Method for optimizing TTC dyeing and evaluating low-temperature cold resistance of wine grape branches

Abstract

The invention discloses a method for optimizing TTC (triphenyltetrazolium chloride) dyeing and evaluating low-temperature cold resistance of wine grape branches, which obviously improves the accuracy and repeatability of experiments by optimizing TTC dyeing conditions (dyeing at 35 DEG C for 72 hours), so that the section dyeing area of the branches reaches 100%. Besides, according to the method, various low-temperature treatment conditions of-10 DEG C,-15 DEG C,-20 DEG C and-30 DEG C are set, and treatment is carried out within 12 hours, 24 hours and 96 hours, so that the low-temperature cold resistance of the wine grape branches is systematically and comprehensively evaluated. Experimental results show that the cold-brew 1301 and the beda show relatively strong cold resistance after being treated at-30 DEG C for 96 hours, the dyeing areas are 79.07% and 76.73% respectively, the dyeing areas of the cabernet sauvignon and the maisseria are 54.53% and 3.45% respectively, and the latter dies basically. The method is simple and convenient to operate and easy to popularize and apply, and also provides a scientific and reliable wine grape variety selection basis.

Description

Technical Field

[0001] The invention relates to the field of plant physiology and agricultural science and technology, and in particular to a method for optimizing TTC staining of wine grape branches and evaluating low temperature resistance. Background Art

[0002] The cold resistance of wine grape varieties is one of the key factors in determining their suitable planting areas and cultivation management. In the existing technology, the cold resistance of grape branches is usually evaluated by low temperature treatment and then using TTC (2,3,5-triphenyltetrazolium chloride) staining method. However, this traditional method has many problems:

[0003] 1. Inconsistent experimental results due to non-optimized dyeing conditions: In the prior art, the selection of TTC dyeing conditions (including temperature and time) is often not precise enough, resulting in poor repeatability and accuracy of dyeing results. Different laboratories or researchers use different dyeing conditions, resulting in poor comparability of experimental results and difficulty in obtaining consistent low temperature resistance evaluation results.

[0004] 2. The low-temperature treatment method is single and the evaluation results are not comprehensive: In the existing technology, the temperature and time settings for low-temperature treatment of grape branches are relatively single, usually only one or several temperature conditions are selected, and the treatment time is also relatively fixed. This single treatment method cannot fully reflect the cold resistance of different varieties of wine grapes under various low-temperature conditions, resulting in one-sidedness and limitations in the evaluation results.

[0005] 3. The experimental process is complicated and the operation is difficult: the existing low-temperature treatment and TTC staining methods usually require more complicated experimental equipment and operation steps, which increases the complexity of the experiment and the difficulty of operation, and is not conducive to promotion and application in actual production and scientific research.

[0006] In order to overcome the above problems, the industry usually tries to improve the dyeing conditions or increase the temperature and time settings of low temperature treatment. However, these improvement methods often lack systematicity and scientificity, and it is still difficult to solve all the above problems. For example, although the dyeing effect can be improved by extending the dyeing time or increasing the dyeing temperature, these improvements often cannot solve the problem of consistency of dyeing results. At the same time, although increasing the temperature and time settings of low temperature treatment can more comprehensively evaluate cold resistance, it will also increase the complexity of the experiment and the difficulty of operation.

[0007] Therefore, how to improve the accuracy and repeatability of TTC staining results of wine grape branches and provide a systematic and comprehensive low temperature resistance evaluation method has become a technical problem to be solved by the present invention. Summary of the invention

[0008] The technical problem solved by the present invention is to provide a method for optimizing TTC dyeing of wine grape branches and evaluating low temperature resistance in response to the defects in the above-mentioned prior art, so as to solve the problems proposed in the above-mentioned background technology that the dyeing conditions are not optimized resulting in inconsistent experimental results, the low temperature treatment method is single resulting in incomplete evaluation results, and the experimental process is complicated and difficult to operate.

[0009] In order to solve the above technical problems, the technical solution adopted by the present invention is as follows:

[0010] A method for optimizing TTC dyeing of wine grape branches and evaluating low temperature resistance comprises the following steps:

[0011] Step 1, taking wine grape branches, placing them under different time and temperature conditions for TTC staining, and soaking them in water as a control;

[0012] Step 2, optimizing the dyeing conditions and determining the optimal dyeing time and temperature, wherein the optimal dyeing condition is dyeing at 35° C. for 72 hours, so that the dyeing area of ​​the branch section reaches 100%;

[0013] Step 3, using the optimized TTC staining conditions to stain the wine grape branches after low temperature treatment, and comparing the staining areas of the branches under different low temperature treatment conditions to evaluate their low temperature resistance;

[0014] Step 4, evaluating the low temperature resistance of the wine grape variety based on the size of the staining area.

