阅读说明：本技术 一种基于led节能光源的马铃薯试管薯诱导方法 (Potato microtuber induction method based on LED energy-saving light source ) 是由 徐志刚 龙家焕 陈松 于 2020-07-28 设计创作，主要内容包括：本发明属于生物技术领域,公开了一种基于LED组合光源的马铃薯试管薯诱导栽培方法,包括：剪取带一片叶的马铃薯脱毒组培苗中间茎段,接种于含3～4％蔗糖和0.7～0.9％琼脂的MS培养基中；先暗适应3～4天,再移至白色LED光源下培养20～30天,得到健壮马铃薯脱毒组培苗；取健壮马铃薯脱毒组培苗,剪取带一片叶的中间茎段,接种于含8％蔗糖和0.7～0.9％琼脂的MS培养基中；先暗适应3～4天,再移至白色LED光源下培养25～30天,最后移至红蓝光LED组合光源进行试管薯诱导处理60天。本发明可使植株形成更有利于试管薯产量形成的生理状态与形态建成,即较高的叶绿素含量、健壮的植株以及薯中更多的干物质分配。(The invention belongs to the technical field of biology, and discloses a test-tube potato induction cultivation method based on an LED combined light source, which comprises the following steps: cutting a middle stem section of the potato virus-free tissue culture seedling with one leaf, and inoculating the middle stem section into an MS culture medium containing 3-4% of sucrose and 0.7-0.9% of agar; dark adaptation is carried out for 3-4 days, and then the potato seedlings are moved to a white LED light source for culture for 20-30 days to obtain robust potato virus-free tissue culture seedlings; taking a strong potato virus-free tissue culture seedling, cutting a middle stem section with one leaf, and inoculating the middle stem section into an MS culture medium containing 8% of sucrose and 0.7-0.9% of agar; dark adaptation is carried out for 3-4 days, then the potato is moved to a white LED light source for culture for 25-30 days, and finally the potato is moved to a red and blue LED combined light source for test tube potato induction treatment for 60 days. The invention can make the plant form a physiological state and a morphological structure which are more beneficial to the formation of the yield of the test-tube potato, namely, higher chlorophyll content, robust plant and more dry matter distribution in the potato.)

1. An LED combined light source-based potato microtuber induction cultivation method is characterized by comprising the following steps:

step (1), basic seedling propagation: under the aseptic condition, cutting a middle stem section of the potato virus-free tissue culture seedling with one leaf, and inoculating the middle stem section into an MS culture medium containing 3-4% of sucrose and 0.7-0.9% of agar; placing the potato seedlings in a dark environment for dark adaptation for 3-4 days, and then transferring the potato seedlings to a white LED light source for culture for 20-30 days to obtain robust potato virus-free tissue culture seedlings;

step (2), light quality treatment: taking the strong potato virus-free tissue culture seedling obtained in the step (1), shearing a middle stem section with one leaf under the aseptic condition, and inoculating the middle stem section into an MS culture medium containing 8% of sucrose and 0.7-0.9% of agar; the method comprises the steps of placing the potato in a dark environment for dark adaptation for 3-4 days, transferring to a white LED light source for culture for 25-30 days, transferring to a red and blue LED combined light source for test tube potato induction treatment, performing induction treatment for 60 days, and harvesting.

2. The LED combined light source-based microtuber potato induction cultivation method according to claim 1, wherein in the step (1), the white LED light source cultivation environment is as follows: the optical density is 60 +/-5 mu mol.m^-2·s^-1(ii) a The photoperiod is 16 h/d; the temperature in the daytime is 20 +/-2 ℃, and the temperature at night is 18 +/-2 ℃; the relative humidity was 70. + -. 5%.

3. The LED combined light source-based microtuber potato induction cultivation method according to claim 1, wherein in the step (2), the white LED light source cultivation environment is as follows: the optical density is 60 +/-5 mu mol.m^-2·s^-1(ii) a The photoperiod is 14-16 h/d; the temperature in the daytime is 20 +/-2 ℃, and the temperature at night is 18 +/-2 ℃; the relative humidity was 70. + -. 5%.

4. The LED combined light source-based potato microtuber induction cultivation method according to claim 1, wherein in the step (2), the red and blue LED combined light source is a combined light source of red light and blue light in a ratio of 1: 1-1: 4.

5. The LED combined light source-based potato microtuber induction cultivation method according to claim 4, wherein the red and blue LED combined light source is a combined light source of red light and blue light in a ratio of 1: 3-1: 4.

6. The LED combined light source-based microtuber induction cultivation method of potato according to claim 5, wherein the red-blue LED combined light source is a combined light source with a red-blue ratio of 1: 3.

