Hindawi Publishing Corporation Spectroscopy: An International Journal Volume 27 (2012), Issue 4, Pages 206–213 doi:10.1155/2012/951064 Chlorophyll Fluorescence Spectra as an Indicator of X-Ray + EMS-Induced Phytotoxicity in Safflower Jitendra Kumar Pandey, 1 Preeti Srivastava, 2 Ram Singh Yadav, 2 and Ram Gopal 1, 3 1 M. N. Saha Center of Space Studies, IIDS, Nehru Science Center, University of Allahabad, Allahabad 211002, India 2 Plant Genetics Laboratory, Department of Botany, University of Allahabad, Allahabad 211002, India 3 Laser Spectroscopy and Nanomaterials Lab, Department of Physics (UGC-CAS), University of Allahabad, Allahabad 211002, India Correspondence should be addressed to Jitendra Kumar Pandey, jkpandey18@gmail.com Copyright © 2012 Jitendra Kumar Pandey et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Abstract. The present investigation deals with the study of in vivo laser-induced chlorophyll florescence spectra (LICF) of safflower leaves (Carthamus tinctorius L.) for X-rays + EMS-treated plants. Seeds were treated with different doses of X- ray + EMS (5, 8, 12, 25, and 30 Kr + 0.5% EMS) and were grown in the green house. The effects of the concerned treatment on chlorophyll (Chl) contents and Chl fluorescence were investigated after 7 days of germination. Results obtained revealed that the values of Chl contents, intensity of Chl fluorescence spectra, and fluorescence intensity ratio (FIR) F685/F730 are directly correlated with the treatment doses monitored. The treatment sets of 8, 12, and 25 Kr + 0.5% EMS doses showed an increase in FIR and thereby a decrease in the Chl contents. However, the lowest treatment dose of 5 Kr + 0.5% showed a decrease in FIR and thereby an increase in chlorophyll contents. Safflower seeds treated with 30 Kr + 0.5% EMS were proved to be lethal as they showed no germination. Thus, our study demonstrates early detection of chlorophyll damage caused by various physical and chemical mutagens through the application of LICF spectra. Keywords: X-ray + EMS treatment, Laser-induced chlorophyll fluorescence, Fluorescence intensity ratio, Photosynthetic pigment contents (chlorophyll a, chlorophyll b, and carotenoids), Safflower (Carthamus tinctorius L.) 1. Introduction Laser-induced fluorescence (LIF) is a powerful tool for plant investigation, and it can illustrate a lot of information about plant health and identity of plants. Leaf pigments emit fluorescence after irradiation with laser light [1]. The in vivo chlorophyll fluorescence spectra of plant leaves shows two fluorescence maxima, one in the spectral region near 685 nm and other in the region near 730 nm [2]. The shape of the fluorescence spectra and the value of the fluorescence intensity ratio (FIR) up to a great extent depend upon the Chl contents and absorbance of the leaves [3]. Fluorescence intensity ratios show a good cor- relation with pigment contents and pigment ratios [4]. The intensity of the red and far-red chlorophyll