THE EFFECT OF Pi OR DEUTERIUM DEPLETED WATER ON FOLIAR STOMATA OF SAINTPAULIA IONANTHA L. EXVITROPLANTLETS SPRINKLED FOLIAR OR RADICULAR

Headnote

ABSTRACT. We studied the effect of Pi water (PiW) or deuterium depleted water (DDW) (with 25 ppm D), treatments applies, during 30 days, administration which were made by spraying on the foliar limbs of Saintpaulia ionantha L. (African violet) exvitroplantlets leafs, or on the substratum, concerning the stomata constitution, identified at the level of inferior epidermis. Both sprinkling with Pi water or deuterium depleted water determined the decrease of stomata density, even to 44% in the case of exvitroplantlets foliar sprinkled with DDW and watered to the base - on the perlite substratum - with distillate water (DW), and just with 19% in treatment with PiW and the stomata cell width with maximum 33%.

Keywords: deuterium depleted water, Pi water, stomata, acclimatization, African violet

INTRODUCTION

The Pi water (PiW) production was elaborated and developed in Japan, in 70 years. Using a bioenergizing installation they obtained a purified drinking water (Fülop, manuscript).

In Romania, in every year takes place an International Symposium namely "Water a miracle". In this manifestations were have been communicated many articles regarding the effects of the Pi water on the plants (cucumbers), in tree growing, rooting on different types of unconventional substratum watered with this type of water (http://www.apa-pi.ro/htmlsursa/simpozion.htm, 2002) and other researches in botanic and vegetal biotechnology is interested by this water (Petrus - Vancea and Cachita, 2008), were describes the positive effects of this type of water in growth plants.

Moreover, interest in exploiting this water type, produced using Bio-Life-Energy System, led to the development of methods with proprietary patent RO 112481 1999 with the authors on Godeanu and collaborators (1999) presented at the salon of invent from Bucharest, in 2005 (http://www.cttecotech.ro/salon/ postere/P330.png, 2007). Technology is refers to the possibility of restoration of land damaged by oil residue with Pi water irrigation and sending back to agriculture, through irrigation with 2 - 100 l/m^sup 2^ for 1 - 30 days (depending on the degree of deterioration).

Deuterium depleted water (DDW) is a water which has a lower deuterium content [10 - 120 ppm D/(D+H)] (Somlyai, 2001; Stefanescu et al., 2004), brevetted and produced for commercialization from 2003 by the Romanian National Research and Development for Cryogenic Technologies Institute, located in Râmnicu Vâlcea (Tamaian and Titescu, 2005 a and b, and 2007).

Recent studies elaborated by Romanian researchers (Berdea et al., 2007) concerning on the development of experimental models for use ASD in studies regarding seeds germination and implementing these models for studying the water action on live cells. The authors cited have found an important effect of ASD in cell division inhibition, both in plants and animals. To study the effect of ASD plants Berdea and colleagues (2007) measured deuterium content of water extracted from plants that have been wetted with ASD, compared with those coming from plants wetted with normal water. The ASD was lowering the rate of germination and growth of the same.

Because, from previous researches (Petrus - Vancea and Cachita, 2004 a), was noticed the fact that after "ex vitro" acclimatization of Saintpaulia (African violet) plantlets, to septic medium, foliar surface sprinkling with PiW, or those administration on the plantlets basis, determined the stimulation of the organogenesis, in the place where it was applied, and in the other hand, the sprinkling of African violet exvitroplantlets with DDW (with 25 ppm D) determined a rhizogenesis intensify, the total amount of leaflets being the same with the one of the control, while applying this type of water at the stalks basis determined a strong inhibition in forming new roots, in this experiment, we decided to continue this studies - concerning the effect of this two types of water - on exvitroplantlets foliar limb stomata formations, its dimension and density.

MATERIALS AND METHODS

African violet exvitroplantlets (Saintpaulia ionantha L.), uniform in size, with 1 - 1.5 cm height, being after 90 days of vitroculture were chosen as experimental model.

For their acclimatization to a septic medium, the propaguls from the vitroshrub were separated, the solidified medium was removed from their rootlets, through washing them with tap water, brought to laboratory temperature. Then, the exvitroplantlets were planted into perlite, place into incubators with the following dimensions 5/22/35 cm, over the tray being placed a transparent lid with two arched and slide walls with a 12 cm high, in the central area (Vancea and Cachita, 2002); initially, the perlite, was watered with 250 ml distillate water (DW), or with Pi water, or deuterium depleted water (25 ppm D) varying with the experimental variant.

During 30 days the exvitroplantlets was sprinkled with different type of water, following this scheme:

V^sub 0^ - exvitroplantlets foliar sprinkled and watered to the base with DW (control);

V^sub 1^ - exvitroplantlets foliar sprinkled with PiW and watered to the base with DW;

V^sub 2^ - exvitroplantlets foliar sprinkled with DW and watered to the base with PiW;

V^sub 3^ - exvitroplantlets foliar sprinkled with DDW, with 25 ppm D and watered to the base with DW;

V^sub 4^ - exvitroplantlets foliar sprinkled with DW and watered to the base with DDW, with 25 ppm D.

