RESEARCH COMMUNICATIONS In conclusion, in the present study we have success- Climatic influence on radial growth of fully induced adventitious roots from the leaf explants of A. paniculata. The adventitious roots were cultured in flask- Pinus wallichiana in Ziro Valley, scale suspension cultures using MS medium supplemented Northeast Himalaya with 2.7 μM NAA and 30 g/l sucrose. Adventitious root cultures showed higher biomass as well as andrographolide Santosh K. Shah1, Amalava Bhattacharyya1,* and accumulation capabilities. Our study demonstrates the Vandana Chaudhary2 possibilities of production of andrographolides for com- 1 mercial purposes in a large scale using bioreactor cultures. Birbal Sahni Institute of Palaeobotany, 53, University Road, Lucknow 226 007, India 2 1. Sharma, A., Singh, R. T., Sehgal, V. and Handa, S. S., Antihepato- Department of Science and Technology, New Mehrauli Road, toxic activity of some plants used in herbal formulation. New Delhi 110 016, India Fitoterapia, 1991, 62, 131–138. 2. Tang, W. and Eisenbrand, G., Chinese Drugs of Plant Origin, An attempt has been made here to study the climatic Springer-Verlag, Berlin, 1992, pp. 97–103. influence on variation of tree-ring width (radial growth) 3. Mishra, P., Pal, N. L., Guru, P. Y., Katiyar, J. C. and Srivastava of Blue Pine (Pinus wallichiana A.B. Jackson) growing Tandon, J. S., Antimalarial activity of Andrographis paniculata in five different sites in and around Ziro Valley, Arun- (Kalmegh) against Plasmodium berghei NK 65 in Mastomys achal Pradesh, Northeast Himalaya. The site chrono- natalensis. Int. J. Pharmacogn., 1992, 30, 263–274. logies have been evaluated to assess inter-site differences 4. Saxena, S., Jain, D. C., Bhakuni, R. A. and Sharma, R. P., Chem- through several statistical analyses, viz. correlation istry and pharmacology of Andrographis species. Indian Drugs, 1998, 35, 458–467. matrices, principal component and hierarchical cluster 5. Kumar, R. A., Sridevi, K., Kumar, N. V., Nanduri, S. and analysis. Analysis of tree growth–climate relationship Rajagopal, S., Anticancer and immunostimulatory compounds suggests that the pre-monsoon precipitation (December– from Andrographis paniculata. J. Ethnopharmacol., 2004, 92, April) is a significant factor influencing the growth of 291–295. Blue Pine in all these sites. 6. Verpoorte, R., van der Heijden, R., ten Hoopen, H. J. G. and Memelink, J., Metabolic engineering of plant secondary metabo- Keywords: Blue Pine, climatic influence, radial growth, lite pathways for the production of fine chemicals. Biotechnol. Lett., 1999, 21, 467–479. tree ring. 7. Dornenburg, H. and Knorr, D., Strategies for improvement of sec- ondary metabolite production in plant cell cultures. Enzyme Mi- BLUE Pine (Pinus wallichiana), a large evergreen conifer crob. Technol., 1995, 17, 674–684. tree, is found all along the Himalayas from west Kashmir 8. Rao, S. R. and Ravishankar, K. A., Plant cell cultures: chemical to east Arunachal Pradesh1, at altitude generally ranging factories of secondary metabolites. Biotechnol. Adv., 2002, 20, 101–153. from 1800 to 3900 m. Typical habitats are mountain screes 9. Butcher, D. N. and Connolly, J. D., An investigation of factors and glacier forelands and appears as a pioneer species, which influence the production of abnormal terpenoids by callus but it also forms old growth forests as a primary species cultures of Andrographis paniculata Nees. J. Exp. Bot., 1971, 22, in mixed forests with deodar (Cedrus deodara), spruce 315–322. (Picea smithiana) and fir (Abies pindrow) in the temperate 10. Murashige, T. and Skoog, F., A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant., 1962, belt. In some places beyond 3000 m, it reaches the tree- 15, 474–497. line and is associated with birch (Betula utilis) and juniper 11. Jain, D. C., Gupta, M. M. and Saxena, S. K., LC analysis of hepa- (Juniperus macropoda)2. Blue Pine growing in several toprotective diterpenoids from Andrographis paniculata. J. sites of the western and central Himalaya has been found Pharm. Biomed. Anal., 2000, 22, 705–709. suitable for various environmental and ecological applica- 12. Klerk, G. J. D., Arnholdt-Schmitt, B., Lieberei, R. and Neumnann, 3–9 K. H., Regeneration of roots, shoots and embryos: physiological, tions in tree-ring analyses , but no such analyses are biochemical and molecular aspects. Biol. Plant., 1997, 39, 53–66. available from the northeastern part of the Himalaya. 13. Gao, X., Zhu, C., Jia, W., Gao, W., Qiu, M., Zhang, Y. and Xiao, In this paper an attempt has been made to analyse tree P., Induction and characterization of adventitious roots directly rings of P. wallichiana growing in and around Ziro Valley, from leaf explants of Panax notoginseng. Biotechnol. Lett., 2005, Lower Subansiri District, Arunachal Pradesh, Northeast 27, 1771–1775. 