatedness among ten wheat landraces and cultivars.

GENETIC ESTIMATION OF SNAIL LYMNAEA STAGNALIS POPULATIONS FROM REGIONS WITH DIFFERENT ANTHROPOGENIC LOAD AS A FIRST STEP IN GENETIC MONITORING

The paper deals with carrying out molecular-genetic typing of populations of snail Lymnaea stagnalis from regions with a different ecological load (the lake Perstok and zatoka of the river Pripyat, Belarus). By means of interpopulation analysis of frequencies of RAPD-fragments, frequencies of zero alleles of RAPD-loci and genetic similarity index we established that these populations are genetically identical because there are no significant distinctions of genetic variability neither between individuals within a population, nor between populations. Results of the cluster analysis and the population analysis also have confirmed that populations of snails are genetically closely related. However the snail population of the lake Perstok is genetically more homogeneous.

Construction and analysis of an intergeneric fusant able to degrade bensulfuron-methyl and butachlor

Rhodococcus sp. BX2 degrades bensulfuron-methyl but not butachlor, and Acinetobacter sp. LYC-1 degrades butachlor but not bensulfuron-methyl. Functional strains were constructed through protoplast fusion of Rhodococcus sp. BX2 and Acinetobacter sp. LYC-1 to generate fusants with an improved ability to simultaneously degrade bensulfuron-methyl and butachlor. Initial identification and stability tests of the fusants were performed. Three fusants with eighth transfer on plates containing two antibiotics and two herbicides were obtained. F1 also grew well in an inorganic salt solution containing bensulfuron-methyl and butachlor. F1 was characterized by its parents' morphological and physio-biochemical features. F1 not only had bands in common with BX2 and LYC-1, but also had its own specific bands analyzed by Random Amplified Polymorphic DNA. The genetic similarity indices between F1 and BX2 and F1 and LYC-1 were 0.507 and 0.470, respectively. The percentages bensulfuron-methyl and butachlor degradation by F1 in an inorganic salt solution supplemented with 100 mg/L bensulfuron-methyl and 100 mg/L butachlor were 65.35 and 62.41 %, respectively, and the percentages in soil contaminated with 10 mg/kg bensulfuron-methyl and 10 mg/kg butachlor with an inoculum size of 5 % at 34 °C and at a pH of 7.5 after 35 days were 63.74 and 61.53 %, respectively. It was demonstrated that F1 could simultaneously degrade bensulfuron-methyl and butachlor.

Molecular characterization of mulberry (Morus spp.) genotypes via RAPD and ISSR

In recent years, DNA-based markers have been used quite extensively because of their many advantages over the traditional morphological and biochemical markers. Many studies have shown that molecular markers are useful in delineating the genetic relationships among closely related mulberry genotypes and cultivars. Thus, in the present study, polymer chain reaction based DNA fingerprinting techniques were used to investigate the genetic relationships among mulberry genotypes growing in different agro-climatic regions of Turkey. 20 RAPD primers generated a total of 173 bands, of which 157 (90.75%) were polymorphic. As for 11 ISSR primers, 124 bands (96.55%) were polymorphic in a total of 128. The similarity index for RAPD technique ranged between 0.24-0.98; 25İs203 with 25İs112 were found to be the closest genotypes, while 24Ke10 and 25İs123 were the most distant ones. According to the ISSR result, the genetic similarity index changed between 0.21-095; 25İs203 with 25İs112 genotypes were the closest, while 25İs08 and 01KaD2 were the most distant ones. The RAPD and ISSR markers were found to be promising for assessing genetic diversity in mulberry genotypes.