7. The LED combined light source-based microtuber potato induction cultivation method according to claim 1, wherein in the step (2), the environmental conditions of the induction treatment are as follows: light sealThe degree is 80 +/-5 mu mol.m^-2·s^-1(ii) a The photoperiod is 8 h/d; the temperature in the daytime is 20 +/-2 ℃, and the temperature at night is 17 +/-2 ℃; relative humidity, 70 ± 5%.

Technical Field

The invention belongs to the technical field of biology, and relates to a test-tube potato induction cultivation method based on an LED combined light source.

Background

The light is one of the essential environmental factors for plant growth and development. The spectrum range of the sunlight is 300-2600 nm, wherein the light (namely visible light) with the wavelength of 390-760 nm is a wave band with physiological activity in the solar radiation spectrum. In the spectrum band of physiological activity, red light (620-700 nm) and blue light (430-490 nm) are the main spectrums absorbed by photosynthesis of plants; in addition, the red light and the blue light are respectively mediated by the photosensitive pigment and the blue light receptor, and the growth and development of the plants are influenced in a signal form.

LED lamps for plant growth have been always regarded as mainstream products in the field of future plant lighting. The photosynthesis of plants is in a visible light spectrum of 380-760 nm, and the absorbed light energy accounts for about 60% -65% of physiological radiation light energy, wherein the absorbed light energy mainly comprises red light and orange light with the wavelengths of 610-720 nm and blue-violet light with the wavelengths of 400-510 nm; the spectral domain width of the LED is about +/-20 nm, and the wavelength is just consistent with the spectral range of plant photosynthesis and morphogenesis. Compared with the traditional light source, the LED lamp can emit monochromatic light spectrum required by plant growth, can realize independent control on different spectra and light output, and is easy to control and manage in plant light supplement. And the LED has the advantages of long service life, high luminous efficiency, low power consumption, short starting time, high color rendering index, low working temperature, small occupied space, firm structure, vibration resistance, good directivity, low working voltage, no ultraviolet radiation, environmental protection and the like. On the premise of meeting the light demand of crops, the energy consumption is saved, and the light energy utilization rate and the space utilization rate are improved.

Potatoes (Solanum tuberosum L.) are annual dicotyledonous herbaceous plants of the solanaceae family, and have been listed as the fourth major food crop in China. Under many years of cultivation, viruses or viroids infect potatoes and are transmitted and accumulated through potato block generations, resulting in severe yield reduction. The infection of potato by viruses or viroids is systemic infection, and no chemical agent capable of effectively controlling potato plant viruses exists at present. The same variety can be infected with a plurality of viruses at the same time, and the potato variety which is resistant to the plurality of viruses at the same time is difficult to breed by any way. The most effective way to solve the problem is to produce qualified detoxified potato seedlings and induce detoxified miniature seed potatoes by using a potato stem tip detoxification technology on the basis of tissue culture. However, the current production of the potato microtubers has the defects of lack of efficient induction spectrum, low induction rate, poor potato expansion effect and low yield, and the stable annual production of the potato microtubers is severely limited.

Disclosure of Invention

The inventor researches and discovers that the addition of red light in the spectrum is beneficial to the increase of the plant height, the leaf area and the induction number of the miniature potatoes, and the number of leaves is more, so that the plants can capture more light energy, but the plants distribute excessive nutrient substances to vegetative growth under monochromatic red light, so that the expansion of the potato in the middle and later stages is not beneficial to the growth of the potato in the test tube; the blue light is added into the spectrum, so that the strong seedling formation of the potato tubepotato plant is facilitated, the enlargement of the miniature potato is promoted, but the induction of the miniature potato is not facilitated due to the overlarge proportion of the blue light, and the premature senility of the nutritive tissue can be caused. Red light and blue light are necessary spectrums needed by potato test tube potato cultivation, but how to balance the application ratio of the red light and the blue light in the spectrums still is a problem that light supplement in potato test tube potato production is restricted. Therefore, the invention aims to find the red-blue spectrum proportion suitable for induction of the tubepotato and provide the potato tubepotato induction cultivation method based on the LED combined light source.

