CC BY
1134
AGRICULTURAL SCIENCES / «ШУШМУМ-ШУГМаУ» 2022
AGRICULTURAL SCIENCES
УДК 634.8.034
Zelenianskaya N. N.,
Doctor of Agricultural Sciences, Senior Research Officer, Deputy Director for Research and Innovation National Scientific Centre «V. Ye. Tairov Institute of Viticulture and Winemaking» NAAS of Ukraine
Samofalov M. A. Graduate student
National Scientific Centre «V. Ye. Tairov Institute of Viticulture and Winemaking» NAAS of Ukraine
DOI: 10.24412/2520-6990-2022-19142-34-37 MANIFESTATION OF THE REGENERATIVE ABILITY OF ROOTSTOCK AND TECHNICAL VARIETY OF GRAPES (VITIS VINIFERA L.) ON MINERAL SUBSTRATES IN VITRO
CONDITIONS
Abstract.
The article presents the results of research on the determination of the regenerative capacity of initial grape explants of grafted and technical grape varieties on different types of substrates. The aim of the work was to determine the regeneration potential of rootstock and technical varieties of grapes in vitro culture on different types of substrates and to establish their influence on further growth and development of the vegetative mass of grape microclones. The mineral substrates perlite, vermiculite and their mixture (1:1) were taken as a basis.
Keywords: in vitro, initial explants, mineral substrates, survival, proliferation, rhizogenesis.
In order to create grape planting material free from viral and bacterial infection, in vitro methods of tissue and organ culture are widely used. This method allows you to shorten the time of reproduction of new grape varieties by 4-5 times, increase the reproduction ratio and use a small amount of raw material.
The basis of this method is the induction of organogenesis from the initial bud on nutrient media or substrates under the conditions of culture rooms. This process takes place in three or more stages. They include: 1 - selection of donor plants; 2 - sterilization; 3 - introduction of explants into in vitro culture; 4 - microclonal reproduction of shoots in vitro culture; 5 - rooting of plants; 6 - planting plants [3].
An important step in microclonal reproduction in vitro is adaptation to non-sterile conditions. It is at this stage that most plants die (up to 70-80%). This is due to a number of their physiological and anatomical features. Microclonal plants are characterized by an underdeveloped waxy cuticle, a damaged stomatal apparatus, weak photosynthetic activity, a vascular connection between the root and shoot, vitrification, an unbranched root system, and the absence of root hairs [2]. Therefore, it is very important to start preparing them for transfer to non-sterile, uncontrolled conditions even when cultivating plants in vitro. One of the ways to eliminate (or minimize) the physiological and anatomical features of microclonal plants listed above is to use nutrient substrates instead of nutrient media.
Substrates should be quite light, loose, moisture-and air-permeable. Many scientific works have been devoted to the issue of optimal substrate selection at the stage of adaptation of microclones [4; 5; 6; 7; 8; 9; 10; 11], but at the same time there are practically no scientific studies on the use of nutrient substrates to intensify the regeneration processes of initial grape explants.
The aim of our work was to determine the regeneration potential of rootstock and technical grape vari-
eties in the culture of tissues and organs in vitro on different types of mineral substrates; to determine the rooting of grape microclones in vivo.
The work was carried out in the Department of Grape Seeding, Reproduction and Biotechnology of the National Research Center "Institute of Viticulture and Winemaking named after V. E. Tairov" during 20192021. The research was conducted on rootstock and technical grape varieties: "Dobrynya", "Garant", "Yarylo", "Zagrey".
All studies were carried out in aseptic conditions of laminar and culture boxes according to physical parameters: temperature = + 24 - +25 oC, lighting - 2500 - 3000 lux, 16-hour photoperiod, air humidity - 60 -70%.
The scheme of experience was as follows:
Control 1 - Murashige and Skoog medium + 0.3 ml/L IOK, 0.2 ml/L 6-BAP;
Control 2 - Murashige and Skoog medium + 0.6 ml/L IOK, 0.5 ml/L 6-BAP;
Variant 1 - Perlite;
Variant 2 - Vermiculite;
Variant 3 - Perlite + Vermiculite (1:1).
In the control variants, the nutrient medium of Mu-rashige and Skoog, prepared according to the prescription, was used.
Mineral substrates were autoclaved at 1 atm. twice for 40 minutes, after which they were saturated with nutrients. Two-bud cuttings were planted on the prepared substrates. Substrates were moistened once a week. Distilled water was used for irrigation.
From the indicators of the manifestation of regeneration properties, the following were determined: the survival of the initial explants (% after 30 days of cultivation), the beginning of the proliferation of axillary buds (days), the number of explants with the proliferation of axillary buds (% after 30 days of cultivation); the number of explants with rudiments of roots (%, after 30 days of cultivation).
