An assessment of the genetic diversity within a collection of<Emphasis Type="Italic">Saccharum spontaneum</Emphasis> L. with RAPD-PCR

A local collection of 33Saccharum spontaneum L. clones and two sugarcane cultivars (LCP 82-89 and LCP 85-384) were assessed for genetic variability using random amplified polymorphic DNA (RAPD)-PCR. A total of 157 polymorphic RAPD-PCR bands were scored with 17 primers. The number of RAPD-PCR products per primer ranged from four to 16. The data were analyzed with two multivariate analysis software programs, NTSYSpc and DNAMAN®. Although these two programs yielded similar results, a bootstrapped phylogenetic tree could only be generated with the DNAMAN® software. A substantial degree of genetic diversity was found within the localS. spontaneum collection. Pairwise genetic homology coefficients ranged from 65% (SES, 196/Tainan 2n = 96) to 88.5% (IND 81-80/IND 81-144). LCP 82-89 and LCP 85-384 shared a greater similarity (82%) than either was to any clone ofS. spontaneum (ranging from 60.5 to 75.2%). The 33S. spontaneum clones were assigned to eight groups independent of their geographic origin or morphology, while the two sugarcane cultivars were assigned to the ninth group. All but two pairs ofS. spontaneum clones could be distinguished by a single RAPD primer OPBB-02. The use of a second primer, either OPBE-04 or Primer 262, separated allS. spontaneum clones. One amplification product from the RAPD primer OPA-11, OPA-11-336, proved to be cultivar-specific and has been adopted for use in our breeding program. Information from this study would help conserve the genetic diversity ofS. spontaneum. Disclaimer: Product names and trademarks are mentioned to report on available data; however, the USDA neither guarantees nor warrants the standard of the product, and the use of the name by USDA does not imply the approval of the product to the exclusion of others that may also be suitable. The experiments reported comply with the current laws of the USA.     

Simple sequence repeats marker polymorphism in emmer wheat (<Emphasis Type="Italic">Triticum dicoccon</Emphasis> Schrank): Analysis of genetic diversity and differentiation

Genetic diversity was investigated in 73 accessions of emmer wheat (Triticum dicoccon Schrank) from 11 geographical regions using a set of 29 simple-sequence repeat (SSR or microsatellite) markers, representing at least two markers for each chromosome. The SSR primers amplified a total of 357 different alleles with an average of 12.31 alleles per locus. The number of fragments detected by each primer ranged between 6 (Xgwm1066) and 21 (Xgwm268). Null alleles were detected in nine of the 29 primers used. A high level of gene diversity index was observed. Across the 29 primers, gene diversity ranged from 0.60 (Xgwm46) to 0.94 (Xgwm655), with a mean of 0.82. There was a highly significant correlation (r=0.882; p<0.01) between gene diversity index and the number of loci, showing the number of loci per se is a strong indicator of diversity. Analysis of genetic diversity within and among eleven geographical regions revealed most of the genetic diversity of the total sample resided within regions. The coefficient of gene differentiation (Gst = 0.27) showed that the genetic variation within and among the 11 geographical regions was 73 and 27%, respectively. High value of mean number of alleles per locus was found in Iran (4.86) followed by Morocco (4.10) and Armenia (4.03). On the contrary, lower mean number of alleles per locus was detected in Yemen (2.83). The average gene diversity index across regions ranged from 0.52 (Slovakia) to 0.67 (Morocco) with an average of 0.60. Multivariate techniques of principal component analysis and clustering were employed to examine genetic relationship among the 73 emmer wheat accessions vis-à-vis geographical regions of collections. The genetic distance coefficients for all possible 55 pairs of regional comparisons ranged from 0.63 (between Iran and Armenia, Georgia and Azerbaijan, Georgia and Slovakia) to 0.97 (between Morocco and Yemen, Spain and Georgia, and Turkey and Iran) with a mean of 0.82. From the PCA results, a two dimensional plot of PC1 versus PC2 was constructed. The scatter plot of the first two principal components which explained altogether 27% of the total variation depicted the presence of a clear pattern of geographical differentiation except in few cases like accessions from Caucasian region. Similar pattern of genetic relationships among accessions was observed in cluster analysis. The study provided genetic information of emmer wheat in relation to geographical regions of origin. The information could be utilized in crop improvement, germplasm conservation programs, and in further investigation.     

