Two-dimensional soil culture method for researching interaction between olive root system and environment

Abstract

The invention discloses a two-dimensional soil culture method for researching interaction between an olive root system and an environment, and belongs to the technical field of olive biology. The two-dimensional soil culture method comprises the following steps: planting germinated olive seeds in a transparent two-dimensional culture container filled with a mixed matrix 1 of quartz sand and acid red soil, and scanning an olive root system by using a camera and an image analysis system in the culture process, so as to realize continuous monitoring on the growth dynamic state of the olive root system. According to the method, the quartz sand and the acid red soil are mixed to be used as the olive growth matrix, the growth condition of olive seedlings in the matrix is good, the growth matrix is further loaded by using a transparent two-dimensional culture container, the growth condition of the olive root system can be clearly observed, and the growth condition of the olive root system can be conveniently analyzed by using an image analysis system. By means of the culture method, environmental influence factors can be applied in the culture process, and the interaction between the olive root system and the environment is researched.

Description

Technical Field

[0001] The invention belongs to the technical field of olive biology, and in particular relates to a two-dimensional soil culture method for studying the interaction between olive root systems and the environment. Background Art

[0002] Olive (Canarium album L.) is a specialty agricultural product with great development potential in Guangdong. It is mainly planted in mountainous and hilly areas with relatively poor soil. However, due to the stress of the soil environment, the yield and quality of olives are significantly limited, so there is an urgent need to cultivate olive rootstock germplasm with broad-spectrum resistance. Studying the growth of olive roots is of great significance for understanding the olive's demand for nutrients and its interaction with the soil environment. At present, the mainstream method for studying the growth of woody plant roots is through in situ observation of soil. However, due to the opacity of the soil medium, in situ observation under natural conditions usually requires large-scale excavation of plant roots, which is a huge workload. The growth of plant roots is affected by a variety of environmental factors and is very sensitive to these factors. Therefore, the data obtained through in situ observation and measurement of soil under field conditions are often difficult to accurately reflect the specific effects of a single environmental factor on root growth. In addition, the root system is easily damaged during the excavation and sampling process, making it difficult to collect the roots completely. At the same time, its spatial configuration in the soil cannot be preserved, resulting in large errors in the acquired data, making it impossible to observe the root system configuration in situ. The acquired data often cannot truly reflect the impact of environmental factors on root growth. Therefore, it is of great significance to explore technologies that can more accurately reflect the interaction between plant roots and a single environmental factor under natural growth conditions. Summary of the invention

[0003] In view of the above problems, the present invention provides a two-dimensional soil culture method for studying the interaction between olive roots and the environment. By using sterilized quartz sand mixed with acidic red soil as a culture medium, a large number of olive seeds are germinated, and olive seedlings with consistent growth conditions are selected to lay the foundation for high-throughput analysis. Next, sterilized quartz sand and acidic red soil are mixed as a growth matrix, and the root growth is observed in situ in a transparent two-dimensional culture device. Finally, the cultivation device is opened to collect the samples intact. A simple, high-throughput and low-error technical solution is established for continuous observation of the growth of olive roots, which successfully realizes the in situ observation of olive root growth under soil conditions without digging.

[0004] To achieve the above object, the present invention provides the following technical solutions:

[0005] A two-dimensional soil culture method for studying the interaction between olive roots and the environment comprises the following steps:

[0006] The germinated olive seeds were planted in a transparent two-dimensional culture container containing a mixed matrix 1 of quartz sand and acidic red soil. During the culture process, the olive roots were scanned using a camera and image analysis system to achieve continuous monitoring of the growth dynamics of the olive roots.

[0007] The present invention selects a mixed matrix of quartz sand and acidic red soil as the culture matrix. First, the matrix is ​​more suitable for the growth of olive seeds. Second, it is easier to observe the growth dynamics of the olive root system using the matrix. Third, the olive root system is not easily damaged when the olive root system is removed at a later stage using the matrix.

[0008] Preferably, the mixed matrix 1 of quartz sand and acidic red soil is a mixture of quartz sand and acidic red soil in a volume ratio of 1:1.

[0009] More preferably, both the quartz sand and the acidic red soil are sterilized.

[0010] Preferably, the root length of the germinated olive seeds is 3 to 4 cm.

