Conservation genetics: beyond the maintenance of marker diversity

One of the major problems faced by conservation biologists is the allocation of scarce resources to an overwhelmingly large number of species in need of preservation efforts. Both demographic and genetic information have been brought to bear on this problem; however, the role of information obtained from genetic markers has largely been limited to the characterization of gene frequencies and patterns of diversity. While the genetic consequences of rarity may be a contributing factor to endangerment, it is widely recognized that demographic factors often may be more important. Because patterns of genetic marker variation are influenced by the same demographic factors of interest to the conservation biologist, it is possible to extract useful demographic information from genetic marker data. Such an approach may be productive for determining plant mating systems, inbreeding depression, effective population size, and metapopulation structure. In many cases, however, data consisting only of marker frequencies are inadequate for these purposes. Development of genealogical based analytical methods coupled with studies of DNA sequence variation within and among populations is likely to yield the most information on demographic processes from genetic marker data. Indeed, in some cases it may be the only means of obtaining information on the long-term demographic properties that may be most useful for determining the future prospects of a species of interest.     

Genetic variability and association of ISSR markers with some biochemical traits in mulberry (Morus spp.) genetic resources available in India

Utilizing intersimple sequence repeat (ISSR) markers, 18 mulberry (Morus spp.) germplasm collections were studied for genetic variability, phylogenetic relationship, and association with protein and sugar content. The genetic polymorphism exhibited by ISSR primers was 100%, and the genetic diversity recorded among the mulberry accessions had an average of 0.263 ± 0.094. Dendrogram (unweighted pair group method analysis) clustered the mulberry accessions into two major groups, one comprised the accessions collected from north or northeast regions of India, and the other comprised three subclusters and one isolate, i.e., Assamjati, a collection from Assam. Another subcluster contained accessions collected from Kerala, which belong to Morus indica. These accessions of M. indica from Kerala were found to be genetically diverse from north and northeast India. Multidimensional scaling of the ISSR data clearly separated the mulberry accessions according to their genetic diversity and protein content. Mulberry accessions were arbitrarily grouped into three classes viz. very low, moderate, and high in terms of protein and sugar content using standard statistical programs. Stepwise multiple regression analysis identified four ISSR markers (835_1,600, 835_5,600, 822_2,500, and 807_2,500) associated with protein content with highly positive correlation (p < 0.001) with linear curves with high F values (18.055 to 48.674; p < 0.001). In case of sugar content, four ISSR markers viz. 812₉₀₀, 817_1,500, 826_1,500, and 810_8,000 showed negative correlation. Hence, DNA markers for proteins seem promising and may be used in marker-assisted breeding program.     

SSR markers for a critically endangered species Euryodendron excelsum and a distantly related species Ternstroemia gymnanthera (Ternstroemiaceae)

Euryodendron excelsum is a critically endangered tertiary relict plant endemic to China. It has only one population remaining in Ba Jia Zhen of Yangchun, Guangdong. In this study, we discovered 25 microsatellite markers from E. excelsumusing a Fast Isolation by Amplified Fragment Length Polymorphism of Sequences COntaining Repeats (FIASCO) protocol. Thirteen loci demonstrated polymorphisms, with the number of alleles per locus ranging from 2 to 13. Values for observed and expected heterozygosity ranged from 0.176 to 1.000 and from 0.261 to 0.889, respectively. Three loci (ZXM-17, ZXM-54, and ZXM-92) were found to significantly deviate from Hardy-Weinberg equilibrium. In addition, five of 13 loci could be successfully cross-amplified in Ternstroemia gymnanthera. These microsatellite loci may help to further survey the adaptive evolution and genetic variation of E. excelsum for guiding its conservation.     

Isolation and characterization of eight compound microsatellite markers in a mangrove tree Kandelia candel (L.) Druce

Kandelia candel is an important mangrove tree species of family Rhizophoraceae. Here we isolated eight codominant compound microsatellite simple sequence repeat (SSR) loci from K. candel. Our isolated loci provided compound SSR markers with polymorphism of three to 11 alleles per locus. The expected and observed heterozygosities ranged from 0.230 to 0.887 and from 0.083 to 1.00, respectively. These markers would be the useful tools for analysing questions concerning population genetic structure and mating system of K. candel.     