Genetic diversity analysis in Boerhavia diffusa L. of different geographic locations in India using RAPD markers

Boerhavia diffusa is extensively used in herbal medicines as well as in the Ayurvedic system, because it contains a set of clinically important compounds. In the present study, the genetic variability in Boerhavia diffusa between accessions of different geographical origin within the Indian territory is assessed through random amplified polymorphic DNA (RAPD) markers. Twenty-eight accessions of Boerhavia were screened with eighteen primers of which nine were found to be the most informative. The degree of polymorphism was found to be high in accessions collected from different places of Uttar Pradesh (Set II) in comparison to other states of India (Set I). A relatively lower level of polymorphism was recorded in accessions collected from diverse locations around Lucknow (Set III). Accessions from neighbouring geographical regions exhibited more similarity than those from distant regions (as revealed by the set I analysis). Certain diagnostic markers may be correlated with morphological character(s) such as plant type. BDL appeared most distinct and divergent from the rest of the accessions and the BDJ plant in set II also showed least similarity estimate. Fragments of 5.62 Kb and 4.47 Kb with primer GN59 was found to be unique for BDP and BD2 having ovate leaf character, whereas ovoid leaf genotype exhibited 0.79 Kb (GN34 primer) fragment. Similarly a unique band type (0.35 Kb) with primer GN83 was present in BDL and BD1 that share light pink flower. Jaccard's, and Nei and Li similarity coefficient values amongst the accessions were in the range of 0.22 to 0.89 and 0.33 to 0.93, respectively. Association of RAPD markers with the leaf characteristics, flower colour as well as with geographical locations has been made. This shows that RAPD markers are also useful for the study of genetic structure of Boerhavia populations.     

Molecular analysis of the DNA polymorphism of wild barley (Hordeum spontaneum) germplasm using the polymerase chain reaction

Summary Hordeum spontaneum is the progenitor of cultivated barley (H. vulgare) and is an important source of genetic variation for barley breeding programs. The genetic diversity ofH. spontaneum in the Australian germplasm collection was investigated using the polymerase chain reaction with random and semi-random primers. This approach was found to be robust in respect of reaction conditions. Genetic dissimilarity values between genotypes were used to produce a phenogram of the relationships among the accessions using the unweighted pair group method with arithmetic mean. The largest divergence was observed among Israeli accessions, whereas the Turkish and Iranian samples clustered as distinct subsets, each apparently related to portion of the Israeli material. The results indicate that the genetic diversity of the wild barleys is broadly correlated with geographic distribution.     

Characterization of intergeneric hybrids of <Emphasis Type="Italic">Erianthus rockii</Emphasis> and <Emphasis Type="Italic">Saccharum</Emphasis> using molecular markers

Erianthus rockii, a wild relative of sugarcane, is drought and cold tolerant, and both are potentially important agronomic traits to the sugarcane industry worldwide. As such it is of interest as a source of parental germplasm to sugarcane breeders and is currently being used in sugarcane introgression programs in both China and Australia. To date morphological characters have been used to verify the putative hybrids produced. Two crosses were generated between two different Saccharum species and two E. rockii accessions. Over 400 AFLP markers were used to identify the intergeneric hybrid progeny as well as determine hybrid diversity. Both crosses generated hybrids but efficiency levels were very different and are probably related to the different Saccharum parent used in each cross. Cross 1 was between a Saccharum officinarum and E. rockii and generated 100% hybrid progeny. Cross 2, however, was between a sugarcane hybrid (S. officinarum × Saccharum spontaneum) and E. rockii and only 10% of the progeny were intergeneric hybrids. Inheritance of markers in the progeny was analysed and for both crosses there were equal numbers of markers from both parents indicating n + n transmission of chromosomes. This is the first verification of E. rockii hybrids with molecular markers. It may now be possible to exploit genes of value from E. rockii in sugarcane breeding programs.     

