CC BY
22
Section 2. Biotechnology
https://doi.org/10.29013/AJT-21-3.4-8-16
Sadoyan Ruzanna Robertovna, Doctor of Biological Sciences, Dean of the Faculty of Biology, Chemistry and Geography, Armenian State Pedagogical University named after Kh. Abovyan, Yerevan, Armenia E-mail: ruzannasad@mail.ru Dupliy Nadejda Gennadievna, Laboratory Assistant of Laboratory of Experimental Mutagenesis, Southern Federal University, Academy of Biology and Biotechnology named after D. I. Ivanovsky, Rostov-on-Don,
Russian Federation E-mail: duplii@rambler.ru Usatov Alexander Vyacheslavovich, Doctor of Biological Sciences, Professor of the Southern Federal University, Academy of Biology and Biotechnology named after D. I. Ivanovsky,
Rostov-on-Don, Russian Federation E-mail: usatova@sfedu.ru Nebish Anna Andreevna, Candidate of Biological Sciences, Head of the Educational Laboratory
at the Department of Genetics and Cytology, Yerevan State University, Yerevan, Armenia E-mail: anna.nebish@mail.ru Piltakyan Armenuhi Armenakovna, PhD student at the Faculty of Biology, Chemistry and Geography, Armenian State Pedagogical University named after Kh. Abovyan,
Yerevan, Armenia E-mail: armenuhipiltakyan@gmail.com Azarov Anatoliy Sergeevich, Design Engineer, Molecular Genetics Laboratory, Southern Federal University, Academy of Biology and Biotechnology named after D. I. Ivanovsky,
Rostov-on-Don, Russian Federation E-mail: bioclon@list.ru
Popov Alexey Sergeevich, Candidate of Agricultural sciences, Director of the FSBSI Agrarian Scientific Center Donskoy, Zernograd, Russian Federation
E-mail: popowaleksey@mail.ru Sirekanyan Inessa Norayrovna, Candidate of Chemical Sciences, Associate Professor at the Faculty of Biology, Chemistry and Geography, Armenian State Pedagogical University named after Kh. Abovyan,
Yerevan, Armenia E-mail: inessasirekanyan@yahoo.com
AGRIMITIN USE EFFICIENCY ON THE CULTIVATION OF SPRING BARLEY UNDER DROUGHT CONDITIONS
Abstract. The effect of the pre-treatment of spring barley (Hordeum vulgare L.) cv 'Schedry' grains by agrimitin or SkQ3 (10- (6'-methylplastoquinonyl) decyltriphenylphosphonium) on the plants growth characteristics and the yield during cultivation in 2017 and 2018 under different drought conditions was studied. For the field experiments the pre-treatment of barley seeds by 2.5nM aqueous solution of agrimitin was used. Two years experiments in the Armavir (Republic of Armenia) and Zernograd (Russia) revealed the stimulation of the plants growth and development and the increasing of the yield of treated plants compared to the untreated plants. The significant differences for the majority of investigated traits were detected in 2018 with less precipitation and higher monthly average temperature during vegetation period than in 2017 especially in Armavir as more arid region than Zernograd (P<0.05). At the same time it was proved that the application of agrimitin had not negative effect on the qualitative traits of the seeds. Our results provide evidence that agrimitin has a great potential and can be recommended as a new effective and safe agrochemical for crop cultivation under drought condition which is especially relevant in face of climate change.
Keywords: Hordeum vulgare L., grain yield, drought stress, Skulachev-ions (SkQ3), agrimitin.
Introduction
Barley (Hordeum vulgare L.) belongs to Poaceae (Graminae) family is the fourth leading cereal crop grown worldwide with the global production at approximately 160 million tons per year [1, 2]. The spring barley is one of the most drought-resistant crop among the cereals and well adapted for cultivation in various soils under the arid climate conditions [3-5]. However, the climate change drastically affects crop productivity and lead to substantial yield losses.