[0015] As a further embodiment of the present invention, the low temperature treatment in step 3 comprises placing the wine grape branches under low temperature conditions of -10°C, -15°C, -20°C and -30°C for 12 hours, 24 hours and 96 hours respectively;

[0016] As a further scheme of the present invention, evaluating the low temperature resistance of wine grape varieties according to the size of the staining area in step 5 means that after treatment at -30°C for 96 hours, the staining area of ​​the branches of Hanniang 1301 is 79.07%, the staining area of ​​the branches of Beda is 76.73%, the staining area of ​​the branches of Cabernet Sauvignon is 54.53%, and the staining area of ​​the branches of Marselan is 3.45%.

[0017] As a further embodiment of the present invention, the wine grape branches include but are not limited to Hanniang 1301, Beda, Cabernet Sauvignon or Marselan varieties.

[0018] As a further embodiment of the present invention, the optimized TTC staining condition is staining at 35°C for 72 hours.

[0019] As a further embodiment of the present invention, the low temperature treatment comprises placing the wine grape branches under low temperature conditions of -10°C, -15°C, and -20°C for 12 hours and 24 hours respectively, and under low temperature conditions of -30°C for 96 hours.

[0020] As a further solution of the present invention, the low temperature resistance of different varieties of wine grape branches is evaluated by comparing the changes in the dyeing area after low temperature treatment.

[0021] As a further embodiment of the present invention, the staining area of ​​the Marselan branches treated at -30°C for 96 hours is 3.45%, indicating that they are basically dead under such conditions.

[0022] As a further solution of the present invention, in step 1, taking wine grape branches is taking annual wine grape branches.

[0023] A method for evaluating the low temperature resistance of wine grape branches, comprising the following steps:

[0024] Step s1, performing TTC staining on wine grape branches according to the method of claim 1;

[0025] Step s2, determining the low temperature resistance of different varieties of wine grapes by comparing the dyeing areas of wine grape branches under different low temperature treatment conditions.

[0026] Compared with the prior art, the present invention has the following beneficial effects:

[0027] 1. Significantly distinguish the low temperature resistance of different varieties. The method of the present invention can significantly distinguish the cold resistance of different wine grape varieties under low temperature conditions. For example, after being treated at -30℃ for 96 hours, Hanniang 1301 and Beida showed strong cold resistance, with staining areas of 79.07% and 76.73%, respectively, while Cabernet Sauvignon and Marselan had staining areas of 54.53% and 3.45%, respectively, and the latter basically died under this condition. This obvious distinction effect provides an important reference for the selection and promotion of wine grape varieties with strong cold resistance.

[0028] 2. In the low temperature treatment evaluation, multiple temperature conditions of -10℃, -15℃, -20℃ and -30℃ were set, and the treatment was carried out in different time periods (12 hours, 24 hours, 96 hours), providing a systematic and comprehensive low temperature resistance evaluation method. This method can distinguish in detail the cold resistance of different varieties of wine grapes under extreme low temperature conditions, providing a scientific basis for the selection of wine grape varieties.

[0029] 3. Optimization of dyeing conditions significantly improves experimental accuracy and repeatability. The present invention has determined that dyeing at 35°C for 72 hours is the optimal condition through TTC dyeing experiments under different temperature and time conditions, so that the dyeing area of ​​the branch section reaches 100%. This optimized dyeing condition not only improves the accuracy and repeatability of the experiment, but also ensures that the physiological state of the branch can be accurately reflected during the evaluation process.

[0030] 4. The method is simple and easy to promote. The dyeing and low-temperature treatment method of the present invention is easy to operate, does not require complicated equipment and conditions, and is easy to promote and apply in actual production and scientific research. This advantage not only reduces the experimental cost, but also improves the feasibility and practicality of actual application.

[0031] Additional aspects and advantages of the present invention will be given in part in the following description and in part will be obvious from the following description, or will be learned through practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative labor.

[0033] Figure 1 The present invention is a flow chart of TTC dyeing optimization and low temperature resistance evaluation of wine grape branches.

[0034] Figure 2 This is an optimization diagram of TTC dyeing conditions for wine grape branches of the present invention.