[0011] Preferably, Hoagland's nutrient solution is used as a nutrient component during the cultivation process.

[0012] Preferably, the transparent two-dimensional culture container is flat, and the width for the growth of olive roots is 10 mm.

[0013] The purpose of the present invention of controlling the width of the transparent two-dimensional culture container for the growth of the olive root system is to directly observe the growth dynamics of the olive root system.

[0014] Preferably, the germination steps of the germinated olive seeds include: first soaking the olive seeds in a sodium hypochlorite solution, then keeping them warm, then soaking them in a gibberellin solution, and then placing them in a mixed matrix 2 of quartz sand and acidic red soil for germination culture.

[0015] The olive seed germination method provided by the invention can greatly reduce the germination time of the olive seeds and improve the germination rate of the olive seeds.

[0016] More preferably, the concentration of the sodium hypochlorite solution is 0.5 g / L; and the immersion time in the sodium hypochlorite solution is 30 min.

[0017] More preferably, the insulation temperature is 55° C. and the insulation time is 30 minutes.

[0018] More preferably, the concentration of the gibberellin solution is 0.5 g / L; and the soaking time in the gibberellin solution is 12 h.

[0019] More preferably, the mixed matrix 2 of quartz sand and acidic red soil is a mixture of quartz sand and acidic red soil in a volume ratio of 1:1.

[0020] Further preferably, both the quartz sand and the acidic red soil are sterilized.

[0021] The beneficial technical effects of the present invention are as follows:

[0022] The present invention mixes quartz sand and acidic red soil as an olive growth matrix, and the olive seedlings grow well in the matrix. The growth matrix is ​​further loaded with a transparent two-dimensional culture container, and the growth of the olive root system can be clearly observed, which is convenient for analyzing the growth of the olive root system using an image analysis system. The culture method can be used to apply environmental influencing factors during the culture process to study the interaction between the olive root system and the environment.

[0023] Another effect of the mixed matrix of quartz sand and acidic red soil provided by the present invention is that the complete olive root system can be easily separated and the olive root system is not easily damaged during the separation process.

[0024] The invention also provides an olive seed germination method with short germination time and high germination rate. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 It is a schematic structural diagram of a detachable two-dimensional cultivation device for root growth of germinating olive seeds in an embodiment of the present invention.

[0026] Figure 2 This is a diagram of the cultivation steps of Example 3.

[0027] Figure 3 This is a two-dimensional culture actual picture of Example 3 using a mixed matrix of quartz sand and acidic red soil in a volume ratio of 1:1; wherein A is the overall picture; and B is an enlarged picture of the root system.

[0028] Figure 4 This is an enlarged view of the two-dimensional cultured root system using different growth matrices in Example 3; wherein A is a growth matrix using acidic red soil; and B is a growth matrix using a mixed matrix of quartz sand and acidic red soil in a volume ratio of 1:1.

[0029] Figure 5 These are the results analyzed by Image J image analysis software and pictures of olive roots sampled after cultivation in two growth matrices, among which A is the average value of the main root length of olive roots cultivated at different time in a mixed matrix of quartz sand and acidic red soil in a volume ratio of 1:1; B is the average value of the number of lateral roots of olive roots cultivated at different time in a mixed matrix of quartz sand and acidic red soil in a volume ratio of 1:1; C is a picture of olive roots sampled after cultivation in two growth matrices, the left side is the sampling result of the mixed matrix of quartz sand and acidic red soil in a volume ratio of 1:1, and the right side is the sampling result of the acidic red soil matrix. DETAILED DESCRIPTION

[0030] Now, various exemplary embodiments of the present invention are described in detail, and this detailed description should not be considered as a limitation of the present invention, but should be understood as a more detailed description of certain aspects, characteristics and embodiments of the present invention. It should be understood that the terms described in the present invention are only for describing specific embodiments and are not used to limit the present invention.

[0031] In addition, for the numerical range in the present invention, it is understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. The intermediate value in any stated value or stated range, and each smaller range between any other stated value or intermediate value in the range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0032] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which the invention pertains. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein may also be used in the practice or testing of the present invention.

