Method for promoting strawberry root development by using K599 agrobacterium rhizogenes

Abstract

The invention discloses a method for promoting strawberry root system development by using K599 agrobacterium rhizogenes. The method comprises an assembly type elevated cultivation device, seedling cultivation management and a method for promoting root system development. And the overhead cultivation device is adopted, so that the problems of gray mold and the like caused by field cultivation can be reduced. Seedling cultivation management is used for helping propagation of strawberry resources, reserving variety characteristics and shortening the seedling period. And promoting root development to help strawberry seedlings to rapidly root, strengthen roots, strengthen seedlings in a short period of time and shorten the seedling raising period. Efficient cultivation management and a rooting promoting method are combined, and the propagation period of the strawberries is shortened. According to the method, the K599 agrobacterium rhizogenes are used for promoting rooting of strawberry seedlings and absorption of plant nutrient substances by root systems, the root length, the root tip number and the root specific surface area are remarkably increased by 19.85%, 39.63% and 68.79% respectively, and the root systems are developed and robust. The strawberry seedling strengthening time can be shortened, cost is low, operation is easy and convenient, and the effect is remarkable.

Description

Technical Field

[0001] The invention discloses a method for promoting strawberry root development by utilizing K599 Agrobacterium rhizogenes, and relates to the technical field of plant cultivation. Background Art

[0002] Strawberry is one of my country's most important cultivated fruits, known for its high nutritional value and delicious flavor, earning it the nickname "Queen of Fruits." However, traditional ground cultivation methods present numerous drawbacks, including susceptibility to pathogens and high labor costs. The use of prefabricated elevated cultivation systems significantly increases yields by optimizing root space and light energy utilization, while also boosting fruit sugar content and firmness. The isolated ridge structure reduces soil-borne diseases and, combined with the greenhouse's rain-proofing function, significantly improves the prevention and control of diseases like gray mold.

[0003] The health of strawberry seedlings' roots is a key factor in determining their quality and subsequent yield. A healthy root system not only improves the plant's resistance to stress and promotes nutrient absorption, but also reduces the risk of disease, thereby ensuring the healthy growth of the strawberry seedlings. Therefore, efficient strawberry planting methods and root growth promotion techniques are crucial for strawberry cultivation. Currently, traditional high-ridge cultivation methods are simple to create but difficult to till the soil. Prolonged use can easily lead to soil compaction, which can affect the root growth of the strawberry seedlings. Furthermore, traditional ridge cultivation requires re-tillage and re-ridgeing after harvest, which requires a significant amount of manual labor. Summary of the Invention

[0004] The technical problem to be solved by the present invention is to provide a method for promoting the development of strawberry roots by using K599 Agrobacterium rhizogenes, thereby solving the problems mentioned in the above background technology.

[0005] To achieve the above objectives, the present invention provides a method for promoting the root growth of plant seedlings using K599 Agrobacterium rhizogenes, wherein the base of the plant seedlings is inserted into a rooting solution containing K599 Agrobacterium rhizogenes for immersion treatment, and the treated seedlings are planted in an assembled elevated cultivation device containing a substrate for post-planting management.

[0006] As an embodiment of the present invention, the immersion depth is from the base of the plant seedling to 0.5 cm below the bud point. After the treated seedlings are dried for 1 to 2 minutes, they are directly planted in an assembled elevated cultivation device containing a substrate for post-planting management.

[0007] As an embodiment of the present invention, the rooting solution containing K599 Agrobacterium rhizogenes has an OD value of 0.4-0.6.

[0008] As an embodiment of the present invention, the immersion time in the rooting solution containing K599 Agrobacterium rhizogenes is 30 seconds.

[0009] As an embodiment of the present invention, the plant seedlings are strawberry seedlings.

[0010] As an embodiment of the present invention, the matrix is composed of peat soil, perlite and vermiculite in a mass ratio of 2:1:1, and the matrix is disinfected by a combined application of pyraclostrobin and mancozeb before planting.

[0011] As an embodiment of the present invention, the specific operation of disinfecting the substrate is: using 1200 times of 250 g / L pyraclostrobin and 800 times of 80% mancozeb wettable powder, ensuring that the agents are in full contact with the substrate, tilling and mixing, and completing filming within 2 hours after application, using an airtight white or silver plastic film with a thickness of ≥0.04 mm to seal the cultivation trough.

