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Nano-agriculture carbon nanotubes enhance tomato seed germination and plant growth
C. Srinivasan and R. Saraswathi
Agriculture is an area where new technologies are often applied to improve the yield of crops. Genetically modified (GM) crops have received mixed reactions from the public, particularly the farmers. The recent approval of Btbrinjal seeds by the government triggered a lot of dissent among farmers and the public quickly prompting the government to withdraw its decision. (For a detailed discussion on Bt-brinjal, see ref. 1.) Nano-agriculture involves the employment of nanoparticles in agriculture with the ambition that these particles will impart some beneficial effects to the crops. Carbon nanotubes (CNTs) have acquired an important status in nanotechnology due to their unique mechanical, electronic and thermal properties, which have led to their exploitation in diverse applications such as sensors, flat panel displays, energy and gas storage2. Also, their application in drug delivery and medical diagnostics is being actively explored. The study on the effects of nanoparticles in plant science is a newly emerging area of research35. Recently, confocal fluorescence image studies have revealed the capacity of singlewalled carbon nanotubes (SWNTs) to traverse across both the plant cell wall and cell membrane6. This study also pointed out that SWNTs can serve as effective nanotransporters to deliver DNA and small dye molecules into intact plant cells. There are also some reports on other nanoparticles as smart treatment-delivery systems in plants7. Compared to plant cell walls and membranes, the penetration of nanoparticles into seeds is expected to be difficult due to the significantly thick seed coat covering the whole seed. In spite of this anticipated hurdle, Khodakovskaya et al.8 demonstrated that CNTs could effectively penetrate seed coat, thereby influencing the seed germination and plant growth. This study commendably established that exposure of seeds to CNTs resulted in the enhanced tomato seed germination and growth rate. The experiment involved placing sterile tomato seeds on standard agar Murashige and Skoog medium (MS medium) with
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different concentrations (10, 20 and 40 g/ml) of added multi-walled carbon nanotubes (MWNTs) and also without nanotubes. It is amazing to note that the seeds placed in the MS medium with the given three concentrations of CNTs sprouted on the third day whereas the seeds in the control experiment (without CNTs) did not germinate in the same period (Figure 1 a). Also, in the next few days the germination rate in CNTs supplemented medium was significantly higher than that observed for the control sample (Figure 1 b). The germination percentage for seeds that were placed on regular medium averaged 71 in 20 days, whereas germination percentage of the seeds placed on medium supple-

mented with CNTs (40 g/ml) averaged 90 during the same period. Tomato plants grown in media supplemented with CNTs not only showed an increase in biomass but also possessed well-developed long stems compared to the control (Figure 2 b). However, the length of the roots was found to be the same in both cases (Figure 2 a) indicating the absence of any toxic effects of CNTs on root development and root elongation in the concentration range studied. Khodakovskaya et al.8 thus demonstrated that CNTs exposed seeds germinated up to two times faster than control seeds and the seedlings weighed more than twice as much as the untreated plants. These desirable effects possibly

Figure 1. Effect of CNTs on tomato seed germination. a, Tomato seeds incubated during 3 days without (left) or with (right) CNTs on MS medium. b, Time of germination and germination percentages of seeds incubated with and without CNTs during 20 days. Re8 printed with permission from Khodakovskaya et al. . Copyright (2010) from American Chemical Society. CURRENT SCIENCE, VOL. 99, NO. 3, 10 AUGUST 2010

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reason suggested for the higher water uptake was that new pores were generated during penetration of seed coat by CNTs, which aided better water permeation. Another possible cause could be the efficient gating of the water channels by the CNTs in the seed coat. Such a hypothesis however requires further substantiation by suitable experimental data. An immediate outcome of the study is the observation of increased biomass, which will have some important economic significance in the production of biofuels. Also the rapid germination and growth rates will yield agriculture products in a short duration. It has to be emphasized that the technology described here cannot be immediately implemented in agriculture. Several issues need to be addressed. It is necessary to know how the plants developed by this technique will fight pests. It has to be investigated whether CNT treated plants can harm other plants in the same field. Finally, it must be definitely established that CNTs are not toxic to plants and the tomatoes obtained by this technology will not harm mankind.
1. Yadugiri, V. T., Curr. Sci., 2010, 98, 1273. 2. Ohashi, T. and Dai, L., Carbon Nanotechnology (ed. Dai, L.), Elsevier, The Netherlands, 2006. 3. Zheng, L., Hong, F., Lu, S. and Liu, C., Biol. Trace Elem. Res., 2005, 104, 83. 4. Lin, D. and Xing, B., Environ. Pollut., 2007, 150, 243. 5. Yang, L. and Watts, D. J., Toxicol. Lett., 2005, 158, 122. 6. Liu, Q., Chen, B., Wang, Q., Shi, X., Xiao, Z., Lin, J. and Fang, X., Nano Lett., 2009, 9, 1007. 7. Gonzales-Melendi, P. et al., Ann. Bot., 2008, 101, 187. 8. Khodakovskaya, M., Dervishi, E., Mahmood, M., Xu, Y., Li, Z., Watanabe, F. and Biris, A. S., ACS Nano, 2009, 3, 3221.

Figure 2. a, 27-day-old tomato seedlings growing on medium with and without CNTs. b, 25-day-old tomato seedlings growing on medium without and with CNTs. Concentration of CNTs in growth medium in g/ml is shown below each figure. Reprinted with permis8 sion from Khodakovskaya et al. . Copyright (2010) from American Chemical Society.

occurred due to the penetration of the nanotubes into the seed coat enhancing water uptake by the seed. Raman spectral study was used to confirm that CNTs penetrate tomato seeds. Tomato seeds incubated in agar media supplemented with CNTs for two days were washed, dried and cut, and the surface was examined by Raman spectroscopy. The observation of the G band at 1569 cm1 in the

Raman spectrum confirmed the presence of CNTs inside the seed. No such band was seen for the control sample. Transmission electron microscopy imaging also confirmed the presence of CNTs inside the seed. Thermogravimetric analysis revealed that the seeds incubated in CNTs for two days possessed a moisture content of 57.6% compared to the value of 38.9% for the control. One

C. Srinivasan* and R. Saraswathi are in the Department of Materials Science, Madurai Kamaraj University, Madurai 625 021, India. *e-mail: ceesri@yahoo.com

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