[0033] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0034] The transparent two-dimensional culture container for the root growth of the germinating olive seeds in Example 3 of the present invention is a detachable two-dimensional cultivation device, and its structural schematic diagram is as shown in FIG. Figure 1 As shown, Figure 1 The unit of the middle number is mm, and the width for olive root growth is 10 mm.

[0035] Example 1

[0036] Study on the effect of different germination culture substrates on the germination rate of olive seeds

[0037] (1) 500 olive seeds of uniform size were selected and sterilized in a 0.5 g / L sodium hypochlorite solution for 30 minutes, and then rinsed three times with sterile water. Subsequently, the seeds were activated in a 55°C water bath for 30 minutes. After the treatment, the sterilized and activated seeds were soaked in a 0.5 g / L gibberellin solution for 12 hours.

[0038] (2) After sterilization and activation, the seeds were divided into five groups and placed in sterilized filter paper, coconut bran, quartz sand, acid red soil, and a mixed matrix of quartz sand and acid red soil in a volume ratio of 1:1 for germination culture. After 48 days of culture, the germination rate was calculated based on the germination of each group. The results are shown in Table 1.

[0039] Table 1 Effects of different germination culture substrates on the germination rate of olive seeds

[0040]

[0041] Table 1 shows that choosing a mixed matrix of quartz sand and acidic red soil in a volume ratio of 1:1 as the culture matrix not only shortens the germination time but also significantly increases the germination rate.

[0042] Example 2

[0043] Study on the effect of different activation temperatures on the germination rate of olive seeds

[0044] (1) 300 olive seeds of uniform size were selected and sterilized in a 0.5 g / L sodium hypochlorite solution for 30 minutes, and then rinsed three times with sterile water. The seeds were then divided into three groups, two of which were activated in a water bath at 55°C and 75°C for 30 minutes, respectively, and the other group omitted the water bath activation step as a control group (CK). After the treatment, the sterilized and activated seeds were soaked in a 0.5 g / L gibberellin solution for 12 hours.

[0045] (2) After the seeds were sterilized and activated, they were placed in a sterilized quartz sand and acidic red soil mixed matrix with a volume ratio of 1:1 for germination culture. After 48 days of culture, the germination rate was calculated based on the germination situation of each group. The results are shown in Table 2.

[0046] Table 2 Effects of different activation temperatures on the germination rate of olive seeds

[0047]

[0048]

[0049] Table 2 shows that the germination of unactivated olive seeds takes a long time and has a low germination rate. The activation temperature is too high and the olive seeds cannot germinate normally. The activation temperature of 55°C can guarantee both the germination rate and the germination rate.

[0050] Example 3

[0051] In situ observation and determination of the morphological architecture of olive roots under different growth media conditions

[0052] (1) The olive seeds were sterilized in a 0.5 g / L sodium hypochlorite solution for 30 minutes, and then rinsed three times with sterile water. Subsequently, the seeds were activated in a 55°C water bath for 30 minutes. After the treatment, the sterilized and activated seeds were soaked in a 0.5 g / L gibberellin solution for 12 hours.

[0053] (2) After the seeds are sterilized and activated, they are placed in a mixed matrix of sterilized quartz sand and acidic red soil in a volume ratio of 1:1 for germination culture.

[0054] (3) In accordance with Figure 1A two-dimensional cultivation device was assembled and filled with sterilized acidic red soil mixed with Hoagland's nutrient solution and a mixed matrix of sterilized quartz sand mixed with Hoagland's nutrient solution and acidic red soil in a volume ratio of 1:1 as a growth matrix.

[0055] (4) Germinated olive seeds with a root length of 3-4 cm and uniform growth were transferred into a two-dimensional cultivation device for cultivation. The device was placed in a light incubator with a light exposure time of 16 hours per day and a relative humidity of 60%, and Hoagland nutrient solution was applied regularly.

[0056] (5) The root system was photographed using a camera system on the 10th, 30th, and 50th days of cultivation, and the root system images were further analyzed using Image J to obtain relevant indicators.

[0057] (6) After cultivation, the two-dimensional cultivation device is opened, the olive roots are taken out, and indicators such as biomass are measured.

[0058] The cultivation steps of Example 3 are shown in FIG. Figure 2 .