[0012] As an embodiment of the present invention, the specific operations of the post-planting management are: watering immediately after planting, the greenhouse management requirements are that the temperature in the greenhouse is 28-30°C and the soil moisture is 70-80%; disinfection is carried out every 3 days; gradually ventilate 7 days after planting, gradually reduce humidity and increase light.

[0013] As an embodiment of the present invention, the assembled elevated cultivation device includes a bracket, a cultivation trough, a mud pusher and an outer frame; the bracket and the outer frame are separate, the outer frame is placed above the bracket and is fixed by a direct cross bar; the cultivation trough and the outer frame are separate, and the bottom of the cultivation trough is 3 cm away from the bottom of the outer frame; the length, width and height of the outer frame are 510 cm*32 cm*48 cm, and the material is a hollow stainless steel plate with a thickness of 1.5 cm; the length, width and height of the cultivation trough are 500 cm*30 cm*45 cm, and the material is a stainless steel plate with a thickness of 1 cm; the mud pusher is built into the cultivation trough, and the material is a stainless steel plate with a thickness of 1 cm; the detachable partition is respectively placed at both ends of the outer frame and the cultivation trough, and the material is PVC with a thickness of 5 mm.

[0014] As an embodiment of the present invention, the assembled elevated cultivation device has a three-layer structure with an overall height of 160 cm. The highest outer frame is 115 cm above the ground, the second outer frame is 70 cm above the ground, and the lowest outer frame is 25 cm above the ground.

[0015] Compared with the prior art, the present invention has the following advantages:

[0016] 1) Compared with traditional elevated cultivation devices, the device of the present invention can be recycled, which greatly reduces the waste of resources caused by repeated construction of elevated platforms, saves time and labor, and is conducive to reducing problems such as gray mold caused by ground planting.

[0017] 2) By setting up the mud pusher, the efficiency of replacing the cultivation medium is greatly improved, which greatly saves labor costs.

[0018] 3) A 3cm height difference is set between the cultivation trough and the bottom of the outer frame to store nutrient solution and avoid waste of resources.

[0019] 4) Treatment of strawberry seedling roots with Agrobacterium rhizogenes K599 effectively promoted root growth, increased the number of root tips, and increased the root specific surface area by 19.85%, 39.63%, and 68.79%, respectively. This achieved the goal of strengthening the seedlings, shortened the strawberry seedling raising cycle, and improved the efficiency of strawberry cultivation. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 This is a picture of the assembled prefabricated elevated cultivation device.

[0021] Figure 2 This is a picture of the prefabricated elevated cultivation device before assembly.

[0022] Figure 1 and Figure 2 Middle: 1-bracket, 2-cultivation trough, 3-mud pusher, 4-outer frame, 5-detachable partitions arranged at both ends of the outer frame (4), 6-detachable partitions arranged at both ends of the cultivation trough (2).

[0023] Figure 3 This is a statistical chart of root growth 30 days after K599 infection of strawberry seedlings with different root treatments. DETAILED DESCRIPTION

[0024] Two exemplary embodiments of the present invention are now described in detail. This description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0025] It should be understood that the terms described herein are intended only to describe particular embodiments and are not intended to limit the present invention. In addition, for numerical ranges herein, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Each smaller range between any intermediate value within a stated value or stated range and any other stated value or intermediate value within the stated range is also encompassed by the present invention. The upper and lower limits of these smaller ranges may be independently included or excluded within the scope.

[0026] Unless otherwise indicated, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art. 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. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of any conflict with any incorporated document, the contents of this specification shall prevail.

[0027] It will be apparent to those skilled in the art that various modifications and variations can be made to the specific embodiments of the present invention without departing from the scope or spirit of the invention. Other embodiments derived from the present invention will be apparent to those skilled in the art.

[0028] The present description and examples are for illustrative purposes only. The terms "include," "comprising," "having," "containing," etc. used herein are open-ended terms, meaning including but not limited to.

[0029] The K599 Agrobacterium rhizogenes strain used in the following examples was derived from the Leisure Agriculture Team of the Jiangsu Yanjiang Regional Agricultural Science Research Institute and is commercially available.

