BR102018073102A2 - SYSTEM FOR THE SELECTION OF GENOTYPES OF TOLERANT PLANTS TO INCREASE THE MECHANICAL IMPEDANCE OF THE SUBSTRATE - Google Patents

Abstract

The system for the selection of plant genotypes tolerant to the increase in the mechanical impedance of the substrate The present invention is a system that allows to evaluate the responses of seeds and seedlings to conditions of physical impediment of the substrate, through the simulation of different levels of mechanical resistance to root penetration at the time of seed germination and seedling emergence. the application of this technique occurs in the areas of seed production and technology, soil management and plant breeding, aiming at evaluating the effects of soil compaction in the establishment of agricultural crops, with consistent results, as well as, and mainly, for the evaluation and comparison of plant genotypes, in order to identify genetic variability regarding the level of tolerance to physical soil impediment, at the time of emergence and growth of plants, which makes the system a useful tool for breeding programs. plants and for the selection of genotypes to be used in the production process.

Description

DESCRIPTIVE REPORT

System for the selection of plant genotypes tolerant of increased substrate mechanical impedance

TECHNICAL FIELD OF THE INVENTION

01. The present invention is a system that allows evaluating the responses of seeds and seedlings to conditions of physical impediment of the substrate, by simulating different levels of mechanical resistance to root penetration at the time of seed germination and emergence seedlings. The application of this technique occurs in the areas of seed production and technology, soil management and plant breeding, aiming at evaluating the effects of soil compaction in the establishment of agricultural crops, with consistent results, as well as, and mainly, for the evaluation and comparison of plant genotypes, in order to identify genetic variability regarding the level of tolerance to the physical impediment of the soil, at the time of emergence and growth of plants, which makes the system a useful tool for breeding programs. plants and for the selection of genotypes to be used in the production process.

TECHNICAL STATUS

02. The intense traffic of agricultural machinery, the modernization of agriculture and the incorrect management of the soil have contributed to the increase of compacted areas. The consequences of soil compaction are the failure of the plant stand due to the death of the plants; unevenness in height / size and in the plant cycle, reduced production, among others. Thus, it is of great importance to study and develop techniques for evaluating the growth and initial development of plants under conditions of physical soil impediment, for

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2/16 to investigate the effects of this stress on the establishment of culture in the field and to support the selection of genotypes for crops and in plant breeding programs.

03. In this sense, the seedling emergency test (NAKAGAWA, J. Vigor tests based on seedling evaluation. In: KRZYZANOWSKI, FC et al. Seed vigor: concepts and tests. Londrina: ABRATES, 1999. p.2.1 -2.21) has been indicated for the evaluation of seed vigor, which has a correlation with seedling emergence in field conditions. By conducting the test, it is possible to infer about the physiological quality of the seed lot by estimating the speed and emergency uniformity. However, in the post-germination stages, plants may have restrictions on their development, with compaction being one of the most common problems in agricultural areas. Thus, the evaluation of seedling emergence under condition of physical impediment of the substrate can be a useful tool in studies to simulate the effects of soil compaction on seedling emergence in field conditions. However, most of the techniques currently used in scientific experimentation have limitations that prevent obtaining consistent results, which makes it necessary, therefore, to create more reliable systems.

04. There are some experimental systems used to investigate the effect of the substrate's mechanical impedance on plant growth. As an example, the one used to evaluate the effect of substrate impedance on the root growth and architecture of the wheat plant (JIN, K .; SHEN, J .; ASHTON, RW; WHITE, RP; DODD, IC; PHILLIPS, AL; PARRY, MAJ; WHALLEY, WR The effect of impedance to root growth on plant architecture in wheat. Plant Soil, v.392, n.1-2, p.323-332, 2015) and the involvement of gibberellin hormone signaling, in response to soil resistance to root penetration in leaf elongation and number of tillers in wheat (COELHO FILHO,

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BAD.; COLEBROOK, E.H .; LLOYD, D.P.A .; WEBSTER, C.P .; MOONEY, S.J .; PHILLIPS, A.L .; HEDDEN, P .; WHALLEY, W.R. The involvement of gibberellin signalling in the effect of soil resistance to root penetration on leaf elongation and tiller number in wheat. Plant Soil, v.371, n.1-2, 81-94, 2013). The systems used in these researches are based on the compression of the sand, placing a weight on it, which increases the mechanical impedance of the root growth environment of the plants. The system is efficient for the evaluation of the effects of the physical impediment of the substrate on the growth of the plants, but it does not allow to estimate the emergence rate of the seedlings of the substrate. The disadvantage of this system is the evaluation of only one seedling, which does not allow obtaining parameters of percentage, speed and emergency uniformity, for which a higher number of seeds / seedlings is required per repetition.