«coyyomum-jmtmal» #mre), mm / agricultural sciences
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When determining the survival rates of initial scion tissues. grape explants on mineral substrates in vitro, we took After 30 days of cultivation, it was established that
into account explants that were characterized by the the survival of the initial grape explants mainly de-presence of a living lateral bud, a green leaf blade, and pended on the type of substrate (Figure-1).
Figure-1
Survival of initial explants of grapes on nutritious mineral substrates (average for 2019 - 2021)
%100 80 60 40 20 0
C1
C2
1
2
iDobrynya H Garant S Yarylo DDZagrey
3
Variants
The most established initial grape explants were found in the controls and the first experimental variant. The number of such explants was equal to 76.7% for the "Dobrynya" variety, 83.3% for the "Garant" variety, 75.0% for the "Yarylo" variety, and 50.0% for the "Zagrey" variety. In other experimental variants, the survival rate of initial grape explants was lower than the control and was equal to: 45.0 - 63.3% for the "Dobrynya" variety, 56.7 - 63.3% for the "Garant" variety, 55.0 - 61.7% for the variety "Yarylo" and 40.0 - 41.0% for the "Zagrey" variety.
One of the main indicators of the development of initial explants of grapes on substrates is the beginning of the proliferation of axillary buds. Acceleration of this process leads to faster formation and development of plants. It was shown that the development of axillary buds of the initial grape explants of all studied varieties started more actively in the plants of the first experimental variant. Thus, on the 10th day of research, 15.0% of the initial explants in the "Dobrynya" variety
were characterized by the beginning of the proliferation of axillary buds, in the "Garant" variety - 30.0% of the initial explants, in the "Yarylo" variety - 32.0%, and in the "Zagrey" variety - 24.0%.
Full accounting of the proliferation of axillary buds, rhizogenesis of initial explants of grapes was carried out on the 30th day of research. The obtained results showed that the most two-bud cuttings with axillary bud proliferation were in controls 1 and 2, as well as in the first and third experimental variants (Figure-2). Thus, in the rootstock grape varieties in the control variants, 62.0 - 71.0% of the initial explants were characterized by a developed axillary bud, in the technical 56.5 - 57.0%. In experimental variants with perlite, the number of such explants was equal to 43.3 - 55.0% in rootstock varieties and 45.0 - 46.7% in technical varieties. In all other experimental variants, the number of initial explants with a developed bud was less and was at the level of 36.7 - 42.3%.
Figure-2
The proliferation of initial grape explants on nutritious mineral substrates (average for 2019 - 2021)
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AGRICULTURAL SCIENCES / «ШУШМУМ-ШУГМаУ» 2022
Determination of the rate of rhizogenesis in the experimental and control variants showed that root formation began earlier and more actively in the control. Thus, 53.7 - 72.0% of the initial explants in the control variants had root primordia on the 30th day of research. After using nutrient substrates, the number of initial explants with primordia or developed roots was 10.2 -
20.4% lower than the control values. Among the experimental options, the best results were obtained in the first and third options. In rootstock varieties, the number of such plants was 46.7 - 55.0%, in technical varieties this indicator was equal to 45.0 - 65.0% (Figure-3).
Figure-3
Rhizogenesis of initial explants of grapes on nutrient mineral substrates (average for 2019-2021)
%100 90 80 70 60 50 40 30 20 10 0
C1
C2 1 2
S Dobrynya В Garant DYrylo HZagrey
3
Variants
But it should be noted that in the initial explants of grapes of the control variants, 2 - 3 roots were formed, which later acquired a large length, but were not branched, in the initial explants of grapes of the experimental variants, on the contrary, more roots were formed (5 - 8 pcs.), they had a greater number of roots of the second- and even third-orders, their length was shorter. This fact is extremely important for transferring grape microclones from in vitro to in vivo conditions.
It should also be noted that despite the fact that in vitro conditions in experimental variants we noted lower rates of survival of initial explants, axillary bud proliferation and rhizogenesis, later, when transferred to in vivo conditions (after 60 days of cultivation), the survival of plants with experimental variants exceeded control values on average by 20.6 - 24.5% for rootstock varieties, for technical varieties - 29.3 - 30.2% (Figure-4).
Figure-4
70,0 70,0,
72,5
66,9
48,0 45,0
45,0
41,0
I
I
-32,4
30,0 30,028 9
I
60,0
58,0
56,0
55052,0
65,6
_Survival of grape microclones under in vivo conditions (average for 2019-2021)
%100 90 80 70 60 50 40 30 20 10 0
74
371,2 Я
C 1 C 2 1 2
□ Dobrynya □ Garant □ Yarylo SZagrey
3
Variants
To increase the adaptation potential of grape mi- on nutritious mineral substrates. croclones in vitro, it is advisable to plant initial grape Optimal substrates are perlite, a mixture of perlite
explants (two-bud cuttings) and cultivate microclones and vermiculite (1:1).
«етушшшм-шишау» #шге)), 2022 / agricultural sciences
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The specified nutritious mineral substrates contributed to the high survival rate of grape microclones in in vivo conditions.
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