Reduced Levels of DNA Polymorphism and Fixed between-Population Differences in the Centromeric Region of Drosophila Ananassae

We have estimated DNA sequence variation within and between two populations of Drosophila ananassae, using six-cutter restriction site variation at vermilion (v) and furrowed (fw). These two gene regions are located close to the centromere on the left and right X chromosome arms, respectively. In the fw region, no DNA polymorphism was detected within each population. In the v region, average heterozygosity per nucleotide was very low in both populations (pi = 0.0005 in the Burma population, and 0.0009 in the India population). These estimates are significantly lower than those from loci in more distal gene regions. The distribution of DNA polymorphisms between both populations was also striking. At fw, three fixed differences between the Burma and India populations were detected (two restriction site differences and one insertion/deletion of approximately 2 kb). At v, each DNA polymorphism in high frequency in the total sample was nearly fixed in one or the other population, although none of them reached complete fixation. The observed pattern of reduced variation within populations and fixed differences between populations appears to correlate with recombination rate. We conclude that recent hitchhiking associated with directional selection is the best explanation for this pattern. The data indicate that different selective sweeps have occurred in the two populations. The possible role of genetic hitchhiking in rapid population differentiation in gene regions of restricted recombination is discussed.     

Phylogeny and biogeography of exacum (gentianaceae): a disjunctive distribution in the Indian ocean basin resulted from long distance dispersal and extensive radiation

Disjunctive distributions across paleotropical regions in the Indian Ocean Basin (IOB) often invoke dispersal/vicariance debates. Exacum (Gentianaceae, tribe Exaceae) species are spread around the IOB, in Africa, Madagascar, Socotra, the Arabian peninsula, Sri Lanka, India, the Himalayas, mainland Southeast Asia including southern China and Malaysia, and northern Australia. The distribution of this genus was suggested to be a typical example of vicariance resulting from the breakup of the Gondwanan supercontinent. The molecular phylogeny of Exacum is in principle congruent with morphological conclusions and shows a pattern that resembles a vicariance scenario with rapid divergence among lineages, but our molecular dating analysis demonstrates that the radiation is too recent to be associated with the Gondwanan continental breakup. We used our dating analysis to test the results of DIVA and found that the program predicted impossible vicariance events. Ancestral area reconstruction suggests that Exacum originated in Madagascar, and divergence dating suggests its origin was not before the Eocene. The Madagascan progenitor, the most recent common ancestor of Exacum, colonized Sri Lanka and southern India via long-distance dispersals. This colonizer underwent an extensive range expansion and spread to Socotra-Arabia, northern India, and mainland Southeast Asia in the northern IOB when it was warm and humid in these regions. This widespread common ancestor retreated subsequently from most parts of these regions and survived in isolation in Socotra-Arabia, southern India-Sri Lanka, and perhaps mainland Southeast Asia, possibly as a consequence of drastic climatic changes, particularly the spreading drought during the Neogene. Secondary diversification from these surviving centers and Madagascar resulted in the extant main lineages of the genus. The vicariance-like pattern shown by the phylogeny appears to have resulted from long-distance dispersals followed by extensive range expansion and subsequent fragmentation. The extant African species E. oldenlandioides is confirmed to be recently dispersed from Madagascar.     

Molecular genetic variation in the centromeric region of the X chromosome in three Drosophila ananassae populations. II. The Om(1D) locus

Restriction-site and sequence-length polymorphism in the Om(1D) locus region on the X chromosome in Drosophila ananassae was investigated for three natural populations (from Burma, India, and Brazil), by using hexanucleotide-recognizing restriction enzymes. The estimates of average heterozygosity per nucleotide (pi) were 0.0085, 0.0043, and 0.0004 for the Burma, India, and Brazil populations, respectively, and the average frequencies of insertions/deletions were 0.078, 0.054, and 0.007/chromosome/kb. While the pi values at this locus are similar to the estimates obtained from other euchromatic loci in D. ananassae or in other Drosophila species, the frequencies of insertions/deletions are much higher than those previously reported from Drosophila. The exceptionally high frequencies of length polymorphisms in the Burmese sample and, to a lesser extent, in the Indian sample indicate that the hypermutability of Om(1D), caused by the frequent insertion of the transposable element tom, may be due to locus-specific rather than to tom element-specific properties. The low level of nucleotide variation in the Brazilian population seems to be due to a recent bottleneck of population size. This population was apparently founded in recent years by a small number of individuals and has been relatively isolated ever since.     