Genetic variation among highly endangered <Emphasis Type="Italic">Bacopa monnieri</Emphasis> (L.) Pennell from Southern India as detected using RAPD analysis

Randomly amplified polymorphic DNA (RAPD) fingerprinting approach was applied to assess genetic diversity in different accessions of rejuvenating and intellect-promoting ancient ayurvedic medicinal herb Bacopa monnieri (L.) Pennell collected from four Southern Indian states, along with in vitro micropropagated samples maintained in the laboratory for 5 years. With the 10 analyzed primers, 110 distinct bands, in the size range of 250–870 bp were observed, and among which 14 (12.72%) were polymorphic. Among the random primers used, only OPD 02 generated highest percentage of polymorphism with 30.77. Whereas OPD 08 generated a maximum of 18 amplified bands, but with only 1% polymorphism. Cluster analysis done on the basis of similarity co-efficient generated from RAPD profiles indicated all the accessions were divided into two sub-groups based on genetic distances under one major cluster. Major cluster is only the lone loose group of the Bm.2 collected from Kerala (KER) and in vitro micropropagated plant (IVMP) maintained in the lab. The other sub-group consisted of Bm.1 and Bm.3 collected from Tamil Nadu (TN) and Andhra Pradesh (AP) respectively, which are genetically similar and showed similarity with accessions from Karnataka Bm.4 (KK). These two sub-groups were joined together at 0.66 genetic distance level. Overall, the levels of genetic similarity within the accessions varied from 0.24 to 0.80, the matrix ranged from 0.36 to 0.80, with a mean value of 0.68 indicating genetic similarity at low level.     

Andrographis paniculata (Burm. f.) Wall. ex Nees (kalmegh), a traditional hepatoprotective drug from India

Andrographis paniculata [(Burm. f.) Wall. ex Nees, family Acanthaceae], commonly known as kalmegh is used both in Ayurvedic and Unani system of medicines for a number of ailments related to digestion, hepatoprotection, hypoglycaemic and as anti-bacterial, analgesic, anti-inflammatory, vermicidal and antipyretic. Although kalmegh is extensively used in Indian Systems of Medicine, but it is not yet cultivated on a large-scale. The supply of herb is met from the wild growing populations which is now under constrain due to over-collection. Considering its value as a drug raw material, genetic variability of A. paniculata was augmented from different geographical regions of India by explorations and was conserved at the National Gene Bank, NBPGR, New Delhi. The therapeutic activity of the herb has been attributed to andrographolide, a bicyclic diterpenoid lactone present in the whole herb. Thirty germplasm collections of A. paniculata were evaluated for bioactive compound andrographolide to find out superior genotype for high drug yield. The andrographolide content in the germplasm was quantitatively analyzed by high performance thin-layer chromatography (HPTLC) at 232 nm. Results indicated wide variability among thirty accessions of A. paniculata for andrographolide content which ranged from 0.72 to 2.99 % on dry weight basis. DIVA-GIS analysis of A. paniculata germplasm for andrographolide content showed that diverse accessions of A. paniculata exist in three Indian states of Orissa, Tamil Nadu and Kerala. Promising accessions having high andrographolide content were identified; IC520361 (2.99 %), IC520395 (2.86 %), IC399125 (2.81 %), IC369404 (2.77 %) and IC520394 (2.61 %). Due to the economic importance of the drug, these promising collections can be exploited for large-scale cultivation for their pharmacologically important constituents mainly andrographolide content and also can serve as potential source for breeding and improvement of cultivars for obtaining higher drug yield.     

