Method of determining content of ethereal oil in leaves of mint plants according to total volume of secretory glands

FIELD: agriculture.

SUBSTANCE: invention relates to the field of agriculture and, in particular, to selection. In the method the content of ethereal oil in leaves of mint plants is determined according to the total volume of the secretory glands. At that the quantity and size of the glands per unit of leaf area of a mint plant is obtained using a light microscope with video-system Lumenera Infinity 2 followed by data processing using the program Infinity camera V:5.02. Also, a light microscope with a digital camera and a scale rule or a binocular with an ocular micrometer is used.

EFFECT: method enables to speed up evaluation of the potential productivity of breeding samples.

3 cl, 2 tbl, 2 ex

 

The invention relates to agriculture, and in particular to selective breeding, and for the creation of a rapid method for the selection of promising hybrids and varieties of mint. There is a method of evaluation of promising hybrids and varieties of mint on the amount of essential oil isolated from the vegetative mass of leaves and inflorescences of a plant, the method hydrodistillation (method Clevenger, modified Loshkareva (GOST 24027.2-80)). The disadvantage of this method is the long time of extraction steam (at least 2 hours), a sufficiently large amount of dry material (not less than 50 g) and the timing of the selection of the analyzed material (the second half of the vegetation period - the phase of budding and mass flowering), so further work with selected breeding material is only possible for the next cropping year.

Other performance indicators of productivity of breeding material can be physiological and biochemical parameters and their complex systems analysis - this method is widely used in forecasting the potential productivity of crops (patent RU 2439873 C2, A01G 7/00, 2009).

The productivity of the essential oil of peppermint is determined by the glandular apparatus of the plant, mainly the number of 10 - cell glands (trichomes) of the spherical shape on the surface of leaves and inflorescence.

Most near them the technical nature of the claimed method is a method of selection of highly productive forms of mint on the amount of essential oil, per I mm2leaf surface (Dissertation for candidate of agricultural Sciences Lysakova POSTGRADUATE, 1984). This method consists in the analysis of chromatography suspension of the cut lamina plant mint in the early stages of vegetation. The disadvantage of this rapid method is the need for high-precision and expensive equipment gas chromatograph.

The problem solved by the invention is the creation of a more simple method for predicting the productivity of the essential oil of interspecific hybrids, intraspecific clones and varieties of mint. The problem is solved by taking into account the number and morphological parameters of trichomes on leaves of mint, selected in the phase of technical maturity, the calculation of the total volume of glandular formations and comparing the obtained data with the results of released varieties old selection of mint. Requires only a light microscope with video Lumenera Infinity 2 or digital camera line to fix the zoom when taking pictures, or binocular microscope with ocular micrometer.

Progress

We selected for evaluation of mint plants are selected 10-30 leaves 5-7 pairs 2-3 storey main flowering shoots in the phase of technical maturity, because in this phase ends with the formation EF is Amalickiah glands and they are experiencing the greatest oil content.

Using video or digital camera attached to a light microscope, photographed at magnification ×500 10 plots each sheet with top and bottom sides, because the glands are often located on both sides of the sheet. On the images, using Infinity camera V:5.02 count the number of glands in the frame, determine the surface area of the sheet in the frame in cm2the number of glands on 1 cm2the upper and lower sides of a sheet. Also measure the diameter and calculate the average value of the diameter of metal.

You can carry out these calculations using scale bars, for which her photograph and take into account that 1 mm =100 μm.

In the absence of a light microscope similar measurements can be performed by using a stereomicroscope. The number of glands in the field of view, the diameter and area of the field of view is done using the ocular micrometer.

Using the data obtained, calculate the total amount of iron per 1000 cm2the leaf area by the formulas:

V1000=Vtop+Vbottom

Vthe top or bottom=4/3πR3×n×1000, where

V1000- the total amount of iron per 1000 cm2the leaf area, cm3;

Vtop- the amount of metal on the upper side, see3;

Vthem- the volume of glands on the underside, see3;

πR=3,14;

R is the radius of the glands, cm;

n is the number of glands on 1 cm2the surface of the sheet pieces

For selection of promising breeding specimens of hybrids and varieties of mint calculated the total amount of iron in the leaves of plants are compared with similar data released varieties of mint or old plants breeding. During the development of this rapid method obtained a positive correlation between the total amount of iron per 1000 cm2the leaf area and collecting the essential oil from the whole plant, that allows to consider this characteristic as an indicator of the productivity of essential oils of mint.

