CN111713407B - A method of encapsulation, transplanting and domestication of Huaiyu Mountain alpine potato with bud stem segments - Google Patents

Abstract

The invention discloses a method for embedding, transplanting and domesticating a dioscorea opposita stem section with buds, which comprises the steps of mixing the dioscorea opposita stem section with buds with a calcium-free ion modified MS culture solution a containing sodium alginate and sucrose to obtain a mixed solution; dripping the mixed solution into MS culture solution b containing chitosan oligosaccharide, graphene quantum dots, carbon nano tubes, calcium chloride and sucrose to obtain the embedding beads of the budded stem segments of the Huaiyoshan high mountain potatoes; soaking the prepared embedded beads in MS culture solution containing chitosan oligosaccharide and graphene quantum dots; soaking the soaked embedding beads in MS culture solution containing chitosan oligosaccharide and carbon nano tubes; transplanting into a substrate, and irrigating an MS culture solution containing chitosan oligosaccharide and carbon nano tubes; pouring MS culture solution containing chitosan oligosaccharide and graphene quantum dots after pouring for 7 days; after the bud stem is grown into a complete plant, the domestication is completed, and the plant is transplanted into a field for cultivation. The method can realize the transplanting and domesticating survival rate of the test-tube plantlets of the Huaiyoshan high-mountain potatoes of more than 98 percent.

Description

A method of encapsulating, transplanting and acclimating Huaiyushan alpine potato with bud stem segments

technical field

The invention relates to the technical field of transplanting and acclimation, and relates to a method for embedding, transplanting and acclimatization of Huaiyu Mountain alpine potato with bud stem segments.

Background technique

Native to the Andes in South America, the potato is an important food and vegetable crop and the most important tuber crop in the world. Potatoes contain a large amount of trace elements, proteins, amino acids, B and C vitamins, and have the reputation of "second bread", "king of nutrition", "underground apple" and so on. Due to the high yield, strong adaptability, easy cultivation, and low water consumption of the growth cycle, potatoes can be supplied throughout the year. Therefore, potatoes can also produce high economic benefits in remote water-scarce areas. It is an industry that helps my country's poverty alleviation. In 2015, with the launch of the potato staple food strategy, potatoes have become China's fourth largest staple food and an important guarantee for food security. Potatoes have low requirements for soil fertility, pH, slope, humidity, and light. Most of them are in alpine areas, with large temperature differences, high altitudes, strong ultraviolet rays, and basically do not use pesticides and other agents that pollute the environment. Therefore, under the background of staple food, the cultivation of alpine potatoes is widely promoted in mountainous areas and hills and gentle slopes, which can save cultivated land and realize the greenness of alpine potatoes. No pollution. Huaiyu Mountain Alpine Potato, also known as hemp seed potato, was approved as a national geographical indication agricultural product in 2013. Huaiyu Mountain Alpine Potatoes are rich in nutrients and do not contain reducing sugars. They are suitable for dieters and people with diabetes.

Long-term growth in relatively harsh environmental conditions often suffers from stresses such as high and low temperature, water shortage, high altitude, strong ultraviolet rays, and nutrient deficiency. Due to the strong plasticity of plant development, plants generally respond to adapt to harsher environmental conditions. unfavorable environment. Huaiyu Mountain alpine potatoes are only suitable for planting in high-altitude alpine areas. Under long-term adversity stress, Huaiyu Mountain alpine potatoes have formed a series of strategies to adapt to the alpine environment. Once Huaiyu Mountain alpine potatoes are moved to low-altitude plains Domestication will break the original mechanism of adapting to adversity and produce mutations. At the same time, the survival rate of domesticated Huaiyushan alpine potato test-tube plantlets transplanted to alpine is low. And this has become a bottleneck restricting the cultivation of alpine potatoes in Huaiyu Mountain.

Contents of the invention

In order to solve the above-mentioned technical problems, the present invention provides a method for embedment, transplanting and acclimatization of potato bud stem segments.

A method for embedding and transplanting domestication of Huaiyu Mountain alpine potato with bud stem segment, comprising the following steps,

(1) Under aseptic conditions, mix the 0.5-1cm budded stem section of Huaiyu Mountain alpine potato with calcium ion-free modified MS culture medium a containing sodium alginate and sucrose to obtain a mixed solution;

(2) Under sterile conditions, drop the mixed solution into MS culture solution b containing chitosan oligosaccharides, graphene quantum dots, carbon nanotubes, calcium chloride, and sucrose for embedding to obtain Huaiyushan alpine potato with bud stem section Embedding beads so that it is not easy to dehydrate and can obtain sufficient nutrition;