Molecular and biochemical characterization of a sand fly population from Sri Lanka: evidence for insecticide resistance due to altered esterases and insensitive acetylcholinesterase

With an increasing incidence of cutaneous leishmaniasis in Sri Lanka, particularly in northern provinces, insecticide-mediated vector control is under consideration. Optimizing such a strategy requires the characterization of sand fly populations in target areas with regard to species composition and extant resistance, among other parameters. Sand flies were collected by human bait and cattle-baited net traps on Delft Island, used as an illegal transit location by many refugees returning to the north of Sri Lanka from southern India where leishmaniasis is endemic. For species identification, genomic DNA was extracted and a fragment of the ribosomal 18S gene amplified. The sequence from all flies analysed matched that of Phlebotomus argentipes Annandale & Brunetti, the primary vector in India and the most likely vector in Sri Lanka. Independent morphological analysis also identified P. argentipes. To establish the current susceptibility status of vector species, data were obtained at the biochemical level, from which potential cross-resistance to alternative insecticides can be predicted. The Delft Island collection was assayed for the activities of four enzyme systems involved in insecticide resistance (acetylcholinesterase, non-specific carboxylesterases, glutathione-S-transferases and cytochrome p450 monooxygenases), establishing baselines against which subsequent collections can be evaluated. There was preliminary evidence for elevated esterases and altered acetylcholinesterase in this population, the first report of these resistance mechanisms in sand flies to our knowledge, which probably arose from the malathion-based spraying regimes of the Anti-Malarial Campaign.     

Burmannia larseniana (Burmanniaceae): a new species from Thailand

A new semi-mycotrophic species, Burmannia larseniana , is described from northern Thailand. The species is related to B. pusilla , a species widely distributed in Cambodia, Vietnam, Burma, India and Sri Lanka, but can be easily distinguished from the latter by its succulent stem, single-veined leaf, and the outer perianth lobes with double margins that encircle the inner perianth lobes. The ecological preferences of the species are also discussed.     

The origin of Indian Star tortoises (Geochelone elegans) based on nuclear and mitochondrial DNA analysis: A story of rescue and repatriation

The Indian Star tortoise (Geochelone elegans) belongs to the family Testunidae and is distributed in southwest India and Sri Lanka. In addition to facing loss of its natural habitat, the species is also illegally traded as food and as an exotic pet internationally. Here we report DNA-based analyses for identification and repatriation of these tortoises into their natural habitat. We have attempted to establish the geographical origin of these tortoises rescued from smugglers, by comparing the microsatellite and mitochondrial markers of rescued animals with animals of known provenance. Star tortoises exhibited strong genetic structure in India. The populations from western India were genetically distinct at microsatellite and mitochondrial loci from southern populations. The rescued individuals had similar multilocus genotypes and mitochondrial DNA haplotypes as the reference individuals from south India. However, the precise geographic origin of many of the rescued samples remains unresolved, because we could not assign them to southern populations and the Neighbor-Joining cluster analysis indicated that some of rescued tortoises formed distinct clusters. These data strongly suggest that the rescued group of tortoises is composed of a mix of individuals from differentiated source populations that are probably located in southern India and possibly Sri Lanka. Our study provides valuable information based on molecular markers for the assessment of genetic diversity in Indian Star tortoises.     

Very low genetic variability in the Indian queenless ant Diacamma indicum

Abstract We developed microsatellite markers and combined them with mitochondrial markers to analyse the population genetic structure of the queenless ant Diacamma indicum. This species, lacking winged queens, is likely to have a restricted female dispersal but exhibits various life history traits suggesting higher dispersal abilities than the other Diacamma species. Only 4 of 11 microsatellites were polymorphic and only 1 had more than 4 alleles over 166 individuals originating from 7 populations from the south of India. Only one mitochondrial DNA (mtDNA) haplotype was detected throughout India (including one population in the north) and Sri Lanka. Such a level of polymorphism is particularly low compared with other Diacamma species having much smaller ranges in the south of India. A strong genetic differentiation was observed between populations separated by more than a few kilometres. We also analysed the genetic differentiation between the Indian populations and two populations from the Japanese island of Okinawa, which are morphologically similar and might belong to the same species. The genetic differentiation was high for both markers, suggesting an absence of ongoing gene flow between these populations.     