05. In this scenario, the present system object of the patent application is proposed. Its main feature is the ability to simultaneously evaluate a larger number of seeds / seedlings in a single system, which is achieved through the use of a perforated disc, with 25 holes (which can be changed to meet the specificities of the species that want to work), in which several seedlings are grown at the same time. With the data obtained it is possible to calculate the percentage and uniformity of seedling emergence, which are important information, especially when it comes to plant breeding in which the development of genotypes with greater tolerance to compacted soils is sought. Thus, genotypes with a higher percentage and uniformity of emergence can be selected using the system, which is relevant for breeding programs and even for the choice of plant material to be cultivated.

06. A sand table system (WHALLEY, W.R .; FINCH-SAVAGE, W.E .; COPE, R.E .; ROWSE, H.R. and BIRD, N.R.A. The response of carrot

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4/16 (Daucus carota L.) and onion (Allium cepa L.) seedlings to mechanical impedance and water stress at sub-optimal temperatures. Plant, Cell & Environment, v.22, p.229-242, 1999) was used to evaluate the responses of carrot and onion seedlings to substrate mechanical impedance and water stress, at sub-optimal temperatures. Different weights were used on the sand to obtain the forces that resulted in the impedance. This was an adaptation of the dead load technique (COLLISGEORGE N .; YOGANATHAN P. The effect of soil strength on germination and emergence of wheat (Triticum aestivum L.) I. Low shear strength conditions. Australian Journal of Soil Research, v.23 , p.577-587, 1985) used to study the growth of wheat plants under different levels of substrate mechanical impedance. In this method, different levels of stress due to mechanical impedance of the substrate are applied to sand using objects of different masses. In the last commented experiment, six seedlings were used in each of the five replicated pots and almost 400 pots were needed to collect the data.

07. Again, the advantage of the system, object of the current patent application, over the one used in the commented works, is the possibility of evaluating a greater number of seedlings simultaneously, with easier estimation of the emergence and uniformity of emergence of the seedlings under mechanical impedance of the substrate.

08. The passage of agricultural machines or compaction rollers over the same place is another methodology used to study the effects of soil compaction on plant growth, allowing to evaluate the emergence, growth and productivity of plants. Each pass of the machine or rollers results in different levels of physical impediment of the soil, which can be measured with the use of penetrometers. Some studies that used this methodology can be cited: BEUTLER, A.N .; CENTURION, J.F .; CENTURION, M.A.P.C .;

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SILVA, A.P. Effect of compaction on the productivity of soybean cultivars in Red Latosol. R. Bras. Ci. Solo, v.30, p.787-794, 2006;

GROTTA, D.C.C .; FURLANI, C.E.A .; SILVA, R.P .; SANTOS, L .; CORTEZ, J.W .; REIS, G.N. Soybean crop as a function of sowing depth and vertical load on the sowing row. Agricultural Engineering, v.27, n.2, p.487-492, 2007; LATIF, N .; KHAN, M.A .; ALI, T. Effects of soil compaction caused by tillage and seed covering techniques on soil physical properties and performance of wheat crop. Soil & Environ., V.27, n.2: p.185-192, 2008; MODOLO, A.J .; FERNANDES, H.C .;

SCHAEFER, C.E.G .; SILVEIRA, J.C.M. Effect of soil compaction on the emergence of soybean seedlings in no-tillage system. Ciênc. Agrotec., V.32, n.4, p.1259-1265, 2008; GIAROLA, N.F.B .;

BRACHTVOGEL, E.L .; FONTANIVA, S .; PEREIRA, R.A .; FIOREZE, S.L. Soy cultivars under no-tillage in compacted Red Latosol. Acta Sci., Agron., V.31, n.4, p.641-646, 2009; ROQUE, A.A.O .; SOUZA, Z.M .; BARBOSA, R.S .; SOUZA, G.S. Control of agricultural traffic and physical attributes of the soil in an area cultivated with sugarcane. Research agropec. bras., v.45, n.7, p.744-750, 2010; MODOLO, A.J .; DEDORDI, G.F .; VARGAS, T.O .; DALLACORT, R .; BAESSO, M.M .; CAMPOS, J.R.R .; TRENTIN, R.G .; ANDRADE, G.S .; TROGELLO, E. Effect of seed furrow openers on the growth of soybean roots in compacted Oxisols. African Journal of Agricultural Research, v.12, n.13, p.1067-1073, 2017; among others.