14. Wu, C. H., Murthy, H. N., Hahn, E. J. and Paek, K. Y., Large- Himalaya, towards understanding its dendroclimatic poten- scale cultivation of adventitious roots of Echinacea purpurea in tial from a new geographical region which is greatly airlift bioreactors for the production of chichoric acid, chlorogenic influenced by the southwest monsoon. acid and caftaric acid. Biotechnol. Lett., 2007, 29, 1179–1182. Tree-ring samples of Blue Pine were collected from five forest sites characterized by open pine mixed broad ACKNOWLEDGEMENTS. This work was partially funded by the University Grants Commission (Special Assistance Programme), leaved forest in and around Ziro Valley. These sites are Department of Science and Technology (FIST Programme), Council of Dobya (DOB), Hari (HAR), Hong (HON), Michi-Rant Scientific and Industrial Research and National Medicinal Plant Board, (MIC) and Raga (RAG; Figure 1). All these sites are New Delhi, India. Received 24 December 2007; revised accepted 23 December 2008 *For correspondence. (e-mail: [email protected]) CURRENT SCIENCE, VOL. 96, NO. 5, 10 MARCH 2009 697 RESEARCH COMMUNICATIONS Figure 1. Map showing sampling sites of Pinus wallichiana at Ziro Valley, Lower Subansiri District, Arunachal Pradesh. Table 1. Details of the tree-ring sampling sites from Ziro Valley there were any errors), were removed from the population Number of prior to chronology development. In order to remove the Site Site code Altitude (m) trees and cores gross growth trend largely unrelated to climate, and to remove effects of differential mean growth rates among Dobya DOB 1320 4 (11) individual trees prior to averaging into the mean chrono- Hari HAR 1260 17 (36) Hong HON 1245 3 (6) logy, standardization of each series was made. It was done Michi-Rant MIC 1255 15 (35) by fitting a growth curve to the ring-width series and then Raga RAG 1582 6 (12) dividing the measured value by the curve value at each year. After applying different detrending options for the standardization to check the improvement of chronology statistics, we have selected cubic spline which is appro- located at latitude and longitude ranging from 27°53′–27°68′ priate for trees growing within the interior of the forests and 93°85′–93°89′ respectively. These sites are close to and not growing in stressed condition. The detrending for human settlements and thus highly disturbed. In HON vil- all the series was done using 30-year spline which retains lage trees are confined to steep, rocky slopes, but most of 50% of the variance contained in the measurement series them are found burnt by forest fire. Generally trees are at a wavelength of 30 years. Two versions of chronolo- young, except in some sites where several isolated, huge, gies, ‘standard’ and ‘residual’, were prepared for each girthed pine trees are recorded which have been preserved site intended to contain a maximum common signal and by the villagers in the memory of their ancestors. Other- minimum amount of noise. In the ‘standard’ chronology, wise, all old trees have been already logged. These pro- detrending of measurement series was done first by fit- tected trees, mentioned to be about 2000 years old by the ting a curve to model biological growth trend to each local people, were planted by their ancestors after migrating series, and dividing out the growth model. The chronology from Tibet, and now these are identified by their names. was computed as a robust estimation of the mean value Hundred cores from 45 trees of Blue Pine have been col- function to remove effects of endogenous stand distur- lected from these five forest sites with the help of Incre- bances; this enhances the common signal contained in the ment borer. Details of the tree-ring samples are given in data. In the ‘residual’ chronology, autoregressive modelling Table 1. was performed on the detrended ring measurement series. All the tree cores of Blue Pine were mounted on a Robust estimation of the mean value function produces a wooden frame. The surface of these cores was cut by the chronology with a strong common signal and without sharp razor blade and polished with the help of different persistence. The pattern of variation is generally similar grades of sandpaper to enhance the clarity of the cells. among trees growing throughout the same region for the Tree rings of these cores were counted under stereo-zoom same time period because of variations in macro climatic microscope and were dated to the calendar year of their factors, which are common to these trees13. Synchroniza- formation using the typical skeleton plot technique10. The tion of growth pattern of Blue Pine in most of these sites width of each dated tree-ring in each core was measured covering a wide area of the Ziro Valley suggests a common using VELMEX measuring system coupled with a com- force controlling the growth in the region. Generally, puter. These measurements and dates were checked using common growth pattern is exhibited in trees where climate the computer program COFECHA11,12. Cores that corre- has a significant role in limiting the tree growth.