Seed germination and seedling growth during vegetation period are the most critical stages which
affected both quality and quantity of crop yields [6]. Environmental stresses can inhibit plant growth and development, and as a result lead to crop yield reduction. Nowadays for sustainable agricultural production and food security new approaches, efforts and techniques are necessary to improve crop stress tolerance [7]. Drought is one of the serious environmental stresses affecting crops including the barley. Seedling tolerance to drought stress is crucial for plant growth and development through the whole vegetation period under water-limited condition. During seed filling to the seed maturity stage
the drought stress causes a negative effect on crops seed production resulting in a significant decrease of the yield [8-10]. In dry years, the yield of barley can decrease up to 5-6 times [11]. The global trends and main challenges facing the world agriculture is to improve and increase the crop yield potential which can be achieved by introduction and/or selection of drought tolerant high-yielding cultivars, as well as rational use of new agrochemicals for seeds pre-treatment [12].
Last decades new agrochemicals were synthesized by the research team of Moscow State University named after M. V. Lomonosov under the leadership of Academician Skulachev called Skulachev-ions (SkQ) (10-(6'-methylplastoquinonyl decyltriphenylphos-phonium) or agrimitin which were demonstrated protective properties in experiments on various types of organisms preventing cell death from oxidative stress [13]. In experiments on the mice the application of SkQimproved impaired dermal wound healing [14]. As an antioxidant component these chemicals include the mitochondria-targeted plastoquinone which is the analog of ubiquinone being involved in plant chloro-plasts [15]. These compounds affect mitochondria directly and prevent the synthesis of dangerous for cells molecules. During the last years mitochondria-targeted antioxidants have been tested on plants [16-17]. By their using the processes of morpho- and rhizogen-esis in vitro cultures of sprout cuttings were stimulated and the regeneration time of initial potato microplants was reduced [18]. It has been shown also that nanomolar concentrations (nM) of the SkQ1 stimulates both the formation of morphogenetic structures and their subsequent stage of the development into plants. The obtained results suggest that SkQs with antioxi-dant properties can accelerate the morphogenesis and cause the yield increasing of wide range of crops [19]. SkQs with nanomolar concentrations prevent the death of plant leaf epidermal or guard cells and improve the wheat crop structure. The agrimitin is of great interest by its high stability in aqueous solutions among the SkQclass compounds [17; 20; 24; 25].
In our previous study the analysis of plants of the spring barley cv 'Shedriy' grown in drought stress imitated by 15% aqueous solution of PEG-6000 revealed the significant reduction in growth rate parameters of seedlings in comparison with control plants [21]. The significant differences in the fresh and dry weight of sprouts and roots in the drought condition compared with untreated control plants were registered in a case of agrimitin 2,5 nM(P < 0.001) which was applied in the present 2-years field experiments.
The aim of our study was to investigate in field experiments the effect of agrimitin on the germination of spring barley grains under drought conditions, morphological and structural indicators of plants cultivation, the grain yield and yield components of spring barley in the arid conditions of the Armavir (Republic of Armenia) and Zernograd (Russian Federation).
Materials and methods
In our experiments the spring barley cv 'Shedriy' obtained by the selective breeding at FSBSI Agrarian Scientific Center Donskoy has been served as the object of study.
Field test and monitoring. The field tests were carried out in Echmiadzin Experimental Field of Scientific Center of Agriculture in Armavir province (Republic of Armenia) and at the experimental farm of FSBSI Agrarian Scientific Center Donskoy in Zernograd province of the Rostov region (Russian Federation). Plants of the spring barley cv 'Shedriy' were evaluated during the 2017 and 2018 crop harvest. The seed treatment was performed in plastic bags after their stirring for 2 min and then drying at room temperature. Next, the seeds were packed in special paper bags and stored in the seed laboratory. The consumption of agrimitin 2.5 nM aqueous solution was calculated at the dose of 1,6 mg/ton of seeds. Plants were grown on 4 plots with area of 50 m2 per plot. Seeds for control without the pre-treatment by agrimitin were maintained for comparative purposes. Field germination was defined as
the production of normal seedlings [26]. Data were calculated by using the following formula according to the Association of Official Seed Analysis:
_ Number of normal seedlings G % — x 1UU
Total number of grans
During our 2-years experiments morphological indicators of barley plants and qualitative indicators of spring barley seeds were studied. Plant height (cm), tillering capacity general and productive, plants survival and the dry weight of roots were measured at the end of vegetation. The effect of agrimitin on the structural traits of spikes, spikelets and grains per plant of spring barley cv 'Shedriy' was examined on the 30 plants with 2 replications for control and experiment per year. After harvest the qualitative indicators of spring barley grains including germination energy, laboratory germination, weight of 1000 seeds and volumetrical weight (g/l) were studied on the 30 plants with 2 replications for control and experiment per year.