(3) Under aseptic conditions, soak the Huaiyushan alpine potato with bud stem segment embedded beads prepared in step (2) with MS culture solution c containing chitosan and graphene quantum dots; use graphene-containing MS culture Liquid immersion can improve the ability of Huaiyu Mountain alpine potato to adapt to the high mountain adversity outside, and improve the survival rate of domestication;

(4) Under sterile conditions, soak the soaked Huaiyushan alpine potato bud stem segment embedded beads in step (3) with MS culture medium d containing chitosan and carbon nanotubes, and use MS containing carbon nanotubes The immersion in the culture solution can further improve the ability of Huaiyu Mountain alpine potatoes to adapt to the high mountain adversity outside, and further improve the survival rate of domestication;

(5) At room temperature, the Huaiyu Mountain alpine potato with bud stem segment embedded beads obtained in step (4) was transplanted into the matrix, and the MS culture solution d containing chitooligosaccharides and carbon nanotubes was poured in the greenhouse every morning and evening; After 7 days, change to MS culture medium c containing chitosan and graphene quantum dots every morning and evening; MS culture medium itself still has enough water, so there is no need to add water during this process;

(6) After the budding stem section grows into a complete plant, the domestication is completed, and it is moved into the field for cultivation. Embedding and soaking can make the plants adapt to the dehydration environment, thereby resisting the stress of high mountains and improving their survival rate.

Preferably, in the step (1), the mass volume ratio of sodium alginate in the modified MS culture medium a without calcium ions is 3%, and the concentration of sucrose is 0.2mol/L.

Preferably, in step ( ₂ ), in MS culture medium b, chitosan oligosaccharide concentration 0.05g/L, graphene quantum dot concentration 0.03-0.05g/L, carbon nanotube concentration 0.03-0.05g/L, CaCl Concentration 0.1mol/L and sucrose concentration 0.2mol/L.

Preferably, in step (3), in MS culture medium c, the chitosan oligosaccharide concentration is 0.05g/L, the graphene quantum dot concentration is 0.03-0.05g/L, and the soaking time is 48h.

Preferably, in step (4), in the MS culture solution d, the chitosan oligosaccharide concentration is 0.05g/L, the carbon nanotube concentration is 0.03-0.05g/L, and the soaking time is 48h.

Preferably, the matrix in step (5) is a mixture of vermiculite and perlite with a mass ratio of 2:1.

Preferably, the amount of each pouring of MS culture solution d is 20 mL/embedded beads.

Preferably, the amount of each pouring of MS culture medium c is 20 mL/embedded beads.

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

The alpine potato of Huaiyu Mountain will mutate when it is transplanted and domesticated in the low-altitude plain area. The test-tube seedlings of alpine potatoes in Huaiyu Mountain are relatively weak, and the survival rate of alpine transplanting is low, only 40-50%. The invention adopts the method of embedding, transplanting and acclimating stem segments with buds, and the survival rate reaches more than 98%. On the one hand, the embedded stem section with buds can ensure sufficient supply of nutrients in the later cultivation process so that it is not easy to dehydrate; The state of domesticated test-tube plantlets is more robust than conventional transplanted test-tube plantlets to ensure the survival rate of material-embedded beads transplanted under alpine adversity.

The invention can realize the transplanting and domestication survival rate of Huaiyu Mountain alpine potato test-tube seedlings to be over 98%, and the operation is simple and convenient, and can realize factory operation.

Detailed ways

Various exemplary embodiments of the present invention will now be described in detail. The detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features and embodiments of the present invention.

It should be understood that the terminology described in the present invention is only used to describe specific embodiments, and is not used to limit the present invention. In addition, regarding the numerical ranges in the present invention, it should be understood that each intermediate value between the upper limit and the lower limit of the range is also specifically disclosed. Each smaller range between any stated value or intervening value in a stated range and any other stated value or intervening value in a stated range is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded from the range.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only the preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can 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 in connection with which the documents are described. In case of conflict with any incorporated document, the contents of this specification control.

It will be apparent to those skilled in the art that various modifications and changes can be made in the specific embodiments of the present invention described herein without departing from the scope or spirit of the present invention. Other embodiments will be apparent to the skilled person from the description of the present invention. The specification and examples in this application are exemplary only.

As used herein, "comprising", "comprising", "having", "comprising" and so on are all open terms, meaning including but not limited to.