A conspectus of Indo‐Burmese Annonaceae

A list of the Annonaceae taxa native to the Indo‐Burmese Region is presented. Species are listed with synonyms and types, and the regional distribution is given by country (India, Sri Lanka, Pakistan, Nepal, Bhutan, Bangladesh and Burma). In total, 195 species from 28 genera are listed. Two new combinations, Monoon nitidum (A. DC.) I. M. Turner and Polyalthia malabarica (Bedd.) I. M. Turner, are included. Polyalthia corticosa (Pierre) Finet & Gagnep. is considered the correct name for P. obliqua in India and Bangladesh. Artabotrys cubittii Chatterjee is reduced to a synonym of A. burmanicus A. DC. Two new records for Burma, Polyalthia bullata King and Uvaria hahnii (Finet & Gagnep.) J. Sinclair, are presented.     

Genetic differentiation among local populations of Asian elephant

Electrohoretically detectable enetic variation for 29 kinds of blood protein encoded by 33 loci was analyzed for 78 Asian eletants (Elephas maximus) which were collected from its four local populations: Sri Lanka, Souti India, Thailand and Nepal. Elehants in Sri Lanka are classified into the subspecies E.m. maximus, and those from the other tlree localities into the subspecies E. m. indicus. Six variable loci were detected, and one of them, the tetrazolium oxidase locus, was observed to show a complete allele substitution between the subspecies. Average heterozgosity within local populations were in a range of 0.0152 ? 0.0303. Whereas the Nei's genetic distance among three local populations of the subspecies indicus were 0.0013 ? 0.0031, the distance between the subspecies indicus and maximus were 0.0328 ? 0.0370, indicating that the two subspecies were well differentiated genetically.     

The role of natural selection in genetic differentiation of worldwide populations of Drosophila ananassae

The main evolutionary forces leading to genetic differentiation between populations are generally considered to be natural selection, random genetic drift, and limited migration. However, little empirical evidence exists to help explain the extent, mechanism, and relative role of these forces. In this study, we make use of the differential migration behavior of genes located in regions of low and high recombination to infer the role and demographic distribution of natural selection in Drosophila ananassae. Sequence data were obtained from 13 populations, representing almost the entire range of cosmopolitan D. ananassae. The pattern of variation at a 5.1-kb fragment of the furrowed gene, located in a region of very low recombination, appears strikingly different from that of 10 noncoding DNA fragments (introns) in regions of normal to high recombination. Most interestingly, two main haplotypes are present at furrowed, one being fixed in northern populations and the other being fixed or in high frequency in more southern populations. A cline in the frequency of one of these haplotypes occurs in parallel latitudinal transects. Taken together, significant clinal variation and a test against alternative models of natural selection provide evidence of two independent selective sweeps restricted to specific regions of the species range.     

Phylogenetic affinities of the Iraqi populations of Saw-scaled vipers of the genus Echis (Serpentes: Viperidae), revealed by sequences of mtDNA genes

The Saw-scaled vipers of the species Echis carinatus range from Sri Lanka and India westwards to Iraq, including the eastern Arabian Peninsula. We collected the species in southern Iraq and compared two mtDNA genes (16S and Cyt b) with other populations of this species and with other species of the genus. Analyses of both Maximum Likelihood and Bayesian Inference confirmed E. carinatus as a monophyletic species. The samples from Iraq cluster with populations of the species from Pakistan and UAE. Populations from India, however, are situated in a separate phylogenetic lineage. This can be explained by the geographic barriers between western (Iraq, Pakistan and UAE) and eastern (India) populations of the species. Soleyman Mountain in southern Pakistan is the main barrier between them and its role is reflected in the genetic distance between populations.     

Morphometric analyses of capsule and seed traits in Scandinavian Papaver radicatum (Papaveraceae) in relation to the bicentric distribution pattern

The bicentric distribution pattern of certain plant species in the southern and northern Scandinavian mountains has been explained in different ways. Either by refugial survival, by late-glacial immigration to the first deglaciated areas in southern and northern Norway or by a successive fragmentation of wide-distributed populations during post-glacial time — or by present-day ecological factors. Even if the bicentric distribution pattern is at least partly explained by present-day ecological conditions, the question about the origin of the bicentric distribution still remains. One way to tackle this problem, is to investigate the morphometric and/or the genetic differentiation between isolated populations of ‘bicentric’ species and its impact on the explanation of the disjunct pattern. The overall morphometric differentiation pattern in seeds and capsules was investigated in populations of Papaver radicatum, a perennial herb with a bicentric distribution in the Scandinavian mountain range. Canonical variates analysis of capsules separates the populations into two groups in accordance with their geographic origin, i.e. the regions of southern and northern Scandinavia. The differentiation pattern indicates a two-step development of the present-day distribution: first separation of the southern and northern Scandinavian occurences of older origin, then separation between populations within each region. The results from canonical variates analysis of seeds shows a weaker variation pattern with a tendency of overlapping inter-region populations. The most northerly situated populations from the southern region are grouped with populations from the northern region. The pattern of differentiation in capsules may be interpretated in terms of refugial survival or late-glacial immigration. However, the variation pattern in seeds rather points to a scenario where a widespread occurrence that previously ranged from southern to northern Scandinavia was successively fragmented, which makes a postglacial development of the differentiation pattern more probable.     