The climate conditions of investigated regions
Climate indicators of the regions of field experiments were obtained at Zernograd and Armavir weather stations. The average decade values of air temperature and precipitation during the vegetation period of spring barley in 2017 and 2018 were compared with the average norm during 1935-2016. Comparative analysis of climatic conditions has shown that the climate in Armavir during 2 observed years was drier than in Zernograd. During vegetation period of both years the average monthly temperature in Armavir was higher than in Zernograd. 2017 characterized by higher precipitation than in 2018 in both regions while the amount of precipitation during the ripening period of grains (June-July) both years was much less in Armavir than in Zernograd with exception of June 2018. Heavy precipitations was registered in Armavir in May 2017 and in Zernograd in July 2018 with almost twice more volume than norm. Very low precipitation was registered in Zernograd in June 2018 with 13 times less than norm and in Armavir in June and July 2017 with 4 times less than in norm.
The design of the experiment. The experimental design was completely randomized. The sowing qualities, grain germination energy, seed germination and the seed index (wei ght of 1000 seeds) were determined according to the GOSTs 20290-74 and 10842-76. The average data of all investigated parameters and their standard deviation (±s.d.) were calculated using Excel software. For all traits the ratio to control was calculated. Obtained data were statistically processed using the STATGRA.PHICS Centurion software.
Results and discussions
Morphological indicators of barley plants. The main morphological indicators of spring barley in 2017 and 2018 after pre-treatment by ag-rimitin are presented in Table 1. The analyses of morphological indicators of plants detected the significant differences in their growth parameters. In the Zernograd with low water stress the application of agrimitin increased the development of all plant parameters up to 10%, and the highest ratio to control was detected in 2018 for the dry weight roots at the end of vegetation period by up to 133,3% (P < 0,01). In the Armavir as more arid region than Zernograd for the majority of the morphological traits the growth of the plants was more intensive than in Zernograd up to 16,1% more for plant height and up to 19,7% more for plants tillering capacity compared to control. As in a case with Zernograd the dry weight of roots in Armavir in 2018 was higher with application of agrimitin than in control and show the significant differences with control (p < 0,001).
Our results showed that grain treatment by agrimitin showed the significant differences for the most parameters in terms of seedling length, plant fresh and dry weight. Data obtained indicate the effectiveness of the application of the agrimitin on spring barley plants grown in the different natural arid regions. These findings are in accordance with the results of other agrochemicals presented byJalilian et al. (2014) and Khafagy et al. (2017) [5; 27; 28].
Table 1.- The effect of agrimitin on the morphological traits of the plants of spring barley in the phase of full ripeness in 2017 and 2018
Variant, year Field germina-tion,% ±s.d. Plant height, cm ±s.d. Tillering capacity, number ±s.d. Dry weight of roots, g ±s.d. Survival, % ±s.d.