09. In these experiments, the results are very variable and difficult to compare, since the field system is complex, involving many variables that can interfere with the results obtained and analyzed, such as, for example, soil type, intensity and frequency of the load applied, influenced by the tension, humidity, texture, structure and initial density of the soil.

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10. However, the system proposed in this patent application exposes a series of advantages regarding the uniformity and standardization of the variables that can interfere with the results, such as texture, moisture and aeration of the substrate, thus allowing greater reliability in the results obtained with the system and obtaining replicable and comparable results, which is essential for the selection of genotypes.

11. Another methodology aimed at studying the mechanical impedance of the substrate consists of the use of vases assembled with overlapping PVC rings (usually three), joined with plastic adhesive tape and closed at the bottom. The moistened soil of the intermediate ring is subjected to a hydraulic press which, by varying the volume of the compacted soil, allows to reach desired levels of compaction. The upper and lower rings of the pots are filled with the fertilized and moistened soil, without using the press. This technique was used to evaluate the growth of the aerial part and root system of soybeans and cover plants in compacted layers of soil (FOLONI, JSS; LIMA, SL; BULL, LT Aerial and root growth of soybeans and cover plants in compacted soil layers Rev. Bras. Ciênc. Solo, v.

30, n.1, p.49-57, 2006). In this system, the type of soil can interfere with the results, since it affects the humidity and densities reached along the PVC column at each intensity and frequency of load applied. In the proposed system, this problem is solved with the use of the capillarity process in sand, which has a standardized texture and structure, which provides uniformity of the substrate moisture throughout the period of conducting the experiment. Also in the proposed system it is guaranteed that the degree of compaction due to the applied load, and that it promotes the mechanical resistance to the penetration of the roots, is constant throughout the study period without alteration of the substrate density.

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12. Different levels of resistance to penetration in PVC columns filled with soil can be obtained by compressing the soil. This system was used for the evaluation of plants grown under different levels of substrate impediment; to quantify the genetic diversity of the early development of the root system of wheat plants in different soil densities and to test whether the number of roots is an adequate trait to evaluate the genotypic tolerance to soil compaction (COLOMBI, T .; KIRCHGESSNER, N .; WALTER, A .; KELLER, T. Root tip shape governs root elongation rate under increased soil strength. Plant Physiology, v.174, p.2289-2301,2017).

13. The system, object of the current patent application, is more suitable to evaluate the tolerance of genotypes in conditions of physical impediment of the substrate, since it presents standardization of factors that can interfere in the results. In the system with the use of soil, factors such as texture, humidity and density can interfere in the standardization of results and comparison with other studies, since these factors interfere in the level of physical impediment imposed on the roots of plants. Thus, different textures and densities of the soil and different humidity will give rise to different levels of mechanical impedance for the same weight used for soil compression. In the system, object of the current patent application, there is the standardization of humidity and aeration of the substrate. The standardization of the levels of mechanical impedance, obtained by the use of standardized weights, is one more advantage in relation to other systems that use soils.

14. Soil compaction tests in pots are also used to assess the effects of impedance on crop development (CENTURION, JF; CENTURION, MAPC; BEUTLER, AN; ROSSINI, LA; FREDDI, OS; SOUZA NETO, EL Compaction of soil in the development and production of soybean cultivars. Científica, v.34, n.2,

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p.203-209, 2006). In this research, the effect of soil compaction on the development of two soybean cultivars in an Oxisol was evaluated. The compaction technique used consisted of compressing the soil through free fall impacts of a plunger with determined weight, in the geometric center of a wooden support with a diameter slightly smaller than the cylindrical PVC vessel used, which had a defined capacity and following indications described in previous research (BEULTER, AN; CENTURION, JF Soil compaction in root development and soybean productivity. Pesquisa Agropecuária Brasileira, v.39, n.6, p.581-588, 2004).