Analysis of clonal germplasm from five Saccharum species: S. barberi , S. robustum, S. officinarum, S. sinense and S. spontaneum . A study of inter- and intra species relationships using microsatellite markers

In order to study relationships within and among species of the highly complex polyploid genus Saccharum, 30 clones from 5 species (S. barberi, S. officinarum, S. robustum, S. sinense, and S. spontaneum) and 6 cultivated clones were analyzed using 15 sugarcane simple sequence repeat (SSR) markers and two multivariate statistical methods. The origins of two species, S. barberi and S. sinense, are unclear, and it has been hypothesized that they arose from crosses of S. officinarum × S. spontaneum. S. robustum is suggested to be ancestral to S. officinarum, which, when crossed with S. spontaneum, principally comprise the genomes of commercial cultivars. Although our analyses supported S. robustum as being an ancestor of S. officinarum, and show the composition of commercial clones as expected, our results concerning the species S. barberi and S. sinense indicated them to be much less related to the main Saccharum germplasm pool than previous molecular marker investigations, suggesting that introgression from other genera may exist in their ancestry. The differing results of our analysis of S. barberi and S. sinense are likely attributable to more equal and larger numbers of clones studied in each species. Given the extremely high ploidy, the use of only 15 SSR markers enabled clear distinction of Saccharum species and clones, and also identified likely mislabeled accessions. In addition, the replication of DNA extraction, polymerase chain reactions, and fragment electrophoresis increased the robustness of our analysis.     

The genus <Emphasis Type="Italic">Corchorus</Emphasis> L. (Malvaceae) in India: species distribution and ethnobotany

Understanding phyto-geographical distribution pattern and ethnobotanical applications are critical for comprehensive conservation and utilisation of native plant genetic resources. Dearth of such scientific reports on Corchorus species in India seriously compromises inclusiveness, precision and penetration of national endeavour to conserve the valuable resources. In this context, an ecogeographical study conducted to analyse geographical distribution, ecological adaptation, species richness and ethnobotanical applications of the genus in the country. The study revealed pandemic distribution of the genus with differential habitat preferences. Combine geographical, edaphic and precipitation based distribution profile of Corchorus spp. demonstrated multilayer interactions between species and environment that led to their differential spatial distribution. The highest species concentration was found in Tamil Nadu followed by Gujarat, Rajasthan, Madhya Pradesh and Maharashtra. Complementarity analysis showed that the major diversity hotspots for the genus present in Tamil Nadu and Gujarat. Besides, wide spectrum ethnobotanical applications of the genus reflect their long association with prevailing ethnic communities across regions. Documentation of such time honoured traditional practices has special bearing on designing comprehensive conservation and management strategies. This information will help to prioritise both ex-situ and in-situ conservation efforts for the genus particularly in ecologically vulnerable sites of India.     

Development of an algorithm identifying maximally diverse core collections

The development of a core collection, one which represents the genetic diversity of a crop with minimal redundancy and increases utility of the collection as a whole, is especially important as the funding for germplasm collections decreases. With limited resources, it is difficult to manage large germplasm collections and disperse genetically diverse germplasm to plant breeders. An algorithm was developed to assist in selection of core collections based on estimates of genetic distance. The criteria for selection of the maximum genetically diverse set were based on rankings of genetic distance between an accession with respect to all other accessions. Depending on the size core which a user wished, a zone around each selected accession was determined and no other accession within these limits was selected. The premise for the algorithm was that the genetic variability represented in the core must be representative of the distribution of genetic distances within the population of interest. In the present study, the algorithm was used with RAPD-marker-based estimates of genetic distance for 270 Theobroma cacao L. accessions and 134 Capsicum accessions that chose a set representing 18.5% of the population and representing the breadth of RAPD-based variation.     