[0030] Example 1

[0031] like Figure 1 and Figure 2 As shown, the assembled elevated cultivation device of the present invention comprises a bracket (1), a cultivation trough (2), a mud pusher (3) and an outer frame (4); the bracket (1) and the outer frame (4) are separated, and the outer frame (4) is placed above the bracket (1) and fixed by a direct cross bar; the cultivation trough (2) and the outer frame (4) are separated, and the bottom of the cultivation trough (2) is 3 cm away from the bottom of the outer frame (4); the outer frame (4) has a length, width and height of 510 cm*32 cm*48 cm, and is made of a hollow stainless steel plate with a thickness of 1.5 cm; the cultivation trough (2) has a length, width and height of 500 cm*30 cm*45 cm, and is made of a stainless steel plate with a thickness of 1 cm; the mud pusher (3) is built into the cultivation trough (2), and is made of a stainless steel plate with a thickness of 1 cm; detachable partitions (5) and (6) are respectively placed at both ends of the outer frame (4) and the cultivation trough (2), and are made of PVC with a thickness of 5 mm.

[0032] The assembled elevated cultivation device has a three-layer structure with an overall height of 160 cm. The highest outer frame (4) is 115 cm above the ground, the second outer frame (4) is 70 cm above the ground, and the lowest outer frame (4) is 25 cm above the ground.

[0033] Example 2

[0034] A method for promoting root growth of strawberry seedlings using K599 Agrobacterium rhizogenes comprises the following steps:

[0035] (1) Seedling collection: Select the secondary runners of the Red Strawberry with basically the same growth height as the test samples. The collection time is selected in mid-February, and retain 2 cm of the runner stem.

[0036] (2) Disinfection of the cultivation substrate: Select a mixed substrate with a mass ratio of peat soil, perlite and vermiculite of 2:1:1 as the cultivation substrate, and disinfect the substrate by applying a combination of pyraclostrobin and mancozeb. The specific concentration is 1200 times of 250 g / L pyraclostrobin and 800 times of 80% mancozeb wettable powder. Ensure that the solution is in full contact with the substrate, plow and mix immediately, and complete the filming within 2 hours after application. Use an airtight white or silver plastic film (thickness ≥ 0.04 mm) to seal the cultivation trough and set aside.

[0037] (3) Seedling treatment and planting: Inoculate K599 rhizogenes in a 15 ml centrifuge tube filled with LB culture medium and shake at 180 r in a constant temperature shaker for 24 h. After 24 h, transfer the centrifuge tube to a low-temperature centrifuge and centrifuge at 5000 r at 4 °C. Discard the supernatant and add MgCl2, 10 mM MES (2-morpholineethanesulfonic acid) and 100 μM AS (acetosyringone) solution at a final concentration of 10 mM. Use MgCl2 to dilute to obtain rooting solutions containing K599 rhizogenes (K599 solutions for short) with OD values of 0, 0.4, 0.6 and 0.8, respectively. Place them in plastic pots for later use, and use pure water as the control group (OD is 0, which is the control group, CK).

[0038] Treatment group 1: Use a sterile knife to make a small cut at 0.5 cm below the base of the seedling to the bud point, insert the seedling into the K599 solution and soak for 30 seconds. Let the treated seedlings dry for 1-2 minutes before planting. Before planting, use a metal rod to punch holes in the treated substrate, and then plant the seedlings into the substrate. The cutting depth should be such that the base of the seedling bud is just exposed to the ground, and the seedlings should be planted vertically.

[0039] Treatment group 2: Insert the base of the seedling to 0.5 cm below the bud point into the K599 solution and soak for 30 seconds. Let the treated seedlings dry for 1-2 minutes before planting. Before planting, use a metal rod to punch holes in the treated substrate, and then plant the seedlings into the substrate. The cutting depth should be such that the base of the seedling's upper bud is just exposed to the ground, and the seedlings should be planted vertically.

[0040] (4) Management after planting: Immediately after planting, water thoroughly and gently press the base of the substrate to ensure that the seedlings are in close contact with the substrate. After planting, the room temperature should be 28-30℃ and the soil humidity should be 70-80%. Disinfect the soil every 3 days. After 7 days of planting, gradually ventilate the soil, gradually reduce the humidity and increase the light.

[0041] After 30 days, the root length, number of root tips and root specific surface area were investigated. Thirty seedlings were planted in each group and the results were averaged.

[0042] The survey results are shown in Tables 1, 2 and Figure 1 As shown:

[0043] Table 1 Root growth of strawberry seedlings in the root ring-cut treatment group infected with K599

[0044]

[0045] Table 2 Root growth of strawberry seedlings in the untreated group with K599 infection

[0046]

[0047] As can be seen from the data of Example 2, the OD value of K599 solution has an important impact on the growth of strawberry seedling root system, and it is not that the higher the OD value, the better. Overall, whether it is root ring cutting or no treatment, K599 infection of each concentration treatment can improve the index of strawberry seedling root length, root tip number, and root specific surface area, but under the same conditions, the infection result of the root no treatment group is significantly better than the root ring cutting group. Wherein, when the OD value is 0.4 and 0.6, the root length and root tip number of the root no treatment group are significantly higher than those of the CK group, and when the OD value is 0.4, it is relatively better, and root length, root tip number, and root specific surface area have increased by 19.85%, 39.63% and 68.79% respectively. When the OD value is 0.8, the effect decreases.