To avoid the mixing of both water types used to sprinkle the exvitroplantlets leafs, and the rooting substratum, we covered the perlite with a thin sheet of aluminium which was pierced to get out trough them the Saintpaulia exvitroplantlets stems, the small orifices having 1 cm diameter. The sprinkling of exvitroplantlets lots was made by foliar spraying or by watering to the base of every plantlet with 5 ml of water/variant, respectively per plantlet at 2 days lapse of time.

The cultures were exposed to a 22 ± 2°C temperature at day and 20 ± 2 °C temperature at night an 16/24 h photoperiod, illuminating them with fluorescent tubes which give out white light with an 1700 lux intensity.

After 30 days of exvitroplantlets placing on the septic medium (Fig. 1) we determined: the survival percent of its and the density, length and the width of stomata. At Saintpaulia leafs the stomata are present only on the inferior epidermis (Petrus - Vancea and Cachita, 2004b). For biometrisations we are used apical leafs, newly formed "ex vitro".

To observe the stoma on the leaves surface we used the imprinting method of superior and inferior epidermis of foliar limbs applied on the youngest leaflets, by applying a colloidal draw (Andrei and Paraschivoiu, 2003). To the level of inferior epidermis we determined - at the optic microscope - the numbers of stomas/microscopic field, and their dimensions (length/width) with 10X object lens and 7X ocular; in the same time, we measured the osteols aperture with 40X object lens, and 7X ocular. The micrometric index was calculated using the method described by Andrei and Paraschivoiu (2003). The photos were realized using a digital camera with 640/480/300 resolution and optic zoom 10X.

Biometrical dates were mathematically [the values recorded at 30 days from exvitroplantlets placing in the septic medium, concordant to the experimental variants V^sub 1^ - V^sub 4^, were related to the homologues dates, biometrisated to the control lot (V^sub 0^), "ex vitro" rooted in aerial part humidification conditions or in substratum watered with distillate water, these being considered as 100%] and statistically (t test) processed.

RESULTS AND DISCUSSIONS

The post acclimatization survival percent - at a 30 day from exvitroplantlets placing in perlite - was 100%, at all experimental lots.

At Saintpaulia the stomata are amaryllidaceous anomocytyc type (Fig. 2). The stomata had a number of three guard cells, in a half moon shape, that had a dimension of 75.6/9.4 µm, in the case of all experimental variants watered with DW, PiW, and DDW (V^sub 0^ - V^sub 4^). To exvitroplantlets, no matter the type of water they were watered with, they were having their closing - opening system of the osteols in function.

The highest density of the stomata - of 16 st/field - was registered at the level of the exvitroplantlets leaflets treated, foliar and basal, with DW, belonging to the control lot (V^sub 0^), and their lowest frequency was noticed at the plantlets leaflets from the lot treated foliar with DDW (V^sub 3^) (Table 1), this one marking a minus of 44%, towards the control V^sub 0^, which was considered 100% (Table 1).

As the dimensions (length and width) of the stomata, the specimens identified of the exvitroplantlets leaflets treated foliar or basal with PiW or DDW (V^sub 1^ - V^sub 4^) have shown identical sizes, of 37.7/18.9 µm (length/width), which presumes - about their length - sizes with 33% (Table 1) higher, towards the ones registered at the stomata on the leaflets watered foliar and basal with DW; about the width of the stomata, in the case of different experimental variants, the noticed average represents reductions of the same proportion (33%), comparatively with the control, which consist in exvitroleaflet stomatas watered only with DW (V^sub ^0) throughout the entire acclimatization period to a septic medium (Table 1).

We didn't observe any changes in the African violet exvitroplantlets thector hairs density, according to the treatment applied during this experiment.

The same experimental protocol described (Petrus - Vancea and Cachita, 2008) in the present article has been followed by us on the chrysanthemum exvitroplantlets (Chrysanthemum morifolium Ramat var. Lamet), after 30 days of acclimatization initiation. It was observed that the foliar applying of deuterium depleted water (DDW), with 25 ppm D, determined a decrease with 50% of the stomata density number.

CONCLUSIONS

The African violet (Saintpaulia ionantha L.) exvitroplantlets watered with Pi water, both foliar and basal, or with deuterium depleted water (with 25 ppm D), in comparison with the parameter determined by the control, have shown in the inferior foliar epidermis 20% less stomata, and to the lot foliar sprinkled with deuterium depleted water (cu 25 ppm D) the number of the stomata dropped to 44%. The width of the stomata was also diminished, at all variants previously mentioned with 37%, in comparison to the control.

References

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