Genetic diversity of the African hexaploid species Solanum scabrum Mill. and Solanum nigrum L. (Solanaceae)

Two hexaploid species of Solanum sect. Solanum are present in Africa: Solanum scabrum and S. nigrum. Solanum scabrum is a widely cultivated species and is used as a leafy vegetable, as a source of medicine and as a source of ink dye. In previous studies a wide range of morphological diversity has been reported in this species and in some studies subspecies have been proposed. Subspecies are also recognized in S. nigrum. However, it has not been established whether or not the morphological differences are reflected at the genomic level. The present study applies AFLPs to study the genetic diversity in S. scabrum and its relationship to geographical provenance, morphological differences and the possible existence of subspecies within S. scabrum and S. nigrum. The data obtained were analyzed with cluster analysis (using UPGMA and NJ). The results indicate that the genetic variation within S. scabrum was higher within accessions than between accessions. Accessions did not cluster according to their geographical provenance, indicating that accessions from different geographical areas were not significantly different genetically. The clustering reflected neither morphological differences nor domestication status (cultivated or wild). The morphological differences exhibited by S. scabrum could be due to selection by farmers for different plant types. The AFLP derived clustering pattern did not segregate the subspecies recognized in S. scabrum and S. nigrum into separate subclusters.     

AFLP Analysis of the Phenetic Organization and Genetic Diversity in the Sugarcane Complex, Saccharum and Erianthus

Amplified fragment length polymorphism (AFLP) markers were evaluated for determining the phylogenetic relationships, and the diversity in the Saccharum complex using 30 clones belonging to S. officinarum, S. robustum, S. spontaneum, S. barberi, S. sinense and the related genus Erianthus. The phenetic tree of the species clones based on AFLP data was consistent with the known taxonomical relationships. AFLP gave higher resolution of closely related species into discrete groups than that by RAPD and RFLP markers, reported earlier. The levels of diversity within the various Saccharum species were also found to be higher than those obtained previously with the same set of clones using RAPD markers. The intraspecies similarity in S. barberi and S. sinense was much higher than interspecies similarity suggesting a clear separation of the two, which are considered ‘horticultural species’. The genetic similarity matrix derived from a single primer combination highly correlated (r = 0.980) with that obtained from all the 12 primer combination used in the study, thus highlighting the efficiency of a single primer combination in delineating species relationships. All the primer combinations could identify markers that are specific to each of the species and the genus Erianthus. Among the species, specific markers were highest in S. spontaneum followed by S. robustum, S. barberi, S. officinarum and S. sinense. Erianthus had a distinct profile with 30% of the total amplified fragments being specific to it. This offers great scope for identifying intergeneric hybrids, which has been very difficult using morphological traits and RAPD markers. High degree of correspondence between the results from the cluster analysis based on Jaccard's similarity index, Neighbour Joining tree based on Sokal and Michener distance matrix and AFTD (Analyses Factorielle on Table of Distances) analysis clearly demonstrated that AFLP markers would be an appropriate tool in providing better information about the relationships among the species, estimation of diversity, and in revealing species and genus specific markers that could be directly applied in sugarcane breeding programmes.     

Assessment of genetic diversity in Solanum trilobatum L., an important medicinal plant from South India using RAPD and ISSR markers