Cytogenetical survey of black rats,Rattus rattus,in southwest and Central Asia,with special regard to the evolutional relationship between three geographical types

A chromosome survey of the black rat, Rattus rattus, was made from animals collected at different localities in Southwest and Central Asia. Asian type black rats (2 n=42) were distributed in northern India, northern Pakistan, while the Oceanian type rats (2 n=38) were found in southern India, southern Pakistan and Central Asia. A border line of distribution of rats with Asian and Oceanian types can be drawn dividing India and Pakistan into northern and southern parts. A hybrid type between Asian and Oceanian types was found in Karachi, Pakistan. Rats with 40 chromosomes, probably a transient type from Asian to Oceanian type, were found in Sri Lanka (Ceylon). It is suggested that these three geographic variants have developed via sequential events of Robertsonian fusion of acrocentric chromosomes in Asian type black rats. This fusion probably took place somewhere in southern India. The Oceanian type black rats that thus developed in southern India migrated widely to the rest of the world through Central Asia and Europe accompanying the movement of mankind.     

Molecular Genetic Variation in the Centromeric Region of the X Chromosome in Three Drosophila Ananassae Populations. I. Contrasts between the Vermilion and Forked Loci

We have surveyed three natural populations of Drosophila ananassae for restriction map variation at the forked (f) and vermilion (v) loci, using 6-cutter restriction enzymes. Both loci are located in the centromeric region of the X chromosome. Two major conclusions can be drawn from the data. First, we found strong evidence for population subdivision, i.e., significant differences in the frequency distributions of polymorphisms and/or haplotypes between the Burma, India, and Brazil populations. Secondly, the pattern of DNA sequence variation between the two loci is unexpectedly different. The level of nucleotide variation in the v locus region is reduced (relative to f), especially in the Burma population. Furthermore, in contrast to v, we found no insertions/deletions larger than 700 bp and no significant linkage disequilibrium at f. The genetic differentiation among subpopulations can readily be attributed to restricted migration as the predominant evolutionary force. According to population genetics theory, the differences in DNA polymorphisms between the two loci are in qualitative agreement with the hypothesis that recombination is reduced in the v locus region ("centromere effect") but not at f. In order to test this hypothesis directly, we determined the cytogenetic positions of several loci in the centromeric region by in situ hybridization and found by comparison with the genetic map that recombination at v is indeed very low, much lower than at f.     

Distribution and Conservation of Slender Lorises (Loris tardigradus lydekkerianus) in Southern Andhra Pradesh,South India

Slender lorises live in forests of southern India and Sri Lanka. Little is known about their distribution patterns and relative densities. We report the results of a survey conducted in 6 forest divisions in the southern parts of the state of Andhra Pradesh, South India. Relatively high densities of lorises occurred in mixed deciduous forests and in adjoining farm lands interspersed with trees. Three distinct populations inhabit the study area. We recommend conservation measures for Loris tardigradus.     

Regional genetic differentiation in Western Australian sandalwood (<Emphasis Type="Italic">Santalum spicatum</Emphasis>) as revealed by nuclear RFLP analysis

Western Australian sandalwood, Santalum spicatum, is widespread in the semi-arid and arid regions of Western Australia, and there is some morphological variation suggestive of two ecotypes. The level and structuring of genetic diversity within the species was investigated using anonymous nuclear RFLP loci. Santalum spicatum showed moderate levels of genetic diversity compared to other Australian tree species. The northern populations in the arid region showed greater levels of diversity and less population differentiation than the southern populations in the semi-arid region due to differences in the distribution of rare alleles. Equilibrium between drift and gene flow in the northern populations indicated that they have been established for a long period of time with stable conditions conducive to gene flow. In contrast, the southern populations showed a relationship between drift and gene flow indicative of a pattern of fragmentation and isolation where drift has greater effect than gene flow. The different patterns of diversity suggest that the ecotypes in the two regions have been subject to differences in the relative influences of drift and gene flow during their evolutionary history.Communicated by D.B. Neale     