The purpose of the invention is realized by the following technical scheme:

an LED combined light source-based potato microtuber induced cultivation method comprises the following steps:

step (1), basic seedling propagation: under the aseptic condition, cutting a middle stem section of the potato virus-free tissue culture seedling with one leaf, and inoculating the middle stem section into an MS culture medium containing 3-4% of sucrose and 0.7-0.9% of agar; placing the potato seedlings in a dark environment for dark adaptation for 3-4 days, and then transferring the potato seedlings to a white LED light source for culture for 20-30 days to obtain robust potato virus-free tissue culture seedlings; and (3) cultivating environment: the optical density is 60 +/-5 mu mol.m^-2.s^-1(ii) a The photoperiod is 14-16 h/d; the temperature in the daytime is 20 +/-2 ℃, and the temperature at night is 18 +/-2 ℃; the relative humidity is 70 +/-5%;

step (2), light quality treatment: taking the strong potato virus-free tissue culture seedling obtained in the step (1), shearing a middle stem section with one leaf under the aseptic condition, and inoculating the middle stem section into an MS culture medium containing 8% of sucrose and 0.7-0.9% of agar; the method comprises the steps of placing the potato in a dark environment for dark adaptation for 3-4 days, transferring to a white LED light source for culture for 25-30 days, transferring to a red and blue LED combined light source for test tube potato induction treatment, performing induction treatment for 60 days, and harvesting.

The middle stem section is a stem section with 3-5 leaves on the upper part after the stem tip of the potato virus-free tissue culture seedling is removed.

In the step (2), the cultivation environment under the white LED light source is the same as that in the step (1): the optical density is 60 +/-5 mu mol.m^{- 2}.s^-1(ii) a The photoperiod is 14-16 h/d; the temperature in the daytime is 20 +/-2 ℃, and the temperature at night is 18 +/-2 ℃; the relative humidity was 70. + -. 5%.

The red and blue LED combined light source is a combined light source of red light and blue light in a ratio of 1: 1-1: 4, and is preferably a combined light source of red light and blue light in a ratio of 1: 3-1: 4. Specifically, the red-blue LED combined light source is selected from a red-blue ratio 1:1 combined light source (RB1), a red-blue ratio 1:2 combined light source (RB2), a red-blue ratio 1:3 combined light source (RB3) and a red-blue ratio 1:4 combined light source (RB4), and preferably the red-blue ratio 1:3 combined light source (RB3) and the red-blue ratio 1:4 combined light source (RB 4).

The environmental conditions of the induction treatment are as follows: the optical density is 80 +/-5 mu mol.m^-2.s^-1(ii) a The photoperiod is 8 h/d; the temperature in the daytime is 20 +/-2 ℃, and the temperature at night is 17 +/-2 ℃; the relative humidity was 70. + -. 5%.

The wavelength of the red light is 660nm, and the wavelength of the blue light is 440 nm.

The light sources are all rectangular lamp panels of 47.5 multiplied by 50cm, 20 light-emitting sets are arranged in the length direction of the rectangular lamp panels, 19 light-emitting sets are arranged in the width direction of the rectangular lamp panels, and each light-emitting set consists of three light-emitting diodes.

The white LED light source or the red and blue LED combined light source is controlled by an adjustable constant current driving power supply, and the light intensity is adjusted and controlled by adjusting and controlling the current.

The invention has the beneficial effects that:

the induced cultivation method of the potato microtuber based on the LED combined light source has the following advantages: the operation is simple; secondly, energy conservation and environmental protection are achieved; and thirdly, the plants can be formed into physiological states and morphologies which are more favorable for the formation of the output of the tubepotato, namely, higher chlorophyll content, robust plants and more dry matters in the potato are distributed, and the tubepotato is efficiently induced.

The invention provides a new way and technical reference for the production of the potato tubepotato and the related biological research, has higher practical application value and is suitable for the factory production of the potato tubepotato.

Drawings

FIG. 1 shows the morphologies of the roots, stems and leaves of three representative tubepotato plants harvested under different red and blue combined lights.

FIG. 2 shows the harvest of tubepotato under different red and blue combined lights.

FIG. 3 shows the effect of different combined red and blue spectra on leaf chlorophyll content of potato tubephrosia plants at 60 d.

FIG. 4 shows the effect of different red and blue combined spectra on the plant height of a potato tuberdyrus plant.

FIG. 5 is a graph showing the effect of different red and blue combined spectra on the stem thickness of a potato tuberdos plant.

FIG. 6 is a graph of the effect of different combined red and blue spectra on the sucrose content in the stems of potato tubephros at harvest.

FIG. 7 is a graph of the effect of different combined red and blue spectra on starch content in the stems of potato tubephros at harvest.

FIG. 8 is a graph of the effect of different combined red and blue spectra on the sucrose content of tubers when potato tubephros were harvested.

FIG. 9 is a graph of the effect of different combined red and blue spectra on starch content in tubers when potato tubephragma plants were harvested.

FIG. 10 is a graph of the effect of different red and blue combined spectra on dry matter partitioning at the time of harvest of potato tubephragma.

Detailed Description

The technical solution of the present invention is further explained by the following embodiments. The methods used in the following examples are conventional unless otherwise specified, and the percentages are by volume unless otherwise specified.