Carbon induce phytotoxicity in plant cells and change the

Carbon
nanotubes (CNTs) are known to have many unique characteristics including
diameter, length, atomic configuration, impurities, defects and functionality,
which allow them were widely used in materials, chemicals, food,
bioengineering, medicine, and other ?elds 1, 2. Because of these physical and
chemical features of CNTs have used in agriculture to increase the crop yield,
mainly in the germination process, root growth, and photosynthesis 1. The
positive, negative and natural effects of CNTs on physiological responses can
variable or even opposite among different plant species 3. For example some
studies showed that multi-walled carbon nanotubes (MWCNTs) did not affect the
growth of wheat 5 and inhibited the growth of rice seedlings 6. But significantly,
enhanced the germination rate of tomatoes 4. Khodakovskaya, et al. (2009) have reported that CNTs could
penetrate plant seed coat and dramatically affect seed germination and plant
growth. However, the penetration, uptake and accumulation of CNTs in plant
cells and tissues are not well documented 8. Plants and plant cells showed
high tendencies to accumulate CNTs 11, 12. Recently, Mariya et al .(2014)  showed that MWCNTs can be absorbed by root
system of tomato plants and reach the leaves and the fruits. Lin  et al. (2009)
reported that adsorption of an extensive amount of MWCNTs on the root surface
may suppress the water in?ux and uptake of nutrients hence inhibiting the plant
growth 14.  Investigations have shown
that CNTs could induce phytotoxicity in plant cells and change the gene
expression of plants 9. Biochymical studies of different plants have
demonstrated that, the use of CNTs can induce the production and accumulation
of oxygen reactive species such as superoxide radical anions and hydroxyl
radicals. ROS generation can lead to protein, lipid, and DNA oxidation and to
cell death 13. Most previous researches of CNTs in the biosciences have
focused on their influence on animal and human cells, but to investigate the
potential effects of CNTs on the plant cells in the natural environment is very
necessary. Liu Q et al. (2009)
reported that CNTs penetrate inside the cells (16). Insertion of MWCNTs into
the wall of epidermal cells and root hairs observed in wheat seedlings (17). Khodakovskaya
(2011) showed
that MWCNTs case enhance the growth of tobacco and regulate cell division via
activating water channels and regulating genes involve d in cell division and
extension (18). According to Serag et al. (2011), short MWCNTs with short length
(in a range of 30 to 100 nm in length) tended to target the nucleus, plastids,
and vacuoles, which further revealed the close relationship between MWCNTs size
and phytotoxicity from the perspective of plant cell biology 19.
Okra (Hibiscus esculents L.)
belongs to the family Malvaceae, and 
grown in all parts of the tropics and during summer in the warmer parts
of the temperate regions. Okra is a popular home garden vegetable and a
good  source  of 
many  nutrients  including 
vitamins  B  and  C,  fiber, 
calcium,  and  folic 
acid (Hegazi and Hamideldin, 2010). In this study, changes of anatomical
structure (thickness and diameter different tissues) and morphological
characterizes of two cultivars of  Okra seedling
 plant under  MWCNT treatment evaluated. Results from
this  research may help to show the responses
and behaviors of family Malvaceae plants 
to MWCNTs. Materials
and methodsPreparation
of MWCNTs

MWCNTs
were obtained from Nanosany Company (Iranian Nanomaterials Pioneers Company,
Mashhad, Iran). Specifications of this carbon nanotubes were detected by using
scanning electron microscope (SEM) (Hitachi S-4160,Tokyo, Japan), and the X-ray
diffraction (XRD) (Philips-X’Pert MPD X-ray refractometer) technique. Also, Raman spectra
of the MWCNTs with OD less than 50 nm was prepared using UV–Vis spectrophotometer
(T80+ UV–VIS spectrophotometer PG instruments Ltd, UK). The metals content of MWCNTs were detected
by energy dispersive X-ray spectroscopy analysis. 

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