Evolutionary relationships and reproductive isolating mechanisms in the rice frog (Fejervarya limnocharis) species complex from Sri Lanka, Thailand, Taiwan and Japan, inferred from mtDNA gene sequences, allozymes, and crossing experiments

The rice frog (Fejervarya limnocharis) species complex is widely distributed, from India to Japan, and most prevalently in Southeast Asia. Conspicuous morphological variation has been reported for this species complex throughout its distribution range. In the present study, we used mtDNA gene sequence and allozyme analyses to infer evolutionary affinities within this species complex using eight populations (Sri Lanka; Bangkok and Ranong in Thailand; Taiwan; and Hiroshima, Okinawa, Ishigaki and Iriomote in Japan). We also conducted crossing experiments among four populations from Japan, Thailand, and Sri Lanka in order to find out more about the reproductive isolating mechanisms that might exist among the East, Southeast, and South Asian populations of this species complex. The crossing experiments revealed that the Sri Lanka population is reproductively isolated from the Hiroshima, Bangkok, and Ranong populations by complete hybrid inviability, and that the Bangkok population may be reproductively isolated from the Hiroshima population by partial hybrid inviability. Thus, it is not unreasonable to regard the Sri Lanka population as a species separated from F. limnocharis. The mtDNA and allozyme data showed that the Ranong population is most closely related to the Bangkok population in nuclear genome, but more similar to the Okinawa and Taiwan populations in mtDNA genome. The present, preliminary survey may raise questions about the species status of these particular populations and also about the nature of the biological species concept.     

Seed-Mediated Gene Flow Promotes Genetic Diversity of Weedy Rice within Populations: Implications for Weed Management

Increased infestation of weedy rice—a noxious agricultural pest has caused significant reduction of grain yield of cultivated rice (Oryza sativa) worldwide. Knowledge on genetic diversity and structure of weedy rice populations will facilitate the design of effective methods to control this weed by tracing its origins and dispersal patterns in a given region. To generate such knowledge, we studied genetic diversity and structure of 21 weedy rice populations from Sri Lanka based on 23 selected microsatellite (SSR) loci. Results indicated an exceptionally high level of within-population genetic diversity (H_e = 0.62) and limited among-population differentiation (F_st = 0.17) for this predominantly self-pollinating weed. UPGMA analysis showed a loose genetic affinity of the weedy rice populations in relation to their geographical locations, and no obvious genetic structure among populations across the country. This phenomenon was associated with the considerable amount of gene flow between populations. Limited admixture from STRUCTURE analyses suggested a very low level of hybridization (pollen-mediated gene flow) between populations. The abundant within-population genetic diversity coupled with limited population genetic structure and differentiation is likely caused by the considerable seed-mediated gene flow of weedy rice along with the long-distance exchange of farmer-saved rice seeds between weedy-rice contaminated regions in Sri Lanka. In addition to other effective weed management strategies, promoting the application of certified rice seeds with no weedy rice contamination should be the immediate action to significantly reduce the proliferation and infestation of this weed in rice ecosystems in countries with similar rice farming styles as in Sri Lanka.     

Development and optimization of sequence-tagged microsatellite site markers to detect genetic diversity within Colletotrichum capsici, a causal agent of chilli pepper anthracnose disease

Genomic libraries enriched for microsatellites from Colletotrichum capsici, one of the major causal agents of anthracnose disease in chilli pepper (Capsicum spp.), were developed using a modified hybridization procedure. Twenty-seven robust primer pairs were designed from microsatellite flanking sequences and were characterized using 52 isolates from three countries India, Sri Lanka and Thailand. Highest gene diversity of 0.857 was observed at the CCSSR1 with up to 18 alleles among all the isolates whereas the differentiation ranged from 0.05 to 0.45. The sequence-tagged microsatellite site markers developed in this study will be useful for genetic analyses of C. capsici populations.     

A revision of the genus Sarcococca (Buxaceae)

SEALY, J. R., 1985. A revision of the genus Sarcococca (Buxaceae). The genus Sarcococca is idely distributed in SE Asia from Afghanistan through the Himalaya to SE Tibet, Assam, Upper Burma and China, southwards in peninsula India, Sri Lanka, Thailand, Indochina, to Java and Sumatra, Taiwan and Luzon. The genus is accepted as comprising 11 species, one with three varieties, two with two varieties each, and another of two forms. These are described in detail. Sarcococca walhchii f. membranacea from Manipur, is a new form and V. zevlanica var. brevifolia is a new combination.