general productive
Zernograd
2017 Control 80.7±2.5 81.2±3.6 2.95±0.5 2.0±0.2 10.8±1.3 78.1±3.6
Agrimitin 85.4±1.7 83.6±4.2 3.27±0.6 2.2±0.2 11.3±1.1 83.9±2.9
Ratio to con-trol,% 105.5 102.9 109.8 109.1 104.4 106.9
2018 Control 82.6±2.8 78.3±1.7 2.87±0.4 2.0±0.2 9.1±0.9 75.2±4.2
Agrimitin 86.1±2.4 86.8±2.1* 3.02±0.6 2.2±0.2 13.6±1.2** 83.5±3.9*
Ratio to con-trol,% 104.1 109.8 105.0 109.9 133.3 110.0
Armavir
2017 Control 78.2±2.5 71.4±2.3 2.89±0.6 2.2±0.2 8.2±1.3 74.5±1.6
Agrimitin 82.3±3.4 80.5±2.1* 3.15±0.7 2.4±0.2 13.6±1.1*** 86.6±1.7*
Ratio to con-trol,% 105.0 111.0 108.3 108.3 139.7 113.0
2018 Control 78.5±0.9 63.9±2.1 3.18±0.2 2.6±0.2 8.0±0.6 75.6±1.0
Agrimitin 83.2±0.7 74.2±2.7* 3.73±0.2* 3.1±0.01* 13.2±0.8*** 82.7±2.5*
Ratio to con-trol,% 106.0 116.1 117.3 119.7 166.5 109.4
* = P<0.05, **= P<0.01, ***= P<0.001
Structural traits of the plants of spring barley.
Pre-sowing treatment of barley seeds by agrimitin effected also on the harvest and its structural elements (Table 2). Data of the structural traits of the barley plants shown differences in investigated parameters of plants during their cultivation in different natural arid regions (controls). The application of agrimitin differently and mostly significantly increased the
spike length, spikelet number and number of grains per spike, and as results the barley yield in investigated regions. In Zernograd the results of parameters after agrimitin pre-treatment were higher from 4% to 13% in 2017, and from 10% to 21% in 2018 compared to control. In Armavir during both years plants data after treatment by agrimitin were higher compared to the control from 11% to 26% (P<0.05 and P<0.001).
Variant, year Spike length, cm ±s.d. Spikelet number per spike ±s.d. Number of grains per spike ±s.d. Yield, C/ha
1 2 3 4 5
Zernograd
2017 Control 6.3±0.4 20.7±1.3 19.1±2.6 47.1
Agrimitin 6.8±0.6 23.8±2.0* 20.4±3.1 49.0*
Ratio to control,% 107.4 113.0 106.4 104.0
2018 Control 6.5±0.7 19.6±1.3 20.1±1.3 39.4
Table 2.- The effect of agrimitin on the structural traits of the plants of spring barley
1 2 3 4 5
Agrimitin 7.2±0.5* 24.8±1.1** 26.2±1.9** 46.8*
Ratio to control,% 109.7 121.0 123.3 115.8
Armavir
2017 Control 6.1±0.2 19.1±1.1 19.6±2.1 41.1
Agrimitin 7.4±0.3* 22.8±1.9* 23.3±2.8* 50.8**
Ratio to control,% 117.6 116.2 115.9 119.1
2018 Control 7.04±0.28 21.5±1.53 21.21±1.68 39.0
Agrimitin 8.59±0.61** 23.36±1.81* 23.58±1.95* 49.0***
Ratio to control,% 122.0 115.2 111.1 125.8
* = P<0.05, **= P<0.01, ***= P<0.001
As regards to interactions between agrimitin treatments and different drought levels, our results show that agrimitin partially counteracted the harmful effects of drought especially in 2018 in both regions, especially at the higher drought stress presented in Armavir. Our results showed that grain treatment by agrimitin showed positive response in terms of structural traits of the plants of spring barley and yield of spring barley cv 'Shedriy'.
Qualitative indicators of spring barley seeds. According to the basic concepts of Food and Agriculture Organization (FAO) and Good Agricultural Practices (GAP) the application of agrichemicals should have both protective and productive inputs to reach maximum productivity. At the same time the main approach of using new chemicals in the agriculture is based on the safety use of their recommended optimal dosage towards an ecologically safe and sus-
tainable agriculture contribute to food security and obtain harmless products of higher quality without any affect on germination characteristic of grains [22].
In our experiment after treatment of seeds by agrimitin and the growth of the plants the seeds from the harvest were analyzed for detection the possible affect from the treatment in comparison to the control. In the Table 3 the qualitative indicators of spring barley seeds are presented. The positive effect of the agrimitin was observed for all investigated traits of the grains.
