CN111374050A - Breeding method of test-tube potatoes - Google Patents

Breeding method of test-tube potatoes Download PDF

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Publication number
CN111374050A
CN111374050A CN201811648452.5A CN201811648452A CN111374050A CN 111374050 A CN111374050 A CN 111374050A CN 201811648452 A CN201811648452 A CN 201811648452A CN 111374050 A CN111374050 A CN 111374050A
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culture
culture medium
steps
liquid
induction
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康潇霄
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Gansu Kangqin Potato Industry Co ltd
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Gansu Kangqin Potato Industry Co ltd
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    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01HNEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H4/00Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
    • A01H4/001Culture apparatus for tissue culture
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01HNEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H4/00Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
    • A01H4/008Methods for regeneration to complete plants

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  • Life Sciences & Earth Sciences (AREA)
  • Developmental Biology & Embryology (AREA)
  • Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Cell Biology (AREA)
  • Botany (AREA)
  • Environmental Sciences (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)

Abstract

The invention belongs to the field of agricultural cultivation production, and particularly relates to a breeding method of a potato microtuber, which comprises the following steps: preparing virus-free seedlings: cutting off terminal buds of the detoxified test-tube plantlet, and cutting stem sections with 3-4 axillary buds; strong seedling culture: the stem section is transferred into a culture bottle filled with a liquid culture medium to culture the stem section into a seedling; and (3) induction culture: after seedlings are formed, pouring out the residual liquid culture solution in the step (2), and adding a liquid induction culture medium into a container to induce the potatoes; and harvesting after 50d of induction culture. The method solves the technical problems of long breeding period, high cost, low efficiency and the like, and has the advantages of high efficiency, high quality, labor saving, easy operation and convenient management.