Assessment of genetic fidelity of micropropagated <Emphasis Type="Italic">Swertia chirayita</Emphasis> plantlets by ISSR marker assay

Inter simple sequence repeat (ISSR) marker assay was employed to validate the genetic fidelity of Swertia chirayita plantlets multiplied in vitro by axillary multiplication upto forty-two passages. Sixteen ISSR primers generated a total of 102 amplicons among the tissue-cultured plants. Forty-eight amplicons were amplified in the outlier (a Swertia species). The outlier (negative control) was employed to rule out the possibility that the invariant fingerprint was due to chance alone and that the ISSR technique employed was not discriminatory enough to detect the off-types. A homogenous amplification profile was observed for all the micropropagated plants. The results confirmed the clonal fidelity of the tissue culture-raised S. chirayita plantlets and corroborated the fact that axillary multiplication is the safest mode for multiplication of true to type plants.     

Development and characterization of SSR markers in lychee (Litchi chinensis)

Sixteen microsatellite loci were isolated from lychee (Litchi chinensis), all of which exhibited polymorphism (two or three alleles per locus), with levels of heterozygosity ranging from 0.021 to 0.900. These loci can help assess the genetic structure of lychee.     

Isolation and characterization of polymorphic microsatellite markers for Alternaria alternata

Alternaria alternata is of major significance as a food and feed contaminant and is able to produce a range of mycotoxins that may elicit adverse effects in both animals and humans. We describe the isolation and characterization of five microsatellite markers for studying A. alternata. Marker polymorphism was screened in 64 isolates of A. alternata. The number of alleles per locus ranged from eight to 24, and allelic diversity ranged from 0.425 to 0.882. These markers will be useful in the study of relationships and population genetics amongst isolates of A. alternata.     

The distinctness and diversity of Ethiopian barleys

 The relative diversity and distinctness of Ethiopian barleys has been investigated using (1) morphology/isozyme/hordein polymorphisms and (2) RFLP markers. In the former a set of 51 landraces from over the whole of Ethiopia was compared with Iranian landraces based on data from previous studies and new hordein data. The two sets of landraces were found to have a comparable diversity. The Ethiopian ones are more diverse morphologically (5 traits), are similar in numbers of alleles per protein locus (17 loci) and in genetic differentiation, but are less diverse in average heterozygosity per locus and degree of polymorphism. However, on the basis of the hordein data the two sources of germplasm are very distinct. The correlation between morphological and protein diversity is very low. In the RFLP study 28 probes evenly distributed across the genome were used to analyse 43 Ethiopian landraces (represented by one single genotype) and to compare them with modern cultivars from North America, Europe and Japan, as well as 3 landraces from Iran, 1 from Nepal, and 1 accession of H. spontaneum from Afghanistan. The major finding was that the Ethiopian germplasm appears to be significantly less diverse than the modern germplasm but that it is genotypically very distinct. The apparent contradiction between a high diversity of useful genes coming from Ethiopia and an apparently low diversity at the DNA level is discussed. Received: 22 July 1996 / Accepted: 26 July 1996     

Comparative effect of different dietary inulin sources and probiotics on growth performance and blood characteristics in growing–finishing pigs

The study was focused on assessment of the effect of an extract of long-chain inulin (LCI) and dried tubers of Jerusalem artichoke (JA) and a multispecies probiotic preparation as well as a combination thereof on growth performance and blood parameters of fattening pigs. In total, 144 pigs (initial body weight 30.0 ± 0.5 kg) were used in a 98-d experiment. The six dietary treatments consisted of the control diet (Con), diet Con supplemented with probiotics (ConP) and four diets supplemented with LCI or JA alone or with probiotics (diets LCIP and JAP). Throughout the fattening period, there was a beneficial effect of the probiotic supplementation to the inulin-containing diets and the average daily gain (ADG) was increased by supplementation of probiotics in combination with inulin sources (p < 0.05). At the end of the fattening period, ADG and feed conversion ratio (FCR) were higher after supplementation of LCI only (p < 0.05). Compared with group ConP, in groups LCI and JA, the ADG and FCR were improved (p < 0.05). Only in the first fattening stage, the addition of the prebiotics and/or probiotics had an impact on the level of white blood cells and some biochemical indices in pigs. In younger animals, probiotic or LCI supplementation increased the IgG level (p < 0.05). There was also an interaction between the probiotics and JA resulting in increased IgG and IgA concentrations (p < 0.05). In the finishing period, LCI addition increased the IgM level (p < 0.05), whereas JA addition increased IgG and IgM levels as well (p < 0.05). In conclusion, both dietary sources of inulin and probiotic supplementation can improve the fattening performance and health status of growing pigs.