Genetic potential analysis of six Tilapia populations by microsatellite DNA markers

Twenty five microsatellite loci were used to analyze two blue tilapia populations ["Xia'ao 1" (ZA), Guangxi population] and four nile tilapia populations [Egypt strain (ZN), 88 strain (XN), Guangxi population (GN), American strain (MN)]. A total of 7775 fragments ranging from 100 bp to 400 bp in length were obtained. Three to eight alleles were amplified in 25 loci and 143 alleles in all the six populations. The average number of alleles in each locus was 5.72. The average values of observed heterozygosity (Ho) ranged from 0.7253 to 0.8160, the average expected heterozygosity (He) ranged from 0.5146 to 0.6834, the average polymorphic information content (PIC) ranged from 0.4212 to 0.6105, and the number of average effective alleles (Ae) ranged from 2.20 to 3.23. The highest genetic similarity index was 0.9130 (between ZA and GA); and the lowest was 0.4352 (between ZA and ZN). The results showed that the four nile tilapia populations contained a high level of genetic potential, and the two blue tilapia populations were moderate.

English

Amplified fragment length polymorphisms (AFLP) were used to evaluate the genetic diversity between 18 date palm (Phoenix dactylifera L.) varieties (11 females and 7 males) collected from the center of Iraq. Six primer pairs were applied to detect polymorphism between varieties. A total of 83 polymorphic AFLP fragments were detected with an average of 13.8 polymorphic fragments/primer combination. Genetic distance was estimated using Jaccard’s genetic similarity index and was ranged from 0.07 to 0.75. Unweighted pair group method with arithmetic mean UPGMA ordered date palm varieties into two main clusters independently of their origin and sex. The first cluster consisted of three sub-clusters. The first one consisted of five female varieties and one male, while the second sub-clusters consisted of five male varieties. The third one consisted of five varieties; four were females and one male. The second main cluster consisted of the remaining two female varieties. Moreover, all primer combinations contributed to the discrimination of date palm varieties, suggesting the efficiency of AFLP method in assessing genetic diversity in date palm. A large range of genetic diversity characterized Iraqi date palm germplasm.   Key words: Genetic diversity, amplified fragment length polymorphisms (AFLP) polymorphisms, molecular characterization, Phoenix dactylifera L.

Genetic diversity analysis by RAPD markers in candidate plus trees of Pongamia pinnata, a promising source of bioenergy

Pongamia pinnata has received much attention in recent years as a source of seed oil that can provide a substitute for diesel fuel. Very little molecular work has been reported on this species. This paper reports our studies on the diversity amongst the CPTs (candidate plus trees) of P. pinnata previously identified on the basis of morphometric traits, particularly pod and seed traits. For this RAPD (random amplified polymorphic DNA) markers were used to determine the genetic diversity among 10 genotypes of P. pinnata CPTs selected for suitability for energy production. For further investigation, 18 primers generating stable band patterns from 40 tested arbitrary primers were selected. A total of 210 amplification products were obtained of which 10.48% were polymorphic. The genetic similarity index ranged from 0.11 to 0.87. Genetic distance values were used to generate a dendrogram (UPGMA) between the genotypes. The Mantel method used for comparing the similarity matrixes produced correlation coefficients that were statistically significant for the RAPD marker. This genomic analysis allows a cost effective characterization of CPTs of P. pinnata. The present investigation supports in future the development of genetic map in Pongamia which are a highly useful tool in breeding and may provide information on the inheritance of features crucial for increase seed yield, oil content and the resistance to key insects and pests. Thus this study warrants Pongamia producers to realize its full potential and contributes for its sustainable production and improvement.