Description

Breeding method of test-tube potatoes
Technical Field
The invention belongs to the field of agricultural cultivation production, and particularly relates to a breeding method of a potato microtuber.
Background
The potato is a few high-yield crop in the world, has the excellent characteristics of comprehensive nutrition, wide adaptability, strong stress resistance, high and stable yield, long industrial chain, dual purposes of grain, vegetable and feed industrial raw materials and the like, and is the fourth crop in the world. An important factor affecting low potato yield is viral degeneration of potatoes. The average yield of the potatoes is reduced by 30 to 80 percent after the potatoes are infected with viruses. The method has the advantages of greatly enhancing the production and seed potato breeding of the detoxified potatoes, improving the supply capacity of the detoxified seed potatoes, accelerating the popularization and application of the detoxified potatoes, stably improving the yield per unit and the total yield of the potatoes, and having significance.
The potato belongs to asexual propagation crops, tubers are usually used for planting, various diseases are easy to transmit and expand damage through tuber generation, and meanwhile, the potato seeds have the particularity of large using amount, easy consumption, long breeding period and the like, so that the breeding of the potato seeds is related to the sustainable development, quality improvement and synergy of the potato industry.
In the traditional method, the breeding of the potato seeds usually takes 4-5 years, and the breeding technical process of the potatoes is as follows: the test-tube plantlet → potato breeder's seed → first generation of breeder's seed → second generation of breeder's seed, the test-tube plantlet is used in the whole breeding process to breed, the survival rate is low, and the planting has big difficulty, is difficult to manage, leads to the low output of breeder's seed, with high costs.
The test tube potato is a detoxified seed potato produced by using modern biotechnology to produce detoxified test tube seedlings in a culture bottle under relatively closed conditions, so that the test tube potato overcomes the defects of field production environment, can be subjected to annual uninterrupted industrial production and is not influenced by climatic conditions; moreover, the production process is easy to control, and the quality of the potato seeds is easier to ensure; and the test tube potato is small in size and light in weight, is convenient to store and transport for a long distance, and can save a large amount of manpower, material resources and financial resources. Therefore, the production of the test-tube potato plays an increasingly important role in the breeding of the detoxified potato and the promotion and application of the detoxified potato.
Disclosure of Invention
Aiming at the problems, the invention provides the breeding method of the test-tube potato, which solves the technical problems of long breeding period, high cost, low efficiency and the like, and has the advantages of high efficiency, high quality, labor saving, easy operation and convenient management.
The breeding method of the test-tube potato for solving the technical problems comprises the following steps:
(1) preparing virus-free seedlings: cutting off terminal buds of the detoxified test-tube plantlet, and cutting stem sections with 3-4 axillary buds;
(2) strong seedling culture: the stem section is transferred into a culture bottle filled with a liquid culture medium; and (5) culturing stem section seedlings by using a liquid strong seedling culture medium.
(3) And (3) induction culture: after seedlings are formed, pouring out the residual liquid culture solution in the step (2), and adding a liquid induction culture medium into a container to induce the potatoes;
(4) and harvesting after 50d of induction culture.
The formula of the liquid culture medium in the step (2) is as follows: 2xPBM macroelement, 1xPBM microelement, 19g/L sucrose, 100mg/L inositol, 0.50mg/L nicotinic acid, 20mg/L glycine.
Wherein the macroelements are: KNO31890mg/L、NH4NO31540mg/L、KH2PO4162mg/L、MgSO7H2O350mg/L、CaCl2H2O 450mg/L;
The trace elements are: KI 0.82mg/L, H3BO45.8mg/L、MnSO4H2O 22.3mg/L、ZnSO7H2O7.5mg/L、Na2MoO2H2O 0.023mg/L、CuSO5H2O 0.028mg/L、CoCl6H2O 0.025mg/L。
The pH value is 5.8-6.0. The pH was adjusted using a 12% by mass hydrochloric acid solution.
Using filter paper as a bridge, uniformly stirring all the raw materials, placing the raw materials in a high-temperature sterilization pot, sterilizing for 23min at 121 ℃, filling about 20mL of culture medium into each bottle, and inoculating 5 stem segments.
The culture conditions in the strong seedling culture are 2000-3000lx of illumination intensity, the illumination time is 16h/d, the temperature is 21-25 ℃, and the subsequent steps are carried out after 5-7 sections of strong seedlings are grown in about 3 weeks.
The induction culture medium adopts a liquid MS culture medium, 8% of sucrose, 5mg/L of hormone BAP and 10mg/L of coumarin, and the pH value is 5.8-6.0.
The processing method comprises the following steps: 20mL of the solution was added to each bottle, and 3 times of treatment were repeated.
The culture conditions in the induction culture are illumination intensity of 1500-.
The number of test tube potatoes (diameter greater than 3mm) formed in each treatment was recorded starting from 10d, observed periodically during induction culture.
The formula of the invention has the advantages of economical and applicable material selection, simple and quick preparation procedure, and convenient and easy-to-learn operation procedures. Solves the problem of insufficient nutrition of the test-tube potato in the later period. The illumination time is adjusted according to the sensitivity of the variety tuber to short sunlight, so as to achieve the induction of different varieties of test tube tuber without adding any growth substance.
The quality of the test tube potatoes prepared by the method is high, the weight of single-seed potatoes is 0.13-0.16 a, the inductivity is more than 87%, and the high-quality potato rate is more than 98.5%; the preparation method has simple preparation steps and low required cost. The yield increase amplitude is between 20 and 58 percent.
Detailed Description
Example 1
A breeding method of a potato microtuber comprises the following steps:
(1) preparing virus-free seedlings: cutting off terminal buds of the detoxified test-tube plantlet, and cutting stem sections with 3-4 axillary buds;
(2) strong seedling culture: the stem section is transferred into a culture bottle filled with a liquid culture medium; and (5) culturing stem section seedlings by using a liquid strong seedling culture medium. The formula of the liquid culture medium is as follows: macroelement 2xPBM, microelement 1xPBM, sucrose 19g/L, inositol 100mg/L, nicotinic acid 0.50mg/L, glycine 20 mg/L. The pH was 6.0. The pH was adjusted using a 12% by mass hydrochloric acid solution.