According to our results the seed treatment by agrimitin has not affected the qualitative indicators of spring barley seeds. Regarding to the interaction between different levels of drought stress and agrimitin application the data indicate that in the most cases all interactions increased grain germination percentage and seed index with significant differences in Armavir in 2018.
Variant, year Germination en-ergy,% ±s.d. Laboratory germi-nation,% ±s.d. 1000 grain weight, g ±s.d. Volumetrical weight, g/l
1 2 3 4 5
Zernograd
2017 Control 85.8±5.6 90.2±2.6 44.3±0.8 718
Agrimitin 94.6±4.8* 94.6±3.8 45.0±1.2 716
Ratio to control,% 109.3 104.7 101.6 100.0
2018 Control 86.3±3.9 90.7±2.8 45.8±1.4 712
Agrimitin 92.7±4.2* 92.3±3.1 47.3±1.8 716
Table 3.- The effect of agrimitin on the qualitative traits of the grains of spring barley
1 2 3 4 5
Ratio to control,% 106.9 101.7 103.2 100.6
Armavir
2017 Control 89.5±4.0 95.0±5.6 46.2±1.7 698
Agrimitin 93.0±3.9 98.5±3.9 48.8±1.1 702
Ratio to control,% 103.8 103.6 105.3 100.6
2018 Control 76.5±5.0 83.0±2.8 48.3±0.1 681.5
Agrimitin 85.0±5.7* 96.5±2.1** 56.1±0.01** 716.6*
Ratio to control,% 111.1 116.3 116.3 105.2
* = P<0.05, **= P<0.01, ***= P<0.001
In Zernograd the data of qualitative traits increased up to 7-9,3% for germination energy compared to the control or normal conditions. In Armavir in 2018 as the driest variant of our experiment the data for qualitative parameters of grains were 11-16% higher than in control. Our study revealed that the pre-treatment of spring barley grains by agrimitin had not negative effect on the qualitative traits of grains.
Conclusion
Nowadays, scientists adopt different technologies and approaches to improve seed viability, plant grow and yield by beating the negative effects of drought stress [5]. Pre-sowing treatment of crop seeds is one of the easiest, low cost and effective techniques to enhance plant tolerance in the different stressful environments [7].
Present study examined the effect of pre-sowing treatment of the spring barley cv 'Schedry' seeds by agrimitin agrochemical on seed germination and seedling growth, as well as morphological and structural characteristics of plants in different arid conditions.
During field experiments the pre-treatment of barley seeds by agrimitin effected on plants morphological and structural traits by increasing of data for all investigated parameters compared to the untreated plants under the same drought stress. For the majority growth traits the significant differences were detected especially in Armavir as more arid region than Zernograd. The highest differences among the
growth parameters of the plants in both regions compared with untreated control were registered for dry weight of roots with significant differences P<0.001. Previous research with similar experiments on the agrimitin treated seeds of winter wheat showed the increasing the plants growth rates and the crop yield [23]. At the same time in presented study it was proved that during 2 years of spring barley cultivation the application of agrimitin had not negative effect on the qualitative traits of seeds.
Thus, our results demonstrated that seed priming treatments by agrimitin as SkQ3 mitochondria-targeted antioxidant efficiently improved the emergence and seedling growth performance under drought stress and can be recommended as a new effective and safe agrochemical for sustainable agriculture and for cultivation the crops in different arid conditions. These effects were related to the inhibition of oxidative stress during water-stress condition which is very important in face of climate change.
Conflicts of interest
The authors declare that they have no conflict of interests.
Acknowledgments
The work was carried out with the financial support of the Foundation for Assistance to Small Innovative Enterprises in the Scientific and Technical Sphere (Innovation Assistance Fund) in the framework of the project № 27000, "Development ofpreparations to increase drought-resistance and yield of crops grown in arid conditions of southern Russia and Armenia based
on the CCU (Center for Collective Usage) Biotechnology, Biomedicine and Environmental monitoring of SFU (Southern Federal University) and the Pro-
gram 10-5/17AR supported by the State Committee of Science (SCS) of the Ministry of Education and Science of the Republic ofArmenia.
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