Example 1. The calculation of V1000for varieties of mint old breeding, zoned for Ukraine and southern regions of Russia, grown on the experimental area of the laboratory in the Main Botanical garden of Russian Academy of Sciences. On selected 30 sheets 5-7 pairs 2-3 storey main flowering shoots in the phase of technical maturity of each variety counted the number of secretory glands, their diameter and V1000(table 1).

Table 1.
The total volume of the glands (V1000) varieties of mint old breeding, cm3.
gradenumber of glands on 1 cm2square does the same piecesthe diameter of the glands, mcmV1000Cm3
the upper side of the leafthe lower side of the leafthe upper side of the leafthe lower side of the leaf
Krasnodar 2334108088,5888,390,509
Kuban 6590125290,6185,990,646
Simferopol 200210850106,6106,20,664
Muscovite65599990,4487,810,605
Uichanco474125792,2195,550,766
The medicament is i.i.d. 4 55189593,2290,940,583

As can be seen from table varieties of mint old breeding have V1000not less than 0.5 cm3(ml). This value (0.5) is taken as the lower limit of the index of V1000in the selection of promising breeding samples interspecific and intraspecific hybrids clones of mint plants for further breeding work.

Example 2. The calculation of V1000For interspecific hybrids, intraspecific mint clones grown on the experimental area of the laboratory in the Main Botanical garden of Russian Academy of Sciences.

On selected 30 sheets 5-7 pairs 2-3 storey main flowering shoots in the phase of technical maturity of each selection of the sample according to the proposed method estimated the number of secretory glands, their diameter and V1000(table 2).

As can be seen from table data samples of mint plants No. 1, 2, 4, 7 and 8 are V1000more than 0.5 cm3that is why they were selected for further breeding work.

Table 2.
The total volume of the glands (V1000) prototypes plant mint, cm3.
No.Intraspecific clones, interspecific hybridsnumber of glands on 1 cm2leaf area, piecethe diameter of the glands, mcmV1000,cm3
the upper side of the leafthe lower side of the leafthe upper side of the leafthe lower side of the leaf
Intraspecific
tetraploid clone M
1arvensis,439785101,7593,090,573
Intraspecific clone M
2arvensis (NV-74)524137387,5787,790,670
Intraspecific clone M
3arvensis (17-Taiwan)41198387,8988.680,502
Intraspecific clone M
4arvensis (18-Japan)385114886,3388,290,541
Intraspecific clone M
5arvensis (24-Syktyvkar)21820477,7873,290,094
Intraspecific clone M
6arvensis (19-Crimea)302100886,288,060,460
7Interspecific hybrid M. piperita and M. crispa539132085,8385,730,611
8Intraspecific clone of M. crispa (Crimea)550174181,7786,440,745

Thus, our proposed method of determining the content of essential oil in the leaves of mint on sumanapala secretory glands is simple and allows you to quickly assess the potential productivity of a selection of samples and to select promising hybrids and clones of plants, further breeding work which will be economically justified. Sources of information

1. GOST 24027.2-80. Raw medicinal plant. Methods for the determination of moisture, ash content, extractive and tannins, essential oils.

2. Lysakova POSTGRADUATE Some morpho-physiological characteristics of interspecific hybrids mint, determine the level of accumulation of essential oil / Dissertation for candidate of agricultural Sciences. Simferopol, 1984.

1. The method of determining the content of essential oil in the leaves of mint on the total volume of the secretory glands, characterized in that the number and size of glands per unit area of the leaf peppermint obtained using a light microscope with video Lumenera Infinity 2 with subsequent data processing using Infinity camera V:5.02.

2. The method according to claim 1, characterized in that the use of light microscope with a digital camera and a scale ruler.

3. The method according to claim 1, characterized in that use binocular with ocular micrometer.



 

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