15. Thus, as discussed for other systems, this compaction technique presents a lack of standardization in the methodology, since it is difficult to standardize variables such as humidity, texture, structure, density, soil type, intensity and frequency of applied load, factors that can interfere with the results and conclusions obtained, in addition to making comparison with other researches difficult. The system, object of the present patent application, is also advantageous in relation to this technique, as it allows for the standardization of humidity, texture of the sand substrate and level of physical impediment imposed.

16. A thin-layer wax system can also be used to assess resistance to root penetration in plant growth substrates. The molten mixture of 60% wax and 40% petrolatum (by weight) is poured into molds and allowed to solidify at room temperature. In this system, PVC tubes were stacked and sealed with adhesive tape, each 130 mm long and with an internal diameter of 10 mm. The tube was capped at the bottom, filled with media (50% sand and 25% vermiculite) and a layer of petrolatum wax, which is responsible for the mechanical impediment to root growth. The seeds were pre-germinated and transplanted into the system. The evaluation consisted of verifying, in the

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9/16 seedlings growing in the system, the number of roots that reached the wax layer (low or no tolerance to the imposed physical impediment) and those that penetrated the layer of wax (tolerant to the imposed physical impediment).

17. This system was used to evaluate the root penetration capacity and biomechanical properties in corn (CHIMUNGU, JG; LOADES, KW; LYNCH, JP Root anatomical phenes predict root penetration ability and biomechanical properties in maize (Zea mays). Journal of Experimetal Botany, v.66, p.3151-3162, 2015). The limitation of this methodology consists in not simulating typical conditions and closer to those verified in the field environment, since it uses a non-edaphic material (layer of wax) to promote resistance to penetration. In addition, it is a laborious technique, since the seeds must be pre-germinated so that, afterwards, the seedlings can be transplanted to the cultivation pot. The planting must be staggered and the upper and lower sections of the substrate must be separated hydraulically with the wax layer and the humidity levels maintained by irrigation of each layer separately. The present system object of the patent application is simple and easy to execute, it does not require pre-germination of the seeds or the transplanting of seedlings, besides simulating conditions closer to that of practice in the field (since layers of different substrates).

18. Tests on agar and based on filter paper are also used in laboratory experiments to evaluate seed germination and seedling establishment in substrate impedance condition (FINCH-SAVAGE1, WE; BASSEL, GW Seed vigor and crop establishment: extending performance beyond adaptation. Journal of Experimental Botany, v.67, n.3 p.567-591, 2016). However, these tests may not actually represent what occurs in the compacted soil, since there are several external and internal influences on the soil matrix

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10/16 that are not properly simulated, such as texture, aeration and moisture.

19. The present system, object of the patent application, is able to represent these agricultural conditions, as there is standardization of the substrate texture, requiring only the use of sand substrate with similar granulometry in different researches. In practice, the physical impediment imposed on root growth is dynamic, as observed in preliminary studies in which the level of impediment varies with the depth of the substrate. On the other hand, in the agar system, the impedance is uniform throughout the substrate. Additionally, the agar system does not allow high levels of physical impediment to be achieved, whereas in the system, object of the current patent application, mechanical impedance values of the substrate of approximately 1.0 MPa can be obtained, which for seedlings has been shown to be efficient for the indication of genotype tolerance to this physical stress (Colombi, T., Kirchgessner, N., Walter, A. & Keller, T. Root tip shape governs root elongation rate under increased soil strength. Plant Physiology. 174, 2289 -2301 (2017)).

20. The present system, object of the current patent application, represents a new technique to evaluate the effects of resistance to the penetration of roots in the substrate during the emergence and initial growth of seedlings through the establishment of different levels of mechanical impedance of the substrate. The system, by means of a perforated disc and applying a load in the center of the disc, allows to impose a constant force on the substrate and also guarantees the germination and emergence of the seedlings that are able to make it through the holes in the disc. The proposed system is useful and applied in experiments and research that deal with the evaluation of the effects of mechanical impedance on plant physiology, morphology and biochemistry, as well as in studies aimed at plant breeding, aiming at the selection of genotypes with greater

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11/16 level of tolerance to the physical impediment of the substrate, overcoming the difficulties of operation and standardization that limits the systems / methodologies available until then and used in the works cited.