Solanum trilobatum L. is an Indian medicinal plant containing rich amount of steroidal glyco-alkoloids that can be used as precursor for commercial steroid production. Two efficient marker systems such as Random Amplified Polymorphic DNA (RAPD) and Inter Simple Sequence Repeats (ISSR) were used for the first time to assess the genetic diversity across 14 S. trilobatum accessions obtained from five South Indian states. Twenty out of 60 RAPD primers generated 189 distinct bands of which 160 were polymorphic with an average of 8 polymorphic bands per primer. A maximum of up to 15 fragments were amplified with an average of 9.45 bands per primer and the amplicons varied in size between 100 and 3,000 bp. The percentage of polymorphism ranged from 55.5 to 100, with an average of 84.6. ISSR profiling using 7 out of 20 primers amplified 83 bands and the number of amplified fragments varied from 2 to 16 with a size range of 200–1,800 bp. Totally 72 polymorphic bands were obtained using 7 ISSR primers at an average of 10.28 polymorphic bands per primer. Polymorphism percentage varied from 50 to 100 among the selected accessions resulting in an average percentage of polymorphism of 86.7. The PIC values ranged from 0.49 to 0.93 for RAPD and 0.16 to 0.90 for ISSR primers. The study pointed out that ISSR markers were more efficient than RAPD markers in evaluating the degree of genetic variation in S. trilobatum. The UPGMA cluster analysis grouped all Tamil Nadu accessions in one cluster and other state accessions in another cluster. The Principal component analysis also substantiates this clustering pattern. Thus the phylogenetic relationship and a high genetic variation revealed in the present study could provide a baseline data for conservation and improvement of this plant in future. Also the molecular markers identified in this study will be helpful in authentication of this species to prevent adulteration in herbal medicine.     

Seed variability in <Emphasis Type="Italic">Madhuca longifolia</Emphasis> (Koenig) J. F. Macbride,a source of oil for use as bio-energy

Madhuca longifolia (Koenig) J. F. Macbride is native to India. Seeds contain 20–50% oil (Mahua oil) and the seed cake is used as manure. Mahua oil is used by rural communities for protection against storage pests. Almost all parts of this tree have medicinal properties. Four explorations and collection missions was conducted during July–October 2007 in Tamil Nadu and the Union Territory of Puducherry. This resulted in the collection of 55 accessions. Characterisation and analysis of 7 seed characters were done. The existence of enormous variability in seed length, thickness and 100 seed weight indicates scope for utilization of these accessions for selection of promising material for extraction of oil. Oil content of kernels has been evaluated. Kernel oil ranged from 44.43 to 61.50%. Three accessions IC556617 with 61.50, IC556632 with 60.80 and IC556632 with 60.55% of kernel oil are superior to the rest. Further collections in the site of these oil rich accessions are promising for more productive Madhuca longifolia genetic resources.     

Evaluation of cultivated and wild barley for resistance to pathotypes ofPuccinia hordei with wide virulence

An isolate ofPuccinia hordei (ND89-3) originally collected in Morocco is virulent on most barley genotypes reported to possess resistance, except cultivar Estate (CI 3410), which possesses theRph3 gene and exhibits a low to intermediate level of resistance (infection type 12). Isolate ND89-3 possesses one of the widest virulence spectrums reported forP. hordei. Accessions ofHordeum vulgare (1,997 in total) andH. spontaneum (885 in total), mostly originating from the Mediterranean region and parts of North Africa, were evaluated with isolate ND89-3 at the seedling stage to identify new sources of leaf rust resistance. Fifty-eight accessions ofH. vulgare, and 222 accessions ofH. spontaneum exhibited low infection types to this isolate. Further evaluations of these resistant accessions with isolates ofP. hordei virulent forRph3,Rph7, andRph12 suggested that most of the resistantH. vulgare accessions possess theRph3 gene. Data suggested additional sources of effective resistance inH. vulgare are rather limited. FiveH. vulgare accessions and 167H. spontaneum accessions were identified as possible sources of new genes for leaf rust resistance. These accessions likely possess resistance genes that are different fromRph1 toRph12, or gene combinations thereof based on their reaction to four leaf rust isolates. Utilization of these accessions in barley breeding will broaden the germplasm resources available for genetic control ofP. hordei. North Dakota Agricultural Experiment Station Journal Series No. 2123. The U.S. Government right to retain a non-exclusive, royalty free licence in and to any copyright is acknowledged.     

