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Random amplified polymorphic DNA (RAPD) and restriction fragment length polymorphism (RFLP) markers were used to estimate intra- and inter-specific variation in three species of an exclusive mangrove genus, Avicennia. Intrapopulation polymorphism among the 10 populations of Avicennia marina, as measured by percentage of polymorphic RAPDs, varied between 17.8 and 38.9%, with a standard deviation of 7.28, and the coefficient of variation was 26.5%. Polymorphism in Avicennia officinalis (Pichavaram population, 32.3%) and Avicennia alba (Coringa population, 37.8%) was greater than the intrapopulation variation observed in the populations of A. marina from each of the respective locations. It was greater than the average percentage of polymorphism at the intrapopulation level (27.47%) but far less than the variation measured at the interpopulation level in A. marina. Interpopulation variation in A. marina (76.7% for RAPDs and 66% for RFLPs) was greater than the variation in any individual population of this species, indicating a high degree of divergence between the populations. Interpopulation variation as revealed by RAPD and RFLP markers did not indicate the existence of more than one distinct entity in this species in India. The implications of these observations in genetic sampling and conservation are discussed. Statistical analysis of 109 RAPDs and 84 RFLPs observed in one representative genotype from each species showed that the widely distributed A. marina was more closely related to A. alba (genetic distance (1 − F) = 0.22) than to A. officinalis (genetic distance (1 − F) = 0.37). RAPD analysis of six randomly selected genotypes in each species and principal component analysis of the data also favoured this observation.

The chloroplast trnS-psbC gene regions from total genomic DNA of 119 accessions from seven small millet species were amplified by polymerase chain reaction (PCR) and digested with eight restriction enzymes individually as well as in combinations of two enzymes to generate restriction fragment length polymorphism (PCR-RFLP). PCR-RFLP with individual enzymes revealed polymorphism between only some species. However, all the species could be distinguished by using a combination of two enzymes, specifically HaeIII and MspI. PCR-RFLP of 11 to 20 accessions with the same enzyme combination showed no intraspecific variation, which established that the differential banding patterns were species specific. In contrast, the same enzyme combination was not useful for differentiating different species of the genera Cajanus, Rhyncosia, Abies, Rhizophora, Ceriops, and Bruguiera, and it also revealed intraspecies variation in three species of Abies. The present study indicated that digestion of trnS-psbC with two four-base recognizing enzymes reveals more variation than with either enzyme alone and that it may be a method of choice for species identification in some genera.

The metastasis associated 18A2/mtsI gene was inserted into the mammalian expression vector pMAMneo placing it under the control of the dexamethasone-inducible MMTV promoter. The construct was transfected into dexamethasone receptor negative F1 and receptor positive F10 cells of the B16 murine melanoma. The transferred gene was switched on in two transfectant clones of F10, by exposure to 10(-6) M dexamethasone, but not in clones of the receptor negative F1 line. One of the F10 transfectant clones (F10-192/10) was characterized further. A 13.5-fold increase in 18A2/mts1 transcripts was found in this clone upon exposure to dexamethasone. There was also a seven-fold increase in lung colonization in an experimental metastasis assay, together with increased expression of depolymerized tubulin and enhanced detection of p53 protein. The number of cells in the S phase increased by 2.5-fold following dexamethasone treatment of the clone. These data suggest a direct involvement of the 18A2/mts1 gene in lung colonization by the tumor cells. The 18A2/mts1 protein promotes tubulin depolymerization, sequesters the p53 phosphoprotein, and induces the cells to enter the S phase, but the relevance of these in the metastatic process remains to be elucidated.

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