[0035] Figure 3 This is a TTC staining diagram of the wine grape branches under low temperature stress of the present invention.

[0036] Figure 4 This is a graph showing the changes in dyeing area of ​​branches treated with different low temperatures according to the present invention. DETAILED DESCRIPTION

[0037] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0038] See also Figure 1 —4. In an embodiment of the present invention, a method for optimizing TTC dyeing of wine grape branches and evaluating low temperature resistance comprises the following steps:

[0039] Step 1, taking wine grape branches, placing them under different time and temperature conditions for TTC staining, and soaking them in water as a control;

[0040] Step 2, optimizing the dyeing conditions and determining the optimal dyeing time and temperature, wherein the optimal dyeing condition is dyeing at 35° C. for 72 hours, so that the dyeing area of ​​the branch section reaches 100%;

[0041] Step 3, using the optimized TTC staining conditions to stain the wine grape branches after low temperature treatment, and comparing the staining areas of the branches under different low temperature treatment conditions to evaluate their low temperature resistance;

[0042] Step 4, evaluating the low temperature resistance of the wine grape variety based on the size of the staining area.

[0043] As a further embodiment of the present invention, the low temperature treatment in step 3 comprises placing the wine grape branches under low temperature conditions of -10°C, -15°C, -20°C and -30°C for 12 hours, 24 hours and 96 hours respectively;

[0044] As a further scheme of the present invention, evaluating the low temperature resistance of wine grape varieties according to the size of the staining area in step 5 means that after treatment at -30°C for 96 hours, the staining area of ​​the branches of Hanniang 1301 is 79.07%, the staining area of ​​the branches of Beda is 76.73%, the staining area of ​​the branches of Cabernet Sauvignon is 54.53%, and the staining area of ​​the branches of Marselan is 3.45%.

[0045] The wine grape branches include but are not limited to Hanniang 1301, Beda, Cabernet Sauvignon or Marselan varieties. The optimized TTC dyeing conditions are dyeing at 35°C for 72 hours. The low temperature treatment includes placing the wine grape branches under low temperature conditions of -10°C, -15°C, and -20°C for 12 hours and 24 hours respectively, and treating them under low temperature conditions of -30°C for 96 hours. By comparing the changes in the dyeing area of ​​different varieties of wine grape branches after low temperature treatment, their low temperature resistance is evaluated. Among them, the dyeing area of ​​the Marselan branches treated at -30°C for 96 hours is 3.45%, indicating that they are basically dead under this condition.

[0046] In step 1, the wine grape branches are annual wine grape branches.

[0047] A method for evaluating the low temperature resistance of wine grape branches, comprising the following steps:

[0048] Step s1, performing TTC staining on wine grape branches according to the method of claim 1;

[0049] Step s2, determining the low temperature resistance of different varieties of wine grapes by comparing the dyeing areas of wine grape branches under different low temperature treatment conditions.

[0050] Embodiment 1:

[0051] In this example, by optimizing TTC dyeing conditions and systematic low temperature treatment, the method of TTC dyeing and low temperature resistance evaluation of wine grape branches is specifically demonstrated. We selected four wine grape varieties: Hanniang 1301, Beda, Cabernet Sauvignon and Marselan.

[0052] First, we took one-year-old wine grape branches, cut them into segments about 10 cm in length, and soaked them in clean water as a control. In order to optimize the TTC staining conditions, we placed these branches at different temperatures (25°C, 30°C, 35°C) and different times (24 hours, 48 ​​hours, 72 hours) for TTC staining experiments. The results showed that after staining at 35°C for 72 hours, the staining area of ​​the branch section reached 100%, so the optimal staining conditions were determined to be 35°C and 72 hours.

[0053] Next, in order to evaluate the cold resistance of different varieties of wine grape branches, the branches were placed under different low temperature conditions. The specific steps are as follows:

[0054] The prepared branches were placed in a low temperature environment of -10°C, -15°C, -20°C and -30°C for 12 hours, 24 hours and 96 hours respectively. After the treatment, the branches were stained using the optimized TTC staining conditions (35°C, 72 hours).

[0055] Observe and record the dyeing area of ​​the branch cut surface after dyeing, and evaluate its cold resistance based on the dyeing area.

[0056] The experimental results are as follows:

[0057] After being treated at -30℃ for 96 hours, the dyeing area of ​​the branches of Hanniang 1301 was 79.07%, and the dyeing area of ​​the branches of Beida was 76.73%, indicating that these two varieties have strong low temperature resistance.