Identification of new sources for resistance to powdery mildew in H. spontaneum-derived winter barley lines

Twenty four barley lines derived from the F ₇generation of crosses between two winter barley cultivars and different accessions of Hordeum spontaneum Koch collected in Israel were tested against a set of ten European and five Israeli powdery mildew cultures, possessing virulence genes which completely match the spectrum of known mildew resistance genes. The comparison of reaction patterns justified the conclusion that new genes for resistance have been conferred from H. spontaneum which also differ from genes previously identified in other wild barley accessions from Israel. Participation in the expression of the resistance reaction of one of the two barley cultivars used in the development of the lines is well documented in some cases. The majority of the lines was found to be highly resistant against all or almost all European isolates, while various kinds of reactions were observed against the Israeli cultures. The effective transfer of novel mildew resistance from H. spontaneum into winter barley confirms similar results with spring barley, indicating natural wild barley populations in Israel as a significant gene pool for yet unexploited mildew resistance in barley.     

Analysis of diversity and distribution of <Emphasis Type="Italic">Jatropha curcas</Emphasis> L. germplasm using Geographic Information System (DIVA-GIS)

Jatropha curcas L., an important biodiesel plant, has been studied for its distribution and diversity in south east coastal zone of India using DIVA-GIS. The Grid maps were generated on the distribution pattern, plant height, number of primary branches, collar length, number of fruits per cluster and oil content. Flowering in the collected accessions were grouped in to very early (35%), early (29%), medium duration (10%), late (20%), and very late (6%) types. Analysis for richness using rarefaction method of DIVA-GIS showed that Ranga Reddy district of Andhra Pradesh is the potential area for germplasm with high oil content. The present study revealed that diverse germplasm accessions of J. curcas are distributed all over the south east coastal zone and enabled us to find out gaps in collection and diversity richness from SEC zone of India for conservation.     

Evaluating landraces of bread wheat <Emphasis Type="Italic">Triticum aestivum</Emphasis> L. for tolerance to aluminium under low pH conditions

Bread wheat (Triticum aestivum L.) landraces held within ex situ collections offer a valuable and largely unexplored genetic resource for wheat improvement programs. To maximise full utilisation of such collections the evaluation of landrace accessions for traits of interest is required. In this study, 250 accessions from 21 countries were screened sequentially for tolerance to aluminium (Al) using haematoxylin staining of root tips and by root regrowth measurement. The staining test indicated tolerance in 35 accessions, with an intermediate response to Al exhibited in a further 21 accessions. Of the 35 accessions classified as tolerant, 33 also exhibited increased root length following exposure to Al. The tolerant genotypes originated from Bulgaria, Croatia, India, Italy, Nepal, Spain, Tunisia, and Turkey. AFLP analysis of the 35 tolerant accessions indicated that these represent diverse genetic backgrounds. These accessions form a valuable set of germplasm for the study of Al tolerance and may be of benefit to breeding programs for expanding the diversity of the gene pool from which tolerant cultivars are developed.     

Isozyme Banding Pattern and Estimation of Genetic Diversity among Guinea Grass Germplasm

Isozyme banding pattern was studied in Guinea grass (Panicum maximum Jacq.), a widely cultivated grass having good fodder value. Similarity among 63 accessions collected from diverse sources was worked out using five enzyme systems (SOD, GOT, ACP, Esterase and Peroxidase) following horizontal starch gel electrophoresis. Biochemical markers such as isozymes are useful supplements in identifying the genetic variation present in any crop. A total of 35 clear and unambiguous bands were used for analysis of which 8 bands were monomorphic. Polymorphism exhibited by 27 bands from all five enzyme systems indicate presence of considerable diversity in this species. The dendrogram generated after UPGMA and SAHN cluster analysis using Jaccard genetic distance showed that 63 accessions from diverse geographical locations could be grouped in three main clusters, of which two could further be divided into sub-clusters. Although the clusters comprised members from different locations, most of the accessions from similar geographical locations tended to cluster in same group.