Example 1

Prepare calcium ion-free MS culture solution a: add sodium alginate and sucrose to make the concentration of sodium alginate 3% (w/v) and the concentration of sucrose 0.2mol/L;

MS culture solution b: Chitooligosaccharide concentration 0.05g/L, graphene quantum dot concentration 0.03g/L, carbon nanotube concentration 0.03g/L, CaCl ₂ concentration 0.1mol/L and sucrose concentration 0.2mol/L;

MS culture medium c: chitosan oligosaccharide concentration 0.05g/L, graphene quantum dot concentration 0.03g/L;

MS culture solution d: chitosan oligosaccharide concentration 0.05g/L, carbon nanotube concentration 0.03g/L;

(1) Under aseptic room temperature conditions, mix the 0.5 cm section of budded stems of Huaiyu Mountain alpine potatoes with the improved MS culture medium a without calcium ions to obtain a mixed solution. The amount of culture medium a: depends on the amount of budded stems, Generally, a stem section with buds is 1-2mL;

(2) drop the mixed solution into MS culture solution b containing oligochitosaccharides, graphene quantum dots, carbon nanotubes, calcium chloride, and sucrose for embedding to obtain Huaiyushan alpine potato with bud stem segment embedded beads;

(3) The prepared Huaiyushan alpine potato with bud stem segment embedded beads is soaked in MS culture solution c containing chitooligosaccharides and graphene quantum dots;

(4) The soaked Huaiyushan alpine potato with bud stem section embedded beads is soaked in MS culture solution d containing chitooligosaccharides and carbon nanotubes;

Under the sterile conditions of the above soaking environment, the temperature is 25±1°C, and the time is 48h;

(5) The obtained Huaiyushan alpine potato with bud stem section embedded beads were transplanted into a mixture matrix of vermiculite and perlite with a mass ratio of 2:1 (20-30°C at room temperature), and watered with shells at 8:00 every morning and evening. The MS culture solution d of oligosaccharides and carbon nanotubes, the amount of each watering is 20mL/bead embedding; after 7 days of watering, change the MS culture solution c containing chitooligosaccharides and graphene quantum dots every morning and evening; The volume is 20mL/bead embedding;

(6) After the stem section with buds grows into a complete plant for 20-25 days, the domestication is completed, and it is moved to the alpine field for planting.

The results showed that the state of the domesticated Huaiyu Mountain alpine potato was more robust than the transplanted test-tube seedlings; 98% of them grew new leaves and survived, and the survival rate was 98%.

Example 2

With embodiment 1, difference is:

MS culture solution b: Chitooligosaccharide concentration 0.05g/L, graphene quantum dot concentration 0.05g/L, carbon nanotube concentration 0.05g/L, CaCl ₂ concentration 0.1mol/L and sucrose concentration 0.2mol/L;

MS culture medium c: chitosan oligosaccharide concentration 0.05g/L, graphene quantum dot concentration 0.05g/L;

MS culture medium d: the concentration of chitosan oligosaccharides is 0.05g/L, and the concentration of carbon nanotubes is 0.05g/L;

The results showed that the state of the domesticated Huaiyu Mountain alpine potato was more robust than the transplanted test-tube seedlings; and 100% of them grew new leaves and survived, and the survival rate was 100%.

Example 3

With embodiment 1, difference is:

MS culture solution b: Chitooligosaccharide concentration 0.05g/L, graphene quantum dot concentration 0.04g/L, carbon nanotube concentration 0.04g/L, CaCl ₂ concentration 0.1mol/L and sucrose concentration 0.2mol/L;

MS culture medium c: chitosan oligosaccharide concentration 0.05g/L, graphene quantum dot concentration 0.04g/L;

MS culture solution d: chitosan oligosaccharide concentration 0.05g/L, carbon nanotube concentration 0.04g/L;

The results showed that the state of the domesticated Huaiyu Mountain alpine potato was more robust than the transplanted test-tube seedlings; and 100% of them grew new leaves and survived, and the survival rate was 100%.

Comparative example 1

With embodiment 1, the difference is that only embedding is carried out, and the two-step soaking of step (3) and step (4) and the pouring of solution c and solution d of step (5) are not carried out;

The result shows that the domesticated Huaiyu Mountain alpine potato, domesticated test-tube plantlet state: more robust than the transplanted test-tube plantlet, but weaker than the test-tube plantlet of Example 1; and 45% grow new leaves to survive, and the survival rate is 45%, which is the The domestication effect of test-tube plantlets is the same.

Comparative example 2

With embodiment 1, difference is, do not carry out the embedding of step (2);

The result shows that the domesticated Huaiyu mountain alpine potato, domesticated test-tube plantlet state: more robust than transplanted test-tube plantlets, but weaker than the test-tube plantlets of Example 1; and 40% grow new leaves to survive, and the survival rate is 40%. The domestication effect of test-tube plantlets is the same.