[0059] Figure 3 This is a two-dimensional culture actual picture of Example 3 using a mixed matrix of quartz sand and acidic red soil in a volume ratio of 1:1; wherein A is the overall picture; and B is an enlarged picture of the root system.

[0060] from Figure 3 The growth of the olive roots can be clearly observed in B.

[0061] Figure 4 This is an enlarged view of the two-dimensional cultured root system using different growth matrices in Example 3; wherein A is a growth matrix using acidic red soil; and B is a growth matrix using a mixed matrix of quartz sand and acidic red soil in a volume ratio of 1:1.

[0062] from Figure 4 It can be seen that compared with the mixed matrix of quartz sand and acidic red soil, the growth of olive roots cannot be observed using pure acidic red soil as the growth matrix, which limits the subsequent use of cameras and software to analyze the growth of olive roots.

[0063] Figure 5 These are the results analyzed by Image J image analysis software and pictures of olive roots sampled after cultivation on two growth matrices, where A is the average taproot length of olive roots at different cultivation times in a mixed matrix of quartz sand and acidic red soil in a volume ratio of 1:1; B is the average number of lateral roots of olive roots at different cultivation times in a mixed matrix of quartz sand and acidic red soil in a volume ratio of 1:1; C is a picture of olive roots sampled after cultivation on two growth matrices, with the left side showing the sampling result of a mixed matrix of quartz sand and acidic red soil in a volume ratio of 1:1, and the right side showing the sampling result of the acidic red soil matrix (the red framed part is the root break).

[0064] Since acidic red soil was used as the growth medium, it was not possible to collect root images through the camera, and there were no root analysis results during the cultivation process. Figure 5 As can be seen in Figure C, the olive root system grows better when the growth matrix is ​​a mixed matrix of quartz sand and acidic red soil in a volume ratio of 1:1. In addition, adding quartz sand to the growth matrix can produce a more complete olive root system.

[0065] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. A two-dimensional soil cultivation method for studying the interaction between olive roots and the environment, characterized in that: The following steps are involved: The germinated olive seeds were planted in a transparent two-dimensional culture container containing a mixed matrix 1 of quartz sand and acidic red soil. During the culture process, the olive roots were scanned using a camera and image analysis system to achieve continuous monitoring of the growth dynamics of the olive roots.

2. The two-dimensional soil culture method for studying the interaction between olive roots and the environment according to claim 1, characterized in that: The mixed matrix 1 of quartz sand and acidic red soil is a mixture of quartz sand and acidic red soil in a volume ratio of 1:

1.

3. The two-dimensional soil cultivation method for studying the interaction between olive roots and the environment according to claim 1, characterized in that: The root length of the germinated olive seeds is 3 to 4 cm.

4. The two-dimensional soil cultivation method for studying the interaction between olive roots and the environment according to claim 1, characterized in that: Hoagland nutrient solution was used as the nutrient component during the cultivation process.

5. The two-dimensional soil cultivation method for studying the interaction between olive roots and the environment according to claim 1, characterized in that: The transparent culture container is flat, and the width for the olive root system to grow is 10 mm.

6. The two-dimensional soil culture method for studying the interaction between olive roots and the environment according to claim 1, characterized in that: The germination steps of the germinated olive seeds include: firstly soaking the olive seeds in a sodium hypochlorite solution, then keeping them warm, then soaking them in a gibberellin solution, and then placing them in a mixed matrix 2 of quartz sand and acidic red soil for germination culture.

7. The two-dimensional soil cultivation method for studying the interaction between olive roots and the environment according to claim 6, characterized in that: The concentration of the sodium hypochlorite solution is 0.5 g / L; the soaking time in the sodium hypochlorite solution is 30 min.

8. The two-dimensional soil cultivation method for studying the interaction between olive roots and the environment according to claim 6, characterized in that: The insulation temperature is 55° C. and the insulation time is 30 minutes.

9. The two-dimensional soil culture method for studying the interaction between olive roots and the environment according to claim 6, characterized in that: The concentration of the gibberellin solution is 0.5 g / L; the soaking time in the gibberellin solution is 12 hours.

10. The two-dimensional soil culture method for studying the interaction between olive roots and the environment according to claim 6, characterized in that: The mixed matrix 2 of quartz sand and acidic red soil is a mixture of quartz sand and acidic red soil in a volume ratio of 1:1.

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