[0048] The embodiments described above are merely descriptions of preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Without departing from the spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by persons skilled in the art should fall within the scope of protection defined by the claims of the present invention.

Claims

1. A method for promoting root growth of plant seedlings using Agrobacterium rhizogenes K599, characterized in that: The base of the plant seedlings is inserted into a rooting solution containing K599 rhizogenes Agrobacterium and immersed therein, and the treated seedlings are planted in an assembled elevated cultivation device containing a substrate for post-planting management.

2. The method for promoting root growth of plant seedlings using Agrobacterium rhizogenes K599 according to claim 1, wherein: The immersion depth is from the base of the plant seedling to 0.5 cm below the bud point. After the treated seedlings are dried for 1 to 2 minutes, they are directly planted in an assembled elevated cultivation device containing a substrate for post-planting management.

3. The method for promoting root growth of plant seedlings using Agrobacterium rhizogenes K599 according to claim 1, wherein: The OD value of the rooting solution containing K599 Agrobacterium rhizogenes is 0.4-0.

6.

4. The method for promoting root growth of plant seedlings using Agrobacterium rhizogenes K599 according to claim 1, wherein: The soaking time in the rooting solution containing K599 Agrobacterium rhizogenes is 30 seconds.

5. The method for promoting root growth of plant seedlings using Agrobacterium rhizogenes K599 according to claim 1, wherein: The plant seedlings are strawberry seedlings.

6. The method for promoting root growth of plant seedlings using Agrobacterium rhizogenes K599 according to claim 1, wherein: The matrix is composed of peat soil, perlite and vermiculite in a mass ratio of 2:1:

1. Before planting, the matrix is disinfected by a method of jointly applying pyraclostrobin and mancozeb.

7. The method for promoting root growth of plant seedlings using Agrobacterium rhizogenes K599 according to claim 6, wherein: The specific operation of disinfecting the substrate is as follows: using 1200 times of 250 g / L pyraclostrobin and 800 times of 80% mancozeb wettable powder, ensuring that the agents are in full contact with the substrate, tilling and mixing, and completing film covering within 2 hours after application. Use an airtight white or silver plastic film with a thickness of ≥0.04 mm to seal the cultivation trough.

8. The method for promoting root growth of plant seedlings using Agrobacterium rhizogenes K599 according to claim 1, wherein: The specific operations of the post-planting management are: water thoroughly immediately after planting, the greenhouse management requirements are that the temperature in the greenhouse is 28-30°C and the soil moisture is 70-80%; disinfection is carried out every 3 days; and gradually ventilate 7 days after planting, gradually reduce humidity and increase light.

9. The method for promoting root growth of plant seedlings using Agrobacterium rhizogenes K599 according to claim 1, wherein: The assembled elevated cultivation device comprises a bracket (1), a cultivation trough (2), a mud pusher (3) and an outer frame (4); the bracket (1) and the outer frame (4) are separated, and the outer frame (4) is placed above the bracket (1) and fixed by a direct cross bar; the cultivation trough (2) and the outer frame (4) are separated, and the bottom of the cultivation trough (2) is 3 cm away from the bottom of the outer frame (4); the outer frame (4) has a length, width and height of 510 cm*32 cm*48 cm, and is made of a hollow stainless steel plate with a thickness of 1.5 cm; the cultivation trough (2) has a length, width and height of 500 cm*30 cm*45 cm, and is made of a stainless steel plate with a thickness of 1 cm; the mud pusher (3) is built into the cultivation trough (2), and is made of a stainless steel plate with a thickness of 1 cm; detachable partitions (5) and (6) are respectively placed at both ends of the outer frame (4) and the cultivation trough (2), and are made of PVC with a thickness of 5 mm.

10. The method for promoting root growth of plant seedlings using Agrobacterium rhizogenes K599 according to claim 9, characterized in that: The assembled elevated cultivation device has a three-layer structure with an overall height of 160 cm. The highest outer frame (4) is 115 cm above the ground, the second outer frame (4) is 70 cm above the ground, and the lowest outer frame (4) is 25 cm above the ground.

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