[0058] The dyeing area of ​​Cabernet Sauvignon under the same conditions is 54.53%, and its cold resistance is moderate.

[0059] The dyeing area of ​​Marselan was only 3.45%, indicating that it was basically dead under such low temperature conditions and had weak cold resistance.

[0060] The method of this embodiment significantly improves the accuracy and repeatability of TTC dyeing results, making the experimental results more reliable and comparable. At the same time, the systematic low-temperature treatment evaluation method can comprehensively and scientifically distinguish the low-temperature cold resistance of different wine grape varieties, providing a strong scientific basis for the selection of wine grape varieties in actual production. This method is easy to operate, does not require complex equipment and conditions, and is easy to promote and apply in production and scientific research. It significantly solves the problems of unoptimized dyeing conditions, single low-temperature treatment methods, and complex experimental processes in the prior art.

[0061] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above and that the present invention can be implemented in other specific forms without departing from the spirit or essential features of the present invention. Therefore, the embodiments should be considered exemplary and non-restrictive in all respects, and the scope of the present invention is defined by the appended claims rather than the above description, and it is intended that all changes falling within the meaning and scope of the equivalent elements of the claims be included in the present invention.

Claims

1. A method for optimizing TTC dyeing of wine grape branches and evaluating low temperature resistance, characterized in that: The following steps are involved: Step 1, taking wine grape branches, placing them under different time and temperature conditions for TTC staining, and soaking them in water as a control; Step 2, optimizing the dyeing conditions and determining the optimal dyeing time and temperature, wherein the optimal dyeing condition is dyeing at 35° C. for 72 hours, so that the dyeing area of ​​the branch section reaches 100%; Step 3, using the optimized TTC staining conditions to stain the wine grape branches after low temperature treatment, and comparing the staining areas of the branches under different low temperature treatment conditions to evaluate their low temperature resistance; Step 4, evaluating the low temperature resistance of the wine grape variety based on the size of the staining area.

2. The method for optimizing TTC dyeing and evaluating low temperature resistance of wine grape branches according to claim 1, characterized in that: The low temperature treatment in step 3 includes placing the wine grape branches under low temperature conditions of -10°C, -15°C, -20°C and -30°C for 12 hours, 24 hours and 96 hours respectively.

3. The method for optimizing TTC dyeing and evaluating low temperature resistance of wine grape branches according to claim 1, characterized in that: In step 5, evaluating the cold resistance of wine grape varieties according to the size of the staining area means that after being treated at -30°C for 96 hours, the staining area of ​​the branches of Hanniang 1301 is 79.07%, the staining area of ​​the branches of Beda is 76.73%, the staining area of ​​the branches of Cabernet Sauvignon is 54.53%, and the staining area of ​​the branches of Marselan is 3.45%.

4. The method for optimizing TTC dyeing and evaluating low temperature resistance of wine grape branches according to claim 1, characterized in that: The wine grape branches include but are not limited to Hanniang 1301, Beda, Cabernet Sauvignon or Marselan varieties.

5. The method for optimizing TTC dyeing and evaluating low temperature resistance of wine grape branches according to claim 1, characterized in that: The optimized TTC staining condition is staining at 35°C for 72 hours.

6. The method for optimizing TTC dyeing and evaluating low temperature resistance of wine grape branches according to claim 1, characterized in that: The low temperature treatment includes placing the wine grape branches under low temperature conditions of -10°C, -15°C, and -20°C for 12 hours and 24 hours respectively, and under low temperature conditions of -30°C for 96 hours.

7. The method for optimizing TTC dyeing and evaluating low temperature resistance of wine grape branches according to claim 1, characterized in that: The cold resistance of different varieties of wine grapes was evaluated by comparing the changes in dyeing area after low temperature treatment.

8. The method for optimizing TTC dyeing and evaluating low temperature resistance of wine grape branches according to claim 1, characterized in that: The staining area of ​​the Marselan branches treated at -30°C for 96 hours was 3.45%, indicating that they were basically dead under this condition.

9. The method for optimizing TTC dyeing and evaluating low temperature resistance of wine grape branches according to claim 1, characterized in that: In step 1, the wine grape branches are annual wine grape branches.

10. A method for evaluating the low temperature resistance of wine grape branches, characterized in that: The following steps are involved: Step s1, performing TTC staining on wine grape branches according to the method of claim 1; Step s2, determining the low temperature resistance of different varieties of wine grapes by comparing the dyeing areas of wine grape branches under different low temperature treatment conditions.

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