Evolutionary trends of microsatellites during the speciation process and phylogenetic relationships within the genusSecale

Eleven weedy or wild species or subspecies of the genus Secale L. were compared with a set of cultivated rye accessions, based on inter-simple sequence repeat (ISSR) markers to analyze their phylogenetic relationships. A total of 846 bands were amplified from reactions using 12 screening primers, including 79 loci with a mean of 10.1 alleles per locus. The number of amplified bands for each primer ranged from 12 to 134, with a mean of 70.5 amplified bands per primer. The presence and distribution of amplified bands in different accessions demonstrate that a rapid evolutionary trend of microsatellite repeats occurred during the speciation process from the perennial wild form to annual cultivated rye. In addition, variation, amplification, and deletion of microsatellites in genomes revealed phylogenetic relationships in the genus Secale. Analysis of the presence, number, and distribution of amplified bands in genomes, as well as the comparison with genetic similarity (GS) indices based on ISSR, indicate that Secale strictum subsp. africanum (Stapf) Hammer, Secale strictum anatolicum (Boiss.) Hammer, Secale sylvestre Host, and Secale strictum subsp. strictum (C. Presl) Hammer emerged in succession from a common ancestor of Secale following geographic separation and genetic differentiation. The annual weedy rye evolved from S. strictum subsp. strictum, which was domesticated as present-day cultivated rye. Data from ISSR analyses separated all investigated accessions of the genus Secale into three distinct groups. These results support the division of the genus Secale into three species: the annual wild species S. sylvestre; the perennial wild species S. strictum, including several differential subspecies forms such as strictum, africanum, and anatolicum; and S. cereale, including cultivated and weedy rye as subspecies forms.

Generic relationship among Cassia L., Senna Mill. and Chamaecrista Moench using RAPD markers

Generic relationships were examined among twenty-four species belonging to genusCassia L., Senna Mill. and Chamaecrista Moench using RAPD marker. Total 80 primers were initially screened, 514 amplification products obtained with 38 informative primers, of which 514 were polymorphic. A vary high degree of polymorphism (100%) was observed among them. UPGMA cluster analysis of genetic similarity indices grouped all the species into three major clusters. Cluster I included four species of Cassia L., Cluster II included eighteen species of Senna Mill. and Cluster III included two species of C. Moench. Highest similarity (0.9%) was observed between Cassia fistula L. and Cassia fistula with nodded filaments and least (0.001%) between Cassia fistula L. and Senna splendida. The Polymorphic information contents (PIC) of the twenty-four species with RAPD marker varied from 0.08 to 0.49 with an average of 0.005. The result confirms the statement of Irwin and Barneby, they divided the genus Cassia L. into three subgenera; Cassia L.,Senna Mill. and C. Moench on the basis of morphological characters. The results obtained from the present study support the previous taxonomic classification of the genus CassiaL. and showed large diversity among the species of three newly created genera. Our results suggested that RAPD marker is a sensitive, precise and efficient tool for genomic analysis of Cassia L. that may be useful in future studies by assigning new unclassified germplasm to specific taxonomic groups and reclassify previously classified species and genera. Key words: Cassia, Senna, Chamaecrista, genetic relationship, RAPD.

Genetic and Chemical Diversity of High Mucilaginous Plants of Sida Complex by ISSR Markers and Chemical Fingerprinting

A method was developed based on multiple approaches wherein DNA and chemical analysis was carried out toward differentiation of important species of Sida complex that is being used for commercial preparation. Isolated DNA samples were successfully performed through PCR amplification using ISSR markers and degree of genetic diversity among the different species of Sida is compared with that of chemical diversity. For genetic fingerprint investigation, selected 10 ISSR primers generating reproducible banding patterns were used. Among the total of 63 amplicons, 62 were recorded as polymorphic, genetic similarity index deduced from ISSR profiles ranged from 12 to 51%. Based on similarity index, S. acuta and S. rhombifolia found to be most similar (51%). High number of species-specific bands played pivotal role to delineate species at genetic level. Investigation based on HPTLC fingerprints analysis revealed 23 bands representing to characteristic chemicals and similarity index ranged from 73 to 91%. Prominent distinguishable bands were observed only in S. acuta, while S. cordifolia and S. rhombifolia shared most bands making them difficult to identify on chemical fingerprint basis. This report summarizes the genotypic and chemotypic diversity and the use of profiles for authentication of species of Sida complex.