The macroelements are: KNO31890mg/L、NH4NO31540mg/L、KH2PO4162mg/L、MgSO7H2O350mg/L、CaCl2H2O 450mg/L;
The trace elements are: KI 0.82mg/L, H3BO45.8mg/L、MnSO4H2O 22.3mg/L、ZnSO7H2O7.5mg/L、Na2MoO2H2O 0.023mg/L、CuSO5H2O 0.028mg/L、CoCl6H2O 0.025mg/L。
Using filter paper as a bridge, uniformly stirring all the raw materials, placing the raw materials in a high-temperature sterilization pot, sterilizing for 23min at 121 ℃, filling about 20mL of culture medium into each bottle, and inoculating 5 stem segments.
The culture conditions are that the illumination intensity is 2000lx, the illumination time is 16h/d, the temperature is 21 ℃, 5-7 sections of strong seedlings can grow after about 3 weeks, and then the subsequent steps are carried out.
(3) And (3) induction culture: after seedlings are formed, pouring out the residual liquid culture solution in the step (2), and adding a liquid induction culture medium into a container to induce the potatoes;
the induction culture medium adopts a liquid MS culture medium, and based on the mass of the MS culture medium, 8 percent of sucrose, 5mg/L of hormone BAP and 10mg/L of coumarin are added, and the pH value is 5.8.
The processing method comprises the following steps: 20mL of the solution was added to each bottle, and 3 times of treatment were repeated.
The culture conditions in the induction culture are 15001x of illumination intensity, 8h/d of illumination time and 18 ℃.
(4) And harvesting after 50d of induction culture.
The number of test tube potatoes (diameter greater than 3mm) formed in each treatment was recorded starting from 10d, observed periodically during induction culture.
Example 2
The rest is as in example 1, wherein:
(1) under aseptic condition, cutting off terminal bud of the virus-free test-tube seedling, and transferring the stem section with 3-4 axillary buds into a culture bottle filled with liquid culture medium. The culture medium adopts a developed liquid culture medium, 2xPBM major elements, 1xPBM trace elements, 19g/L of cane sugar and MS of organic matters) and has a pH value of 5.9, filter paper is used as a bridge, the culture medium is sterilized at 121 ℃ for 23min, about 20mL of the culture medium is filled in each bottle, 5 stem segments are inoculated, and the culture conditions are illumination intensity of 2500lx, illumination of 16h/d, temperature of 24 ℃ and about 3 weeks, so that 5-7 strong seedlings can be grown.
Cutting terminal buds of virus-free potato test-tube seedlings cultured by solid strong seedlings, and directly adding the cut terminal buds into a liquid culture medium to produce test-tube potatoes; and the cut terminal buds can be inoculated into a solid culture medium for strong seedling culture and used for breeding and producing the next test-tube potato, and the test-tube potato production is carried out in a cycle of year to year. The terminal buds cut each time are used as the next generation of basic seedlings for breeding, the terminal buds grow fast and well in the solid strong seedling culture medium and grow regularly and consistently, and a solid foundation is laid for the production yield of the next generation of test tube potatoes.
The solid medium may be:
(2) induced culture
After the seedling is formed, the culture solution is basically absorbed completely, if the culture solution is remained, the culture solution is poured out, and then the autoclaved liquid induction culture medium is added. The induction culture medium is treated by adopting a liquid MS culture medium, 8% of sucrose and different hormones, the pH value is 5.9, 20mL of the induction culture medium is added into each bottle, and the treatment is repeated for 3 times after each bottle is treated. The culture conditions comprise illumination intensity of 20001x, illumination time of 8h/d and temperature of 20 ℃.
(3) The number of test tube potatoes (diameter greater than 3mm) formed in each treatment was recorded starting from 10d, observed periodically during the culture. And harvesting after 50d of induction culture.
Example 3
Other contents are as in example 1, the breeding method of potato tuberdos comprises the following steps:
(1) preparing virus-free seedlings: cutting off terminal buds of the detoxified test-tube plantlet, and cutting stem sections with 3-4 axillary buds;
(2) strong seedling culture: the stem section is transferred into a culture bottle filled with a liquid culture medium; and (5) culturing stem section seedlings by using a liquid strong seedling culture medium. The formula of the liquid culture medium is as follows: macroelement 2xPBM, microelement 1xPBM, sucrose 19g/L, organic matter MS, and pH value 5.8. The pH was adjusted using a 12% by mass hydrochloric acid solution.
Using filter paper as a bridge, uniformly stirring all the raw materials, placing the raw materials in a high-temperature sterilization pot, sterilizing for 23min at 121 ℃, filling about 20mL of culture medium into each bottle, and inoculating 5 stem segments.
The culture conditions are that the illumination intensity is 2000-3000lx, the illumination time is 16h/d, the temperature is 21-25 ℃, and 5-7 sections of strong seedlings can grow after about 3 weeks, and then the subsequent steps are carried out.
(3) And (3) induction culture: after seedlings are formed, pouring out the residual liquid culture solution in the step (2), and adding a liquid induction culture medium into a container to induce the potatoes;
the induction culture medium adopts a liquid MS culture medium, sucrose is 8 percent, different hormones are used for treatment, and the PH value is 6.0.
The processing method comprises the following steps: 20mL of the solution was added to each bottle, and 3 times of treatment were repeated.
The culture conditions in the induction culture are 25001x of illumination intensity, 8h/d of illumination time and 23 ℃.
(4) And harvesting after 50d of induction culture.
The number of test tube potatoes (diameter greater than 3mm) formed in each treatment was recorded starting from 10d, observed periodically during induction culture.
The breeding method of the invention is adopted to carry out field potato breeding tests on Longshu No. 3 and Longshu No. 7 respectively, and the results show that: the test-tube potato emergence rate, the growth potential, the single-plant potato bearing number and the single-plant potato weight average of the method are increased compared with those of the traditional method, and the difference is obvious.
While the foregoing shows and describes the fundamental principles and principal features of the invention, together with the advantages thereof, the foregoing embodiments and description are illustrative only of the principles of the invention, and various changes and modifications can be made therein without departing from the spirit and scope of the invention, which will fall within the scope of the invention as claimed. The scope of the invention is defined by the appended claims and equivalents thereof.

