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Since the first report on Camptothecin detection in Nothapodytes nimmoniana by Govindachari and Viswanathan (Phytochem 11:35–29, 1972), considerable work has been done on biotechnology and its applications on the species. Plant tissue culture techniques have applications in clonal propagation, CPT production, and conservation of N. nimmoniana. Discovery of CPT production by endophytes existing in symbiotic association with N. nimmoniana has provided new insights into finding alternative sources of the alkaloid. Development of molecular markers such as RFLP, RAPD, ISSR, and AFLP has facilitated understanding of population ecology and genetics of the species. Molecular information generated from these studies is promising in establishing strategies for conservation and sustainable use of N. nimmoniana populations under overexploitation pressure. The advances in instrumentation in the 20th century, such as desorption electrospray ionization mass spectrometry allowed CPT analysis in tissues without sample pretreatment. Other ancient techniques for qualitative and quantitative analysis such as chromatography, spectroscopy, and H1-NMR are applied in the detection of CPT due to variable sensitivity to the alkaloid. The review covers work on plant tissue culture for clonal propagation and CPT production in N. nimmoniana. Besides symbiotic endophyte sources of CPT in N. nimmoniana,population genetics studies and instrumentation analysis of the alkaloids are reviewed.
Among the alternative plant sources of camptothecin (CPT), Nothapodytes nimmoniana is regarded as the most convenient source for large-scale isolation of the monoterpenoid indole anticancer alkaloid. As a result, CPT annual trade value has grossed over billion US Dollars in recent years. Somatic embryogenesis (SE) offers potential application in the rapid clonal propagation of the tree and production of the alkaloid, so as to mitigate indiscriminate harvest of its endangered natural population to meet industrial demand. However, response to the production of embryogenic callus (EC) in the in vitro cultures of N. nimmoniana is poor to scant. In the present study, two-dimensional electrophoresis (2-DE) and mass spectrometry (MaSp/MaSp) were employed in studying proteome expression changes between EC and non-embryogenic callus (NEC) of the forest tree. The results of the study showed higher metabolic and physiological processes associated with embryogenic competence acquisition in the callus cultures; high cellular oxidative stress, energy metabolism, protein synthesis, and other metabolic processes played a key role in upregulated expression of the identified proteins in EC over NEC. Putative role of the expressed proteins during embryogenic competence acquisition by N. nimmoniana callus cultures has provided some insight into the physiology of the competence acquisition through cellular roles played by oxidative stress and metabolic processes. Further studies on metabolic physiological processes associated with EC production could have application in optimizing culture conditions for mass propagation through the SE, so as to mitigate the indiscriminate harvest of endangered N. Nimmoniana natural population for CPT.
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