Comparative example 3

With embodiment 1, the difference is that step (3) is not carried out;

The results show that the domesticated Huaiyu Mountain alpine potato, domesticated test-tube plantlet state: more robust than the transplanted test-tube plantlet, but weaker than the test-tube plantlet of Example 1; The domestication effect of test-tube plantlets is the same.

Comparative example 4

With embodiment 1, the difference is that step (4) is not carried out;

The results show that the domesticated Huaiyu Mountain alpine potato, domesticated test-tube plantlet state: more robust than the transplanted test-tube plantlet, but weaker than the test-tube plantlet of Example 1; The domestication effect of test-tube plantlets is the same.

Comparative example 5

With embodiment 1, the difference is that step (5) only waters culture solution c;

The result shows that the domesticated Huaiyu mountain alpine potato, domesticated test-tube plantlet state: more robust than the transplanted test-tube plantlet, but weaker than the test-tube plantlet of embodiment 1; The domestication effect of test-tube plantlets is the same.

Comparative example 6

With embodiment 1, the difference is that step (5) only carries out once culture solution watering in the morning;

The results show that the domesticated Huaiyu mountain alpine potato, the state of domesticated test-tube plantlets: more robust than the transplanted test-tube plantlets, but weaker than the test-tube plantlets of Example 1; and 60% grow new leaves and survive, and the survival rate is 60%.

The above descriptions are only preferred embodiments of the present invention, and are not intended to limit the present invention. Any modifications, equivalent replacements and improvements made within the spirit and principles of the present invention should be included in the protection scope of the present invention within.

Claims (4)

1. A method for embedding, transplanting and domesticating a stem section of a Huaiyoshan high mountain potato with buds is characterized by comprising the following steps,

(1) Mixing 0.5-1cm stem segment with buds of rhizoma Solani Tuber osi of Mount Huaiyehne with calcium ion-free modified MS culture solution a containing sodium alginate and sucrose under aseptic condition to obtain mixed solution;

(2) Under the aseptic condition, dropping the mixed solution into MS culture solution b containing chitosan oligosaccharide, graphene quantum dots, carbon nano tubes, calcium chloride and sucrose for embedding to obtain the embedding beads of the potato stem segments with the buds in the mountains of the Huaiyanshan;

(3) Under the aseptic condition, soaking the bud stem embedded beads of the Huaiyoshan high mountain potatoes prepared in the step (2) in MS culture solution c containing chitosan oligosaccharide and graphene quantum dots;

(4) Under the aseptic condition, soaking the stem embedded beads of the potatoes with buds in the mountainous Huaiyeh, which are soaked in the step (3), in MS culture solution d containing chitosan oligosaccharide and carbon nano tubes;

(5) Transplanting the budding stem embedded beads of the mountain potatoes in the Huaiyan mountain, which are obtained in the step (4), into a matrix at room temperature, and irrigating an MS culture solution d containing chitosan oligosaccharide and carbon nano tubes in the morning and evening every day; pouring the MS culture solution c containing chitosan oligosaccharide and graphene quantum dots in the morning and evening every day after 7 days of pouring;

(6) After the stem section with the buds grows into a complete plant, domesticating the plant, and transplanting the plant into a field for cultivation;

in the step (1), the mass volume ratio of sodium alginate in the calcium ion-free modified MS culture solution a is 3%, and the sucrose concentration is 0.2mol/L;

in the step (2), in the MS culture solution b, the concentration of chitosan oligosaccharide is 0.05g/L, the concentration of graphene quantum dots is 0.03-0.05g/L, the concentration of carbon nano tubes is 0.03-0.05g/L, caCl is added ₂ The concentration is 0.1mol/L and the sucrose concentration is 0.2mol/L;

in the step (3), in the MS culture solution c, the concentration of chitosan oligosaccharide is 0.05g/L, the concentration of graphene quantum dots is 0.03-0.05g/L, and the soaking time is 48h;

in the step (4), in the MS culture solution d, the concentration of the chitosan oligosaccharide is 0.05g/L, the concentration of the carbon nano tube is 0.03-0.05g/L, and the soaking time is 48h.

2. The method for embedding, transplanting and domesticating the stem segments of the Huaiyoshan high mountain potatoes as claimed in claim 1, wherein the substrate in the step (5) is a mixture of vermiculite and perlite in a mass ratio of 2: 1.

3. The method for embedding, transplanting and domesticating the stem segments with the buds of the Huaiyshan mountain potatoes as claimed in claim 1, wherein the amount of MS culture solution d poured per time is 20 mL/embedded bead.

4. The method for embedding, transplanting and acclimatizing a wye mountain alpine potato stem segment with buds as claimed in claim 1, wherein the amount of MS culture solution c poured each time is 40 mL/embedded bead.