Claims (8)

1. A breeding method of a potato microtuber is characterized in that: the method comprises the following steps:
(1) preparing virus-free seedlings: cutting off terminal buds of the detoxified test-tube plantlet, and cutting stem sections with 3-4 axillary buds;
(2) strong seedling culture: the stem section is transferred into a culture bottle filled with a liquid culture medium to culture the stem section into a seedling;
(3) and (3) induction culture: after seedlings are formed, pouring out the residual liquid culture solution in the step (2), and adding a liquid induction culture medium into a container to induce the potatoes;
(4) and harvesting after 50d of induction culture.
2. The method according to claim 1, wherein the method comprises the following steps: the formula of the liquid culture medium in the step (2) is as follows: 2xPBM macroelement, 1xPBM microelement, 19g/L sucrose, 100mg/L inositol, 0.50mg/L nicotinic acid, 20mg/L glycine.
3. The method according to claim 2, wherein the method comprises the following steps: the pH value of the liquid culture medium is 5.8-6.0.
4. A method as claimed in claim 3, wherein the method comprises the steps of: the pH was adjusted using a 12% by mass hydrochloric acid solution.
5. The method according to claim 1, wherein the method comprises the following steps: the culture conditions in the strong seedling culture are illumination intensity of 2000-3000lx, illumination time of 16h/d and temperature of 21-25 ℃.
6. The method according to claim 1, wherein the method comprises the following steps: the induction culture medium adopts a liquid MS culture medium, 8 percent of sucrose, 5mg/L of hormone BAP and 10mg/L of coumarin.
7. The method according to claim 6, wherein the method comprises the following steps: the pH value of the induction culture medium is 5.8-6.0.
8. The method according to claim 1, wherein the method comprises the following steps: the culture conditions in the induction culture are illumination intensity of 1500-.
CN201811648452.5A 2018-12-30 2018-12-30 Breeding method of test-tube potatoes Pending CN111374050A (en)

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111374051A (en) * 2018-12-31 2020-07-07 甘肃康勤薯业有限公司 Cultivation method of potatoes in net shed
CN111374049A (en) * 2018-12-30 2020-07-07 甘肃康勤薯业有限公司 Culture medium for virus-free seedlings of hydroponic potatoes and application of culture medium
CN116114600A (en) * 2022-12-06 2023-05-16 江苏宝德农业科技有限公司 Application of paper in potato detoxification mini-potato production and rapid propagation and field planting method of tissue culture seedlings for potato detoxification mini-potato production

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CN111374051A (en) * 2018-12-31 2020-07-07 甘肃康勤薯业有限公司 Cultivation method of potatoes in net shed
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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111374049A (en) * 2018-12-30 2020-07-07 甘肃康勤薯业有限公司 Culture medium for virus-free seedlings of hydroponic potatoes and application of culture medium
CN111374051A (en) * 2018-12-31 2020-07-07 甘肃康勤薯业有限公司 Cultivation method of potatoes in net shed
CN116114600A (en) * 2022-12-06 2023-05-16 江苏宝德农业科技有限公司 Application of paper in potato detoxification mini-potato production and rapid propagation and field planting method of tissue culture seedlings for potato detoxification mini-potato production

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Application publication date: 20200707