Online Resource File

Title: Roles of hydrogen sulfide and nitric oxide in the alleviation of cadmium-induced oxidative damage in alfalfa seedling roots

Journal: BioMetals

Authors: Le Li · Yanqin Wang · Wenbiao Shen*

Affiliations: College of Life Sciences, Cooperative Demonstration Laboratory of Centrifuge Technique of Nanjing Agricultural University and Beckman Coulter Ltd. Co., Nanjing Agricultural University, Nanjing 210095, PR China

*Correspondence to

Wenbiao Shen, Ph. D., College of Life Sciences, Nanjing Agricultural University, Nanjing 210095, PR China; E-mail: ; Fax(Tel), +86-025-84396542


Fig. S1 The colors of the increasing concentrations of NaHS plus CdCl2 for 30 min. From left to right: water; 100 μΜ NaHS + 100 μΜ CdCl2; 1 mΜ NaHS + 100 μΜ CdCl2; 100 μΜ NaHS + 1 mΜ CdCl2; 1 mΜ NaHS + 1 mΜ CdCl2


Fig. S2

Fig. S2 Effects of treatments of alfalfa seedling roots with NaHS and Cd simultaneously or separately on NO production. Seedling roots were incubated in quarter-strength Hoagland’s solution for 5 days, then transferred to the same solution containing 0 or 100 µM NaHS for 6 h, then exposed to 0 or 100 μM CdCl2 or the combination treatment with 100 µM NaHS for another 3 h. Sample without chemicals was the control (Con). NO production in alfalfa seedling root tips was detected by fluorescence probe DAF-FM DA, after different treatments under fluorescence and bright field microscopy (TCS-SP2 confocal laser scanning microscope; Leica Lasertechnik GmbH). Photographs showing the identical results. Bars, 200 μm (a). Mean relative DAF-FM DA fluorescence densities corresponding to (a), were also given (b), taking control (lane 1) as 100%. Lanes 1-5 represent various treatments as follows: 1, Cd-free pretreatment followed by Cd-free treatment; 2, NaHS pretreatment followed by Cd-free treatment; 3, Cd-free pretreatment followed by Cd treatment; 4, NaHS pretreatment followed by Cd treatment; 5, Cd-free pretreatment followed by NaHS plus Cd treatment. Values are the mean ± SE of six independent experiments. Bars denoted by the same letter did not differ significantly at P < 0.05 according to Duncan’s multiple range test

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