21. The present system, object of the current patent application, allows to evaluate the responses of the seeds in condition of mechanical impedance of the substrate, standardizing disturbing factors that could interfere in the results, such as texture, humidity, aeration and level of physical impediment of the substrate. The major obstacle found in the current methodologies, previously mentioned, is the fact that there is no standardization of all variables that can interfere with the results obtained. Thus, in general, the advantages of the present system object of the patent application consists of: i) standardization of humidity and aeration of the substrate, since water is uniformly distributed throughout the substrate due to its upward flow through capillarity; ii) standardization of the level of physical impediment with the use of the same weight and, consequently, the same level of force imposed on the system, which allows to standardize the level of mechanical impediment of the substrate; iii) the standardization of temperature and light, when the system is mounted in a growth room with all these conditions controlled, which is advantageous compared to experiments in a greenhouse or field, in which there is no control of the environment; finally, iv) the substrate used is an inert material (sand), with the same granulometry, which allows the standardization of the substrate, which is not possible with the use of soils.

22. The present system, object of the current patent application, also allows the evaluation of several seedlings simultaneously, different from some schemes found in the literature. Additionally, the system favors the evaluation of roots that, in soils suffer much breakage and disruption and, in the sand, it is easily separated for quantification purposes

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12/16 and evaluation. Thus, it has a relatively low cost and high yield. Therefore, it can be used to analyze the effects of resistance to substrate penetration on seedling growth and development, as well as it can be used for research purposes in plant breeding in order to verify the genotypic variability and the selection of genotypes more tolerant to the compaction of the plant. ground.

DESCRIPTION OF THE FIGURES

23. Figure 1: Illustration of the System for the selection of plant genotypes tolerant to the increase of the mechanical impedance of the substrate.

24. Figure 2: Length (I), area (J) volume (K) and average root diameter (L) of soybean seedlings under different levels of substrate compression.

25. Figure 3: Dry root matter (M) and part area (N) of soybean seedlings under different levels of substrate compression.

26. Figure 4: Visual view of soybean seedlings under different levels of substrate compression. (O) control, (P) compressed.

DETAILED DESCRIPTION OF THE INVENTION

27. The invention consists of a system that aims to evaluate the responses of seeds and seedlings to conditions of physical impediment of the substrate by simulating different levels of soil compaction at the time of seedling emergence. The proposed system is based on the principle that the sand, when subjected to the application of a force (weight) reproduces, without deformation of its particles, a level of compaction, which can be used to simulate the impedance verified in soil of areas of cultivation. Different forces (weights) can be used to simulate different values of mechanical impedance, which will vary according to the characteristics of the sand, but which can be easily measured with a penetrometer. Thus, it is possible to evaluate the effect of the mechanical resistance of the substrate on the germination of

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13/16 seeds, seedling emergence and on the development of seedling roots in this condition.

28. The system comprises a container (A), of variable shape, provided that it has an area larger than the recipe (B), open at the top and closed at the bottom, which can be a plastic tray or similar; circular container (B), open at both ends, which can be a PVC pipe or pipe or similar; perforated disc (F); sand with standardized granulometry (C); fabric or net made of nylon, plastic, or similar material, which prevents sand from leaving the container (B) and allows free flow of water (D) and weight (G)

29. The container (A) has an area larger than the diameter of the circular container (B), allowing the accommodation of the container (B) inside. Larger containers (A) allow to accommodate larger diameters of receivers (B) and larger perforated discs (F), with a greater number of perforations, or with a smaller number of perforations, but with larger dimensions. These parameters can be adjusted according to the plant species of interest. The container (B) is placed upright inside the container (A), both of which are filled with sand with standardized granulometry (C). At the bottom of the container (B) is placed a fabric that can be of organza or similar material or nylon, plastic, or similar material (D), which has the function of preventing the leakage of sand (C). To moisten the sand (C) contained in the container (B), water (E) is added to the container (A) until the sand (C) contained in the container (A) is saturated. Inside the container (B), at the top, a perforated disk (F) of metal, or similar material, is placed, with a diameter slightly smaller than the diameter of the container (B), which allows the internal fitting of the disk (F) to the container (B). Example of dimensions of the perforated metal disc (F): 19 cm in diameter, drilled with 25 holes, homogeneously distributed over the surface, at

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14/16 except for the central part, and with 2 cm in diameter each hole, disposed inside the container (B) of 20 cm in diameter. These dimensions, disc diameter, number of holes and hole diameter, can be adapted to fit the plant species of interest.

30. The system also has a weight (G), which is placed on the perforated disc (F) in the central position (free of holes), which can be changed to simulate different loads and, therefore, different impedances on the substrate. For the purposes of standardization and comparison of results from different studies, it is necessary to standardize the applied force and sand granulometry to impose the physical impediment. Seedlings (H) develop in this system with substrate impedance and emerge in the holes in the disc. The system allows the daily assessment of the number of seedlings emerged, which allows estimating the emergency uniformity, in addition to the evolution of their development, by the height they reach. The system can be disassembled at predetermined periods, or after the end of seedling emergence, that is, when there is no longer a change in the number of seedlings that emerged in daily assessments. Afterwards, the morphological, physiological and / or biochemical characteristics of the seedlings can be evaluated.

DEMONSTRATION EXPERIMENTS

31. An experiment was carried out with the system proposed in this patent application, in which there is uniformity and standardization of the intensity and frequency of the applied load (weight applied on the system), use of container, pipe and disc with similar dimensions in all treatments evaluated, and use of sand with defined granulometry (> 1 mm = 20%; 1-0.5 mm = 39%; <0.5 mm = 41%). Two treatments were used, which consisted of a system without mechanical impedance (control), on the system only the perforated disc was used, and another with physical impediment of the substrate, where a weight of 11 kg was placed on the sand surface. A 515 x size tray was used

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300 x 95mm (length, width and height, respectively) with a capacity of 11 liters, 200mm diameter PVC pipe and 190mm diameter perforated metal disc containing 25 holes. Soybean seeds were sown (Glycime max), and after seven days, seedlings were removed from the system. With the aid of WinRhizo®, software for image analysis designed specifically for root evaluation, there was a reduction in the length, area and volume of roots, in addition to an increase in the root diameter of seedlings subjected to the physical impediment of the substrate (Figure 2). In addition, there was a reduction in root dry matter when subjected to mechanical impedance, although the dry matter of the aerial part did not differ statistically (Figure 3). Thus, the physical impediment of the substrate changed the morphology, mainly of the roots, of the seedlings, as can be seen in Figure 4. The seedlings submitted to this treatment presented a shorter root system and thicker roots, compared to the control, without physical impediment. Therefore, it was possible to diagnose differences between the treatments tested in this system, validating the proposal to verify the effects of the mechanical impedance of the substrate on germination, emergence and seedling growth.

CONCLUSION

32. The developed system is efficient for the evaluation of the effects of the compaction of the substrate on the germination, emergence and development of the seedlings. It can be used for various agricultural crops, requiring only adjustments regarding the dimensions of the containers A and B and the holes in the metal disk F, as well as definition of the standardization of the sand granulometry and the weight (s) used. The system can be very useful for morphological, physiological and biochemical studies of plant development under conditions of mechanical impedance of the substrate and has practical applicability for

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16/16 plant breeding companies, which can be used in the characterization and selection of genotypes in terms of tolerance to stress due to soil compaction.

Claims (5)

Claim 1. System for the selection of plant genotypes tolerant to increased substrate mechanical impedance, characterized by comprising container (A), open at the top and closed at the bottom; circular container (B), open at both ends; perforated disc (F), sand with standardized granulometry (C), nylon or plastic fabric or net, or similar material (D) and weight (G). 2. System for the selection of plant genotypes tolerant to the increase of the substrate's mechanical impedance, according to claim 1, characterized in that the container (A) comprises an area larger than the diameter of the container (B), allowing the accommodation of the container ( B) inside. 3. System for the selection of plant genotypes tolerant to the increase of the mechanical impedance of the substrate, according to claim 1, characterized in that the perforated disc (F) comprises a diameter smaller than the diameter of the circular container (B), so that allow its internal fit, and holes evenly distributed over the surface, except for the central part. 4. System for the selection of plant genotypes tolerant to the increase of the mechanical impedance of the substrate, according to claim 1, characterized in that the container (B) comprises at its lower end a fabric or net made of nylon, plastic, or similar material (D) that prevents sand from leaking (C). 5. System for the selection of plant genotypes tolerant to the increase of the mechanical impedance of the substrate, according to claims 1 to 4, characterized in that the container (B) with the fabric or mesh of nylon, plastic, or similar material (D ) be placed in a vertical position inside the container (A); the containers (A) and (B) are Petition 870180149835, of 11/09/2018, p. 26/30 2/2 filled with sand with standardized granulometry (C), with the sand (C) contained in the container (A) moistened with water (E) until saturation; the perforated disc (F) is inserted into the container (B), on the top and the weight (G) is placed on the perforated disc (F), in the central position, free of holes.